molecules

Review Collective Locomotion of Human Cells, Wound Healing and Their Control by Extracts and Isolated Compounds from Marine Invertebrates

Claudio Luparello * , Manuela Mauro , Valentina Lazzara and Mirella Vazzana

Department of Biological, Chemical and Pharmaceutical Sciences and Technologies (STEBICEF), University of Palermo, 90128 Palermo, Italy; [email protected] (M.M.); [email protected] (V.L.); [email protected] (M.V.) * Correspondence: [email protected]; Tel.: +39-91-238-97405



Received: 10 May 2020; Accepted: 25 May 2020; Published: 26 May 2020


Abstract: The collective migration of cells is a complex integrated process that represents a common theme joining morphogenesis, tissue regeneration, and tumor biology. It is known that a remarkable amount of secondary metabolites produced by aquatic invertebrates displays active pharmacological properties against a variety of diseases. The aim of this review is to pick up selected studies that report the extraction and identification of crude extracts or isolated compounds that exert a modulatory effect on collective cell locomotion and/or skin tissue reconstitution and recapitulate the molecular, biochemical, and/or physiological aspects, where available, which are associated to the substances under examination, grouping the producing species according to their taxonomic hierarchy. Taken all of the collected data into account, marine invertebrates emerge as a still poorly-exploited valuable resource of natural products that may significantly improve the process of skin regeneration and restrain tumor cell migration, as documented by in vitro and in vivo studies. Therefore, the identification of the most promising invertebrate-derived extracts/molecules for the utilization as new targets for biomedical translation merits further and more detailed investigations.

Keywords: marine invertebrates; cell migration; wound healing

1. Introduction The collective migration of cells is a complex integrated process at the basis of several different physiological events, such as embryo development, tissue shaping, and wound healing, whereas, in the course of diseases, like carcinogenesis, it is strongly implied in the step of tumor cell metastatization. In the recent years, a great deal of literature papers has focused on the molecular, biophysical and cytological aspects of cell locomotory patterns in the awareness of the clinical relevance shown by the advancement of knowledge on the mechanism and dynamism of collective cell migration. These studies shed light upon the mechanobiological bases of movement generation and speed/orientation setting and unveiled some of the signal transduction properties that are linked to the biochemical features of the migratory process. As a result of this research, the roles played in epithelial-mesenchymal transition (EMT) by integrin, junction protein and cytoskeletal dynamics, growth factor/cytokine and metalloprotease secretion, as well as cell interactions with components of the extracellular matrix (ECM) have been increasingly elucidated. A comprehensive overview of these topics can be found in several previously-published reviews [1–3]. Among the events of tissue repair involving collective cell motility, skin wound healing represents a multi-step coordinated process, which, following the initial inflammatory and hemostatic stages, associates re-epithelization, i.e., epidermal cell locomotion and growth on top of the granulation tissue,

Molecules 2020, 25, 2471; doi:10.3390/molecules25112471 www.mdpi.com/journal/molecules Molecules 2020, 25, x FOR PEER REVIEW 2 of 42 Molecules 2020, 25, 2471 2 of 40 hemostatic stages, associates re-epithelization, i.e., epidermal cell locomotion and growth on top of the granulation tissue, with tissue recovery implying the restoration of vascularization via withangiogenesis tissue recovery and fibroplasia, implying the which restoration is the ofrecruitment vascularization of activated via angiogenesis fibroblasts and and fibroplasia, newly- whichdifferentiated is the recruitment myofibroblasts of activated for ECM fibroblasts reconsti andtution newly-di and woundfferentiated contraction myofibroblasts [4–8]. Once for ECMthe reconstitutionwound damage and is woundrepaired, contraction a final tissue [4– 8remodeling]. Once the is woundundertaken damage to restore is repaired, skin integrity a final tissueand remodelingstructure [9,10]. is undertaken During towound restore re-epithelization, skin integrity and human structure keratinocytes [9,10]. During move wound in a re-epithelization,spatially- and humandirectionally-organized keratinocytes move way, in a spatially- whose andmolecular directionally-organized basis can be found way, whose in specific molecular cell-ECM basis can beinteractions, found in specific like the cell-ECM binding interactions, of type XVII like thecolla bindinggen by of both type intermediate XVII collagen filament by both intermediateand actin filamentcytoskeleton and actinvia hemidesmosomal cytoskeleton via actinin hemidesmosomal 4, and the creation actinin 4,of andgradients the creation of epidermal of gradients growthof epidermalfactor (EGF) growth immobilized factor (EGF) within immobilized the ECM and within presen theted ECM to andcell receptors presented [11,12]. to cell receptorsIndeed, EGF [11 ,is12 ]. Indeed,one of EGFthe most is one potent of the regulators most potent of keratinocy regulatorste of locomotory keratinocyte behavior locomotory along behaviorwith transforming along with transforminggrowth factor growth (TGF)- factorβ, whose (TGF)- prominentβ, whose prominentregulatory regulatoryeffect on epidermal effect on epidermalcell motility cell during motility duringwound wound repair repairvia ERK via signaling ERK signaling pathways pathways is widely is widely acknowledged acknowledged [13]. Scientific [13]. Scientific interest interest has hasalso also focused focused upon upon the the transcriptional transcriptional factors factors that that are are up-regulated up-regulated and and activated activated in inmigrating migrating keratinocyteskeratinocytes during during the the repair repair process, process, leading leading to theto the identification, identification, among among others, others, of Smad7, of Smad7, Stat3, andStat3, c-Jun, and which c-Jun, are which functionally are functionally involved ininvolved the promotion in the promotion of cutaneous of woundcutaneous healing, wound as extensively healing, reviewedas extensively by Boudra reviewed and Ramsey by Boudra [14 ].and Ramsey [14]. CollectiveCollective cell cell motility motility can can be studiedbe studied both both in culture in culture and in and vivo in. Invivo. the firstIn the case, first the case, migratory the behaviourmigratory of behaviour cell monolayers of cell canmonolayers be evaluated can be via evaluated the “in vitro via woundthe “in healing”vitro wound or “scratch” healing” assay, or in“scratch” which a assay, gap is in created which bya gap scratching is created a pipetteby scratching tip through a pipette a confluent tip through sheet a confluent of cells insheet culture of (Figurecells in1A), culture and the(Figure eventually-occurring 1A), and the eventually-occurring bidimensional cell bidimensional migration leading cell migration to the closure leading of to the scratchthe closure being of recorded the scratch by eitherbeing recorded phase-contrast by either microscopic phase-contrast images microscopic captured at images regular captured intervals at or time-lapseregular intervals videomicroscopy or time-lapse [15]. videomicroscopy [15].

FigureFigure 1. 1.(A A)) Phase-contrastPhase-contrast microscopic image of of a a scratch scratch created created in in a amonolayer monolayer of of HEPG2 HEPG2 liver liver carcinomacarcinoma cells. cells. (B )B Light) Light microscopic microscopic image image of MDA-MB231 of MDA-MB breast231 breast carcinoma carcinoma cells migratedcells migrated through thethrough pores of the a polycarbonate pores of a polycarbonate filter in a Boyden filter chamberin a Boyden assay, cham fixedber with assay, 95% fixed ethanol, with and 95% stained ethanol, with 0.02and g stained toluidine with blue 0.02/mL. g Bartoluidine= 10 µ blue/mL.m. Bar = 10 μm.

WhenWhen the the model model system system studied studied is a tumor is a celltumor line, cell the scratchline, the assay scratch is commonly assay is complemented commonly bycomplemented either the “Boyden by either chamber” the “Boyden or the “transwell chamber” migration or the “transwell/invasion” migration/invasion” assays, as originally assays, developed as byoriginally Albini and developed coworkers by [ 16Albini], which and measure coworkers the [16], ability which of a populationmeasure the of ability neoplastic of a population cells to migrate of individuallyneoplastic cells through to migrate a physical individually barrier, i.e.,through an untreated a physical or barrier, basement i.e., membrane an untreated (Matrigel)-coated or basement porousmembrane polyester (Matrigel)-coated membrane, in porous response polyester to a chemoattractant membrane, in response or a motility-promoting to a chemoattractant compound or a (Figuremotility-promoting1B) [17,18]. compound (Figure 1B) [17,18]. DealingDealing with within vivoin vivoexperimental experimental systems, systems, the group the group locomotion locomotion of tumor of cellstumor might cells be might examined be throughexamined the through chick embryo the chick chorioallantoic embryo chorioallantoic membrane membrane (CAM) assay, (CAM) which assay, implies which the implies grafting the of neoplasticgrafting of cells neoplastic on the CAM,cells on their the subsequentCAM, their interactionsubsequent withinteraction the blood with vessels, the blood and vessels, the invasion and ofthe the invasion adjacent mesenchymalof the adjacent tissue, mesenchymal which can betiss studiedue, which by microscopical,can be studied immunohistochemical, by microscopical, andimmunohistochemical, molecular techniques and [19 ].molecu CAMlar might techniques be used [19]. also CAM to study might the be angiogenetic used also processto study that the is characterizedangiogenetic by process migration that ofis mesodermalcharacterized blood by migration vessels into of themesodermal basement membraneblood vessels and into the ECMthe leadingbasement to their membrane compartmentalization and the ECM leading into new to vesselstheir compartmentalization in the ectoderm [20]. Theinto incisionnew vessels of the in rodent the dorsumectoderm has [20]. been The commonly incision usedof the as rodent an in vivodorsummodel has tobeen study commonly the effect used of molecules as an in vivo and model extracts

Molecules 2020, 25, 2471 3 of 40 on the whole process of skin repair. Using this technique, the evolution of wound closure can be macroscopically assessed at different time points and, once the animals are sacrificed, histological observations, immunohistochemical analyses, biophysical evaluations, such as the quantitation of tensile strength, and biochemical/molecular assays can be performed on the biopsies. This enables collecting additional multi-perspective information on the fine tuning of tissue repair and also to evaluating the ability of supplemented substance(s) to promote a quick, robust, and histologically-shaped healing aimed to obtain the most correct skin replacement [21]. In vivo burn injury models may be created by pressing for few seconds a shaved and stretched area of the rodent dorsum skin with a metal block pre-heated at 100 ◦C or more, thus enabling studying the effect of novel compounds for topical therapeutics that are designed for the treatment of burn lesions [22]. Progresses in the knowledge of the molecular bases of wound repair and in the design of novel more efficacious medicaments are of basic importance to develop effective treatments for compromised healing occurring in pathologies, such as diabetes, arteriosclerosis, malnourishment, and malignant tumors. However, it is worth-mentioning that, although small animal models are commonly used for wound healing research, they display some biological limitations due to the differences in the histological architecture of rodent vs. human skin, and in the primary physiological act of healing, i.e., myofibroblast contraction for rodents vs. re-epithelization for humans. Other experimental models, such as the porcine one, have been proven to be more linkable with the human healing process, but the costs of housing and the difficulties in handling have limited their usability for first-line exploratory research [23,24]. Another aspect to consider is that “pure” scarring-free regeneration, which only sees new cells completely replacing the lost ones, is very uncommon in humans. In fact, regeneration is a well-known phenomenon in which animals restore lost parts of their body through four overlapping steps: (a) wound healing and histolysis, (b) blastema formation, (c) recognition of positional information, and (d) repatterning of the missing portion. As discussed by Krafts [25], the repair process in the context of the healing of human tissues consists of both regeneration and replacement, with the latter seeing the intervention of inflammatory cells and the sequential formation of granulation and fibrous tissues, which make the biological system more multifaceted. It is widely recognized that seas and oceans, which account for about 97% of the whole water in the world, have a still little exploited asset in terms of biodiversity and, consequently,marine species-derived biological products. Much research interest has been focused on marine invertebrates, which represent more than 50% of the species inhabiting the aquatic environment in Europe. A remarkable amount of secondary metabolites is produced by aquatic invertebrates to aid in both the inter-individual signalization and the defense from predators, competitors, infective agents and UV radiation damages due to the adaptation mechanisms to the specific life conditions in the considerably differing marine ecosystems. An ever increasing number of these substances displays active pharmacological properties with an extensive spectrum of indications, making them prospective drugs of marine origin to be developed for therapeutic applications [26]. Within this context, the aim of this review is to pick up selected studies that report the extraction and identification of mixed fractions or isolated chemicals which exert a modulatory effect on collective cell migration and/or skin tissue reconstitution and recapitulate the molecular, biochemical, and/or physiological aspects, where available, which are associated to the metabolites or extracts under examination. Such compounds will be listed respecting the taxonomic hierarchy of the producing invertebrate species in dedicated paragraphs, in which the structural aspects of each organism and their contribution to “blue biotech” applications, if any, will also be briefly outlined.

2. Porifera Porifera is the oldest metazoan group that is still widespread on the planet. They are very simple animals, which are devoid of real tissues or organs, and only consist of a few types of specialized cells that are surrounded by a collagen-rich extracellular matrix [27]. Molecules 2020, 25, 2471 4 of 40 Molecules 2020, 25, x FOR PEER REVIEW 4 of 42

ChondrosiaChondrosia reniformis reniformis(Nardo, (Nardo, 1847; 1847; Demospongiae,Demospongiae, Chondros Chondrosiida:iida: Chondrosiida Chondrosiidae)e) is is a asessile sessile speciesspecies that that lives lives in in the the shallowshallow coastscoasts of the Mediterranean Sea Sea and and the the South-West South-West coast coast of of the the AtlanticAtlantic Ocean, Ocean, which which can can reach reach a considerablea considerable body body thickness thickness (Figure (Figure2). 2).

FigureFigure 2. 2.A A specimen specimen ofofChondrosia Chondrosia reniformis demosponge. ©© BernardBernard Picton Picton (CC (CC BY-NC-SA BY-NC-SA 4.0); 4.0); http:http://www.marinespecies.org/.//www.marinespecies.org/.

ThisThis species species shows shows remarkable remarkable regenerativeregenerative properties and, for for this this reason, reason, has has been been chosen chosen asas a a model model system system for for studiesstudies onon the molecular pa pathwaysthways of of tissue tissue restoration restoration in in lower lower metazoan, metazoan, highlightinghighlighting the the involvement involvement of TGF of superfamilyTGF superfamily proteins proteins [28]. Unlike [28]. otherUnlike sponges, other thissponges, invertebrate this lacksinvertebrate the silica spiculeslacks the that silica commonly spicules strengthenthat commo thenly other strengthen species’ body,the other instead species’ obtaining body, its instead stiffness throughobtaining a greater its stiffness amount through of collagen a greater fibers amount that are of tightlycollagen packed fibers in that a complex are tightly fibrillar packed network. in a Collagencomplex extracts fibrillar from network.C. reniformis Collagenhave extracts shown considerablefrom C. reniformis potentiality have inshown carrying considerable out the role ofpotentiality nanoparticle in carrying vector and out drug-coating the role of nanoparticle material [ 29vector,30] inand the drug-coating design of biocompatible material [29,30] sca inff theolds fordesign regenerative of biocompatible medicine scaffolds [31]. Moreover, for regenerative sticky hydrogels medicine with[31]. Moreover, self-healing sticky properties hydrogels have with been obtainedself-healing from properties collagen extracted have been from obtained the ectosome from andcollagen choanosome extracted of thisfrom demosponge the ectosome showing and remarkablechoanosome rheological of this demosponge properties thatshowing make remark them suitableable rheological as injectable properties matrices that for make biomedical them applicationssuitable as injectable [32]. Particular matrices interest for biomedical has been appl givenications to the [32]. cosmetic Particular and interest wound has healing been e givenffect of trypsin-digestedto the cosmetic andC. reniformis wound healingcollagen effect hydrolysates. of trypsin-digested In their C. paper, reniformis Pozzolini collagen et al.hydrolysates. [33] isolated diInfferent their paper, peptide Pozzolini fractions et byal. [33] HPLC isolated and demonstrateddifferent peptide that fractions those onesby HPLC particularly and demonstrated enriched in hydroxyproline-containingthat those ones particularly collagenous enriched peptides in hydroxyproline-containing promoted the migration of HaCaTcollagenous keratinocytes peptides and L929promoted fibroblasts the migration in “scratch” of assays,HaCaT also keratinocytes accelerating and cell L929 proliferation fibroblasts at thein “scratch” site of the assays, wound also with respectaccelerating to the untreatedcell proliferation controls at at the the site same of timethe wound points. with In addition, respect theseto the preparationsuntreated controls appeared at the able tosame trigger time type points. I collagen In addition, synthesis these and extracellular preparations release appeared by fibroblasts able to tr andigger protect type keratinocytes I collagen fromsynthesis UV damage and extracellular by enhancing release cell by viability fibroblast afters and exposure protect and keratinocytes down-regulating from UV the damage UV-induced by expressionenhancing of cellKRT1 viabilityand KRT10 after exposurekeratin genes and responsibledown-regulating of skin the photoaging UV-induced via expression epidermal thickeningof KRT1 andand decrement KRT10 keratin of elasticity. genes responsible When also of considering skin photoagi theng radical via epidermal scavenging thickening activity and shown decrement by these collagenof elasticity. hydrolysates When also and considering their lack of the toxicity radica towardsl scavenging different activity cultured shown cytotypes, by these the collagen peptide fractionshydrolysates from C. and reniformis their lackcollagen of toxicity show towards considerable different biotechnological cultured cytotype potentials, the inpeptide the development fractions offrom formulations C. reniformis that collagen are designed show to considerable mitigate the damagesbiotechnolo ingical UV-exposed potential or in wounded the development skin. of formulationsJaspis stellifera that[Carter, are designed 1879; Demospongiae,to mitigate the damages Tetractinellida: in UV-exposed Ancorinidae; or wounded Figure3 skin.A] is another sessileJaspis species stellifera typical [Carter, of tropical 1879; and Demospongiae, Indo-West Pacific Tetractinellida: environments. Ancorinidae; A number Figure of anti-cancer 3A] is bioactiveanother moleculessessile species have typical been isolated of tropical from and this In marinedo-West invertebrate, Pacific environments. such as jaspiferins, A number jaspiferals, of anti- stelliferins,cancer bioactive and stellettin molecules [34 –have41]. been In their isolated publication, from this Cheng marine et invertebrate, al. [42] produced such additionalas jaspiferins, data regardingjaspiferals, the stelliferins, mobility controland stel exercisedlettin [34–41]. by theIn their triterpenoid publicatio stellettinn, Cheng B et (3 Zal.,3a [42]S,5a producedR,9aR,9bS )- 3a,6,6,9a-tetramethyl-3-[(3additional data regardingE,5 E)-6-(5-methyl-6-oxopyran-2-yl)hepta-3,5-dien-2-ylidene]-4,5,5a,8,9,9b-the mobility control exercised by the triterpenoid stellettin B hexahydro-1Hcyclopenta(3Z,3aS,5aR,9aR,9bS)-3a,6,6,9a-tetramethyl-3-[(3 [a]naphthalene-2,7-dione;E,5E Figure)-6-(5-methyl-6-oxopyran-2-yl)hepta-3,5-3B on human U87MG and GBM8401 glioblastomadien-2-ylidene]-4,5,5a,8,9,9b-hexahydro-1Hcyclop cells. In particular, the migration andenta invasion [a]naphthalene-2,7 of tumor cells-dione; was inhibitedFigure 3B in bothon scratchhuman wound U87MG healing and andGBM8401 transwell glioblastoma chamber assays cells. whenIn particular, exposed the to anmigration increasing and concentration invasion of of thetumor triterpene cells was in the inhibited range between in both 1 scratch and 10 µwounM. Thisd healing appeared and to transwell be based, atchamber least in assays part, upon when the exposed to an increasing concentration of the triterpene in the range between 1 and 10 μM. This

Molecules 2020, 25, x FOR PEER REVIEW 5 of 42 Molecules 2020, 25, 2471 5 of 40 appeared to be based, at least in part, upon the p-Akt downsignalling-mediated inhibition of the interaction between F-actin and the actin-binding protein P-girdin, responsible for pseudopoda p-Aktextension downsignalling-mediated at the cell leading edge, inhibition and the expressi of the interactionon of matrix between metallopro F-actintease and 2, phosphorylated the actin-binding proteinintegrin P-girdin, β1 and responsible phosphorylated for pseudopoda focal adhesion extension kinase at the by cell this leading cancer edge, cytotype and the expression[43]. In β ofconsideration matrix metalloprotease that glioblastoma 2, phosphorylated cell survival integrin and the angiogenetic1 and phosphorylated processes focalin models adhesion in vitro kinase byand this in cancervivo are cytotype also remarkably [43]. In consideration reduced in the that presence glioblastoma of stellettin cell survival B [42], andthis themolecule angiogenetic is a processesstrong candidate in models asin anti-cancer vitro and inagen vivot againstare also malignant remarkably brain reduced tumors. in the presence of stellettin B [42], this molecule is a strong candidate as anti-cancer agent against malignant brain tumors.

Figure 3. (A) Image of a paraffin section of Jaspis stellifera demosponge. Photo by D. Drew (YPM IZ 087028).Figure 3. Courtesy A) Image of of the a paraffin Peabody section Museum of Jaspis of Natural stellifera History, demosponge. Division Photo of Vertebrate by D. Drew Paleontology, (YPM YaleIZ 087028). University; Courtesy peabody.yale.edu. of the Peabody (B) 2D Museum structure ofof stellettinNatural B;History, https:// pubchem.ncbi.nlm.nih.govDivision of Vertebrate / compoundPaleontology,/5352082#section Yale University;=2D-Structure peabody.yale.edu.. B) 2D structure of stellettin B; https://pubchem.ncbi.nlm. nih.gov/compound/ 5352082#section=2D-Structure. Negombata magnifica (Keller, 1889; Demospongiae, Poecilosclerida: Podospongiidae), formerly namedNegombata Latrunculia magnifica magnifica, (Keller, is also 1889; known Demospongiae, as toxic finger-sponge Poecilosclerida: and it typically Podospongiidae), has a brilliant orange-redformerly named coloration Latrunculia and a magnifica, digitate or is leafyalso known morphology as toxic with finger-sponge crooked branchesand it typically (Figure has4A). Ita isbrilliant distributed orange-red in the coloration Indian Ocean and anda digitate the Red or Sea,leafy where morphology it intensively with crooked populates branches the coral reefs.(Figure When 4A). It theseis distributed sponges in form the Indian colonies, Ocea theyn and protect the Red themselves Sea, where viait intensively chemical populates defences, in particularthe coral reefs. releasing When a these reddish sponges fluid form containing colonies latrunculins,, they protect toxicthemselves substances via chemical that are defences, known to bindin particular G-actinin releasing a reversible a reddish manner fluid and containing 1:1 stoichiometric latrunculins, ratio toxic [44]. substances Latrunculin that B (4 areR)-4-[(1 knownR,4 Zto,8 Z, 10bindS,13 RG-actin,15R)-15-hydroxy-5,10-dimethyl-3-oxo-2,14-dioxabicyclo[1 in a reversible manner and 1:1 stoichiometric ratio 1.3.1] [44]. heptadeca-4,8-dien-15-yl]-1,3- Latrunculin B (4R)-4- thiazolidin-2-one;[(1R,4Z,8Z,10S,13 FigureR,15R)-15-hydroxy-5,10-dimethyl-3-oxo-2,14-dioxabicyclo[14B is a member of this family that is a promising agent for1.3.1] the heptadeca- development of4,8-dien-15-yl]-1,3-thiazolidin-2-one; anti-glaucoma and anti-arrythmia Figure therapies 4B is [ 45a ,member46]. This of macrolidethis family andthat itsis a semi-synthetic promising derivativesagent for the 15- developmentO-methyl-latrunculin of anti-glaucoma B, N-hydroxymethyl- and anti-arrythmia latrunculin therapies B [45,46]. and N-acetyl-latrunculin This macrolide Band were its foundsemi-synthetic to restrain derivatives the migration 15-O of-methyl-latrunculin the murine brain-metastatic B, N-hydroxymethyl- melanomacell latrunculin line B16B15b B inand both N-acetyl-latrunculin transwell chamber B were and scratchfound to wound restrain healing the migration assays [of47 ].the Latrunculin murine brain-metastatic A ((4R)-4-[(1 R, 4melanomaZ,8E,10Z,12 cellS,15 lineR,17 B16B15bR)-17-hydroxy- in both 5,12-dimethyl-3-oxo-2,16dioxabicyclo[13.3.1]transwell chamber and scratch wound healing nonadeca-4,8,10- assays [47]. trien-17-yl]-1,3-thiazolidin-2-one;Latrunculin A ((4R)-4-[(1R,4 FigureZ,8E,104CZ,12 andS,15 itsR,17 carbamateR)-17-hydroxy- derivatives 5,12-dimethyl-3-oxo- 17- O-[N-(phenyl) carbamoyl]-latrunculin2,16dioxabicyclo[13.3.1] A, nonadeca-4,8,10-trien-17-yl]-1,3-thiazolidin-2-one; 17-O-[N-(3-chloropropyl)carbamoyl] latrunculin AFigure and 17- 4CO-[ Nand-(benzyl) its carbamoyl]-latrunculincarbamate derivatives A appeared 17-O to-[N be-(phenyl)carbamoyl]-latrunculin able to impair the invasive behavior A, of prostate17-O-[N cancer-(3- cellschloropropyl)carbamoyl] in Boyden chambers latrunculin in the presence A and of 17- Matrigel,O-[N-(benzyl)carbamoyl]-la as well as the disaggregationtrunculin A ofappeared spheroids into vitro be able[48 ].to Inimpair this paper,the invasive a putative behavior inhibitory of prostate effect cancer of these cells molecules in Boyden on chambers the activation in the and signalizationpresence of Matrigel, of hypoxia-inducible as well as the factor disaggregation (HIF)-1, a of transcription spheroids in factor vitro that[48]. isIn involvedthis paper, in a the metastaticputative inhibitory attitude of effect prostate of these cancer molecules cells [49 ],on was the presumed. activation Afterwards,and signalization the in of vitro hypoxia-invasive phenotypeinducible factor of MDA-MB231 (HIF)-1, a transcription breast cancer factor cell that was is alsoinvolved found in tothe be metastatic impaired attitude in Boyden of prostate chamber assayscancer bycells derivatives [49], was ofpresumed latrunculin. Afterwards, A, the most the potent in vitro being invasive 17-O-phenylethyl-latrunculin phenotype of MDA-MB231 A and Nbreast-p-methoxyphenylacetyl-15- cancer cell was also foundO-methyl-latrunculin to be impaired A. in Interestingly, Boyden chamber the latter assays one by is likely derivatives to recognize of dilatrunculinfferent and yetA, unknownthe most target(s) potent than being actin microfilaments.17-O-phenylethyl-latrunculin Of note, 15-O -methyl-latrunculinA and N-p- Bmethoxyphenylacetyl-15- also exhibited a powerfulO-methyl-latrunculin anti-angiogenic activity, A. Interestingl whereas 17-y,O -phenylethyl-latrunculinthe latter one is likely Ato was recognize different and yet unknown target(s) than actin microfilaments. Of note, 15-O-methyl-

Molecules 2020, 25, x FOR PEER REVIEW 6 of 42 Molecules 2020, 25, 2471 6 of 40 latrunculin B also exhibited a powerful anti-angiogenic activity, whereas 17-O-phenylethyl- significantlylatrunculin cytotoxic A was againstsignificantly MCF-7 cytotoxic breast cancer agains cellst MCF-7 [50], with breast both cancer properties cells being [50], appropriate with both to potentialproperties anti-breast being appropriate cancer drugs. to potential anti-breast cancer drugs.

Figure 4. (A) A specimen of Negombata magnifica demosponge. Photo © Alexander Vasenin (CC Figure 4. A) A specimen of Negombata magnifica demosponge. Photo © Alexander Vasenin (CC BY-SA 3.0) https://en.wikipedia.org/.(B) Two-dimensional (2D) structure of latrunculin B; https: BY-SA 3.0) https://en.wikipedia.org/. B) Two-dimensional (2D) structure of latrunculin B; //pubchem.ncbi.nlm.nih.gov/compound/Latrunculin-B#section=2D-Structure.(C) 2D structure of https://pubchem.ncbi. nlm.nih.gov/compound/Latrunculin-B#section=2D-Structure. C) 2D latrunculin A; https://pubchem.ncbi.nlm.nih.gov/compound/Latrunculin-a#section=2D-Structure. structure of latrunculin A; https:// pubchem.ncbi.nlm.nih.gov/compound/Latrunculin-a#section= Pseudoceratina2D-Structure. arabica (Keller, 1889; Demospongiae, Verongiida: Pseudoceratinidae; Figure5A is distributed in the shallow sandy lagoons and seagrass beds of the southern Red Sea. It appearsPseudoceratina as a sprawling arabica and (Keller, branching 1889;sponge Demospongiae, and his body Vero mightngiida: show Pseudocera varyingtinidae; characteristics, Figure appearing5A is distributed dense, rubbery,in the shallow flexible, sandy or waxy. lagoons A and number seagrass of literaturebeds of the data southern indicate Red that Sea. these It spongesappears produce as a sprawling several and compounds branching with sponge varied and biological his body activities,might show e.g., varying the antimicrobial characteristics, and antifungalappearing parasympatholytic dense, rubbery, flexible, moloka’iamine or waxy. andA nu moloka’iakitamide,mber of literature data and theindicate skeletal thatα these-chitin ofsponges particular produce interest several for dicompoundsfferent biotechnological with varied biological applications, activiti suches, e.g., as the electrochemistry antimicrobial and and α regenerativeantifungal parasympatholytic medicine [51,52]. moloka’iamine The bromotyrosine and moloka’iakitamide, derivative (1’R,5’ andS,6’ theS)-2-(3’,5’-dibromo-1’,6’ skeletal -chitin of -dihydroxy-4’-oxocyclohex-2’-enyl)particular interest for different biotechnological acetonitrile obtained applications, from P. arabicasuch aswas electrochemistry found to impair and the TGFregenerativeβ-stimulated medicine epithelial-mesenchymal [51,52]. The bromotyrosine transition derivative of A549 (1’ lungR,5’S cancer,6’S)-2-(3’,5’-dibromo-1’,6’ cells and the related - celldihydroxy-4’-oxocyclohex-2’-enyl migration in scratch wound closure) acetonitrile assays obtained through from the P. inhibition arabica was of found both theto impair activity the of β typeTGF I TGF--stimulatedβ receptor epithelial-mesenchymal kinase ALK5 and the transition consequent of A549 motility-addressed lung cancer cells and actin the microfilament related cell reorganizationmigration in scratch thereby wound being a closure promising assays anti-tumoral through the pharmacological inhibition of both tool the [53 ].activity Noteworthy, of type theI β bidimensionalTGF- receptor locomotion kinase ofALK5 the highly-malignant and the consequent MDA-MB231 motility-addressed breast cancer cellsactin inmicrofilament wound closure assayreorganization was consistently thereby restrained being a bypromising the already-mentioned anti-tumoral pharmacological moloka’iamine tool (3-[4-(2-aminoethyl)-2,6- [53]. Noteworthy, dibromophenoxy]propan-1-amine;the bidimensional locomotion of Figurethe highly-mal5B and itsignantP. arabica MDA-MB231-obtainedderivative breast cancer ceratinin cells in A, wound whereas theclosurein vitro assayinvasive was activityconsistently of the restrained same cell lineby inthe transwell already-mentioned assays was decreasedmoloka’iamine by exposure (3-[4-(2- to twoaminoethyl)-2,6- other P. arabica-obtained dibromophenoxy]propan-1-amine; moloka’iamine derivatives, Figure i.e., ceratinin 5B and D and its hydroxymoloka’iamineP. arabica-obtained (2-amino-1-derivative [4-(3-aminopropoxy)-3,5-ceratinin A, whereas the dibromophenyl] in vitro invasive ethanol; activity Figure of the5C. same In the cell same line paper, in transwell evidence wasassays shown was that decreased also the alkaloid by exposure subereamolline to two other A produced P. arabica-obtained by another verongidmoloka’iamine sponge derivatives, distributed in thei.e., Southern ceratinin Red andD Westernand Arabianhydroxymoloka’iamine Sea, i.e., Suberea mollis (2-amino-1-(Row, 1911; Demospongiae,[4-(3-aminopropoxy)-3,5 Verongiida:- Aplysinellidae;dibromophenyl] Figure ethanol;5D) wasFigure able 5C. to In inhibit the same the paper, scratch evidence wound was healing shownin that vitro alsoby MDA-MB231the alkaloid breastsubereamolline cancer cells A [54 produced]. Of note, byS. another mollis is verongid a good producer sponge distributed of a number in of the compounds Southern withRed and overt anti-microbialWestern Arabian and anti-oxidantSea, i.e., Suberea hepatoprotective mollis (Row, 1911; functions, Demospongiae, such as aeroplysinins, Verongiida: Aplysinellidae; the brominated Figure 5D) was able to inhibit the scratch wound healing in vitro by MDA-MB231 breast cancer

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Molecules 2020, 25, 2471 7 of 40 cells [54]. Of note, S. mollis is a good producer of a number of compounds with overt anti- microbial and anti-oxidant hepatoprotective functions, such as aeroplysinins, the brominated arginine-derivedarginine-derived alkaloids alkaloids subereamines, subereamines, and the brominatedand the phenolicbrominated molecules phenolic subereaphenols, molecules amongsubereaphenols, others [55–57 among]. others [55–57].

Figure 5. (A) A specimen of Pseudoceratina arabica demosponge © Danièle Heitz; https://nomadica. jimdofree.comFigure 5. /.(AB) )A 2D structurespecimen ofof moloka’iamine; Pseudoceratina https: arabica//pubchem.ncbi.nlm.nih.gov demosponge © Danièle/compound Heitz;/ Moloka_Iamine#sectionhttps://nomadica.jimdofree.=Structures com/..(C B)) 2D 2D structure structure of of hydroxymoloka’iamine; moloka’iamine; https://pubchem.ncbi.nlm. https://pubchem.ncbi. nlm.nih.govnih.gov/compound/Moloka_Iam/compound/25016152.(ine#section=Structures.D) A specimen of Suberea mollisC) demosponge;2D Abbasstructure et al., 2014of (CChydroxymoloka’iamine; BY 3.0). https://pubchem.ncbi.nlm.nih.gov/compound/25016152. D) A specimen of Suberea mollis demosponge; Abbas et al., 2014 (CC BY 3.0). Siphonocalina (Callyspongia) siphonella (Lévi, 1965; Demospongiae, Haplosclerida: Callyspongiidae; Figure6A),Siphonocalina also named (Callyspongia colonial tube-sponge) siphonella for its(Lévi, appearance 1965; asDemospongiae, a bunch of pale lavenderHaplosclerida: long tubes,Callyspongiidae; is a species typicalFigure of6A), the also Red named Sea. colonial This organism tube-sponge is a sourcefor its appearance of secondary as metabolitesa bunch of endowedpale lavender with anti-infective long tubes, is a and species anti-inflammatory typical of the Red properties, Sea. This organism and cytotoxicity is a source against of secondary cancer cellsmetabolites [58–60]. Sipholenol endowed A with ((3R ,5aanti-infectiveR,6R, 7S,9aR )-6-and [2-[(1anti-inflammatoryR,3aR,5S,8aR)-1-hydroxy-1,4,4,6-tetramethyl- properties, and cytotoxicity 2,3,3a,5,8,8a-hexahydroazulen-5-yl]ethyl]-2,2,5a,7-tetramethyl-4,5,6,8,9,9a-hexahydro-3H-benzo[b]against cancer cells [58–60]. Sipholenol A ((3R,5aR,6R, 7S,9aR)-6- [2-[(1R,3aR,5S,8aR)-1-hydroxy- oxepine-3,7-diol;1,4,4,6-tetramethyl-2,3,3a,5,8,8a-hexahydroazulen-5-yl]ethyl]-2,2,5a,7-tetramethyl-4,5,6,8,9,9a- Figure6B is a triterpenoid that is produced by this demosponge displaying an anti-cancerhexahydro-3H-benzo[b]oxepine-3,7-diol; activity that is based upon its ability Figure to counteract 6B is a triterpenoid the multidrug that resistance is produced of neoplastic by this cellsdemosponge via direct inhibitiondisplaying of an P-glycoprotein anti-cancer activity [61]. Aliphaticthat is based polar upon and its aromatic ability to ester counteract derivatives the of thismultidrug molecules resistance exhibited of neoplastic a strong cells anti-migratory via direct inhibition and anti-invasive of P-glycoprotein effect towards [61]. Aliphatic MDA-MB231 polar malignantand aromatic breast ester cells derivatives in scratch woundof this healingmolecules and exhibited Boyden a chamber strong anti-migratory assay in the presence and anti- of basementinvasive membrane effect towards extract MDA-MB231 [62]. The down- malignant regulation breast ofcells cell in motility scratch wound appeared healing to be and due, Boyden at least in part,chamber to the assay inhibition in the presence of phosphorylation of basement of membra proteinne tyrosine extract kinase-6 [62]. The (PTK6), down- anregulation anti-apoptotic of cell andmotility pro-metastatic appeared factor to be for due, this at cell least line in [63part,]. Aklto the et al.inhibition [64] demonstrated of phosphorylation that the sipholenolof protein A-4-tyrosineO-30,40-dichlorobenzoate kinase-6 (PTK6), an derivative anti-apoptotic was alsoand activepro-metastatic as potential factor anti-breast for this cell cancer line [63]. compound. Akl et In particular,al. [64] demonstrated they found that that the this sipholenol molecule A-4- inhibitedO-3′,4′-dichlorobenzoate the proliferation of derivative a panel ofwas breast also tumoractive cellas lines potential with dianti-breastfferent neoplastic cancer compound. phenotypes, In impairingparticular, cellthey cycle found progression, that this molecule and it restrainedinhibited boththe cell proliferation motility in transwellof a panel chamber of breast assays tumorin vitrocell linesand tumorwith different mass expansion neoplastic in MDA-MB231phenotypes, xenograftimpairing nude cell mice cycle model progression,in vivo .and Such it restrained effects appeared both cell to motility be based, in transwell at least in chamber part, upon assays the interferencein vitro and with tumor the metastasis-associated mass expansion in breastMDA-MB231 cancer kinase xenograft (BRK) nude and focalmice adhesionmodel in kinase vivo. (FAK)Such signaleffects cascades, appeared in turn to be inhibiting based, at the least activation in part, up ofon some the downstream interference signallingwith the metastasis-associated factors, such as AKT, mitogen-activatedbreast cancer kinase protein (BRK) kinase and (MAPK), focal adhesion and paxillin. kinaseThus, (FAK) sipholenol signal cascades, A appears in turn to be inhibiting a promising the scaffactivationold for the of developmentsome downstream of novel signalling anti-malignant factors, breastsuch as cancer AKT, drugs.mitogen-activated protein kinase (MAPK), and paxillin. Thus, sipholenol A appears to be a promising scaffold for the development of novel anti-malignant breast cancer drugs.

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Figure 6. A) A specimen of Siphonocalina siphonella demosponge © Alexander Vasenin (CC BY-SA FigureFigure 6. ( 6.A )A A) A specimen specimen of ofSiphonocalina Siphonocalina siphonella siphonellademosponge demosponge© Alexander© Alexander Vasenin Vasenin (CC (CC BY-SA BY-SA 3.0); 3.0); https://commons.wikimedia.org/. B) 2D structure of sipholenol A; https://pubchem.ncbi. https:3.0);//commons.wikimedia.org https://commons.wikimedia.org/./.(B) 2D structure B) 2D structure of sipholenol of sipholenol A; https://pubchem.ncbi.nlm.nih.gov A; https://pubchem.ncbi. / nlm.nih.gov/compound/Sipholenol-A#section=Structures. compoundnlm.nih.gov/compound/Siphole/Sipholenol-A#section=Structuresnol-A#section=Structures..

XestospongiaXestospongiasp. sp. (Demospongiae, (Demospongiae, Haplosclerida: Haplosclerida: Petrosiidae; Petrosiidae; Figure Figure7 is widely7 is widely distributed distributed in the Xestospongia sp. (Demospongiae, Haplosclerida: Petrosiidae; Figure 7 is widely distributed Indianin the and Indian South and Pacific South Oceans. Pacific These Oceans. animals These represent animals a rich represent source ofa rich secondary source metabolites of secondary with in the Indian and South Pacific Oceans. These animals represent a rich source of secondary verymetabolites different biologicalwith very properties different bi thatological have beenproperties shown that effective have asbeen enzymatic shown effective inhibitors, as anti-malarial, enzymatic metabolites with very different biological properties that have been shown effective as enzymatic anti-bacterial,inhibitors, anti-malarial, vasodilatory, andanti-bacterial, anti-cancer vaso agentsdilatory, [65–70 and]. Among anti-cancer them, ranieramycinsagents [65–70]. are Among a group inhibitors, anti-malarial, anti-bacterial, vasodilatory, and anti-cancer agents [65–70]. Among ofthem, biologically-active ranieramycins tetrahydroisoquinoline are a group of biologically-active compounds. tetrahydroisoquinoline Halim et al. [71] reported compounds. a significant Halim them, ranieramycins are a group of biologically-active tetrahydroisoquinoline compounds. Halim inhibitoryet al. [71] ereportedffect exerted a significant by a member inhibitory of this effect family, exerted ranieramycin by a member M, of on this the family, motile ranieramycin and invasive et al. [71] reported a significant inhibitory effect exerted by a member of this family, ranieramycin behaviorM, on the of H460motile lung and cancer invasive cells behavior in scratch of woundH460 lung healing cancer and cells matrigel-containing in scratch wound transwell healing assays and M, on the motile and invasive behavior of H460 lung cancer cells in scratch wound healing and atmatrigel-containing non-toxic or growth-impairing transwell assays concentrations. at non-toxic or growth-impairing concentrations. matrigel-containing transwell assays at non-toxic or growth-impairing concentrations.

FigureFigure 7. Examples7. Examples of specimens of specimens of Xestospongia of Xestospongiasp. (A) X. mutasp. A©) RenaudX. muta Houdinet; © Renaud https: Houdinet;//flickr.com /; Figure 7. Examples of specimens of Xestospongia sp. A) X. muta © Renaud Houdinet; (B)https://flickr.com/;X. testudinaria, photo B by) AlbertX. testudinaria Kok (Public, domain)photo https:by //commons.wikimedia.orgAlbert Kok (Public /. domain) https://flickr.com/; B) X. testudinaria, photo by Albert Kok (Public domain) https://commons.wikimedia.org/. 3. Cnidariahttps://commons.wikimedia.org/.

3. CnidariaCladiella australis (Macfadyen, 1936; Anthozoa, Alcyonacea: Alcyoniidae; Figure8A), also known 3. Cnidaria as the finger blanching soft coral, is a species distributed in the Western Pacific ocean, displaying Cladiella australis (Macfadyen, 1936; Anthozoa, Alcyonacea: Alcyoniidae; Figure 8A), also brown orCladiella white color,australis depending (Macfadyen, on whether 1936; theAnthozoa, polyps areAlcyonacea: extended orAlcyoniidae; retracted. It Figure is know 8A), that also this known as the finger blanching soft coral, is a species distributed in the Western Pacific ocean, organismknown producesas the finger several blanching secondary soft metabolites coral, is a withspecies promising distributed biological in the activities Western as Pacific hypoglicemic, ocean, displaying brown or white color, depending on whether the polyps are extended or retracted. It anti-cancerdisplaying and brown anti-melanogenic or white color, compounds, depending on the whether latter having the polyps potential are extended applications or retracted. in medical It is know that this organism produces several secondary metabolites with promising biological cosmetologyis know that [72 –this74]. organism Among theproduces molecules several that secondary were obtained metabolites from C. with australis promising, research biological interest activities as hypoglicemic, anti-cancer and anti-melanogenic compounds, the latter having hasactivities been focused as hypoglicemic, on the neuroprotective anti-cancer austrasulfone and anti-melanogenic (4-ethenylsulfonylbutan-2-one; compounds, the latter Figure having8B[ 75]. potential applications in medical cosmetology [72–74]. Among the molecules that were obtained potential applications in medical cosmetology [72–74]. Among the molecules that were obtained from C. australis, research interest has been focused on the neuroprotective austrasulfone (4- from C. australis, research interest has been focused on the neuroprotective austrasulfone (4-

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Molecules 2020, 25, 2471 9 of 40 ethenylsulfonylbutan-2-one; Figure 8B [75]. Its synthetic precursor, dihydroaustrasulfone alcohol, exhibited the ability to restrain the migratory behaviour of A549 lung tumor cells in both wound Itshealing synthetic and precursor, transwell dihydroaustrasulfonechamber assays in vitro. alcohol, This inhibitory exhibited theactivity ability appeared to restrain to be the the migratory result behaviourof the inactivation of A549 lung of extracellular tumor cells in signal-regulated both wound healing kinase and (ERK) transwell 1/2 intracellular chamber assays signalling,in vitro . Thiswhich, inhibitory in turn, activity down-regulated appeared tothe be levels the result of activated of the inactivation focal adhesion of extracellularkinase (FAK), signal-regulated PI3K, and p- kinaseAKT. (ERK)Such derangement 1/2 intracellular of signalling,transduction which, mechan inisms turn, also down-regulated determined thethe levelsdecrease of activatedof the focalproduction adhesion of kinase matrix (FAK), metalloprot PI3K, andeinase p-AKT. (MMP)-2 Such and derangement -9 enzymes, of responsible transduction for mechanisms the observed also determinedreduced tumor the decrease size ofin the a productionA549 xenograft of matrix nude metalloproteinase mice model (MMP)-2 [76]. andInterestingly, -9 enzymes, responsibledihydroaustrasulfone for the observed alcohol reduced was tumor also sizeproven in a A549to restrain xenograft the nude platelet-derived mice model [ 76growth]. Interestingly, factor dihydroaustrasulfone(PDGF)-BB promoted alcohol the wasprolifer alsoation proven and to restrainmigration the platelet-derivedof vascular smooth growth muscle factor (PDGF)-BBcells, as promoteddocumented the proliferation by wound andhealing, migration Boyden of vascularchamber, smooth and transwell muscle cells, assays, as documented thereby playing by wound a healing,vasoprotective Boyden role chamber, and being and transwella good candidate assays, thereby for the playingtreatment a vasoprotectiveof vascular occlusive role and diseases being a good[77,78]. candidate for the treatment of vascular occlusive diseases [77,78].

Figure 8. (A) A specimen of Cladiella australis soft coral © David (CC BY-NC-SA 4.0) https://www. Figure 8. A) A specimen of Cladiella australis soft coral © David (CC BY-NC-SA 4.0) https:// marinespecies.org/.(B) 2D structure of austrasulfone; https://pubchem.ncbi.nlm.nih.gov/compound/ www.marinespecies.org/. B) 2D structure of austrasulfone; https://pubchem.ncbi.nlm.nih.gov/ 50907657#section=Structures. compound/50907657#section=Structures. Cladiella pachyclados (Klunzinger, 1877; Anthozoa, Alcyonacea: Alcyoniidae; Figure9A) is a soft coralCladiella typical pachyclados of reef-associated (Klunzinger, and 1877; subtropical, Anthozoa, in Alcyon particularacea: Western Alcyoniidae; Pacific, Figure environments. 9A) is a Itsoft is known coral typical that this of marinereef-associated invertebrate and subtropical, produces eunicellin in particular (i.e., Western [(1R,2R,3 Pacific,R,5S,6S environments.,7S,8R,11S,14R )- 5,6,14-triacetyloxy-6,14-dimethyl-10-methylidene-3-propan-2-yl-15-oxatricyclo[6.6.1.02,7]pentadecan-It is known that this marine invertebrate produces eunicellin (i.e., 11-yl]acetate)-based[(1R,2R,3R,5S,6S,7S diterpenes,8R,11S,14R with)-5,6,14-triacetyloxy-6, anti-inflammatory 14-dimethyl-10-methylidene-3-propan-2- and anti-cancer properties. In particular, Hassanyl-15-oxatricyclo[6.6.1.02,7]pentadecan-11-yl]acetate)-based et al. [79] showed the powerful anti-migratory and anti-invasive diterpenes rolewith played anti-inflammatoryin vitro on PC3 prostateand anti-cancer cancer cells properties. by some In of theseparticular, compounds, Hassan withet al. the[79] most showed effective the powerful in both experimental anti-migratory tests beingand pachycladinanti-invasive A role ([(1 playedR,2S,3R in,6R vitro,7R,8 Ron,9 RPC3,12 Spros,13State)-9-acetyloxy-12,13-dihydroxy-3,9,13-trimethyl-6- cancer cells by some of these compounds, propan-2-yl-15with the oxatrimost cyclo[6.6.1.02,7]pentadecan-3-yl]effective in both experimental butanoate; Figuretests 9B,being sclerophytin pachycladin F methyl etherA ((1R,2R,6R([(1R,2S,3,7R,8R,9S,12S,13SR,6R,7R,8R,9R,12)-12-methoxy-9,13-dimethyl-3-S,13S)-9-acetyloxy-12,13-dihydroxy-3,9,13- methylidene-6-propan-2-yl-15oxatricyclo trimethyl-6-propan-2-yl- [6.6.1.02,7]pentadecane-9,13-diol;15 oxatri cyclo[6.6.1.02,7]pentadecan-3-yl] Figure9C), polyanthelinbutanoate; Figure A ([( 1R,2R,6S,8R,9S,10R,13R,14R9B, sclerophytin F methyl )-2,6,10-ether trimethyl-13-propan-2-yl-((1R,2R,6R,7R,8R,9S,12S,13S 15,16-)-12-methoxy-9,13-dimethyl-3- dioxatetracyclo [6.6.1.12,6.09,14]hexadecan-10-yl] methylidene-6-propan-2-yl-15 acetate; Figure9D, andoxatricyclo[6.6.1.02,7]pentadecane-9,13-diol; sclerophytin A ((1R,2R,6R,7R,8R,9R,12S,13S )-9,13-dimethyl-3-methylidene-6-propan-2-yl-15Figure 9C), polyanthelin A oxatricyclo[6.6.1.02,7]([(1R,2R,6S,8R,9S,10R,13R,14R pentadecane-9,12,13-triol;)-2,6,10-trimethyl-13-propan-2-yl- Figure9E[79]. 15,16- dioxatetracyclo [6.6.1.12,6.09,14]hexadecan-10-yl] acetate; Figure 9D, and sclerophytin A ((1R,2R,6R,7R,8R,9R,12S,13S)-9,13-dimethyl-3-methylidene-6-propan-2-yl- 15oxatricyclo[6.6.1.02,7] pentadecane-9,12,13-triol; Figure 9E [79].

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Figure 9. (A) A specimen of Cladiella pachyclados soft coral © Danièle Heitz; https://nomadica. Figure 9. A) A specimen of Cladiella pachyclados soft coral © Danièle Heitz; https:// jimdofree.com/.(B) Two-dimensional (2D) structure of pachycladin A; https://pubchem.ncbi.nlm. nomadica.jimdofree.com/. B) Two-dimensional (2D) structure of pachycladin A; nih.gov/compound/Pachycladin-A#section=2D-Structure.(C) 2D structure of sclerophytin F methyl https://pubchem.ncbi.nlm. nih.gov/compound/Pachycladin-A#section=2D-Structure. C) 2D ether; https://pubchem.ncbi.nlm.nih.govcompound/21635655.(D) 2D structure of polyanthelin A; structure of sclerophytin F methyl ether; https://pubchem.ncbi.nlm.nih.gov compound/21635655. https://pubchem.ncbi.nlm.nih.gov/compound/12168107.(E) 2D structure of sclerophytin A; https: D) 2D structure of polyanthelin A; https:// pubchem.ncbi.nlm.nih.gov/compound/12168107. E) 2D //pubchem.ncbi.nlm.nih.gov/compound/Sclerophytin-A#section=2D-Structure. structure of sclerophytin A; https:// pubchem.ncbi.nlm.nih.gov/compound/Sclerophytin-A Sarcophyton#section=2D-Structure. crassocaule (Moser, 1919, Anthozoa, Alcyonacea: Alcyoniidae; Figure 10A is a subtropical species of sessile soft corals distributed in the Western Pacific area, New Caledonia, Taiwan and RyukyuSarcophyton Island. crassocaule A number (Moser, of terpenoids 1919, Anthozoa, with anti-tumoral Alcyonacea properties: Alcyoniidae; have been Figure extracted 10A is bya thissubtropical marine invertebrate species of sessile [80]. Studies soft coralsin vitro distributedon the anti-migratoryin the Western ePacificffects ofarea, compounds New Caledonia, that are producedTaiwan byand S. Ryukyu crassocaule Island. have A beennumber focussed of terpenoi on bladderds with and anti-tumoral gastric tumor properties cell models. have Inbeen the firstextracted case, the by cembrenolide this marine diterpeneinvertebrate 13-acetoxysarcocrassolide [80]. Studies in vitro ([(on1S,3S,5S,8E,12E,14S,15S the anti-migratory effects)-5,9,13- of trimethyl-18-methylidene-17-oxo-4,16-dioxatricyclo[13.3.0.03,compounds that are produced by S. crassocaule have been focussed 5]octadeca-8,12-dien-14-yl] on bladder and gastric acetate;tumor Figurecell 10models.B) was provenIn the to impairfirst case, locomotion the cembre in scratchnolide wound diterpene healing assays,13-acetoxysarcocrassolide apart from inducing apoptotic([(1S,3S,5S,8E,12E,14S,15S cell death after 24 h)-5,9,13- of exposure. trimethyl-18-methylidene-17-oxo-4,16-dioxatricyclo[13.3.0.03, Western blot and proteomic analyses demonstrated the up- or down-regulation5]octadeca-8,12-dien-14-yl] of a number acetate; of proteins, fig 10B) including was proven members to ofimpair the group locomotion of heat shockin scratch proteins, wound key enzymeshealing of assays, the Krebs apart cycle, from and inducing proteins apoptotic that are associated cell death toafter apoptosis 24 h of andexposure. DNA repair,Western although blot and no correlationproteomic between analyses the demonstrated biochemical data the andup- or the down-r anti-migratoryegulation e ffofect a number was shown of proteins, [81]. Similar including results onmembers 13- acetoxysarcocrassolide-induced of the group of heat shock motility proteins, restraining key enzymes were of obtained the Krebs with cycle, gastric and carcinoma proteins that cells, in whichare associated the up-regulation to apoptosis of p38-and DNA and c-Jun repair, N-terminal although kinaseno correlation (JNK)-transduction between the pathways biochemical and down-regulationdata and the anti-migratory of phosphoinositide effect was 3-kinase shown (PI3K) [81]./ AKTSimilar signalization results on 13- were acetoxysarcocrassolide- also observed [82]. inducedSinularia motility nanolobata restraining(Verseveldt, were 1977;obtained Anthozoa, with gastric Alcyonacea: carcinoma Alcyoniidae) cells, in which populates the up- the coralregulation reefs or of rocks p38- in and shallows c-Jun fromN-terminal East Africakinase to (JNK)-tra the Westernnsduction Pacific pathways area. The and bioactive down- compoundregulation 11-dehydrosinulariolideof phosphoinositide 3-kinase (1R,3S,5S,8E,13R (PI3K)/AKT signalization)-5,9,13-trimethyl-16-methylene-4,14-dioxa were also observed [82]. tricyclo[11.3.2.03,5] octadec-8-ene-12,15-dione11-dehydrosinulariolide; Figure 11A that is produced by this soft coral has been tested for the control of the migratory behavior of CAL-27 oral squamous cancer cells in scratch wound healing assays. The obtained results demonstrate that the wound repair function is down-regulated by the molecule, as well as cell growth and survival, whereas apoptotic death is promoted. Proteomic analyses showed that exposure to 11-dehydrosinulariolide determined the modification of the levels of proteins that are associated with cell growth, apoptosis, translation and protein folding, and energy metabolism [83]. Subsequent investigations on another bioactive molecule extracted from S. nanolobata, i.e., sinularin

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Figure 9. A) A specimen of Cladiella pachyclados soft coral © Danièle Heitz; https:// nomadica.jimdofree.com/. B) Two-dimensional (2D) structure of pachycladin A; https://pubchem.ncbi.nlm. nih.gov/compound/Pachycladin-A#section=2D-Structure. C) 2D structure of sclerophytin F methyl ether; https://pubchem.ncbi.nlm.nih.gov compound/21635655. D) 2D structure of polyanthelin A; https:// pubchem.ncbi.nlm.nih.gov/compound/12168107. E) 2D structure of sclerophytin A; https:// pubchem.ncbi.nlm.nih.gov/compound/Sclerophytin-A #section=2D-Structure.

Sarcophyton crassocaule (Moser, 1919, Anthozoa, Alcyonacea: Alcyoniidae; Figure 10A is a subtropical species of sessile soft corals distributed in the Western Pacific area, New Caledonia, Taiwan and Ryukyu Island. A number of terpenoids with anti-tumoral properties have been extracted by this marine invertebrate [80]. Studies in vitro on the anti-migratory effects of compounds that are produced by S. crassocaule have been focussed on bladder and gastric tumor cell models. In the first case, the cembrenolide diterpene 13-acetoxysarcocrassolide ([(1S,3S,5S,8E,12E,14S,15S)-5,9,13- trimethyl-18-methylidene-17-oxo-4,16-dioxatricyclo[13.3.0.03, Molecules 2020, 25, x FOR PEER REVIEW 11 of 42 Molecules5]octadeca-8,12-dien-14-yl]2020, 25, 2471 acetate; fig 10B) was proven to impair locomotion in scratch wound11 of 40 healing assays, apart from inducing apoptotic cell death after 24 h of exposure. Western blot and Figure 10. A) A specimen of Sarcophyton crassocaule soft coral © Leen Van Ofwegen (CC BY-NC- proteomic analyses demonstrated the up- or down-regulation of a number of proteins, including SA 4.0); https://www.marinespecies.org/ B) 2D structure of 13-acetoxysarcocrassolide; ((9E)-13-hydroxy-4,9,13-trimethyl-17-methylidene-5,15-dioxatricyclo[12.3.1.04,6] octadec-9-en-16-one; membershttps://pubchem.ncbi.nlm.nih.gov/compound/21775908 of the group of heat shock proteins, key enzymes. of the Krebs cycle, and proteins that Figureare associated11B), confirmed to apoptosis this compound and DNA asrepair, an additional although potentialno correlation anti-cancer between molecule. the biochemical In fact, sinularindataSinularia and promoted the nanolobataanti-migratory the inhibition (Verseveldt, effect of was growth 1977; shown Anthozoa, and [8 motility,1]. Similar Alcy the onacea:results block on atAlcyoniidae) 13- G2/ Macetoxysarcocrassolide- cell cycle populates phase, the and thecoralinduced onset reefs of apoptosismotilityor rocks restraining inin A2058shallows melanoma were from obtained East cells Af and ricawith proteomic to gastric the Western carcinoma analyses Pacific found cells, area. modifications in Thewhich bioactive the in up- the levelscompoundregulation of oncosuppressor 11-dehydrosinulariolideof p38- and and c-Ju apoptosis-associatedn N-terminal (1R,3S,5S,8E,13R kinase proteins )-5,9,13-trimethyl-16-methylene-4,14-dioxa(JNK)-tra consistentnsduction with thepathways observed and phenotypic down- changestricyclo[11.3.2.03,5]regulation [84]. of phosphoinositide octadec-8-ene-12,15-dione11-dehydrosinulariolide; 3-kinase (PI3K)/AKT signalization were alsoFigure observed 11A [82].that is produced by this soft coral has been tested for the control of the migratory behavior of CAL-27 oral squamous cancer cells in scratch wound healing assays. The obtained results demonstrate that the wound repair function is down-regulated by the molecule, as well as cell growth and survival, whereas apoptotic death is promoted. Proteomic analyses showed that exposure to 11- dehydrosinulariolide determined the modification of the levels of proteins that are associated with cell growth, apoptosis, translation and protein folding, and energy metabolism [83]. Subsequent investigations on another bioactive molecule extracted from S. nanolobata, i.e., sinularin ((9E)-13-hydroxy-4,9,13-trimethyl-17-methylidene-5,15-dioxatricyclo[12.3.1.04,6] octadec-9-en-16-one; Figure 11B), confirmed this compound as an additional potential anti-cancer molecule. In fact, sinularin promoted the inhibition of growth and motility, the block at G2/M cell cycle phase, and the onset of apoptosis in A2058 melanoma cells and proteomic analyses found

modificationsFigure 10. (A in) Athe specimen levels of of Sarcophytononcosuppressor crassocaule and softapoptosis-associ coral © Leen Vanated Ofwegen proteins (CC consistent BY-NC-SA with the 4.0);observed https:// phenotypicwww.marinespecies.org changes [84]./ (B ) 2D structure of 13-acetoxysarcocrassolide; https://pubchem. ncbi.nlm.nih.gov/compound/21775908.

FigureFigure 11. 11. (AA)) 2D 2D structure structure of of 11-dehydrosinulariolid 11-dehydrosinulariolide;e; http://www.chemspider.com/Chemical- http://www.chemspider.com/Chemical- Structure.10470882.htmlStructure.10470882.html..(B )B 2D) structure2D structure of sinularin; of sinularin; https://pubchem.ncbi.nlm.nih.gov https://pubchem.ncbi.nlm.nih.gov//compound / Sinularin#sectioncompound/ Sinularin#section=2D-Structure.=2D-Structure. Clavularia koellikeri (Dean, 1927; Anthozoa, Alcyonacea, Clavulariidae; Figure 12) is a widespread Clavularia koellikeri (Dean, 1927; Anthozoa, Alcyonacea, Clavulariidae; Figure 12) is a species throughout the Indian and Pacific oceans, from the Red Sea and East Africa across the widespread species throughout the Indian and Pacific oceans, from the Red Sea and East Africa Indo-Pacific to Micronesia, being a typical inhabitant of the Great Barrier Reef. Soft corals belonging to across the Indo-Pacific to Micronesia, being a typical inhabitant of the Great Barrier Reef. Soft this taxonomic genus are an important resource of secondary metabolites, mainly terpenoids, which corals belonging to this taxonomic genus are an important resource of secondary metabolites, are endowed with unique structures and a wide range of biological activities [85,86]. Among them, mainly terpenoids, which are endowed with unique structures and a wide range of biological clavukoellian A and clavukoellian D-related 4-O-deacetylparalemnolin D exerted an anti-locomotory activities [85,86]. Among them, clavukoellian A and clavukoellian D-related 4-O- effect on human umbilical vein endothelial (HUVEC) cells in wound healing assays, thus showing deacetylparalemnolin D exerted an anti-locomotory effect on human umbilical vein endothelial interesting anti-angiogenetic activity that merits further investigation [87]. (HUVEC) cells in wound healing assays, thus showing interesting anti-angiogenetic activity that merits further investigation [87].

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Figure 12. An image of Clavularia koellikeri; Wang et al. (2019) [87].

Figure 12. An image of Clavularia koellikeri ; Wang etet al.al. (2019) [87]. [87]. Cyanea capillata (Linnaeus, 1758; Scyphozoa, Semaeostomeae: Cyaneidae; Figure 13), commonly known as lion’s mane jellyfish, populates the cold waters of the more northerly areas CyaneaCyanea capillata capillata(Linnaeus, (Linnaeus, 1758; 1758; Scyphozoa, Scyphozoa, Semaeostomeae: Semaeostomeae: Cyaneidae; Cyaneidae; Figure 13Figure), commonly 13), of the North and Arctic seas and the Pacific and Atlantic oceans, also finding itself along the coasts knowncommonly as lion’s known mane as jellyfish,lion’s mane populates jellyfish, the populates cold waters the cold of the waters more of northerly the more areas northerly of the areas North of Southeast China [88–90]. This organism represents the largest jellyfish known to date, with an andof the Arctic North seas and and Arctic the Pacific seas and and the Atlantic Pacific and oceans, Atlantic also findingoceans, also itself finding along theitself coasts along of the Southeast coasts umbrella from which originate a large number of reddish-brown or yellowish tentacles long up Chinaof Southeast [88–90]. China This organism [88–90]. This represents organism the represen largest jellyfishts the largest known jellyfish to date, known with anto date, umbrella with froman to 30 m and wide up to one meter, endowed with considerable amounts of nematocysts that whichumbrella originate from a which large numberoriginate of a reddish-brown large number orof yellowishreddish-brown tentacles or yellowish long up to tentacles 30 m and long wide up up release toxins dangerous for humans [91]. It is widely acknowledged that jellyfishes are important toto one 30 meter,m and endowed wide up with to one considerable meter, endowed amounts with of nematocystsconsiderable that amounts release of toxins nematocysts dangerous that for sources of many different bioactive molecules. This species, in particular, is a good producer of humansrelease [ 91toxins]. It isdangerous widely acknowledged for humans [91]. that jellyfishesIt is widely are acknowledged important sources that jellyfishes of many di ffareerent important bioactive antioxidants conceivably aimed to protect its supercoiled DNA from oxidative damage. In fact, molecules.sources of This many species, different in particular, bioactive ismolecules. a good producer This species, of antioxidants in particular, conceivably is a good aimed producer to protect of the molecular cloning and recombinant protein production of peroxiredoxin-, thioredoxin-, and itsantioxidants supercoiled conceivably DNA from oxidative aimed to damage.protect its In supercoiled fact, the molecular DNA from cloning oxidative and recombinant damage. In protein fact, Cu/Zn superoxide dismutase (SOD) homologues was successfully performed starting from cDNA productionthe molecular of peroxiredoxin-, cloning and recombinant thioredoxin-, protein and Cuproduction/Zn superoxide of peroxiredoxin-, dismutase (SOD)thioredoxin-, homologues and libraries of C. capillata tentacles [92–94]. In addition, while using the same molecular source a wasCu/Zn successfully superoxide performed dismutase starting (SOD) from homologues cDNA libraries was successfully of C. capillata performedtentacles starting [92–94 ].from In addition,cDNA recombinant protein with potent anti-bacterial activity was expressed and purified [95]. Wang et whilelibraries using of the C. samecapillata molecular tentacles source [92–94]. a recombinant In addition, protein while withusing potent the same anti-bacterial molecular activity source was a al. [96] demonstrated the promoting effect of C. capillata’s tentacle extracts on the migratory expressedrecombinant and purifiedprotein with [95]. potent Wang et anti-bacterial al. [96] demonstrated activity was the promotingexpressed and effect purified of C. capillata [95]. Wang’s tentacle et behavior of human umbilical vein endothelial cells in scratch wound healing assays, thus extractsal. [96] on demonstrated the migratory the behavior promoting of human effect umbilical of C. capillata vein endothelial’s tentacle cells extracts in scratch on the wound migratory healing considering this mixture a good candidate as wound healing regulator, improving tissue repair assays,behavior thus of considering human umbilical this mixture vein a goodendothelial candidate cells as in wound scratch healing wound regulator, healing improving assays, thus tissue in unfavorable conditions. In a following study, they isolated and purified a C. capillata-specific repairconsidering in unfavorable this mixture conditions. a good In candidate a following as study,wound they healing isolated regulator, and purified improving a C. capillata tissue -specificrepair granulin, the 5782 Da-bioactive component of the extract, able to trigger cell locomotion, and also granulin,in unfavorable the 5782 conditions. Da-bioactive In acomponent following study, of the they extract, isolated able toand trigger purified cell alocomotion, C. capillata-specific and also up-regulation of cyclin B1 and D1 and cell growth, via the activation of ERK1/2 and MAPK signal up-regulationgranulin, the of5782 cyclin Da-bioactive B1 and D1 component and cell growth, of the ex viatract, the able activation to trigger of ERK1cell locomotion,/2 and MAPK and signalalso cascade [97]. cascadeup-regulation [97]. of cyclin B1 and D1 and cell growth, via the activation of ERK1/2 and MAPK signal cascade [97].

Figure 13. An indivividual of Cyanea capillata Alexander Semenov; https://www.flickr.com/. Figure 13. An indivividual of Cyanea capillata © Alexander Semenov; https://www.flickr.com/.

RhopilemaFigure 13 esculentum. An indivividual(Kishinouye, of Cyanea 1891; capillata Scyphozoa, © Alexander Rhizostomeae: Semenov; https://www.flickr.com/. Rhizostomatidae; Figure 14) Rhopilema esculentum (Kishinouye, 1891; Scyphozoa, Rhizostomeae: Rhizostomatidae; Figure is a jellyfish species that inhabits pelagic and tropical environments, in particular populating the 14) is a jellyfish species that inhabits pelagic and tropical environments, in particular populating NorthwesternRhopilema Pacific esculentum area, the(Kishinouye, coasts of China1891; Scyphozoa, and Japan, Rhizostomeae: the Yellow Sea Rhizostomatidae; and the Bohai Sea. Figure It can the Northwestern Pacific area, the coasts of China and Japan, the Yellow Sea and the Bohai Sea. It achieve14) is a the jellyfish diameter species of several that inhabits cm and thepelagic weight and of tropical 30 kg. This environments, organism is commonlyin particular known populating as flame can achieve the diameter of several cm and the weight of 30 kg. This organism is commonly the Northwestern Pacific area, the coasts of China and Japan, the Yellow Sea and the Bohai Sea. It can achieve the diameter of several cm and the weight of 30 kg. This organism is commonly

Molecules 2020, 25, x FOR PEER REVIEW 13 of 42 Molecules 2020, 25, 2471 13 of 40 known as flame jellyfish due to its mouth-arms that are endowed with abundant filamentous and large spindle-shaped appendices bearing undefined terminal clubs. R. esculentum is an eatable jellyfishspecies, due widespread to its mouth-arms in the market that are for endowed its pleasant with flavour abundant and filamentous its lipid profile and large rich spindle-shapedof fatty acids appendicesand glycerophospholipids bearing undefined that terminal makes it clubs.a potentR.ial esculentum healthy foodis an for eatable human species, consumption widespread and an in thealiment market that for is its suitable pleasant for flavour some commercial and its lipid fishes profile and rich crustaceans of fattyacids [98,99]. and In glycerophospholipidsaddition, significant thatinsecticidal makes it properties a potential were healthy demonstrated food for human in th consumptione full proteinous and anvenom aliment of this that invertebrate, is suitable for somethereby commercial being a fishespotential and component crustaceans of [98environmentally-friendly,99]. In addition, significant insecticides insecticidal [100], propertieswhereas werepeptides demonstrated that were obtained in the full from proteinous R. esculentum venom’s hydrolysates of this invertebrate, exhibited therebyangiotensin being I-converting a potential componentenzyme (ACE)-inhibitory of environmentally-friendly and, therefore, insecticides anti-hyp [ertensive100], whereas and anti-oxidant peptides that properties were obtained [101,102]. from R. esculentumResearch’s interest hydrolysates has recently exhibited been angiotensin focused I-converting on the tissue enzyme repair (ACE)-inhibitory properties of the and, collagen therefore, anti-hypertensivecomponent extracted and anti-oxidant from this propertiesorganism. [101 Felician,102]. Researchet al. [103] interest demonstrated has recently the been ability focused of on thepreparations tissue repair of properties collagen ofpeptides, the collagen which component were obtained extracted through from thiseither organism. collagenaseFelician II- or et papain al. [103 ] demonstrateddigestion, to theinduce ability the ofmotility preparations of human of collagenvein umbilical peptides, cells which in scratch were wound obtained healing through assays. either collagenaseMoreover, II-the or same papain preparations digestion, were to induce applied the to motility the back of of human wounded vein mice umbilical and the cells results in scratch that woundwere obtained healing assays. in vivo Moreover,confirmed thetheir same regenerative preparations properties were applied determining to the a back significant of wounded decrease mice andof the resultswounded that area, were especially obtained inin vivocase confirmedof treatment their with regenerative collagenase properties II-digested determining samples. a significantImmunohistochemical decrease of the analyses wounded of area,the areas especially of the in in-progress case of treatment tissue reconstitution with collagenase showed II-digested the samples.up-regulation Immunohistochemical of TGFβ1 and β analyses-FGF, known of the factors areas of that the are in-progress involved tissue in the reconstitution inflammatory showed and theangiogenetic up-regulation phases of TGF ofβ skin1 and repairβ-FGF, whose known acceleration factors that by are the involved peptide in themixture inflammatory makes it anda angiogeneticpromising product phases offor skin wound repair repair. whose Noteworthy, acceleration the by thecollagen peptide component mixture makes extracted it a promisingfrom R. productesculentum for wound has also repair. been Noteworthy, the object theof collagenstudies sh componentowing its extractedremarkable from anti-photoagingR. esculentum has and also beenhemostatic the object properties, of studies showingand its itssuitability remarkable and anti-photoaging safety for use andas hemostatica scaffold properties,supporting and the its suitabilitychondrogenic and safety differentiation for use as of a sca adultffold stem supporting cells in the cartilage chondrogenic tissue engineering, differentiation which of adult merits stem cellsfurther in cartilage exploitation tissue [104–106]. engineering, which merits further exploitation [104–106].

FigureFigure 14. A14. specimen A specimen of Rhopilema of esculentumRhopilema. ©esculentumBill Abbott. (CC© BY-SABill Abbott 2.0); https: (CC//en.wikipedia.org BY-SA 2.0); /. https://en.wikipedia.org/. 4. Mollusca 4. MolluscaUroteuthis (Doryteuthis) singhalensis (Ortmann, 1891; Cephalopoda, Myopsida: Loliginidae, FigureUroteuthis 15A) is a ( demersalDoryteuthis species,) singhalensis which (Ortmann, is typical 1891; of subtropical Cephalopod anda, tropicalMyopsida: areas, Loliginidae, especially ofFigure the Indo-West 15A) is a demersal Pacific zone, species, representing which is typical one of subtropical the commercially-important and tropical areas, especially squid species of inthe India. Indo-West This cephalopodPacific zone, hasrepresenting been used one as of a sourcethe commercially-important of the polysaccharide squid chitosan, species which in exhibitsIndia. This powerful cephalopod antioxidant has been and usedanticoagulant as a source of the properties, polysaccharide and ofchitosan, invertebrate which exhibits collagen, whichpowerful is a antioxidant good alternative and anticoagulant to the mammalian properties, counterpart and of invertebrate being the collagen, animal’s which skin is availablea good inalternative large amount to the as mammalian fish markets’ counterpart left-over [being107,108 the]. animal’s Veeruraj skin et al.available [109] recently in large reportedamount as the applicationfish markets’ of aleft-over biocomposite [107,108]. matrix Veeruraj that iset constitutedal. [109] recently by the reported radical scavenging-carotenoidthe application of a astaxhantinbiocomposite ((6S matrix)-6-hydroxy-3-[( that is constituted1E,3E,5E,7E,9E,11E,13E,15E,17E by the radical scavenging-carotenoid)-18-[(4S)-4-hydroxy-2,6,6- astaxhantin trimethyl-3- ((6S)-6- oxocyclohexen-1-yl]3,7„12,16-tetramethyloctadeca-1,3,5,7,9,11,13,15,17-nonaenyl]-2,4,4-trimethylcyclohydroxy-3-[(1E,3E,5E,7E,9E,11E,13E,15E,17E)-18-[(4S)-4-hydroxy-2,6,6- trimethyl-3- hex-2-en-1-one;oxocyclohexen-1-yl]3,7,,12,16-tetramethyloct Figure 15B incorporated into a scaadeca-1,3,5,7,9,11,13,15,17-nonaenyl]-2,4,4-ffold of collagen, both extracted from U. singhalensis, fortrimethylcyclo the healing hex-2-en-1-one; of dermal wounds Figure of 15B rats incorporatedin vivo. The into obtained a scaffold data of collagen, demonstrated both extracted a higher woundfrom U. contraction singhalensis, rate for and the tensile healing strenght of dermal and a wounds faster epithelization of rats in vivo. time The at theobtained excision data site, withdemonstrated an increased a higher deposition wound of contraction newly synthesized rate and tensile collagen strenght in the and damaged a faster epithelization area following fibroblasttime at the proliferation excision site, and with activation. an increased Histopathological deposition observationsof newly synthe confirmedsized collagen the remarkable in the

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damaged area following fibroblast proliferation and activation. Histopathological observations healingconfirmed properties the remarkable of the material, healing additionally properties puttingof the material, the onset additionally of a pronounced putting and the accelerated onset of a revascularizationpronounced and of theaccelerated tissue in evidence.revascularization The obtained of the cumulative tissue in evidence. results highlighted The obtained the cumulative potentiality of thisresults composite highlighted sponge the in potentiality providing aof superior-quality this composite woundsponge dressing, in providing also ina termssuperior-quality of stability, low-cost,wound biodegradability, dressing, also in terms and biocompatibility, of stability, low-cost, which biodegradability, may have additional and biocompatibility, applications in which tissue engineeringmay have and additional regenerative applications medicine. in tissue engineering and regenerative medicine.

FigureFigure 15. (15.A) AA) specimen A specimen of Uroteuthis of Uroteuthis singhalensis singhalensis; http://;www.borneomolluscs.com http://www.borneomolluscs.com..(B) 2D structure B) 2D of astaxanthin;structure of https:astaxanthin;//pubchem.ncbi.nlm.nih.gov https://pubchem.ncbi./compoundnlm.nih.gov/compound/Ast/Astaxanthin#sectionaxanthin#section==2D-Structure .2D- Structure. Sepia kobiensis (Hoyle, 1885; Cephalopoda, Sepiida: Sepiidae; Figure 16), also known as kobi cuttlefish,Sepia is a kobiensis benthic species(Hoyle, distributed1885; Cephalopoda, in the Indo-West Sepiida: Pacific Sepiidae; area Figure from Japan 16), also to the known Persian as kobi Gulf. It iscuttlefish, an eatable is organism a benthic with species a nutritional distributed value in thatthe Indo-West is very close Pacific to that area of fin from fish, Japan although to the the Persian various parametersGulf. It is tested an eatable (i.e., protein,organism moisture with a nutritional and ash contents, value that and is total very lipid, close cholesterolto that of fin and fish, triglyceride although levels)the various vary in theparameters different tested parts of (i.e., the protein, body, head, moisture arm,tentacle, and ash andconten coatts, [ 110and]. total Additionally, lipid, cholesterol chitosan andand collagen triglyceride of this levels) cephalopod vary in attractedthe different keen parts research of the interest.body, head, The arm, first tentacle, component and coat showed [110]. a numberAdditionally, of therapeutically-interesting chitosan and collagen properties,of this cephalopod i.e., anti-bacterial attracted keen and research anti-microbial, interest. as The well first as anti-oxidantcomponent and showed anti-lipidemic a number activities, of therapeutically-inte the latter being successfullyresting properties, applied toi.e.,in anti-bacterial vivo rat models and of carbonanti-microbial, tetrachloride-induced as well as anti-oxidant hepatic damage and [anti-lipidemic111–114]. Ramasamy activities, and the Shanmugan latter being [115 successfully] prepared a bioscaappliedffold to with in vivo collagen rat andmodels chitosan of carbon extracted tetrachloride-induced from S. kobiensis andhepatic tested damage it in an [111–114].in vivo rat woundRamasamy healing and model. Shanmugan They found [115] that prepared the hybrid a bioscaffold film determined with collagen an increased and chitosan wound reductionextracted andfrom fibroblast S. kobiensis proliferation and tested and it in di ffanerentiation in vivo rat towardswound healing myofibroblasts, model. They thereby found determining that the hybrid the up-regulationfilm determined of collagen an increased deposition wound in the reduction damaged and tissue. fibroblast Moreover, proliferation the presence and ofdifferentiation chitosan also guaranteedtowards amyofibroblasts, free radical-scavenging thereby determining action that protected the up-regulation the proliferating of collagen cytotypes, deposition i.e., epithelial in the cellsdamaged and fibroblasts, tissue. Moreover, from the cytotoxic the presence influence of chit of reactiveosan also oxygen guaranteed species a (ROS). free radical-scavenging Thus, the merging of theaction beneficial that protected effects of the the proliferating two macromolecules cytotypes, makes i.e., epithelial this material cells a and promising fibroblasts, candidate from asthe a dressingcytotoxic that influence supports of a fasterreactive wound oxygen repair species process. (ROS). Thus, the merging of the beneficial effects of the two macromolecules makes this material a promising candidate as a dressing that supports a faster wound repair process.

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Figure 16. A specimen of Sepia kobiensis. © Maynard Hogg (CC BY-NC-SA 2.0); https://www.flickr.com/. FigureFigure 16. 16.A specimen A specimen of Sepia kobiensisof Sepia. © kobiensisMaynard. Hogg© Maynard (CC BY-NC-SA Hogg 2.0); (CC https: BY-NC-SA//www.flickr.com 2.0); /. https://www.flickr.com/.Successful results were also achieved utilizing collagen preparations from the local cuttlefish SepiaSuccessful officinalis results (Linnaeus, were also1758; achieved Cephalopoda, utilizing Sepiida: collagen Sepiidae; preparations Figure from 17), the anlocal invertebrate cuttlefish thatSepia offiiscinalis widelySuccessful(Linnaeus, distributed results 1758; in were the Cephalopoda, Western also achieved Mediterran Sepiida: utilizingean Sepiidae; collagen Sea, which Figure preparations can 17 ),be an also invertebrate fr omfound the along local that cuttlefishthe is West widely distributedAfricanSepia officinalis coasts. in the (Linnaeus,Collagen Western Mediterraneanhydrogels 1758; Cephalopoda, were Sea, applied which Sepiida: to can the beSepiidae; excised also found Figureskin along of 17),female the an West invertebrate Wistar African rats coasts.thatand Collagenprovenis widely to hydrogels distributed be a good were qualityin the applied Western dressing to the Mediterran promoting excised skinean tissue ofSea, female restoration which Wistar can tobe rats levelsalso and found that proven arealong comparable to the be aWest good qualitytoAfrican those dressing coasts.of a commercial promotingCollagen hydrogels tissuevegetable restoration wereextract-based applied to levels toemulsion thatthe excised are used comparable skin as reference.of female to those InWistar of particular, a commercial rats and S. vegetableofficinalisproven to extract-based’s becollagen a good films quality emulsion induced dressing used a quick as promoting reference. contraction tissue In particular, and restoration closureS. oofffi to cinalisthe levels wound,’s collagenthat withare comparable films an absence induced aofto quick inflammationthose contraction of a commercial signs and and, closure vegetable on ofthe the other extract-based wound, hand, with evidence emulsion an absence of neo-vesselused of inflammationas reference. growth andIn signs particular, active and, ECM on S. the otherofficinalisdeposition hand,’s evidence collagenat the histological offilms neo-vessel induced analysis growth a quick [116]. and contraction Of active note, ECM collagen and deposition closure preparations of at the the wound, histological of this with cephalopodanalysis an absence [are116 ]. Ofofwidely note, inflammation collagenused for preparations signsdiverse and, application, on of the this other cephalopod such hand, as evidencegelling are widely agent,of neo-vessel used thickener, for diverse growth foaming application, and activeagent, suchECM and as gellingemulsifierdeposition agent, atin thickener, thefood histological industries foaming analysis or agent, natural [116]. and bioa emulsifierOf note,ctive collagenfiller in food promoting preparations industries im orplant of natural this osteointegration cephalopod bioactive are filler promotingwidely[117,118]. used implant for osteointegrationdiverse application, [117 ,118such]. as gelling agent, thickener, foaming agent, and emulsifier in food industries or natural bioactive filler promoting implant osteointegration [117,118].

FigureFigure 17. A17. specimen A specimen of Sepia offiofcinalis Sepia. © Hansofficinalis Hillewaert. © Hans (CC BY-SA Hillewaert 4.0); http: (CC//www.marinespecies. BY-SA 4.0); orghttp://www.marinespecies.org/.Figure/. 17. A specimen of Sepia officinalis. © Hans Hillewaert (CC BY-SA 4.0); http://www.marinespecies.org/. MytilusMytilus galloprovincialis galloprovincialis(Lamarck, (Lamarck, 1819; 1819; Bivalvia,Bivalvia, Mytilida: Mytilidae; Mytilidae; Figure Figure 18A) 18A) isis a widely a widely distributeddistributedMytilus bivalve bivalve galloprovincialis mollusk mollusk populating populating(Lamarck, 1819; thethe MediterraneanMedite Bivalvia,rranean Mytilida: and and theMytilidae; the Black Black Sea, Sea,Figure the the 18A)eastern eastern is aAtlantic widely Atlantic oceandistributedocean from from Morocco bivalve Morocco tomollusk theto the British populating British Isles, Isles, but the also butMedite presentalsorranean present in Japan, and in theJapan, California, Black California, Sea, South the eastern Africa,South Atlantic southernAfrica, Australia,oceansouthern from andAustralia, Morocco New Zealand. and to Newthe ItBritish Zealand. is a lamellibranch Isles, It isbut a lamellibranchalso mollusk, present which in mollusk, Japan, is, equipped California,which is, with equipped South lamellar Africa, with gills thatsouthernlamellar absorb gills Australia, oxygen that andabsorb and simultaneously Newoxygen Zealand. and retainsimultaneously It is fooda lamellibranch for feeding. retain food Therefore,mollusk, for feeding. which being Therefore,is, a filtering equipped organism,being with a itlamellarfiltering plays an organism,gills indispensable that absorb it plays role oxygen atan ecological indispensable and simultaneously level, role as marine at ecological retain purifier food level, and for it asfeeding. is widelymarine Therefore, used purifier to evaluate and being it is thea efffilteringwidelyects of stress usedorganism, environmentalto evaluate it plays an conditionthe indispensable effects [119 of,120 strerole]. Thesess at environmentalecological invertebrates level, arecondit as endowedmarineion [119,120].purifier with an and excellentThese it is nutritionalinvertebrateswidely used value, aretoowing endowedevaluate to their withthe remarkable aneffects excellent of richness nutritstressional environmental in proteins,value, owing carbohydrates, conditto theirion remarkable [119,120]. essential richness vitamins,These minerals,invertebratesin proteins, and carbohydrates, di arefferent endowed varieties withessential of an fatty excellent vitamins, acids thatnutrit minerals, playional beneficial value, and different owing roles forto varieties their human remarkable of health fatty and,acids richness for that this reason,inplay proteins, beneficial they are carbohydrates, in roles great for demand human essential on health the vitamins, international and, forminerals, this market reason, and [ 121different they–123 are]. varietiesRapana in great venosa of demand fatty(Valenciennes, acids on thatthe 1846;play Gastropoda, beneficial roles Neogastropoda: for human health Muricidae; and, Figurefor this 18 reason,B) is a predatorythey are in benthic great speciesdemand of on tropical the

international market [121–123]. Rapana venosa (Valenciennes, 1846; Gastropoda, Neogastropoda: Muricidae; Figure 18B) is a predatory benthic species of tropical environments typical of the

Molecules 2020, 25, 2471 16 of 40 environments typical of the Northwest Pacific area and introduced in the Atlantic, Mediterranean, and Black Sea [124,125]. It shows a variable color from grey to orange-brown or blonde, with darker brown dashes on the spiral ribs, and it is a widely consumed and commercially valuable mollusk in China. Recently, considerable interest has been paid to the concentrations of metals or pollutants, even radioactive, which R. venosa might contain, for an assessment of environmental monitoring and also the protection of human health, since this species is eatable and frequently consumed [126,127]. The anti-viral, anti-bacterial, and anti-cancer potentials of the hemocyanins obtained from the snail hemolymph have been extensively analyzed and demonstrated, thereby being promising components for formulations of anti-microbial and anti-tumoral vaccines [128–130]. Moreover, R. venosa’s shell was utilized to fabricate bionic coupling-layered ceramic/Al composites with improved lightweight and mechanical strength properties [131]. Being the gastropod a common predator of the mollusk M. galloprovincialis, Badiu and coworkers [132,133] examined in parallel the effect of lipid extracts and aminoacidMolecules 2020 preparations, 25, x FOR PEER from REVIEWM. galloprovincialis and R. venosa in in vivo wound healing tests. 17 of 42

FigureFigure 18. 18.( AA)) AA groupgroup ofof Mytilus galloprovincialis bivalves. ©© PillonPillon Robe Robertrt (CC (CC BY-NC-SA BY-NC-SA 4.0); 4.0); http:http://www.marinespecies.org///www.marinespecies.org/.( BB)) ImagesImages of the shell shell of of RapanaRapana venosa venosa. .©© H.H. Zell Zell (CC (CC BY-SA BY-SA 3.0); 3.0); https:https://en.wikipedia.org/.//en.wikipedia.org/.( C) 2DC structure) 2D of eicosapentaenoicstructure of acid; https:eicosapentaenoic//pubchem.ncbi.nlm.nih. acid; govhttps://pubchem.ncbi.nlm.nih./compound/Eicosapentaenoic-acid#section gov/compound/E=2D-Structureicosapentaenoic-acid#section=2D-Structure..(D) 2D structure of docosahexenoic D) acid;2D https:structure//pubchem.ncbi.nlm.nih.gov of docosahexenoic/compound acid; /Docosahexaenoic-acid#sectionhttps://pubchem.ncbi.nlm.nih.gov/compound/=Structures.(E) 2D structureDocosahexaenoic-acid#section=Structures. of arachidonic acid; https://pubchem.ncbi.nlm.nih.gov E) 2D structure/compound of/Arachidonic-acid#section arachidonic acid; = 2D-Structurehttps://pubchem.ncbi.nlm.nih.gov/compound. /Arachidonic- acid#section=2D-Structure.

Pinctada martensii (Dunker, 1880; Bivalvia, Ostreida: Margaritidae; Figure 19), also known as Pinctada imbricata (Röding, 1798), is a mollusk that is characteristic of benthic environments in the Atlantic and Indo-West Pacific areas and in the intertidal rocky regions of the Mumbai coast, to date listed among the most invasive species in the Mediterranean Sea [134]. It is one of the most important bivalves for the production of marine pearls all over the world and especially of the round «South China Sea pearls» that are typical of southern China. Despite the remarkable performance observed in the 90s [135], the rearing of this species subsequently decreased also due to environmental deterioration [136]. For this reason, this organism is commonly used to study the levels of environmental pollution, the effects of pollutants on the health of marine organisms, and the influence of various environmental stressors, such as changes in water temperature due to climate modification, on aquaculture yields. Proteins extracted from the meat and the nacre of this invertebrate have been shown to exert various functions, such as anti-microbial, anti- hypertensive, and osteogenetic [137–139]. Yang et al. [140] studied the effect of preparations of small molecular peptides from P. martensii’s mantle, i.e., the shell-producing organ, on skin wound repair in in vivo rat models. They demonstrated that the treatment was able to promote

Molecules 2020, 25, 2471 17 of 40

Lipid extracts, containing slightly different complex mixtures with a high cholesterol content and good although varying amounts of several saturated and polyunsaturated fatty acids (PUFAs), including ω-3 eicosapentaenoic ((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17- pentaenoic acid, Figure 18C) and docosahexenoic acids, ((4Z,7Z,10Z,13Z,16Z,19Z)-docosa- 4,7,10,13,16,19-hexaenoic acid, Figure 18D) and, only for R.venosa, ω-6 arachidonic acid ((5Z,8Z,11Z,14Z)-icosa-5,8,11,14-tetraenoic acid, Figure 18E), were applied to skin burns in in vivo rat models in order to examine their potential healing activity. The results showed an acceleration of healing time when compared to a commercial fish oil-containing ointment used as reference, with complete re-epithelization and the deposition of dense ECM, re-vascularization, and absence of edema and inflammation Preparations from M. galloprovincialis exhibited a slightly faster recovery, thus suggesting the more active involvement of eicosapentanoic and docosahexenoic acids, which are abundant in its extract, in the wound healing process [132]. The same experimental study was also performed after application of aminoacid mixtures from protein extracts of M. galloprovincialis and R. venosa, which showed significant differences regarding aminoacidic distribution and content. Additionally, in this case, the correct formation of new epithelium and well-structured vessels, the improved collagen deposition and the reduced burn edema and leukocyte infiltration were observed [133]. This is likely due to the preponderance of thiol-containing aminoacids in the mixture, which exert anti-oxidant action preventing excessive lipid peroxidation and other aspects of the oxidative stress that are linked to the healing process. Of note, the damaged rat tissues healed faster when treated with R. venosa’s extracts, probably because of the beneficial effects of Leu, Lys, Thr, and Pro essential aminoacids, only present in these preparations. Pinctada martensii (Dunker, 1880; Bivalvia, Ostreida: Margaritidae; Figure 19), also known as Pinctada imbricata (Röding, 1798), is a mollusk that is characteristic of benthic environments in the Atlantic and Indo-West Pacific areas and in the intertidal rocky regions of the Mumbai coast, to date listed among the most invasive species in the Mediterranean Sea [134]. It is one of the most important bivalves for the production of marine pearls all over the world and especially of the round «South China Sea pearls» that are typical of southern China. Despite the remarkable performance observed in the 90s [135], the rearing of this species subsequently decreased also due to environmental deterioration [136]. For this reason, this organism is commonly used to study the levels of environmental pollution, the effects of pollutants on the health of marine organisms, and the influence of various environmental stressors, such as changes in water temperature due to climate modification, on aquaculture yields. Proteins extracted from the meat and the nacre of this invertebrate have been shown to exert various functions, such as anti-microbial, anti-hypertensive, and osteogenetic [137–139]. Yang et al. [140] studied the effect of preparations of small molecular peptides from P. martensii’s mantle, i.e., the shell-producing organ, on skin wound repair in in vivo rat models. They demonstrated that the treatment was able to promote robust collagen deposition and re-epithelialization, thus accelerating wound healing, via the up-regulation of EGF, TGF-β1, and cyclin D1. The increase of CD31+ cytotypes in the tissue was indicative of enhanced neovascularization, whereas the up-regulation of IL-10 and the down-regulation of IL-6 was a sign of inhibition of the inflammatory response. A hypothetical explanation of the observed effect was based upon the high content in glutamic acid, glycine, cysteine, and phenylalanine of the preparations, which guarantee the nitrogen balance that is necessary for the continuous protein synthesis during angiogenesis, fibroblast growth, collagen deposition, and wound remodeling. In addition, Gln-Leu and Asp-Leu pairs, which are very common in the peptides, might act as radical scavengers, and Arg is known to promote collagen accumulation. Additionally, hydrophobic aminoacids, that improve immunity, accounted for more than half of the total aminoacids in the peptides. Interestingly, a similar ameliorated healing quality was obtained by the gastric administration of P. martensii’s mantle peptide preparations to mouse skin wound models [140]. Molecules 2020, 25, x FOR PEER REVIEW 18 of 42

robust collagen deposition and re-epithelialization, thus accelerating wound healing, via the up- regulation of EGF, TGF-β1, and cyclin D1. The increase of CD31+ cytotypes in the tissue was indicative of enhanced neovascularization, whereas the up-regulation of IL-10 and the down- regulation of IL-6 was a sign of inhibition of the inflammatory response. A hypothetical explanation of the observed effect was based upon the high content in glutamic acid, glycine, cysteine, and phenylalanine of the preparations, which guarantee the nitrogen balance that is necessary for the continuous protein synthesis during angiogenesis, fibroblast growth, collagen deposition, and wound remodeling. In addition, Gln-Leu and Asp-Leu pairs, which are very common in the peptides, might act as radical scavengers, and Arg is known to promote collagen accumulation. Additionally, hydrophobic aminoacids, that improve immunity, accounted for more than half of the total aminoacids in the peptides. Interestingly, a similar ameliorated healing quality was obtained by the gastric administration of P. martensii’s mantle peptide preparations Molecules 2020, 25, 2471 18 of 40 to mouse skin wound models [140].

FigureFigure 19. 19.Images Images of of the the shellshell ofof PinctadaPinctada imbricataimbricata.. ©© NaturalNatural History History Museum Museum Rotterdam Rotterdam (CC (CC BY-NC-SABY-NC-SA 4.0); 4.0); http: http://www.marinespecies.org/.//www.marinespecies.org/. Euchelus asper (Gmelin, 1791; Gastropoda, Seguenziida: Chilodontaidae; Figure20) is an eatable Euchelus asper (Gmelin, 1791; Gastropoda, Seguenziida: Chilodontaidae; Figure 20) is an and shell-oriented benthic mollusc that is typical of tropical areas and distributed in the Indo-West eatable and shell-oriented benthic mollusc that is typical of tropical areas and distributed in the Pacific region, populating the west coast and south-east coast of India and the rocky beaches near the Indo-West Pacific region, populating the west coast and south-east coast of India and the rocky low tide points [141]. It is characterized by a thick and conoidal dull shell with brown, rosy, and black beaches near the low tide points [141]. It is characterized by a thick and conoidal dull shell with dots. Ether-soluble fractions of this organism were able to induce immunostimulated phagocytosis brown, rosy, and black dots. Ether-soluble fractions of this organism were able to induce in vitro and immunosuppressive activity in vivo [142,143], whereas methanol extracts have been immunostimulated phagocytosis in vitro and immunosuppressive activity in vivo [142,143], shown to exert an anti-osteoporotic effect in vivo, as indicated by the prominent anti-osteoclastogenic whereas methanol extracts have been shown to exert an anti-osteoporotic effect in vivo, as activity and reduction in bone loss induced by the lack of estrogen in a ovariectomised mice indicated by the prominent anti-osteoclastogenic activity and reduction in bone loss induced by model [144]. Agrawal et al. [145] demonstrated that methanol preparations from E. asper displayed the lack of estrogen in a ovariectomised mice model [144]. Agrawal et al. [145] demonstrated that strong anti-angiogenic potential in CAM assays due to their inhibitory role on mesodermal blood vessel methanol preparations from E. asper displayed strong anti-angiogenic potential in CAM assays migration and compartmentalization resulted, at least in part, from a decreased ECM remodelling by due to their inhibitory role on mesodermal blood vessel migration and compartmentalization MMPs. As a confirmation to this hypothesis, the methanol extracts were also found to down-regulate resulted, at least in part, from a decreased ECM remodelling by MMPs. As a confirmation to this the activity of MMP-2 and -9 in A549 lung cancer cells and, since these enzymes are involved in hypothesis, the methanol extracts were also found to down-regulate the activity of MMP-2 and - the process of cell migration and invasion, A549 cell motility in scratch wound healing assays was 9 in A549 lung cancer cells and, since these enzymes are involved in the process of cell migration consistently reduced. Moleculesand invasion, 2020, 25 ,A549 x FOR cell PEER motility REVIEW in scratch wound healing assays was consistently reduced. 19 of 42

FigureFigure 20. 20Images. Images of theof the shell shell of Euchelus of Euchelus asper .asper© H.. Zell© H. (CC Zell BY-SA (CC 3.0);BY-SA https: 3.0);//commons.wikimedia. https://commons. orgwikimedia.org..

HaliotisHaliotis diversicolor diversicolor(Reeve, (Reeve, 1846; 1846; Gastropoda,Gastropoda, Lepetellida:Lepetellida: Haliotidae; Haliotidae; Figure Figure 21) 21 )is is a asea sea snail snail thatthat can can be be found found in in temperatetemperate north-westernnorth-western Pa Pacificcific Ocean Ocean off off Japan,Japan, Taiwan, Taiwan, Vietnam, Vietnam, western western Australia,Australia, Indonesian Indonesian waters, waters, and and off Newoff New Caledonia Caledo [146nia]. [146]. Its coloration Its coloration is very is variable, very variable, normally reddish-brown,normally reddish-brown, scarlet, and scarlet, green inand the green external in the part, external while part, the while inner the surface inner is surface silvery is with silvery light greenwith andlight red green reflections. and red It isreflections. one of the It most is one important of the commercialmost important species commercial of molluscs, species especially of inmolluscs, the southern especially areas ofin China.the southern Like most areas mollusks, of China. H.Like diversicolor most mollusks, is widely H. diversicolor used as a bioindicator is widely forused the studyas a bioindicator of the effects for of the environmental study of the pollutantseffects of environmental or other types ofpollutants environmental or other stressors types of on environmental stressors on marine organisms at the cellular and/or molecular level [147–149]. The shell of this species, also called “shijueming”, has been considered to be a traditional Chinese medicine used in the past for eye diseases and skin lesions. Xu et al. reported an effective action of this traditional medicine towards the survival rate of human lens epithelial cells damaged by condition of oxidative stress [150], demonstrating its antioxidant and anti-inflammatory properties. Based on those, more recently Chen et al. [151] tested the effect of H. diversicolor’s shell powder on inflammation and healing rates of burn injured rats. Their study revealed that the preparation induced a noticeable decrease in neutrophil infiltration, probably due to the down- regulation of nitric oxid synthase, and a significant improvement in the wound healing process with the formation of a well-arranged collagen matrix stimulated by the increase of TGF-β1 content during the tissue remodeling phase.

Figure 21. Images of the shell of Haliotis diversicolor squamata. © Natural History Museum Rotterdam (CC BY-NC-SA 4.0); http://www.marinespecies.org/.

Euphausia superba (Dana, 1850; Malacostraca, Euphausiacea: Euphausiidae; Figure 22) is a pelagic krill that is distributed in the circumpolar belt between the Antarctic continent and the Polar Front [152]. This crustacean is considered to be an economic and ecological keystone [153]

Molecules 2020, 25, x FOR PEER REVIEW 19 of 42

Figure 20. Images of the shell of Euchelus asper. © H. Zell (CC BY-SA 3.0); https://commons. wikimedia.org.

Haliotis diversicolor (Reeve, 1846; Gastropoda, Lepetellida: Haliotidae; Figure 21) is a sea snail that can be found in temperate north-western Pacific Ocean off Japan, Taiwan, Vietnam, western Australia, Indonesian waters, and off New Caledonia [146]. Its coloration is very variable, normally reddish-brown, scarlet, and green in the external part, while the inner surface is silvery with light green and red reflections. It is one of the most important commercial species of

Moleculesmolluscs,2020 ,especially25, 2471 in the southern areas of China. Like most mollusks, H. diversicolor is widely19 of 40 used as a bioindicator for the study of the effects of environmental pollutants or other types of environmental stressors on marine organisms at the cellular and/or molecular level [147–149]. The marineshell of organisms this species, at the also cellular called and “shijueming”,/or molecular has level been [147 –considered149]. The shell to be of a this traditional species, alsoChinese called “shijueming”,medicine used has in been the consideredpast for eye to diseases be a traditional and skin Chinese lesions. medicine Xu et al. used reported in the an past effective for eye diseasesaction andof skinthis traditional lesions. Xu medicine et al. reported towards an e theffective survival action rate of thisof human traditional lens medicineepithelial towards cells damaged the survival by ratecondition of human of lensoxidative epithelial stress cells damaged[150], demonstrating by condition ofits oxidativeantioxidant stress and [150 anti-inflammatory], demonstrating its antioxidantproperties. and Based anti-inflammatory on those, more recently properties. Chen Based et al. on[151] those, tested more the recently effect of ChenH. diversicolor et al. [151’s] shell tested thepowder effect of onH. inflammation diversicolor’s shell and powderhealing onrates inflammation of burn injured and healing rats. Their rates study of burn revealed injured rats.that Theirthe studypreparation revealed induced that the a preparation noticeable induceddecrease ain noticeable neutrophil decrease infiltration, in neutrophil probably infiltration, due to the probablydown- dueregulation to the down-regulation of nitric oxid synthase, of nitric and oxid a significant synthase, andimprovement a significant in the improvement wound healing in the process wound healingwith the process formation with theof formationa well-arranged of a well-arranged collagen matrix collagen stimulated matrix by stimulated the increase by the of increase TGF-β1 of TGF-contentβ1 content during during the tissue the remodeling tissue remodeling phase. phase.

FigureFigure 21. 21.Images Images of theof shellthe shell of Haliotis of Haliotis diversicolor diversicolorsquamata. squamata.© Natural © Natural History MuseumHistory Museum Rotterdam (CCRotterdam BY-NC-SA (CC 4.0); BY-NC-SA http://www.marinespecies.org 4.0); http://www.marinespecies.org/./.

EuphausiaEuphausia superba superba(Dana, (Dana, 1850; 1850; Malacostraca, Malacostraca, Euphausiacea: Euphausiacea: Euphausiidae; Euphausiidae; Figure Figure 22) is 22) a pelagic is a krillpelagic that iskrill distributed that is distributed in the circumpolar in the circumpolar belt between belt the Antarcticbetween continentthe Antarctic and thecontinent Polar Front and [the152 ]. ThisPolar crustacean Front [152]. is considered This crustacean to be an is economic considered and to ecological be an economic keystone and [153 ecological] for its active keystone involvement [153] in carbon sequestration, iron and nitrogen recycling, ammonium absorption, and conversion of phytoplankton into animal proteins by connecting the different trophic levels [154,155]. The study of the cold adaptation mechanisms of E. superba at a transcriptome level has given interesting results that are useful for the design of new and improved recombinant enzyme variants that are endowed with adaptability to cold temperatures [156]. Much interest has been focused on the high nutritional content of this organism, due to the presence of proteins showing a higher biological value than milk or other animal counterparts, a remarkable amount of long-chain, in particular, ω-3, PUFAs, which makes it the second most popular source after fish oil, and all nine essential amino acids thus meeting the FAO/WHO requirements for human health [157,158]. For this reason, krill oil, which is rich in eicosapentaenoic and docosahexaenoic acids, vitamin A and E, and the carotenoid astaxanthin [159], is widely used as a food supplement also in light of its ability to reduce the risks of different pathologies, such as coronary heart diseases, diabetes, hypertension, osteoporosis, inflammation, autoimmune diseases, metabolic syndrome, tumors, and neurological diseases [160–163]. Costanzo et al. [164] have tested the ability of krill oil to ameliorate the state of intestinal inflammation in an in vitro model while using HT-29 and Caco-2 cell lines. In particular, when confluent cells, mimicking the intact barrier of the gut epithelium, were submitted to scratch wound healing assay in the presence of a mixture of the pro-inflammatory tumour necrosis factor-α and interferon-γ cytokines, the co-exposure to krill oil determined an increase of the rate of cell layer reconstitution. This result, complemented by the evidence of a better recovery of intercellular adhesion systems with a reduction of inflammation-related actin stress fibers, a decrease of cell death, and an impairment of adhesiveness and invasiveness of Molecules 2020, 25, x FOR PEER REVIEW 20 of 42

for its active involvement in carbon sequestration, iron and nitrogen recycling, ammonium absorption, and conversion of phytoplankton into animal proteins by connecting the different trophic levels [154,155]. The study of the cold adaptation mechanisms of E. superba at a transcriptome level has given interesting results that are useful for the design of new and improved recombinant enzyme variants that are endowed with adaptability to cold temperatures [156]. Much interest has been focused on the high nutritional content of this organism, due to the presence of proteins showing a higher biological value than milk or other animal counterparts, a remarkable amount of long-chain, in particular, ω-3, PUFAs, which makes it the second most popular source after fish oil, and all nine essential amino acids thus meeting the FAO/WHO requirements for human health [157,158]. For this reason, krill oil, which is rich in eicosapentaenoic and docosahexaenoic acids, vitamin A and E, and the carotenoid astaxanthin [159], is widely used as a food supplement also in light of its ability to reduce the risks of different pathologies, such as coronary heart diseases, diabetes, hypertension, osteoporosis, inflammation, autoimmune diseases, metabolic syndrome, tumors, and neurological diseases [160–163]. Costanzo et al. [164] have tested the ability of krill oil to ameliorate the state of intestinal inflammation in an in vitro model while using HT-29 and Caco-2 cell lines. In particular, when confluent cells, mimicking the intact barrier of the gut epithelium, were submitted to scratch wound healing assay in the presence of a mixture of the pro-inflammatory tumour necrosis factor- α and interferon-γ cytokines, the co-exposure to krill oil determined an increase of the rate of cell Moleculeslayer 2020reconstitution., 25, 2471 This result, complemented by the evidence of a better recovery20 of 40 intercellular adhesion systems with a reduction of inflammation-related actin stress fibers, a decrease of cell death, and an impairment of adhesiveness and invasiveness of luminal bacteria luminalon cells, bacteria strongly on cells,supports strongly the value supports of krill the valueoil as of a krillprospective oil as a prospective drug against drug human against chronic human chronicintestinal intestinal inflammation. inflammation. The beneficial The beneficial effect e ffofect krill of krilloil was oil was supposed supposed to be to bemostly mostly based based upon upon itsits richness richness in in this this lipid lipid component component due due toto thethe absenceabsence of ω-3-3 PUFAs PUFAs in in corn corn oil, oil, used used as as a anegative negative controlcontrol for for the the study. study.

Figure 22 22.. SpecimensSpecimens of of EuphausiaEuphausia superba superba.. ©© BrettBrett Wilks; Wilks; http://www.antarctica.gov.au/. http://www.antarctica.gov.au/.

5.5. Echinodermata Echinodermata TheThe phylum phylum Echinodermata, Echinodermata, gathering gathering invertebrates invertebrates that display that adisplay radial symmetry,a radial symmetry, incorporates aboutincorporates 6000 living about marine 6000 species livingaround marine the species globe. ar Inound recent the years, globe. echinoderms In recent years, have attractedechinoderms much attentionhave attracted as producers much ofattention biomolecules as producers with promising of biomolecules anti-microbial, with promising anti-protozoal, anti-microbial, anti-fungal, anti- and anticancerprotozoal, properties, anti-fungal, making and themanticancer potential properti candidateses, making for drug them development potential candidates and future therapeuticfor drug applicationsdevelopment [165 and–168 future]. Astropecten therapeutic indicus applications(Döderlein, [165–168]. 1888; Asteroidea, Astropecten Paxillosida: indicus (Döderlein, Astropectinidae; 1888; FigureAsteroidea, 23), also Paxillosida: called comb Astropectinidae; star or sand star, Figure is a starfish 23), al thatso called is distributed comb star in or the sand tropical star, environmentsis a starfish ofthat the Indo-Pacificis distributed area, in the and tropical in particular environments of the United of the Arab Indo-Pacific Emirates, Qatar,area, and Thailand, in particular Gulf of Mannar,of the andUnited Palk BayArab region. Emirates, It is aQatar, benthic Thailand, echinoderm Gulf that of isMannar, typically and found Palk in theBay sediments region. It of is the a benthic intertidal andechinoderm subtidal regions,that is typically where itfound feeds in using the sediments infaunal molluscs,of the intertidal and represents and subtidal a prototypic regions, where marine organismit feeds using to study infaunal the eff molluscs,ects of environmental and represents pollution a prototypic [169,170 marine]. Recently, organism Baveja to etstudy al. [171 the] effects isolated the AiP1 metalloproteinase from the coelomic fluid of A. indicus and demonstrated its fibrinolytic potential that is likely mediated by the von Willebrand factor (vWf)-like domain located in its central β sheet. Moreover, the ability of AiP1 to modulate the motile attitude of HaCaT keratinocytes and HEK293 embryonic kidney epithelial cells was checked through scratch wound healing assays demonstrating the additional AiP1-promoting effect on cell migration since vWf is known to sustain cell locomotory behavior. This result was confirmed by the evidence of the up-regulation of N-cadherin, indicative of EMT, in HEK293 cells. The wound healing activity exerted by AiP1, coupled with the increased cell proliferation reported in the same paper, prompt a more detailed study of the still unidentified mechanism of action of this component of A. indicus’s coelomic fluid to devise a novel treatment agent for tissue repair. Molecules 2020, 25, x FOR PEER REVIEW 21 of 42

of environmental pollution [169,170]. Recently, Baveja et al. [171] isolated the AiP1 metalloproteinase from the coelomic fluid of A. indicus and demonstrated its fibrinolytic potential that is likely mediated by the von Willebrand factor (vWf)-like domain located in its central β sheet. Moreover, the ability of AiP1 to modulate the motile attitude of HaCaT keratinocytes and HEK293 embryonic kidney epithelial cells was checked through scratch wound healing assays demonstrating the additional AiP1-promoting effect on cell migration since vWf is known to sustain cell locomotory behavior. This result was confirmed by the evidence of the up-regulation of N-cadherin, indicative of EMT, in HEK293 cells. The wound healing activity exerted by AiP1, coupled with the increased cell proliferation reported in the same paper, prompt a more detailed

Moleculesstudy 2020of the, 25 still, 2471 unidentified mechanism of action of this component of A. indicus’s coelomic fluid21 of 40 to devise a novel treatment agent for tissue repair.

FigureFigure 23. 23An. An individual individual of ofAstropecten Astropecten indicus indicus. ©. ©Ria Ria Tan Tan (CC (CC BY-NC-ND BY-NC-ND 2.0); 2.0); https: https://www.flickr.//www.flickr.com /. com/. Himerometra magnipinna (AH Clark, 1908; Crinoidea, Comatulida: Himerometridae; Figure 24A), also knownHimerometra as Himerometra magnipinna robustipinna (AH Clark,(Carpenter, 1908; Crinoidea, 1881) or red Comatulida: feather star, Himerometridae; is a sessile comatulid Figure that lives24A), on also hard known corals and as Himerometra hard substrates robustipinna in the Indian (Carpenter, and Pacific 1881) Oceans, or red being feather abundant star, inis thea sessile coastal areascomatulid of Vietnam that lives and inon Nha hard Trang corals Bay and in hard the South substrates China in Sea the [172 Indian]. Usually, and Pacific it has reddishOceans, maroonbeing armsabundant and feeds in the with coastal phytoplankton areas of Vietnam and zooplankton and in Nha on detritus. Trang Bay Given in thatthe crinoidsSouth China can regrow Sea [172]. arms, internalUsually, organs, it has andreddish entire maroon viscera, arms the and species feeds of with this order,phytoplankton including andH. magnipinna zooplankton, were on detritus. the object ofGiven considerable that crinoids studies can on regrow the cellular arms, basisinternal of theorgans regeneration, and entire process viscera, with the particularspecies of focusthis order, on the mechanismsincluding H. of transdimagnipinnafferentiation, were the [173 object–177]. Aof recentconsiderable paper by studies Tseng eton al. the [178 cellular] focused basis on theof etheffect ofregeneration the H. magnipinna process-derived with particular anthraquinone focus ( +on)-rhodoptilometrin the mechanisms (1,3,8-trihydroxy-6-[(1of transdifferentiationS [173–177].)-1-hydroxy propyl]anthracene-9,10-A recent paper by Tseng dione; et Figure al. [178] 24B) focused on the migratoryon the effect behavior of the of gingivalH. magnipinna and oral-derived mucosa fibroblastsanthraquinone through (+)-rhodoptilometrin scratch wound healing (1,3,8-trihydroxy-6-[(1 and transwell migrationS)-1-hydroxy assays. propyl]anthracene-9,10- In particular, they found andione; increased Figure number 24B) on of the migrating migratory cells behavior and improved of gingival wound and oral repair mucosa rate in fibroblasts the presence through of the compound,scratch wound albeit healing only limited and transwell to gingival migration fibroblasts. assays. This wasIn particular, coupled to they the up-regulationfound an increased of FAK, fibronectin,number of and migrating type I collagen, cells and selected improved as migration-associatedwound repair rate in ECM-related the presence markers. of the compound, In the same paper,albeit increased only limited cell viability to gingival and glycolysis fibroblasts./oxidative This was phosphorylation coupled to activity,the up-regulation with the latter of FAK, known tofibronectin, sustain cell mobilityand type functions, I collagen, were selected found inas rhodoptilometrin-treatedmigration-associated ECM-related human gingival markers. cells, In thereby the strongly suggesting the potential of the molecule as a beneficial additive in formulations for dental sameMolecules paper, 2020, 25increased, x FOR PEER cell REVIEW viability and glycolysis/oxidative phosphorylation activity, with 22 ofthe 42 healthlattercare known and to gingival sustain recession cell mobility treatment. functions, were found in rhodoptilometrin-treated human gingival cells, thereby strongly suggesting the potential of the molecule as a beneficial additive in formulations for dental health care and gingival recession treatment.

Figure 24. (A) An individual of Himerometra magnipinna. © Andrew Green; https://reeflifesurvey.com/. Figure 24. A) An individual of Himerometra magnipinna. © Andrew Green; (B) 2D structure of rhodoptilometrin; https://pubchem.ncbi.nlm.nih.gov/compound/Rhodoptilometrin# https://reeflifesurvey.com/. B) 2D structure of rhodoptilometrin; section=2D-Structure. https://pubchem.ncbi.nlm.nih.gov/ compound/ Rhodoptilometrin#section=2D-Structure.

Stichopus japonicus (Selenka, 1867; Holothuroidea, Synallactida: Stichopodidae; Figure 25), also known as Apostichopus japonicus (Selenka, 1867) or Japanese sea cucumber, is a benthic species distributed in the Northwestern Pacific area. It may show three different colors, either red, green, or black, and it populates the shallow coasts from the intertidal zone to depths greater than 100 m. This echinoderm is one of the most important maricultured species in Korea, China, Japan, and several Southeast Asian countries, due to its high market value for human feeding [179]. It is widely recognized that most bioactive properties of sea cucumbers derive from components extracted from their body wall. For example, in the case of S. japonicus total extracts exhibited anti- oxidant characteristics, and isolated molecules, such as unsaturated fatty acids and a fucosylated chondroitin sulfate glycosaminoglycan, were proven to exert α-glucosidase inhibitory, immunostimulatory, anti-coagulant, and hypolipidemic activities [180–184]. Collagen, which is the major component of the body wall of S. japonicus [185], has been studied for its potential as an additive and functional food ingredient that is capable of maintaining food quality thanks to its gelling properties [186,187]. Park et al. [188] examined the effect of S. japonicus’s collagen on keratinocyte biology in vitro. When submitted to scratch wound healing assay, in the presence of this collagen preparation, the HaCaT cell migration rate was higher than that obtained in both bovine gelatin-coated and non-coated plates, being similar to that shown when the cells were seeded onto rat type I collagen substrates. This was accompanied by an up-regulation of fibronectin, as shown by RT-PCR, whose synthesis and extracellular deposition is a recognized fundamental event in the first steps of wound repair. In addition, S. japonicus’s collagen increased keratinocyte proliferation positively acting on cell entry in the mitotic phase. Thus, this sea cucumber appears as a valid alternative source with respect to other fish and mammalian species for the extraction of bulky amounts of collagen for biomedical applications, also in light of the documented ability of this collagen to improve skin barrier functions and inhibit melanogenesis, and to play anti-aging, anti-allergic, and anti-wrinkle roles [189–191].

Molecules 2020, 25, 2471 22 of 40

Stichopus japonicus (Selenka, 1867; Holothuroidea, Synallactida: Stichopodidae; Figure 25), also known as Apostichopus japonicus (Selenka, 1867) or Japanese sea cucumber, is a benthic species distributed in the Northwestern Pacific area. It may show three different colors, either red, green, or black, and it populates the shallow coasts from the intertidal zone to depths greater than 100 m. This echinoderm is one of the most important maricultured species in Korea, China, Japan, and several Southeast Asian countries, due to its high market value for human feeding [179]. It is widely recognized that most bioactive properties of sea cucumbers derive from components extracted from their body wall. For example, in the case of S. japonicus total extracts exhibited anti-oxidant characteristics, and isolated molecules, such as unsaturated fatty acids and a fucosylated chondroitin sulfate glycosaminoglycan, were proven to exert α-glucosidase inhibitory, immunostimulatory, anti-coagulant, and hypolipidemic activities [180–184]. Collagen, which is the major component of the body wall of S. japonicus [185], has been studied for its potential as an additive and functional food ingredient that is capable of maintaining food quality thanks to its gelling properties [186,187]. Park et al. [188] examined the effect of S. japonicus’s collagen on keratinocyte biology in vitro. When submitted to scratch wound healing assay, in the presence of this collagen preparation, the HaCaT cell migration rate was higher than that obtained in both bovine gelatin-coated and non-coated plates, being similar to that shown when the cells were seeded onto rat type I collagen substrates. This was accompanied by an up-regulation of fibronectin, as shown by RT-PCR, whose synthesis and extracellular deposition is a recognized fundamental event in the first steps of wound repair. In addition, S. japonicus’s collagen increased keratinocyte proliferation positively acting on cell entry in the mitotic phase. Thus, this sea cucumber appears as a valid alternative source with respect to other fish and mammalian species for the extraction of bulky amounts of collagen for biomedical applications, also in light of the documented ability of this collagen to improve skin barrier functions and inhibit melanogenesis, and to play anti-aging, anti-allergic,Molecules 2020 and, 25, x anti-wrinkle FOR PEER REVIEW roles [189–191]. 23 of 42

Figure 25. A specimen of Stichopus japonicus .. ©© AlexanderAlexander Semenov; Semenov; https://www.flickr.com. https://www.flickr.com.

IsostichopusIsostichopus badionotus badionotus(Selenka, (Selenka, 1867; 1867; Holothuroidea, Holothuroidea, Synallactida: Synallactida: Stichopodidae; Stichopodidae; FigureFigure 2626)) is ais resource a resource of great of great economic economic importance importance in the in south-east the south-east of the of Gulf the of Gulf Mexico, of Mexico, Central Central America andAmerica the Caribbean and the Caribbean zone. In fact, zone. being In fact, rich being in fatty rich acids in fatty important acids important for human for health human [192 health], it is used[192], as it a is functional used as a functional food capable food of capable sustaining of sustaining growth, improving growth, improving the lipid the profile, lipid andprofile, reducing and thereducing levels of the cholesterol, levels of cholesterol, lipoproteins, lipoproteins, and triglycerides and triglycerides in both serum in both and theserum liver and where the liver it was provenwhere to it down-regulate was proven theto expressiondown-regulate of genes the that expr areession associated of genes with dyslipidemiathat are associated [193]. The with most importantdyslipidemia molecules [193]. for The biomedical most important purposes molecule obtaineds for from biomedical this sea cucumber, purposes also obtained responsible from ofthis the mentionedsea cucumber, effects, also are responsible fucosylate chondroitinof the mentioned sulphate effects, and fucoidanare fucosylate [194] whosechondroitin additional sulphate properties and asfucoidan anti-coagulant, [194] whose anti-thrombotic, additional andproperties anti-malarial as anti-coagulant, compounds anti-thrombotic, have been acknowledged and anti-malarial [195–197 ]. Licompounds et al. [198] reported have been the abilityacknowledged of small molecule [195–197]. oligopeptides Li et al. [198] that werereported obtained the fromability I. badionotusof small bymolecule enzymatic oligopeptides hydrolysis tothat induce were obtained the healing from of woundsI. badionotus in a diabeticby enzymatic rat model hydrolysisin vivo toafter induce back the healing of wounds in a diabetic rat model in vivo after back skin section. In particular, in the peptide-treated individuals the wound status displayed a better histological appearance with a higher tensile strength, and the microscopic observations showed increased angiogenesis, collagen deposition, and epithelial reconstitution confirmed by the higher expression of vascular endothelial growth factor (VEGF) in the tissue and more elevated levels of NO and stromal cell- derived factor 1α, both being involved in the secretion of angiogenic factors, in animal sera. Moreover, the peptide treatment favoured the enhancement of antioxidant levels and the reduction of the pro-inflammatory interleukin-6 and -8, tumor necrosis factor-α, chemokine (C-C motif) ligand 2 and C-reactive protein, counteracted by the up-regulation of the anti- inflammatory interleukin-10. This indicates that the natural peptide mixture from I. badionotus sea cucumber might have promising applications for the treatment of diabetic wound healing, which still represents a significant clinical problem.

Molecules 2020, 25, x FOR PEER REVIEW 23 of 42

Figure 25. A specimen of Stichopus japonicus. © Alexander Semenov; https://www.flickr.com.

Isostichopus badionotus (Selenka, 1867; Holothuroidea, Synallactida: Stichopodidae; Figure 26) is a resource of great economic importance in the south-east of the Gulf of Mexico, Central America and the Caribbean zone. In fact, being rich in fatty acids important for human health [192], it is used as a functional food capable of sustaining growth, improving the lipid profile, and reducing the levels of cholesterol, lipoproteins, and triglycerides in both serum and the liver where it was proven to down-regulate the expression of genes that are associated with dyslipidemia [193]. The most important molecules for biomedical purposes obtained from this sea cucumber, also responsible of the mentioned effects, are fucosylate chondroitin sulphate and fucoidan [194] whose additional properties as anti-coagulant, anti-thrombotic, and anti-malarial compounds have been acknowledged [195–197]. Li et al. [198] reported the ability of small Molecules 2020, 25, 2471 23 of 40 molecule oligopeptides that were obtained from I. badionotus by enzymatic hydrolysis to induce the healing of wounds in a diabetic rat model in vivo after back skin section. In particular, in the skinpeptide-treated section. In particular, individuals in the the wound peptide-treated status displayed individuals a better the woundhistological status appearance displayed with a better a histologicalhigher tensile appearance strength, with and a higherthe microscopic tensile strength, observations and the sh microscopicowed increased observations angiogenesis, showed increasedcollagen angiogenesis,deposition, and collagen epithelial deposition, reconstitution and epithelial confirmed reconstitution by the higher confirmedexpression by of thevascular higher expressionendothelial of growth vascular factor endothelial (VEGF) growthin the tissue factor an (VEGF)d more inelevated the tissue levels and of moreNO and elevated stromal levels cell- of NOderived and stromal factor cell-derived1α, both being factor involved 1α, both in being the secretion involved of in theangiogenic secretion factors, of angiogenic in animal factors, sera. in animalMoreover, sera. Moreover,the peptide the treatment peptide treatmentfavoured favouredthe enhancement the enhancement of antioxidant of antioxidant levels levelsand the and thereduction reduction of ofthe the pro-inflammatory pro-inflammatory interleukin-6 interleukin-6 and and -8, -8, tumor tumor necrosis necrosis factor- factor-α,α chemokine, chemokine (C-C (C-C motif)motif) ligand ligand 2 and2 and C-reactive C-reactive protein, protein, counteracted counteracted by the up-regulationby the up-regulation of the anti-inflammatory of the anti- interleukin-10.inflammatory This interleukin-10. indicates that This the indicates natural peptide that the mixture natural from peptide I. badionotus mixture from sea cucumber I. badionotus might havesea promisingcucumber might applications have promising for the treatment applications of diabetic for thewound treatment healing, of diabetic which wound still represents healing, a significantwhich still clinical represents problem. a significant clinical problem.

Figure 26. A specimen of Isostichopus badionotus. © Dan Hershman (CC BY 2.0); https://commons. wikimedia.org/.

Ophiocoma erinaceus (Müller & Troschel, 1842; Ophiuroidea, Ophiacanthida: Ophiocomidae; Figure 27), also known as brittle star, is typical of tropical- and reef-associated environments and is distributed in the whole Indo-Pacific area (except for Pakistan and western India) being dominant in the Persian Gulf and in the coast of South Africa [172,199]. In the search of novel anti-angiogenic compounds, O. erinaceus’s alcoholic extracts, which were endowed with anti-oxidant properties, were administered to A2780cp ovarian tumor cells in culture and an in vivo CAM model, demonstrating a significant inhibition of cell growth and a down-regulation of formation of new blood vessels, coupled to the decrease of VEGF, TGF-β, and FGF-β mRNA expression in endothelial cells [200,201]. Among the components of the extract, an isolated polysaccharide and a saponin fraction were subjected to further studies. In fact, the first compound was found able to exert the previously-mentioned effects, and also decrease the viability of HUVEC cells, their expression of paxillin and MMP-9 mRNAs, whose products are involved in metastatization, and migration in scratch wound healing assays [202]. More recently, the brittle star polysaccharide was also proven to accelerate the wound healing in a male Wistar rat model in vivo [203]. On the other hand, the brittle star saponins appeared to be responsible for the radical scavenging function of the extract via increase of the SOD level in exposed cervical cancer HeLa cells. The survival rate of this cell line was also decreased due to the triggering of the intrinsic apoptotic pathway, differently from that found with non-tumoral fibroblasts. Moreover, saponins impaired cell locomotion in wound healing assay in vitro, parallelly reducing the adhesion of cells to matrigel substrates and inducing the down-regulation of MMP-2 and MMP-9 [204,205]. Of note, brittle star saponins also exhibited an anti-inflammatory ability both in in vitro assays and on macrophages that were differentiated from THP-1/M monocytes, as evidenced by the observed suppression of pro-inflammatory cytokines gene expression and production [206]. Thus, the active Molecules 2020, 25, x FOR PEER REVIEW 24 of 42

Figure 26. A specimen of Isostichopus badionotus. © Dan Hershman (CC BY 2.0); https://commons. wikimedia.org/.

Ophiocoma erinaceus (Müller & Troschel, 1842; Ophiuroidea, Ophiacanthida: Ophiocomidae; Figure 27), also known as brittle star, is typical of tropical- and reef-associated environments and is distributed in the whole Indo-Pacific area (except for Pakistan and western India) being dominant in the Persian Gulf and in the coast of South Africa [172,199]. In the search of novel anti- angiogenic compounds, O. erinaceus’s alcoholic extracts, which were endowed with anti-oxidant properties, were administered to A2780cp ovarian tumor cells in culture and an in vivo CAM model, demonstrating a significant inhibition of cell growth and a down-regulation of formation of new blood vessels, coupled to the decrease of VEGF, TGF-β, and FGF-β mRNA expression in endothelial cells [200,201]. Among the components of the extract, an isolated polysaccharide and a saponin fraction were subjected to further studies. In fact, the first compound was found able to exert the previously-mentioned effects, and also decrease the viability of HUVEC cells, their expression of paxillin and MMP-9 mRNAs, whose products are involved in metastatization, and migration in scratch wound healing assays [202]. More recently, the brittle star polysaccharide was also proven to accelerate the wound healing in a male Wistar rat model in vivo [203]. On the other hand, the brittle star saponins appeared to be responsible for the radical scavenging function of the extract via increase of the SOD level in exposed cervical cancer HeLa cells. The survival rate of this cell line was also decreased due to the triggering of the intrinsic apoptotic pathway, differently from that found with non-tumoral fibroblasts. Moreover, saponins impaired cell locomotion in wound healing assay in vitro, parallelly reducing the adhesion of cells to matrigel substrates and inducing the down-regulation of MMP-2 and MMP-9 [204,205]. Of note, brittle star saponins also exhibited an anti-inflammatory ability both in in vitro assays and on macrophages Molecules 2020, 25, 2471 24 of 40 that were differentiated from THP-1/M monocytes, as evidenced by the observed suppression of pro-inflammatory cytokines gene expression and production [206]. Thus, the active principles principlespresent in present extracts in from extracts O. erinaceus from O. appear erinaceus toappear be promising to be promising candidates candidates against human against cervical human cervicalcarcinoma carcinoma and in andthe prevention in the prevention of angiogenesis-related of angiogenesis-related disorders. disorders.

FigureFigure 27. 27. AnAn individualindividual of Ophiocoma erinaceus erinaceus. .© ©PatrickPatrick Randall Randall (CC (CC BY-NC-SA BY-NC-SA 2.0); 2.0); https:// https: //www.flickr.comwww.flickr.com/./.

StichopusStichopus herrmanni herrmanni(Semper, (Semper, 1868; 1868; Olothuroidea, Olothuroidea, Synallactida: Synallactida: Stichopodidae; Stichopodidae; Figure Figure 28 ),28), also knownalso known as curryfish as curryfish herrmanni, herrmanni, is a benthic is a speciesbenthic distributed species distributed in the Indo-West in the PacificIndo-West region, Pacific being dominantregion, being in tropical dominant shallow in waters, tropical which shallow represents waters, an important which represents fishery commodity. an important This orgamism,fishery previouslycommodity. known This as orgamism,Stichopus variegatus previouslyin Indonesiaknown as andStichopus Malaysia, variegatus has found in Indonesia use for food and consumption Malaysia, andhasMolecules preparation found 2020 use, 25 , offor x FOR traditional food PEER consumption REVIEW medicines and in Asian preparatio countries,n of traditional and for environmental medicines in monitoring, Asian countries, 25 e.g., of 42 the quantificationand for environmental of marine pollution monitoring, by polycyclic e.g., the aromatic quantification hydrocarbons of marine [207 ,pollution208]. The tegumentby polycyclic of this invertebratearomaticBiocomposites hydrocarbons is known of hydroxyapatite to contain [207,208]. high andThe amounts S.tegument herrmannii of collagen of’s this collagen and invertebrate glycosaminoglycans have been is known proven thatto to contain increase have been high the the objectamountsnumber of researches. ofof neo-formed collagen Biocomposites andosteoblasts glycosaminoglycans of hydroxyapatiteduring bone healing andthat S.have herrmanniiand beenregeneration,’s the collagen object with have of no beenresearches. toxic proven side to increaseeffects the[209]. number Zohdi of et neo-formed al. [210] prepared osteoblasts a S. during hermannii bone’s healingextract-containing and regeneration, hydrogel with dressing no toxic side (“Gamat effects Hydrogel”), [209]. Zohdi which, et al. [210 when] prepared applied a S.to hermanniiburn skin’s extract-containinglesions in rats, improved hydrogel wound dressing (“Gamatcontraction Hydrogel”), and closure, which, aided when by applied a reduced to burn inflam skinmatory lesions reaction, in rats, with improved signs woundof more contraction efficient andre-epithelization closure, aided byat athe reduced histological inflammatory observat reaction,ion. The with signsglycosaminoglycan of more efficient component re-epithelization in S. at theherrmannii histological extracts observation. was shown The glycosaminoglycan to play a role as a positive component modulator in S. herrmannii of inflammation extracts also was in shown the tohealing play a roleprocess as aof positive oral mucous modulator traumatic of inflammation ulcers in rats also[211]. in More the healingrecently, process Arundina of oral et al. mucous [212] traumaticdemonstrated ulcers inthe rats stimulating [211]. More effects recently, of Arundinathe extracts, et al. which [212] demonstratedare also rich thesources stimulating of soluble effects ofmediators, the extracts, on which the growth are also behavior rich sources and of osteogen soluble mediators,ic differentiation on the growth of mesenchymal behavior and stem osteogenic cells, ditherebyfferentiation confirming of mesenchymal that preparations stem cells, from thereby S. herrmannii confirming may that preparationshave beneficial from effects S. herrmannii on the mayacceleration have beneficial of the ewoundffects on healing the acceleration and tissue of regeneration the wound healing processes. and tissue regeneration processes.

Figure 28.28. A specimen of Stichopus herrmanni . ©© AndrewAndrew Green; Green; httphttps:s://reeflifesurvey.com/.//reeflifesurvey.com/.

6. Tunicata Tunicates (subphylum Chordata) are marine organisms that are covered by a ”tunic”, i.e., a tough body shield which mainly incorporates a highly crystalline cellulose nanofiber called tunicin [213,214], and proteins that contain 3,4,5-trihydroxyphenylalanine (TOPA) and pyrogallol amino acid [215,216]. Tunicates are known to be among the richest sources of biologically-active compounds together with sponges and bryozoans [217,218] and possess a very efficient mechanism of healing allowing for the repair of tunic injuries in sea water, a wet and salty environment similar to human body fluid [213]. In particular, at the pH of marine water (8.2) pyrogallol is oxidized and provides the key adhesion functionality forming covalent cross-links with tunicin, which has 120 gigapascals of stiffness and contributes to the mechanical support of the tunic, other TOPAs, or proteins [214]. These pyrogallol-mediated bonds favor the seal of the torn tunic and the control of bleeding. In light of such observations, a limited number of studies have been addressed to the identification of tunicate compounds active on cell migration in vitro and in vivo and tissue repair. Phallusia arabica (Savigny, 1816; Ascidiacea, Phlebobranchia: Ascidiidae; Figure 29A) is a dark grey large solitary ascidian that populates the Red Sea, and the North Atlantic and South Pacific oceans. Ethanolic extracts of P. arabica exhibited a positive activity on the skin repair rate in excision rat models in vivo and a significant enhancement of the contraction in incision wounds with a faster re-epithelization and collagen deposition period [219]. It is supposed that the beneficial effect of the ascidian extract lies in the abundant presence of flavonoids, whose anti- inflammatory, anti-fungal, anti-oxidant, and wound healing properties are acknowledged, and the anti-oxidant n-hexadecanoic (palmitic) acid [220] (Figure 29B).

Molecules 2020, 25, 2471 25 of 40

6. Tunicata Tunicates (subphylum Chordata) are marine organisms that are covered by a “tunic”, i.e., a tough body shield which mainly incorporates a highly crystalline cellulose nanofiber called tunicin [213,214], and proteins that contain 3,4,5-trihydroxyphenylalanine (TOPA) and pyrogallol amino acid [215,216]. Tunicates are known to be among the richest sources of biologically-active compounds together with sponges and bryozoans [217,218] and possess a very efficient mechanism of healing allowing for the repair of tunic injuries in sea water, a wet and salty environment similar to human body fluid [213]. In particular, at the pH of marine water (8.2) pyrogallol is oxidized and provides the key adhesion functionality forming covalent cross-links with tunicin, which has 120 gigapascals of stiffness and contributes to the mechanical support of the tunic, other TOPAs, or proteins [214]. These pyrogallol-mediated bonds favor the seal of the torn tunic and the control of bleeding. In light of such observations, a limited number of studies have been addressed to the identification of tunicate compounds active on cell migration in vitro and in vivo and tissue repair. Phallusia arabica (Savigny, 1816; Ascidiacea, Phlebobranchia: Ascidiidae; Figure 29A) is a dark grey large solitary ascidian that populates the Red Sea, and the North Atlantic and South Pacific oceans. Ethanolic extracts of P. arabica exhibited a positive activity on the skin repair rate in excision rat models in vivo and a significant enhancement of the contraction in incision wounds with a faster re-epithelization and collagen deposition period [219]. It is supposed that the beneficial effect of the ascidian extract lies in the abundant presence of flavonoids, whose anti-inflammatory, anti-fungal, anti-oxidant, and wound healing properties are acknowledged, and the anti-oxidant n-hexadecanoic (palmitic)Molecules 2020 acid, 25 [220, x FOR] (Figure PEER REVIEW 29B). 26 of 42

Figure 29. (A) An image of Phallusia arabica. © Frédéric André; https://doris.ffessm.fr/.(B) 2D Figure 29. A) An image of Phallusia arabica. © Frédéric André; https://doris.ffessm.fr/. B) 2D structure of n-hexadecanoic acid; https://pubchem.ncbi.nlm.nih.gov/compound/Palmitic-acid#section= structure of n-hexadecanoic acid; https://pubchem.ncbi.nlm.nih.gov/compound/Palmitic-acid# 2D-Structure. section=2D-Structure. Phallusia nigra (Savigny, 1816; Ascidiacea, Phlebobranchia: Ascidiidae; Figure 30) is a velvet black Phallusia nigra (Savigny, 1816; Ascidiacea, Phlebobranchia: Ascidiidae; Figure 30) is a velvet large solitary ascidian, very common in the warm waters of the western Atlantic, ranging from Bermuda black large solitary ascidian, very common in the warm waters of the western Atlantic, ranging to Brazil, as well as in other seas like the Red Sea, Gulf of Aden, and Gulf of Guinea. It occurs as the from Bermuda to Brazil, as well as in other seas like the Red Sea, Gulf of Aden, and Gulf of Guinea. major component of fouling communities on the hull of ships, piers, pilings and harbour installations. It occurs as the major component of fouling communities on the hull of ships, piers, pilings and Also the methanolic extract of P. nigra exhibited a beneficial effect towards wound healing in in vivo harbour installations. Also the methanolic extract of P. nigra exhibited a beneficial effect towards rat models, similar to those that were obtained with P. arabica extracts, and also in this case the positive wound healing in in vivo rat models, similar to those that were obtained with P. arabica extracts, influence on tissue repair speed and quality was ascribed to the flavonoid and n-hexadecanoic acid and also in this case the positive influence on tissue repair speed and quality was ascribed to the components of the preparation [221,222]. flavonoid and n-hexadecanoic acid components of the preparation [221,222].

Figure 30. An image of Phallusia nigra. © Rosana M. Rocha; http://invasions.si.edu/.

Styela clava (Herdman, 1881; Ascidiacea, Stolidobranchia: Styelidae; Figure 31), also known as stalked sea squirt, is a solitary ascidian displaying a brown or yellow rough and wrinkled surface which is native of the Northwestern Pacific area and it has become a conspicuous member of fouling communities in Northwestern European waters. The cellulose films manufactured from S. clava’s tunic (SCT) have been the object of research [223]. These films exhibited good biocompatibility and degradability [224] and, when regeneration and angiogenesis in the surgical wounds of Sprague–Dawley rats were examined after the application of three different preparations of SCT-cellulose hydrogels, the one regenerated from freeze-dried membranes

Molecules 2020, 25, x FOR PEER REVIEW 26 of 42

Figure 29. A) An image of Phallusia arabica. © Frédéric André; https://doris.ffessm.fr/. B) 2D structure of n-hexadecanoic acid; https://pubchem.ncbi.nlm.nih.gov/compound/Palmitic-acid# section=2D-Structure.

Phallusia nigra (Savigny, 1816; Ascidiacea, Phlebobranchia: Ascidiidae; Figure 30) is a velvet black large solitary ascidian, very common in the warm waters of the western Atlantic, ranging from Bermuda to Brazil, as well as in other seas like the Red Sea, Gulf of Aden, and Gulf of Guinea. It occurs as the major component of fouling communities on the hull of ships, piers, pilings and harbour installations. Also the methanolic extract of P. nigra exhibited a beneficial effect towards wound healing in in vivo rat models, similar to those that were obtained with P. arabica extracts, and also in this case the positive influence on tissue repair speed and quality was ascribed to the Molecules 2020, 25, 2471 26 of 40 flavonoid and n-hexadecanoic acid components of the preparation [221,222].

Figure 30. An image of Phallusia nigra . © RosanaRosana M. M. Rocha; Rocha; http://invasions.si.edu/.http://invasions.si.edu/.

Styela clava Styela clava(Herdman, (Herdman, 1881; 1881; Ascidiacea, Ascidiacea, Stolidobranchia: Stolidobranchia: Styelidae; Styelidae; Figure Figure 31 31),), also also known known as stalkedas stalked sea squirt,sea squirt, is a solitaryis a solitary ascidian ascidian displaying displaying a brown a brown or yellow or yellow rough rough and wrinkled and wrinkled surface whichsurface is nativewhich ofis native the Northwestern of the Northwestern Pacific area Paci andfic area it has and become it has abecome conspicuous a conspicuous member member of fouling S. clava communitiesof fouling communities in Northwestern in Northwestern European European waters. The waters. cellulose The cellulose films manufactured films manufactured from from ’s Molecules 2020, 25, x FOR PEER REVIEW 27 of 42 tunicS. clava (SCT)’s havetunic been(SCT) the have object been of research the object [223 ].of These research films [223]. exhibited These good films biocompatibility exhibited good and degradabilitybiocompatibility [224] and and, degradability when regeneration [224] andand, angiogenesis when regeneration in the surgical and angiogenesis wounds of Sprague–Dawley in the surgical appeared to be a better dressing support when compared to regenerated plain and alginate- ratswounds were examinedof Sprague–Dawley after the application rats were of threeexamined different after preparations the application of SCT-cellulose of three different hydrogels, supplemented hydrogels [225]. This was due to the highest strength and lowest elongation level thepreparations one regenerated of SCT-cellulose from freeze-dried hydrogels, membranes the one appeared regenerated to be from a better freeze-dried dressing supportmembranes when comparedbeing exhibited to regenerated by this plainpreparation and alginate-supplemented and the consequent hydrogelsamelioration [225 of]. Thisgranulation was due tissue to the highest formation, strength angiogenesis, and lowest and elongation skin reconstitution level being in exhibited the absence by this of preparationtoxic effects.and At the the molecular consequent ameliorationlevel, the reconstituted of granulation freeze-dried tissue formation, membranes angiogenesis, appeared and to skin up-regulate reconstitution the levels in the of absence VEGF, of β toxiccollagen effects. and At TGF- the molecular1, with level,activation the reconstituted of p-Smad 2/3 freeze-dried signalization, membranes in the damaged appeared tissue. to up-regulate More therecently, levels of Song VEGF, et al. collagen [226] studied and TGF- theβ 1,effect with of activation selenium-loaded of p-Smad STC-cell 2/3 signalization,ulose films inon the cutaneous damaged tissue.wounds More of diabetic recently, rats, Song showing, et al. [226 also] studied in this ex theperimental effect of selenium-loaded system, the acceleration STC-cellulose of the healing films on cutaneousprocess with wounds the complete of diabetic recovery rats, showing, of re-epithe also inlization this experimental and skin repair system, following the acceleration the activation of the healingof the process downstream with the signaling complete recoverypathways of re-epithelizationof VEGF, angiopoietin-2/1, and skin repair and following insulin, the and activation the ofrestoration the downstream of anti-oxidative signaling pathwaysstatus. These of VEGF,data demonstrate angiopoietin-2 a potential/1, and insulin,use of the and S. clava the restoration-derived ofmaterial anti-oxidative as a beneficial status. These supplem dataentary demonstrate compound a potential in formulations use of the S. for clava topical-derived application material asto a beneficialcutaneous supplementary wounds. compound in formulations for topical application to cutaneous wounds.

FigureFigure 31. An31. image An of Styelaimage clava. of© ClaudeStyela Nozclava.ères (CC© BY-NC-SAClaude 4.0);Nozères http://www.marinespecies.org (CC BY-NC-SA /4.0);. 7. Conclusionshttp://www.marinespecies.org/.

7. ConclusionsAn assortment of key events in development, health, and disease depends on collective cell motility which represents a common theme joining morphogenesis, tissue regeneration, and tumor biology [1]. An assortment of key events in development, health, and disease depends on collective cell motility which represents a common theme joining morphogenesis, tissue regeneration, and tumor biology [1]. It is widely acknowledged that several signal transduction and molecular mechanisms underlying and critical for cell migration and re-epithelization in wound repair are highly similar to those that are involved in epithelial tumor ingrowth and metastasis. As for medical treatments, it is known that, in some cases, anticancer agents may show interference with wound healing and, in turn, drugs used for non-healing ulcers of diabetic patients may promote carcinogenesis. Thus, there are many molecular and physiologic points in common between wound closure and tumor progression [227]. On the other hand, over the years, the literature showed the rapid rising of the prevalence, severity, and complexity of both cancer and wound healing pathologies, with the consequent remarkable increase in healthcare costs. It is imperative for modern society the procurement of alternative sources of bioactive compounds to replace overexploited resources and, on the other hand, improve treatments and the quality of life of patients via novel and more efficacious drugs. Marine invertebrates are a still poorly exploited valuable resource of natural products that may significantly improve the process of skin regeneration and restrain tumor cell migration, as documented by the in vitro and in vivo studies presented in this review and summarized in a synoptic way in Table 1. Only a very limited number of compounds and methods of extraction have been patented and/or introduced for treatment schemes to date, i.e., krill oil preparations [228], cellulose films from S. clava (patent KR20150117748A South Korea), S. herrmanni’s collagen for biocomposites [229], and S. japonicus’s fucosylated chondroitin sulfate (patent CN102367285A China). In conclusion, there is still an enormous potential offered by marine invertebrates towards a variety of applications that may significantly improve human health and the well-being of future generations. Therefore, the

Molecules 2020, 25, 2471 27 of 40

It is widely acknowledged that several signal transduction and molecular mechanisms underlying and critical for cell migration and re-epithelization in wound repair are highly similar to those that are involved in epithelial tumor ingrowth and metastasis. As for medical treatments, it is known that, in some cases, anticancer agents may show interference with wound healing and, in turn, drugs used for non-healing ulcers of diabetic patients may promote carcinogenesis. Thus, there are many molecular and physiologic points in common between wound closure and tumor progression [227]. On the other hand, over the years, the literature showed the rapid rising of the prevalence, severity, and complexity of both cancer and wound healing pathologies, with the consequent remarkable increase in healthcare costs. It is imperative for modern society the procurement of alternative sources of bioactive compounds to replace overexploited resources and, on the other hand, improve treatments and the quality of life of patients via novel and more efficacious drugs. Marine invertebrates are a still poorly exploited valuable resource of natural products that may significantly improve the process of skin regeneration and restrain tumor cell migration, as documented by the in vitro and in vivo studies presented in this review and summarized in a synoptic way in Table1. Only a very limited number of compounds and methods of extraction have been patented and/or introduced for treatment schemes to date, i.e., krill oil preparations [228], cellulose films from S. clava (patent KR20150117748A South Korea), S. herrmanni’s collagen for biocomposites [229], and S. japonicus’s fucosylated chondroitin sulfate (patent CN102367285A China). In conclusion, there is still an enormous potential offered by marine invertebrates towards a variety of applications that may significantly improve human health and the well-being of future generations. Therefore, the advancement of knowledge in this direction will result in new approaches that involve the identification of the most promising candidates for the utilization as new extracts/molecules, also in combination with existing drugs, which are targeted for biomedical translation.

Table 1. Anti-cancer and wound healing-promoting properties of molecules and crude extracts from marine invertebrates.

Effects Phylum Species Compounds References Stellettin B Latrunculin B and 15-O-methyl-latrunculin B Latrunculin A Bromotyrosine derivative (1’R,5’S,6’S)-2-(3’,5’-dibromo-1’,6’ Jaspis stellifera [42] -dihydroxy-4’-oxocyclohex-2’-enyl) Negombata magnifica [47] acetonitrile Anti-cell Pseudoceratina arabica [48] Moloka’iamine migration/invasion Porifera Suberea mollis [53] Ceratinin A properties Siphonocalina [54] Ceratinin D hydroxymoloka’iamine (Callyspongia) siphonella [62,64] subereamolline A Xestospongia sp. [71] Sipholenol A and its aliphatic polar and aromatic ester derivatives Sipholenol A-4-O-30,40-dichlorobenzoate derivative Ranieramycins (i.e ranieramycin M) Dihydroaustrasulfone alcohol (synthetic precursor of Austrasulfone) Pachycladin A Sclerophytin F methyl ether Cladiella australis [76–78] Polyanthelin A Cladiella pachyclados [79] Sclerophytin A Cnidaria Sarcophyton crassocaule [81,82] Cembrenolide diterpene Sinularia nanolobata [83,84] 13-acetoxysarcocrassolide Clavularia koellikeri [87] 11-dehydrosinulariolide Sinularin Clavukoellian A and clavukoellian D-related 4-O-Deacetylparalemnolin D Mollusca Euchelus asper Methanol extracts [145] Echinodermata Ophiocoma erinaceus Saponins [204,205] Molecules 2020, 25, 2471 28 of 40

Table 1. Cont.

Effects Phylum Species Compounds References Pro wound-healing Fraction enriched in hydroxyproline- Porifera Chondrosia reniformis [33] properties containing collagenous peptides Cyanea capillata Tentacle extracts, Granulin [96,97] Cnidaria Rhopilema esculentum Collagen peptides [103] Uroteuthis (Doryteuthis) Radical scavenging-carotenoid astaxhantin singhalensis [109] Chitosan and collagen Sepia kobiensis [115] Collagen Sepia officinalis [116] Lipid extracts; Aminoacid mixture from Mollusca Mytilus galloprovincialis [132,133] protein extract and Rapana venosa [140] Peptides extracted from P. martensii’s mantle Pinctada martensii [151] H. diversicolor’s shell powder Haliotis diversicolor [164] Krill oil Euphausia superba Astropecten indicus AiP1 metalloproteinase [171] Himerometra Anthraquinone (+)-rhodoptilometrin [178] magnipinna Collagen [188] Echinodermata Stichopus japonicus Oligopeptides [198] Isostichopus badionotus Polysaccharides [203] Ophiocoma erinaceus “Gamat Hydrogel” (S. hermannii’s [210] Stichopus herrmanni extract-containing hydrogel dressing) Ethanolic extracts Phallusia arabica Methanolic extract [219] Tunicata Phallusia nigra Cellulose films manufactured from S. clava’s [221,222] Styela clava tunic (SCT) [225,226] selenium-loaded STC-cellulose films

Author Contributions: C.L., M.M., V.L. and M.V. collected the references; C.L. and M.V. wrote the paper. All authors have read and agreed to the published version of the manuscript. Funding: This research was funded by The University of Palermo (Italy), grant Fondo Finalizzato alla Ricerca (FFR) 2018 to C.L. and M.V. Conflicts of Interest: The authors declare no conflict of interest.

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