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Fabrication of Biopolymer Based Nanoparticles for the Entrapment of Chromium and Iron Supplements

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Fabrication of Biopolymer Based Nanoparticles for the Entrapment of Chromium and Iron Supplements processes Article Fabrication of Biopolymer Based Nanoparticles for the Entrapment of Chromium and Iron Supplements Nishay Patel 1, Mohammed Gulrez Zariwala 2 and Hisham Al-Obaidi 1,* 1 The School of Pharmacy, University of Reading, Reading RG6 6AD, UK; [email protected] 2 Faculty of Science & Technology, University of Westminster, 115 New Cavendish Street, London W1W 6UW, UK; [email protected] * Correspondence: [email protected]; Tel.: +44-11-8378-6261 Received: 20 May 2020; Accepted: 15 June 2020; Published: 19 June 2020 Abstract: The objective of this study was to encapsulate iron and chromium into novel nanoparticles formulated using chitosan (CS), dextran sulfate (DS) and whey protein isolate (WPI) for oral drug delivery. Empty and loaded CS-DS nanoparticles were prepared via complex coacervation whilst whey protein nanocarriers were produced by a modified thermal processing method using chitosan. The physiochemical properties of the particles were characterized to determine the effects of formulation variables, including biopolymer ratio on particle size and zeta potential. Permeability studies were also undertaken on the most stable whey protein–iron nanoparticles by measuring Caco-2 ferritin formation. A particle size analysis revealed that the majority of samples were sub-micron sized, ranging from 420–2400 nm for CS-DS particles and 220–1000 nm for WPI-CS samples. As expected, a higher chitosan concentration conferred a 17% more positive zeta potential on chromium-entrapped WPI nanoparticles, whilst a higher dextran volume decreased the size of CS-DS nanoparticles by 32%. The addition of iron also caused a significant increase in size for all samples, as seen where the loaded WPI samples were 296 nm larger than the empty particles. Caco-2 iron absorption revealed that one formulation, which had the lowest particle size (226 10 nm), caused a 64% greater iron ± absorption compared to the ferrous sulfate standard. This study describes, for the first time, the novel design of chromium- and iron-entrapped nanoparticles, which could act as novel systems for oral drug delivery. Keywords: iron; chromium; whey protein isolate; nanoparticles; oral drug delivery 1. Introduction Iron and chromium have major roles in the metabolism and oxygen exchange processes [1]. Chromium is a trace element that has received a significant interest in recent years. It aids in the metabolism of carbohydrates and is thought to impair glucose tolerance in those with chromium deficiencies. This can lead to the development of maturity-onset diabetes-a chronic condition costing the national health service (NHS) over nine billion Pounds sterling annually [2,3]. Chromium supplements can be used to prevent insulin resistance and may be seen in the future as a cost effective strategy to reduce the onset of type 2 diabetes [4]. Iron is an essential mineral that plays a vital role in oxygen transport and DNA replication [5]. Almost 20% of the world’s population is iron deficient—a condition that is prevalent in most countries [6]. Iron is lost from the body through the natural shedding of skin, during pregnancy and heavy menstruation but cannot be synthesized in vivo [7]. Iron is also poorly absorbed from dietary sources meaning that supplementation may be needed to maintain healthy levels. In previous studies, iron-entrapped nanoparticles were formulated using ascorbic acid which augments the absorption of dietary non-haem iron by reducing ferric iron to the bioavailable ferrous form [8–10]. Processes 2020, 8, 707; doi:10.3390/pr8060707 www.mdpi.com/journal/processes Processes 2020, 8, 707 2 of 16 Processes 2020, 8, x FOR PEER REVIEW 2 of 17 Nanoparticles areare defineddefined as as solid solid colloidal colloidal structures structures ranging ranging in sizein size from from 1 to 1 1000 to 1000 nm. nm. They They can becan formed be formed of natural of natural polymers polymers such as such chitosan as chitosan and have and a number have a ofnumber advantages of advantages including enhancingincluding theenhancing stability the of drugs,stability increasing of drugs, encapsulation increasing encaps efficiencyulation and efficiency having better and uptakehaving inbetter the small uptake intestine. in the Thissmall is intestine. because nanoparticlesThis is because can nanoparticles permeate enterocyte can permeate membranes enterocyte easier thanmembranes microparticles easier than and canmicroparticles facilitate the and controlled can facilitate release the of thecontrolled encapsulated release drug of [11the,12 encapsulat]. These qualitiesed drug are [11,12]. of particular These importancequalities are for of particular this study importance because chromium for this stud is poorlyy because absorbed chromium (<2%) is [13 poor]. Therefore,ly absorbed nanoparticles (<2%) [13]. wereTherefore, formulated nanoparticles because were they canformulated improve because drug bioavailability they can improve and reduce drug toxicitybioavailability whilst maintaining and reduce therapeutictoxicity whilst eff ectsmaintaining [14]. therapeutic effects [14]. Biopolymers are naturally abundant polymers and possess a broad range of attributes which can be used to optimize the properties of nanoparticles. In In this this study, study, chitosan (CS), dextran sulfate (DS) and wheywhey protein protein isolate isolate (WPI) (WPI) were were chosen chosen due todu theme to being them cost being effective cost andeffective generally and recognized generally asrecognized safe (GRAS) as safe [15]. (GRAS) CS is a polysaccharide[15]. CS is a polysaccharide produced from produced the exoskeleton from the of exoskeleton crustaceans of by crustaceans the partial deacetylationby the partial ofdeacetylation chitin [16]. It of is composedchitin [16]. of It repeatingis composed glucosamine of repeating units glucosamine and is a biodegradable units and andis a pharmaceuticallybiodegradable and acceptable pharmaceutically excipient acceptable (Figure1). CSexcipient has a pKa (Figure of 6.5 1). and CS possesses has a pKa a high of ratio6.5 and of aminopossesses groups a high that ratio undergo of amino protonation groups in that acidic undergo conditions, protonation making itin soluble acidic inconditions, water (Figure making1)[ 17 it]. Itsoluble has mucoadhesive in water (Figure properties 1) [17]. It and has can mucoadhesi complexve with properties chromium and to can produce complex nanoparticles. with chromium DS to is aproduce polyanionic nanoparticles. polymer which DS is possesses a polyanionic negatively poly chargedmer which sulfate possesses groups, negatively dictating its charged ability tosulfate form stablegroups, coacervates dictating its with ability CS [to18 form]. It is stable highly coacervates hydrophilic with and CS has [18]. been It is used highly to control hydrophilic the release and has of drugsbeen used such to as control doxorubicin the release [19]. Complexof drugs coacervationsuch as doxorubicin was chosen [19]. as Complex opposed coacervation to ionic gelation was because chosen theas opposed nanoparticles to ionic have gelation enhanced because mechanical the nanopa strengthrticles and stabilityhave enhanced than those mechanical traditionally strength produced and usingstability CS than and those sodium traditionally triphosphate produced [20]. using CS and sodium triphosphate [20]. Figure 1. 1. ChemicalChemical structures structures of (a of) chitosan (a) chitosan and (b and) dextran (b) dextran sulfate sulfatewith indication with indication of their ionizable of their ionizablegroups. groups. Whey protein is a by-product of the cheesecheese manufacturingmanufacturing process and consists of globular proteins, mainly β-lactoglobulin. It It has has excellent gelation gelation pr propertiesoperties and is widely used by athletes to stimulate muscle muscle growth. growth. Hence, Hence, it it can can work work synergistically synergistically with with chromium, chromium, which which is issuggested suggested it itcan can increase increase muscle muscle mass, mass, to to improve improve nutritional nutritional health health and and athletic athletic performance performance [21]. [21]. Therefore, chromium-entrapped nanoparticles were produced using whey protein isolate through thermal processing. Y. Y. Chen was the firstfirst to fabricate and entrap an arginine-richarginine-rich hexapeptide into CS-DS nanoparticles [[22].22]. ThisThis increasedincreased thethe popularitypopularity of of biopolymer-based biopolymer-based nanoparticle nanoparticle research research and and led led to furtherto further developments developments in the in fieldthe field such such as the as encapsulation the encapsulation of insulin of insulin into CS-based into CS-based nanoparticles nanoparticles [23,24]. [23,24].The aim of this study was to build on previous research by fabricating four types of novel chromium-The aim and of iron-entrappedthis study was nanoparticlesto build on previous using di researchfferent ratios by fabricating of biopolymers four types for oral of novel drug delivery.chromium- The and properties iron-entrapped of the nanoparticlesnanoparticlessuch using as di size,fferent charge ratios and of stability biopolymers were thenfor oral assessed drug todelivery. find the The optimum properties formulations. of the nanoparticles Permeability such studies as size, werecharge also and performed stability were using then Caco-2 assessed cells to determinefind the optimum the differences formulations. in ironabsorption Permeability between studies
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