Natural Product Research Formerly Natural Product Letters

ISSN: 1478-6419 (Print) 1478-6427 (Online) Journal homepage: http://www.tandfonline.com/loi/gnpl20

Multiple-physiological benefits of bird’s nest ( australasicum) frond extract for dermatological applications

Wen-Wen Zeng & Lih-Shiuh Lai

To cite this article: Wen-Wen Zeng & Lih-Shiuh Lai (2017): Multiple-physiological benefits of bird’s nest fern () frond extract for dermatological applications, Natural Product Research, DOI: 10.1080/14786419.2017.1405400 To link to this article: https://doi.org/10.1080/14786419.2017.1405400

View supplementary material

Published online: 22 Nov 2017.

Submit your article to this journal

Article views: 28

View related articles

View Crossmark data

Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=gnpl20 Natural Product Research, 2017 https://doi.org/10.1080/14786419.2017.1405400

SHORT COMMUNICATION Multiple-physiological benefits of bird’s nest fern (Asplenium australasicum) frond extract for dermatological applications

Wen-Wen Zeng and Lih-Shiuh Lai

Department of Food Science and Biotechnology, National Chung Hsing University, Taichung, Taiwan

ABSTRACT ARTICLE HISTORY p-coumaric acid and fucose-rich polysaccharide have been studied Received 19 July 2017 for many bio-functions in skin including cutaneous protection from Accepted 3 November 2017 oxidative damage and antiageing, respectively, as well as wound KEYWORDS healing. The physiological activities of various bird’s nest fern extracts Bird’s nest fern; fibroblast; (BNFE), containing considerable fucose-rich mucilage and p-coumaric skin ageing; collagen; acid, on fibroblast and human skin were first investigated. BNFE with fucose-rich mucilage; p- higher polysaccharide content generally contributed to a better coumaric acid moisture holding capability. Furthermore, BNFE showed pronouncedly enhancing effect on collagen production and growth of fibroblast (NIH-3T3), clinical trial results revealed that the emulsions with 1% BNFE showed good moisturising effect and improved the elasticity of human skins effectively. The potential of BNFE for cosmetics and medical applications such as natural moisturiser, antiageing and wound repairing was possibly related to the fucose-rich mucilage and various phenolic compounds including p-coumaric acid in BNFE.

1. Introduction Asplenium australasicum (J. Sm.) Hook and Asplenium nidus L. were recognised to be similar but different according to the of chloroplast DNA sequence data in Asplenium taxa (Perrie and Brownsey 2005). The differentiating appearance between the

CONTACT Lih-Shiuh Lai [email protected] supplemental data for this article can be accessed at https://doi.org/10.1080/14786419.2017.1405400. © 2017 Informa UK Limited, trading as Taylor & Francis Group 2 W.-W. ZENG AND L.-S. LAI two species have also been reported by Holttum (1974) that the midrib of A. australasicum fronds is strongly and acutely keeled on lower surface, while the costa of A. nidus fronds is broad but only slightly prominent (not keeled) on lower surface. The young frond of A. australasicum is a popular fern vegetable in Taiwan for its slimy texture, which may relate to abundant mucilage in its midrib. It was also called ‘bird’s nest fern’ (BNF) since it usually forms funnel-shaped nest on tree trunk for receiving more rain and organic materials. A. australasicum and A. nidus could be utilised as medicinal for their physiological activities such as contraception and alleviate headache (Petard 1985; Bourdy et al. 1996). The aqueous ethanol extracts of A. australasicum fronds also demonstrated antioxidant activities including reducing power and scavenging reactive oxygen species, and the main phenolic compounds in the extracts was found to be p-coumaric acid (p-CoA) (Chang 2012), which has been reported as one of the powerful antioxidants to protect cultured skin cells (including human fibroblasts and keratinocytes) from oxidative damage (Phan et al. 2001). The mucilage isolated from BNF have been reported to show good moisture holding capability and quite high intrinsic viscosity due to the strong interaction between its large molecular weight (4.87 MDa) polysaccharide (mainly consisted of galactose and fucose) and water molecules (Zeng and Lai 2016). Many researches have pointed out that L-fucose and fucose-rich oligo- and polysaccharides (FROP-s), composed of fucose, galactose and acetyl galacturonic acid with high molecular weight (Péterszegi et al. 2003), influence several cel- lular and tissue reactions known to undergo age-dependent modifications such as improving collagen fibrillogenesis by fibroblasts and increasing elastin biosynthesis in vitro and in vivo (Isnard et al. 2004). Research about skin-caring influences of extracts ofA. australasicum fronds, containing both p-CoA and fucose-rich polysaccharide, is quite limited. Therefore, the aim of this study is to assess the relation between the skin health-related activities and the potential bioactive compounds in various BNFE.

2. Results and discussions 2.1. Potential active compositions and moisture holding capability of BNFE As compared with the water-extracted mucilage (WM) which contained 27.9% protein, 52.8% carbohydrate and showed 27.0% moisture holding capability (MHC) (Zeng and Lai 2016), BNFE had much lower amounts of hydrophilic components such as proteins and carbohy- drates, leading to lower MHC (Table S1). However, significantly higher amount of other bio- active components in BNFE, including phenolics and flavonoids, would be possibly beneficial for skincare-related applications. p-coumaric acid (p-CoA) was determined as the main phe- nolic compounds in BNFE as shown in Figure S1. Phan et al. (2001) pointed out that a mixture of powerful antioxidant compounds, including the mainly phenolic acids like p-coumaric acid and protocatechuic acid as well as some flavonoid aglycones, in Chromolaena odorata ethanol extract may contribute to enhanced wound healing through protecting skin cell from oxidative damage. Moreover, the much higher extraction yield (about 26.8~29.4%) of BNFE than WM (about 4.8%) may show economic advantages. Monosaccharide composition analysis revealed that the mucilage in WE was mainly composed of 29.0% Gal, 17.9% Fuc, 15.1% Xyl, 12.5% Ara, 11.4% Man (Figure S2), and is consistent with the findings of Zeng and Lai (2016) for WM extracted at 70 °C. NATURAL PRODUCT RESEARCH 3

2.2. Enhanced proliferation activity of NIH-3T3 by BNFE All BNFE not only show no cytotoxicity to fibroblast NIH-3T3 at the addition level (0–400 μg/ mL) but also showed enhanced cell proliferation (Table 1). The increase of mitochondrial dehydrogenase activity proved the stimulatory effect of BNFE on cell metabolism. The via- bility of NIH-3T3 generally increased in a dose-dependent and time-dependent model, in which the viability got higher than that of one-day treatment at the same concentration level, and was generally raised significantly with the increasing BNFE concentration up to 300 μg/mL, then declined. It was thought that the fucose-rich mucilage and phenolic com- pounds in BNFE may contribute to the enhancing proliferation of fibroblast, therefore, p-CoA and WM were further tested in parallel. The viability of fibroblast followed a bell-shape way in enhancing proliferation for one-day cultivation by reaching a maximum at 50 μg/mL WM (Table S2), which was approximately equivalent to the mucilage content in 300 μg/mL of WE due to the yield ratio of WM to WE was about 4.8/26.8. Similarly, the enhanced prolifer- ation effect of p-CoA (Table S2) seemed to reach a maximum under a concentration of 1–2 μg/mL which were also approximately equivalent to the p-CoA content in 300 μg/mL of WE or AE40 (Table S1), then levelled off. These results implied that both WM and p-CoA contributed to the proliferative activity of BNFE, though the synergistic contribution of other phenolics cannot be excluded.

2.3. Collagen production of fibroblasts stimulated by BNFE Age-related thinning of dermis and epidermis is the major driving force for the ageing-look skin in elderly, largely due to the loss of collagen, the major structural protein in skin (Shao et al. 2017). On the other hand, it also reported that age-related thinning of the dermis leads to poor wound healing (Thomas and Burkemper 2013), hence the regulation of collagen production is critical in skin health. The relative percentage of collagen content was 306, 334 and 217% for treatment with WE, AE25 and AE40 at 50 μg/mL, respectively, which were much higher than that of the control and comparable to that of L-ascorbate (known for stimulating collagen synthesis through prolyl hydroxylation (Murad et al. 1981)) at the same concentration (Figure1(a)). By the treatment of BNFE, the collagen accumulation generally followed a bell-shape way, which was essentially consistent with their enhancing effects on NIH-3T3 proliferation since collagens interact with cells to regulate their proliferation, migra- tion and differentiation (Ricard-Blum 2011). WM and p-CoA treatment could also stimulate

Table 1. Effect of bird’s nest fern extracts on the viability of NIH-3T3 mouse embryonic fibroblasts cultivated in DMEM plus CS medium.

Concentration of aqueous extracts (μg/mL) days 50 100 200 300 400 WE 1 d 100.2 ± 1.0 101.1 ± 1.6 109.4 ± 3.7* 114.6 ± 1.9** 104.6 ± 5.2 3 d 120.8 ± 4.3* 129.0 ± 3.5** 143.8 ± 6.4** 208.4 ± 4.4*** 189.1 ± 10.2** AE25 1 d 104.7 ± 3.6 109.7 ± 2.1* 103.4 ± 0.6* 115.6 ± 4.2* 102.7 ± 3.9 3 d 110.0 ± 8.6 120.6 ± 6.3* 159.4 ± 2.6*** 253.2 ± 4.5*** 121.8 ± 2.4** AE40 1 d 115.3 ± 0.7*** 117.8 ± 1.9** 121.8 ± 4.2* 131.1 ± 4.6** 129.6 ± 3.3** 3 d 116.9 ± 0.9*** 142.8 ± 4.6** 193.8 ± 4.2*** 223.2 ± 2.4*** 141.9 ± 2.1*** *Differ significantly p < 0.05 with control (viability=100). **Differ significantlyp < 0.01 with control (viability=100). ***Differ significantly p < 0.001 with control (viability=100). Note: Each data was expressed as the mean of three replications ± standard deviation. 4 W.-W. ZENG AND L.-S. LAI

Figure 1. Relative collagen production of NIH-3T3 mouse embryonic fibroblasts stimulated by (a) bird’s nest fern extracts (BNFE), (b) bird’s nest fern mucilage (WM) and (c) p-coumaric acid. Bars represent mean ± S.D. of three cultured wells. Means with *, ** and *** differ significantly (p < 0.05, p < 0.01 and p < 0.001, respectively) with control. NATURAL PRODUCT RESEARCH 5 fibroblast to produce collagen under a quite low concentration range (Figure 1(b) and (c)). The phenolic acids in Symphytum officinale L. were also suggested to positively affect skin healing process because excess of reactive oxygen species present can cause damage of healthy cells (Sowa et al. 2017). For wound healing purposes, a cell growth-enhancing activity on fibroblasts or endothelial cells may help rehabilitate cutaneous tissues and tissue regen- eration in a shorter time (Chen et al. 2014). In preliminary, BNFE have shown the beneficial effects of retarding skin ageing and accelerating the rehabilitation of wound by promoting fibroblast growth and constructing the major ECM in skin leading to maintain skin thickness (Table 1 and Figure 1).

2.4. Cutaneous properties of human skin during 28-day clinical trial WE and AE40 were formulated into a model skincare emulsion and examined for a place- bo-controlled clinical trial with intra-individual comparison of two formulations on the fore- arm. No allergic reaction was noticed in 46 participated volunteers. A higher increase in skin hydration by applying emulsions with 1% BNFE than that of vehicle (control) in each week (Figure S3), implied the underlying moisturising and retarding ageing effect of WE and AE40, because a disrupted balance between water content in stratum corneum and skin surface lipids would affect the appearance and mechanical properties of the skin such as irregular skin dryness, elasticity loss, atrophy or wrinkling (Sator et al. 2003; Kapoor and Saraf 2010). Formulations with 1% BNFE, particularly for AE40, also showed better improvement of dry and flaky shin than control for the same individual (Figure S4). Such properties were not only good for aesthetic purposes but also for maintenance of normal skin conditions and pre- vention of dry skin alterations. The formulation with 1% WE was proven to have potentials of retarding ageing of human skin by the evidences of increasing skin elasticity (Figure S5a), which may result from the high enhancing activity on collagen production because the expression level of collagen (especially type I), alignment and organisation of collagen fibres primarily contribute to the tensile strength of skin (Kwan et al. 2011).

3. Experimental The experimental details, Table S1–S2 and Figures S1–S5 were provided in the supplementary material.

4. Conclusion The cellular proliferative activity and promotion of collagen production of BNFE made them applicable as multiple functional ingredients in wound healing or cosmeceutical treatments. From the in vivo experiments, WE and AE40 in a vehicle of emulsion were quite effective in terms of improving skin hydration and further increasing elasticity. The bio-properties of BNFE on skin were possibly attributed to the cooperative effects of their active compositions including the fucose-rich mucilage and p-coumaric acid.

Acknowledgments Sincere appreciation to the kind sponsorship of National Science Council, Taiwan (project number: NSC 101-2313-B-005 -048 -MY3), without which this work would not have been possible. 6 W.-W. ZENG AND L.-S. LAI

Disclosure statement No potential conflict of interest was reported by the authors.

Funding This work was supported by the National Science Council [grant number NSC 101-2313-B-005 -048 -MY3].

References Bourdy G, François C, Andary C, Boucard M. 1996. Maternity and medicinal in Vanuatu II. Pharmacological screening of five selected species. J Ethnopharmacol. 52:139–143. Chang CH. 2012. Effect of hot-air drying and extraction conditions on the antioxidant and functional properties of Asplenium australasicum (J Sm) Hook [master thesis]. Taichung: National Chung Hsing University. Chen YS, Lee SM, Lin YJ, Chiang SH, Lin CC. 2014. Effects of danshensu and salvianolic acid B from Salvia miltiorrhiza Bunge (Lamiaceae) on cell proliferation and collagen and melanin production. Molecules. 19:2029–2041. Holttum RE. 1974. Asplenium Linn., sect. Thamnopteris Presl. Gardens’ Bulletin, Singapore. 27:143–154. Isnard N, Fodil-Bourahla I, Robert AM, Robert L. 2004. Pharmacology of skin aging. Stimulation of glycosaminoglycan biosynthesis by L-fucose and fucose-rich polysaccharides, effect of in vitro aging of fibroblasts. Biomed Pharmacother. 58:202–204. Kapoor S, Saraf S. 2010. Assessment of viscoelasticity and hydration effect of herbal moisturizers using bioengineering techniques. Pharmacogn Mag. Oct;6:298–304. Kwan KH, Liu X, To MK, Yeung KW, Ho C-M, Wong KK. 2011. Modulation of collagen alignment by silver nanoparticles results in better mechanical properties in wound healing. Nanomed Nanotechnol Biol Med. 7:497–504. Murad S, Grove D, Lindberg K, Reynolds G, Sivarajah A, Pinnell S. 1981. Regulation of collagen synthesis by ascorbic acid. Proceedings of the National Academy of Sciences. 78:2879–2882. Perrie LR, Brownsey PJ. 2005. Insights into the biogeography and polyploid evolution of New Zealand Asplenium from chloroplast DNA sequence data. Am Fern J. 95:1–21. Petard P. 1985. Plantes Utiles de Polynesie Raau Tahiti [Useful Plants of Polynesie Raau Tahiti]. J Soc Ocean. 41:148–149. Péterszegi G, Isnard N, Robert AM, Robert L. 2003. Studies on skin aging. Preparation and properties of fucose-rich oligo-and polysaccharides. Effect on fibroblast proliferation and survival. Biomed Pharmacother. 57:187–194. Phan T-T, Wang L, See P, Grayer RJ, Chan S-Y, Lee ST. 2001. Phenolic compounds of Chromolaena odorata protect cultured skin cells from oxidative damage: implication for cutaneous wound healing. Biol Pharm Bull. 24:1373–1379. Ricard-Blum S. 2011. The collagen family. Cold Spring Harb Perspect Biol. 3:a004978. Sator PG, Schmidt JB, Hönigsmann H. 2003. Comparison of epidermal hydration and skin surface lipids in healthy individuals and in patients with atopic dermatitis. J Am Acad Dermatol. Mar;48:352–358. Shao Y, He T, Fisher G, Voorhees J, Quan T. 2017. Molecular basis of retinol anti-ageing properties in naturally aged human skin in vivo. Int J Cosmetic Sci. 39:56–65. Sowa I, Paduch R, Strzemski M, Zielińska S, Rydzik-Strzemska E, Sawicki J, Kocjan R, Polkowski J, Matkowski A, Latalski M. 2017. Proliferative and antioxidant activity of Symphytum officinale root extract. Nat Prod Res. 10:1–5. Thomas DR, Burkemper NM. 2013. Aging skin and wound healing. Clin Geriatr Med. 29:xi–xx. Zeng W-W, Lai L-S. 2016. Characterization of the mucilage extracted from the edible fronds of bird’s nest fern (Asplenium australasicum) with enzymatic modifications. Food Hydrocoll. 53:84–92. 本文献由“学霸图书馆-文献云下载”收集自网络，仅供学习交流使用。

学霸图书馆（www.xuebalib.com）是一个“整合众多图书馆数据库资源，

提供一站式文献检索和下载服务”的24 小时在线不限IP 图书馆。 图书馆致力于便利、促进学习与科研，提供最强文献下载服务。

图书馆导航：

图书馆首页 文献云下载 图书馆入口 外文数据库大全 疑难文献辅助工具