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 fern (Asplenium 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 (Asplenium australasicum) 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 species according to the taxonomy 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 W.-W. ZENG AND L.-S. LAI 2 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 ferns 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).