Developement of Natural Multifunctional Cosmetic Ingredients Using Biotechnological Approach

Materials based on renewable sources are becoming increasingly important to the cosmetic industry. The use of biotechnology to produce natural cosmetic ingredients has the potential to address a number of environmental and sustainability concerns. This study explores the use of mushroom Grifola frondosa, of the Basidiomycetes phylum, for the production of new extracellular polysaccharide for cosmetic use. The study focuses on the isolation, purification and characterisation of Grifo/a frondosa extracellular polysaccharide (GFEPS), along with the evaluation of its in vitro and in vivo efficacy as multifunctional cosmetic ingredient.

GFEPS was obtained using the biotechnological method of submerged fermentation. Purification and structural characterisation of a newly isolated polysaccharide was carried out by a range of analytical methods. Ion exchange chromatography was used to separate GFEPS into neutral (nEPS) and charged (cEPS) fractions. Molecular weight of GFPS, nEPS and cEPS was determined by Gel permeation chromatography as 216 KOa, 34 KOa and 251 KOa, respectively. Composition and linkage analysis were explored by Gas chromatography - Mass spectrometry (GC - MS) method, which revealed GFEPS to be a heterogenous extracellular polysaccharide consisting of D-mannose, O-glucose, O-galactose and Lfucose. Cytotoxicity analysis of GFEPS showed no significant toxic effects on endothelial (HMEC-1) and keratinocyte (HaCaTs) cell lines.

An in vitro assay to measure reactive oxygen species production by human white blood cells was used as a marker for immunological activity of GFEPS. Antioxidant assays revealed a considerable scavenging activity upon peroxynitrite and superoxide, suggesting that GFEPS is a freeradical scavenging agent. In addition, it was shown that GFEPS significantly inhibited the production of reactive oxygen species from white blood cells in a dose dependant manner, suggesting that the new polysaccharide has a high antioxidant potential.

An evaluation of the in vivo effects of GFEPS on human skin was carried out in the form of short term instrumental study, assessing skin hydration, trans-epidermal water loss (TEWL) , skin erythema and skin topography. Polyacrylic acid hydrogel was used as a vehicle for GFEPS (1 % and 5% wlw), as well as for five other commercially available polysaccharides. The results have shown a significant increase in skin hydration produced by GFEPS. It also outperformed all tested commercial polysaccharide actives in improving the skin barrier, shown by the highest decrease in TEWL values. A positive but moderate effect on reduction in skin erythema was also observed, as well as an improvement in the skin topography parameters.

Overall, this thesis has shown that extracellular polysaccharide derived from mushroom G. frondosa presents a naturally produced, multifunctional and effective new material for cosmetic use.

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