Development of cosmeceutical formulations based on mushroom extracts and their individual compounds

Resumen

The cosmetic industry is constantly in search of ingredients from natural sources because of their multifunctional properties, competitive effectiveness, and lower toxicity effects. Otherwise, mushrooms have been an important part of our diet for years due to their rich nutritional and abundant content in biomolecules; however, their cosmeceutical and nutricosmetic potential have not been fully exploited. This study aimed at optimizing the extraction of bioactive compounds from Ganoderma lucidum and Agaricus blazei by Response Surface Methodology (RSM) and comparing conventional extraction techniques with more sustainable methods. Further studies regarding their nutritional and chemical composition (e.g., sugars, organic acids, tocopherols, and fatty acids), were conducted. Also, extracts obtained from those mushrooms, but also from Agaricus bisporus, Pleurotus ostreatus and Lentinula edodes extracts, using the Soxhlet extraction system, were analysed in terms of antiinflammatory, anti-tyrosinase, antioxidant, and antimicrobial activities followed by their characterization in terms of phenolic acids and related compounds, ergosterol, and triterpenoids. Furthermore, the obtained extracts were incorporated in a base cosmetic cream to ascertain bioactive properties sustainment. Moreover, to evaluate the individual cosmeceutical contribution of p-hydroxybenzoic acid, p-coumaric acid, protocatechuic acid, cinnamic acid and ergosterol, their anti-inflammatory, anti-tyrosinase, and antimicrobial activities were studied, followed by incorporation of these compounds in a base cosmetic cream. Because of the possibility of the extracts and the individual compounds to lose their potency over time either due to oxidation or degradation, microparticles were prepared using the atomization/coagulation technique. The microparticles were characterized in terms of morphology, particle size distribution, and encapsulation efficiency. This is followed by incorporation of the produced microparticles into a base cosmetic cream and comparison of their performance with the free forms (formulations containing free extracts/individual compounds). The most promising extracts and their corresponding formulations were submitted to in vitro safety evaluation using the MTT and LDH assays in keratinocyte (HaCaT) and fibroblast (HFF-1) cell lines. Moreover, they were submitted to ex vivo skin permeation studies using a Franz diffusion apparatus with pig ear skin as permeation membrane to determine topical bioavailability of the bioactive compounds present in the extracts. The optimization study on bioactive compounds recovery from G. lucidum and A. blazei using RSM showed that the values predicted by the models were in close agreement with experimental observations. Furthermore, it was concluded that the extraction yield of bioactive compounds from the studied mushrooms was highly dependent on the extraction variables (time, temperature, solvent proportion, solid-liquid ratio, and power). A. blazei and G. ludidum also showed high nutritional contribution. α-Tocopherol, oxalic acid and twenty-one fatty acids (mainly PUFA) were quantified in A. blazei. G. lucidum showed a similar fatty acids’ profile (PUFA˃SFA˃MUFA) and with the presence of α-tocopherol and δ-tocopherol. Comparatively, A. blazei revealed lower fat content, but higher energy contribution. The cell viability effect of the extract prepared from A. blazei on Caco-2 and HT29-MTX cells was found to be maintained in a concentration-dependent manner. The ethanolic extracts obtained from the studied mushrooms showed antioxidant, antiinflammatory, anti-tyrosinase and antimicrobial activities, and after incorporation into the base cosmetic cream, the final cosmeceutical formulations preserved some of these bioactivities. G. lucidum extract was found to be a rich source of triterpenoids mainly due to the high contribution of ganoderic acid A, C2, and H, while phenolic acids (p-hydroxybenzoic acid, pcoumaric acid and protocatechuic acid), the related compound cinnamic acid, and ergosterol were identified in A. bisporus, P. ostreatus, L. edodes and A. blazei extracts. Each one of these compounds also showed anti-inflammatory, anti-tyrosinase and antimicrobial activities. Furthermore, microparticles were successfully used to encapsulate extract/individual compounds with high encapsulation efficiency and yield. After incorporation of the free extracts/individual compounds and the produced microparticles into the base cosmetic cream, the encapsulated forms allowed for a controlled release leading to bioactivity preservation when compared with the free forms. Because they presented the most interesting cosmeceutical potential, P. ostreatus and G. lucidum extracts were submitted to toxicological studies, revealing no toxicity in keratinocytes and fibroblast cell lines, which is indicative of their potential safety to be used as cosmeceutical ingredients. Protocatechuic acid and syringic acid were the only compounds permeating from the G. lucidum extract, while very low detection was observed for compounds present in the P. ostreatus extract. Similar results were also obtained for the formulation prepared using both extracts. The results showed that tritepenoids in G. lucidum might be natural permeation enhancers and these results pointed out for the suitability of using these mushroom extracts as skin care ingredients. Overall, the results showed that skin care products can be formulated using mushroom extracts and that these extracts have the potential to be utilised at large scale use. Hence, integration of wastes generated by the mushroom processing industry, which can serve as raw materials for the obtainment of bioactive extracts, can be intensified.