Formulation Containing Phytosomes of Carotenoids from Nyctanthes arbor-tristis and Tagetes patula Protect D-galactose Induced Skin Aging in Mice

Background:Carotenoids play important role in delay of aging process.Orange coloured tubular calyx of flowers of Nyctanthes arbor-tristis contains crocin,an apocarotenoid which forms a major component of stigma of saffron.Due to presence of crocin in orange coloured tubular calyx of Nyctanthes arbor-tristis,it can be used as an economical substitute to saffron for its medicinal and cosmetic utilities.Lutein from flower petals of Tagetes patula L.,is another popular carotenoid which has antioxidant effect and many health benefits.The carotenoids are highly unstable when exposed to atmosphere.One of the ways to improve stability of these phytoconstituents,is their entrapment in phytosomes.Preparation of phytosomes will have dual advantages of improving stability as well as bioavailability of molecule.Objective:In the present study crocin and lutein rich extracts were entrapped into phytosomes to improve stability and efficacy of topical preparation.Methods:The phytosomes of Carotenoid rich extract of tubular calyx of Nyctanthes arbor-tristis L.and the petals of Tagetes patula L.(standardized for crocin and lutein content)were prepared using lipid film hydration technique and these phytosomes were then incorporated into gel base.The gel formulation was evaluated for stability as per ICH guidelines.Efficacy of formulation was evaluated by d-galactose induced aging model.Aging in skin was induced by administration of d-galactose(100 mg/kg bwsc.)to albino mice for 42 days.The gel formulation was applied topically for 42 days.Then the effect of formulation on skin aging was evaluated by estimation of biochemical parameters viz.glutathione and malondialdehyde(MDA)and histopathological studies of treated skin samples.Expression of COL type I and elastin genes in skin samples was also carried out by RT-PCR.Results:Percent entrapment(%w/w)of crocin and lutein in phytosomes were found to be 60.20%and 50.81%,respectively.Accelerated stability studies showed improvement in stability of carotenoids viz.crocin and lutein and the content of crocin and lutein in formulation was found in the range of 99.98%w/w to 99.85%w/w at the end of three months.The formulation containing extract of Phytosomes of carotenoid rich extracts of Nyctanthes arbor-tristis L.and the petals of Tagetes patula L.exhibited potent antiaging activity through significant(P<0.05)increase in dermal and epidermal layers,and increase in GSH levels of skin as compared to the untreated group.The treatment with the gel formulation revealed upregulation of collagen type I and elastin genes.There was significant reduction in lipid peroxidation as revealed through reduction in MDA levels as compared to untreated group.Conclusion:Crocin and lutein have potential to prevent skin aging via upregulation of collagen type I gene and elastin gene.Upregulation of genes resulted into increase in the thickness of epidermal and dermal layer along with reduction in oxidative stress in skin.Entrapment of carotenoid rich extracts of Nyctanthes arbor-tristis and Tagetes patula in phytosomes enhanced the stability and efficacy of the formulation.