Evaluation of Skin Depigmentation on The Melanocytes of Zebrafish Using Biomolecules of Aloe vera and Punica granatum

Abstract

Depigmentation is a process where the synthesis of melanin pigment is inhibited in the pigment cells of the melanocyte resulting in discolouration of the skin. Thus, whitening or lightening cosmetics are utilized extensively for the prevention and treatment of irregular hyperpigmentation. Recently, many topical depigmenting agents have been developed and widely used for the treatment of pigmentary diseases. Hydroquinone is the main component of topical pharmacological agents, which exhibits great efficacy, but has a relatively high rate of adverse side effects. During the period of present study in this thesis, different techniques related to collection of plant material and animal identification and authentication of plant species, extraction, analytical High Performance Liquid Chromatography (HPLC) has been done. Further, to identified percentage of active compounds in the crude extracts of Aloe vera leaves and Punica granatum fruits has been analysed by Liquid Chromatography- Electrospray Ionization mass spectrometry/Mass Spectrometry (LC-ESI/MS). In vivo and in vitro pharmacological studies have been investigated of same plants and their active compounds for the development of novel skin lightening agents using zebrafish (Danio rerio) animal model. Our findings concluded that crude extract of Aloe vera and Punica granatum and pure aloin and gallic acid showed anti-melanogenic property in vivo and in vitro. This was confirmed by using pure aloin and gallic acid as standards. Crude extract of Aloe vera leaves and Punica granatum fruits inhibits the activity of tyrosinase enzyme and tyrosinase protein expression thus inhibiting melanogenesis in the embryos of zebrafish. Further depigmentation effect of these extract was visualized over the scale s melanocytes of zebrafish. It was observed that crude extract of Aloe vera leaves and Punica granatum fruits significantly aggregates melanin and make the zebrafish skin pale. LC50 (Probit analysis) further confirms the non-toxic nature of these extracts. All these f