Formulation Development And Pharmacological Evaluation Of Topical Hyalurosomes Nanogel Of Calotropis Gigantea For The Treatment Of Vitiligo

Abstract

Objective: The objective of the paper is preparation and pharmacological evaluation of topical hyalurosomes nanogel of Calotropis Gigantea for the treatment of Vitiligo.

Material and methods: Fresh milky white latex of Calotropis Gigantea was collected by cutting of stem twigs and identified by FT-IR for the presence of active phytoconstituents. Hyalurosomes as nanogel formulation was prepared by solvent evaporation method. The nanogel for anti-vitiligo activity was evaluated and performed by utilising cell viability, melanin content, tyrosinase activity assay and Fontana-Masson silver staining on B16F10 cell line.

Result: The FT-IR analysis has shown the broad peak at 3253 cm^-1 to OH-str. frequencies of uscharin, calotropin, lubeol, and b-amyrin. The Petroleum ether latex of Calotropis Gigantea contain i.e., lupeol, beta-amyrin, alpha calotropeol, and beta-calotropeol were utilized as drug for the topical nanogel delivery. The Cell viability assay stated that CGHGF showed 68.71±2.76% cytotoxicity, as compared to CG-Carbopol Gel (52.10±9.63%) at 50μM concentration. The MTT assay stated the CG-Carbopol gel show 56.11±11.55% and CGHGF has shown 82.95±11.61% cytotoxicity at 50 µM concentration. Melanin and tyrosinase activity stated that petroleum latex of Calotropis Gigantea solution showed an increase in melanin content by 1.20 fold upon first treatment (p<0.05) and 1.20 fold upon second treatment (p<0.05). CGHGF is showed a 2.25 fold increase in melanin with first treatment (p<0.0001) and a 2.89 fold increase with second treatment (p<0.0001). Fontana-Masson silver staining is performed to visualize melanin pigment in cells exposed to Petroleum latex of Calotropis Gigantea CGHGF formulations. Compared to the control, the amount of melanin granules was significantly increased and stained in petroleum latex of Calotropis Gigantea based nanogel.

Discussion: Unique deformable nature of hyalurosomes and CG Carbopol gel also assists in skin penetration enhancement of latex containing active constituents. Cell uptake studies confirmed higher penetration of encapsulated moiety through prepared CGHGF.