Fisetin loaded binary ethosomes for management of skin cancer by dermal application on UV exposed mice

Fisetin loaded binary ethosomes were prepared and optimized using Box-Behnken design for dermal application to alleviate skin cancer. The prepared formulations were evaluated for vesicle size, entrapment efficiency and flux of fisetin. Additionally, the optimized formulation was further evaluated by transmission electron microscopy, confocal laser microscopy, vesicles-skin interaction, dermatokinetic study and DPPH (2, 2-diphenyl-1-picryl-hydrazyl) assay. The in vivo study was carried out for the evaluation of tumor incidence, pro-inflammatory cytokines such as TNF-alpha and IL-1 alpha, lipid peroxidation values, glutathione content and catalase activity in mice. The optimized binary ethosomes formulation presented sealed unilamellar shaped vesicles, with vesicles size, entrapment efficiency and flux of 99.89 +/- 3.24 nm, 89.23 +/- 2.13% and 1.01 +/- 0.03 mu g/cm(2)/h respectively. The confocal images of rat skin clearly illustrated the deeper penetration of rhodamine B loaded binary ethosomes formulation. Further, the binary ethosomes gel treated rat skin showed substantial increase in C-Skin max and AUC(0-8) in comparison to rat skin treated with conventional gel. In vivo study revealed that the mice pre-treated with fisetin binary ethosomes gel caused marked decrease in the levels of TNF-alpha and IL-1 alpha as compared to the mice exposed to UV only. Further the binary ethosomes gel treated mice group had demonstrated less percentage of tumour incidences (49%) as compared to mice treated with UV only (96% tumour incidence). The novelty of the work lies in successful optimization of formulation using Box-Behnken design and characterization of binary ethosomes carrier of fisetin and demonstration of improved dermal delivery of the same. The overall data suggest that the fisetin loaded binary ethosomes formulation is a potential dermal delivery system for the management of skin cancer.