Ethosomal Nano-carriers Encapsulating Novel Green Synthesized AgNPs and Sericin Protein as Potential Therapeutics against Skin Carcinoma

Abstract

UV radiations are the main etiological factor that contributes to the induction of skin carcinoma. The incidence of skin carcinoma is increasing at an alarming rate both in the Indian and worldwide scenario. Due to the ignorance and socioeconomic conditions, most of the non-melanoma skin cancer (NMSC) cases are not registered in the national and state cancer registry. Therefore, development of new therapeutics having higher efficacy along with skin regenerative properties is one of the important issues in today’s rapid and progressive world. In this study, novel and economic resources were selected for developing patient-friendly non-toxic therapeutics against skin carcinoma. Four chitosan formulations were prepared using silver nanoparticles (AgNPs) synthesized through biological routes and silk sericin protein. Similarly, four ethosomal formulations were prepared by encapsulating the previously prepared chitosan formulations to overcome the barrier function provided by the stratum corneum. The prepared formulations were characterized through high throughput techniques such as DLS, FE-SEM, TEM, ATR-FT-IR, and XRD. The hemolysis and hemagglutination assays proved the hemocompatibility of the prepared formulations upon incubation with RBCs. The MTT assay on A431 skin carcinoma and normal HaCaT cells showed their dose-dependent cytotoxicity. DCFH-DA, Rhodamine 123 staining provided an insight into the in vitro ROS production and mitochondrial membrane depolarizing capabilities of the prepared formulations. The morphological changes in the cell membrane, depolarization of mitochondrial membrane potential, DNA fragmentation, and annexin V-FITC staining confirms apoptosis being the primary cause of death in the chitosan and ethosomes treated samples. The time-dependent increase in intracellular production of superoxide and nitric oxide radicals confirmed that ROS are the primary source for triggering the cascade for the destruction of the cancer cells. The in vivo immune response studies in mice model showed the triggering of T cell markers in response to the exposure of prepared chitosan and ethosomal formulation.