Chitosan Assisted Immunotherapy for Intervention of Experimental Leishmaniasis via Amphotericin B Loaded Solid Lipid Nanoparticles

Abstract

Solid lipid nanoparticles (SLNs) have emerged as an excellent substitute over polymeric nanoparticles and when incorporated with chitosan which activates the macrophage to impart an immune response, produces excellent results to fight against the deleterious disease like leishmania where its parasite diminishes the immunity of host to induce resistance. Based upon these hypothesis chitosan coated SLNs were developed and loaded with amphotericin B (AmB) for immunoadjuvant chemotherapy of leishmania infection. Both uncoated and chitosan coated AmB loaded SLNs (AmB‐SLNs) were fabricated using solvent emulsification and evaporation method. The various process and formulation parameters involved in AmB‐SLNs preparation were optimized in respect to particle size, and stability of the particles. In vitro hemolytic test credited the formulations to be safe when injected in veins. The cellular uptake analysis demonstrated that the chitosan coated AmB‐SLN was more efficiently internalized into the J774A.1 cells. The in vitro antileishmanial activity revealed their high potency against leishmania infected cells in which chitosan coated AmB‐SLNs were distinguishly efficacious over commercial formulations (Ambisome and Fungizone). In vitro cytokine estimation study revealed that chitosan coated AmB‐SLNs activated the macrophages to impart a specific immune response through enhanced production of TNF‐α and IL‐12 with respect to normal control. Furthermore, cytotoxic studies in macrophages and acute toxicity studies in mice evidenced the better safety profile of developed formulation in comparison to marketed formulations. This study indicates that the AmB‐SLNs are a safe and efficacious drug delivery system which promises strong competence in antileishmanial chemo and immunotherapy.