Chitosan nanoparticles as a delivery platform for neurotoxin II from Androctonus australis hector scorpion venom: Assessment of toxicity and immunogenicity.

In recent years, biodegradable polymers based nanoparticles received high interest for the development of vaccine delivery vehicles. In this study, chitosan nanoparticles encapsulating Aah II toxin (AahII-CNPs) isolated from Androctonus australis hector venom, were investigated as vaccine delivery system. Particles obtained by ionotropic gelation were characterized for their size, surface charge, morphology and toxin release profile from Aah II-CNPs. Toxin-nanoparticles interactions were assessed by Fourier Transform Infrared Spectrometry and X-Ray Diffraction. An immunization protocol was designed in mice to investigate anti-toxin immunity and the protective status induced by different Aah II immune formulations. Unloaded chitosan nanoparticles presenting a spherical shape and smooth surface, were characterized by a size of 185 nm, a dispersion index (PDI) of 0.257 and a zeta potential of +34.6 mV. Aah II toxin was successfully entrapped into chitosan nanoparticles as revealed by FTIR and XRD data. Entrapment efficiency (EE) and Loading capacity (LC) were respectively of 96.66 and 33.5%. Aah II-CNPs had a diameter of 208 nm, a PDI of 0.23 and a zeta potential of +30 mV. Encapsulation of Aah II reduced its toxicity and protected mice until 10 LD. Mice were immunized via a dual prime-boost scheme. Nanoentrapped Aah II immunogen elicited systemic innate and humoral immune responses as well as local spleen parenchyma hyperplasic alterations. Aah II-CNPs immunized mice withstood high lethal doses of native Aah II, one-month post-boost inoculation. This study provided encouraging and promising results for the development of preventive therapies against scorpion envenoming mainly for the populations at-risk.