In vitro evaluation of the suppressive effect of chitosan/poly(vinyl alcohol) microspheres on attachment of C. parvum to enterocytic cells

Abstract

We present a new strategy to suppress the attachment of Cryptosporidium parvum to the enterocytes cell surface by bioadhesive microspheres. An optimized microsphere system based on chitosan/poly(vinyl alcohol) was prepared by experimental design for the delivery of Diloxanide Furoate–cyclodextrin complex. Formulations were characterized in terms of size, surface charge, drug release, IR spectroscopy and morphology. Bioadhesion properties of chitosan/poly(vinyl alcohol) microspheres, evaluated in the human enterocytic HCT-8 model, were concentration and time dependent. In vitro efficacy of chitosan/poly(vinyl alcohol) microspheres against Cryptosporidium was tested in infected cultures and stages of parasite were assessed by immunofluorescence. The degree of adherence to cells and the inhibition of infectivity were directly related with the lowest level of cross-linking. The C. parvum attachment to cells surface was efficiently suppressed by a concentration of 100μg/ml of microspheres. TEM observations showed no epithelial-cell damage when microspheres were co-incubated in infected cultures. These results were coincident with the lack of toxicity in cytocompatibility studies. Microspheres remained adhered after 72h to the apical area of enterocytes. The results suggest that chitosan/poly(vinyl alcohol) with adequate size and appropriate surface characteristics suppress by impairment the attachment of sporozoites to enterocytes and may have a great potential in the oral chemotherapy of Cryptosporidium infections.