Intravaginal mucoadhesive microspheres of tenofovir disoproxil fumarate (TDF) has been formulated using chitosan as the matrix-forming mucoadhesive polymer and evaluated. Ionotropic gelation method was used to produce the microspheres. Drug-polymer compatibility, percentage yield, particle size determination by optical microscopy, drug content & encapsulation efficiency, ex vivo mucoadhesion and in vitro release were evaluated. Based on the evaluation studies, F6 was selected as the best formulation and evaluated further for release kinetics and curve fitting analysis. F6 followed Korsmeyer-Peppas and Higuchi models. Accelerated stability study was conducted as per ICH guidelines. Drug content analysis, FTIR, XRD, SEM analysis showed that it remained stable for a period of six months.