Objective: The efficiency of acyclovir, a potent antiviral agent, is mainly limited by its low permeability.The purpose of this investigation was to formulate and assess nanoparticle impregnated buccal films to improve acyclovir permeability and thereby enhance bioavailability. Methods: Acyclovir was incorporated into the polymeric materials and formulated as nanoparticles. The prepared nanoparticles were then loaded into various films (F5-F7) prepared with varying quantities of hydroxyethyl cellulose and Eudragit RL 100. The prepared films were evaluated for physico-mechanical characters, mucoadhesion, swelling, in vitro acyclovir release and ex vivo diffusion. Results: The films so prepared showed adequate mucoadhesive strength and excellent physico-mechanical properties. Rapid hydration of films (>35%) were observed in 15 min, followed by a relatively slower phase of hydration which continued till 60 min. In vitro release profile exhibited biphasic pattern, while the rate of acyclovir release was higher with film F7. Prepared films exhibited either first order (F5, control) or Higuchi model (F6, F7) kinetics. The enhancement in acyclovir permeation in F5, F6 and F7 loaded films were 2.2, 4.1 and 6.7 folds, respectively, when compared to control. Conclusion: This study concludes that the drug loaded nanoparticles impregnated buccal film could be an alternative approach to enhance the oral bioavailability of acyclovir, and need to be proved in vivo.