Conventional formulations can not achieve wound healing efficiently and fail to accelerate wound regeneration. To overcome these problems, it was planned to develop nanoformulations that perform a positive effect on the wound healing duration and are suitable for topical use. In this study, liposomal film formulations that encapsulated d-panthenyl triacetate (PTA) and coenzyme Q10 (CoQ10) were optimized by using response surface methodology (RSM) and were analyzed for their wound healing efficacy and cytotoxicity on fibroblast (CCD1079 Sk) and keratinocyte (HEKa) cells. Swelling index, puncture strength, and puncture deformation values, which were choosen as dependent variables for the liposomal film formulation were found as 556.9% +/- 21.3, 3.98 +/- 0.98 N/mm(2), and 6.57% +/- 1.12, respectively. Cumulative release of 65.32% for PTA and 12.23% for CoQ10 was obtained after 24 hours of in vitro release study in sink conditions. The in vitro cytotoxicity and wound healing assay results suggested that optimum formulation could be used safely on fibroblast and keratinocyte cells and provided wound closure entirely after 24 h. Consequently, the optimum liposomal film containing PTA and CoQ10 formulations could be proposed as an innovative approach in wound healing treatment, considering their release, mechanical properties, stability, and effectiveness.