In this study, vitamin A palmitate (VAP)-loaded poly(lactic-co-glycolic acid) (PLGA)/chitosan-coated PLGA nanoparticle (NP) systems were prepared by the nanoprecipitation technique. The prepared systems were characterized by parameters such as particle size, polydispersity index (PDI), ζ-potential, encapsulation efficiency, in vitro dissolution, and release kinetic study. Then, the cytotoxicity and wound healing profiles of the designed NP formulations in HaCaT (human keratinocyte skin cell lines) were determined. The particle size of VAP-loaded NPs was obtained between 196.33 ± 0.65 and 669.23 ± 5.49 nm. PDI data proved that all NPs were prepared as high quality and monodisperse. While negative ζ-potential values of Blank-NP-1 and NP-1 encoded PLGA NP formulations were obtained, positive ζ-potential was obtained in chitosan-coated NPs. In vitro release studies of NPs observed rapid dissolution in the first 1–6 h, but prolonged dissolution of VAP after rapid dissolution. As a result of cell culture studies and wound healing activity studies, it was determined that NP-7 was the most effective. It was thought that the reason for this was that the NP-7 coded formulation was a chitosan-coated PLGA nanoparticle with the smallest particle size, and it was concluded that the efficiency of VAP was increased with its nanoparticle structure. This study demonstrated the similar wound healing effects of VAP-loaded nanoparticle systems, in particular NP-7, which increases keratinocyte cell proliferation at lower concentrations (10 μg·mL–1) than vitamin A alone (100 μg·mL–1). VAP-loaded nanocarriers that can be used in the pharmaceutical industry have been successfully produced and the results obtained have been evaluated as promising for this industry.