Tissue regeneration in many skin defects is progressing with new treatments in recent years. Tissue engineering with the use of scaffolds offers more versatile and faster solutions in treatment. Extracellular matrix (ECM) and its three-dimensional (3D) network structure as a biological bond by imitating the tissue microstructure has been used for tissue repair, which can answer many existing challenges. Vitamin A, which comes in several forms such as retinols, retinals, and retinoic acids, is a necessary vitamin that is crucial for wound healing. In this research, sheep kidney capsule tissue decellularized with sodium dodecyl sulfate (SDS) containing different doses of vitamin A has been used as an ECM in skin tissue engineering. The above scaffold was evaluated in terms of properties such as biocompatibility, analysis of mechanical properties, attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR), hydrophilicity, antibacterial, and cell adhesion. The findings reported suitable properties for wound dressing, especially at a dose of 15,000 U/ml vitamin A for this scaffold. Then, the above scaffold was evaluated on the full-thickness wound model in rat, which showed good wound contraction, and increased VEGF factor. It showed a decrease in IL-1β level. Therefore, the use of the above-mentioned decellularized scaffold in combination with medicinal agents effective in wound healing can be introduced for further pre-clinical studies.