Background: Tissue-engineered scaffolds for skin wound healing have undergone marvelous progress. The recognition that a three-dimensional scaffold more closely mimics the biomechanical environment of wounds and advancing knowledge of cell biology has led to the next-generation of engineered bioscaffolds with nanotechnology. A unifying approach is required for the translational success of bioscaffolds, involving clinicians, biologists and chemists. The decellularized materials were expanding their clinical utility due to high clinical results ahead of the ones with autografts. They are gradually gaining market space due to their ease of standardized production, constant availability for grafting and biomechanical/ biochemical advantage. Hence, the present study aimed to develop biobased decellularized extracellular matrix (dECM) impregnated with eco-friendly synthesized silver nanoparticles and then to evaluate their wound healing activity the excision wound model of rats. Methods: The dECM was prepared to achieve acellularity, intactness and adequate tensile strength. The same was confirmed by morphological (histology, scanning electron microscopy and Fourier transform infrared spectroscopy), mechanical (tensile strength) and biological (DNA quantification) analyses. Further, the dECM was impregnated with the eco-friendly synthesized keratin-chitosan-silver nanoparticles to produce ‘NanoBioscaffolds’ for extended biocompatibility and were evaluated for wound healing activity in rats. Result: The findings of the histopathology (H and E staining), immunohistochemistry (the proliferative activity of keratinocytes by Ki67 staining) and biochemical analysis (anti oxidative status by catalase estimation) revealed that wound healing activity was promoted by inducing proliferation and migration of the keratinocytes and detoxification of reactive oxygen species activity (ROS) in NanoBioscaffold treated group.