Kaolinite nanocomposites (NCs) could be utilized as agents for wound healing owing to their efficiency and low toxicity. The present study was conducted to synthesize a novel silver/kaolinite NCs (Ag/Kaol NCs) and investigate their chitosan derivation (Ag/Kaol/Chit NCs) using oak extract. XRD, SEM, EDX, FT-IR, and DLS were employed for the investigation of structural and physio-chemical properties of the synthesized NCs. The obtained results revealed that synthesized Ag/Kaol NCs were mesoporous and spherical with sizes ranging from 7–11 nm. They also demonstrated successful synthesis between silver and kaolinite using the extract. Cytotoxicity and in vitro antibacterial activity were also investigated. The clinical effects of ointments containing the NCs for improving wound healing were studied on the wound area, total bacterial count, histological parameters, and protein expression of some genes. Nanocomposites were safe up to 0.50 mg/mL. The results of in vivo and in vitro antibacterial activity showed that Ag/Kaol NCs, were of antibacterial activity ( ). The results of antioxidant activity indicated that Ag/Kaol NCs have antioxidant structures. Our findings concerning molecular mechanism implied that Ag/Kaol/Chit increased the expression of Wnt/ -catenin and collagen ( ). In sum, Ag/Kaol/Chit showed antibacterial activity and improved wound healing by decreasing the inflammation and promoting the proliferative phase. The novel NCs showed wound healing properties by decreasing inflammation and total bacterial count and increasing proliferative phase. The application of Ag/Kaol/Chit was suggested as a green agent for improving infected wound healing.