Silver nanoparticles (AgNPs) and regenerated silk fibroin (RSF) have recently attracted significant interests for their potential applications in preventing wound-related infections and in tissue engineering. Indeed, nano-silver has long been recognized as one of the most effective antimicrobial agents, and silk fibroin is well known for its capability of stimulating cell activities and facilitating tissue regeneration. In this study, a green synthesis approach was used to create a composite hydrogel (CoHy) of RSF stabilized with CarboxymethylCellulose-Na (CMC-Na) and loaded with AgNPs. Their swelling ratios were up to 59 g/g when tested in different physiologically relevant fluids. Material characterizations by Scanning electron microscopy (SEM) with Energy Dispersive X-ray Spectroscopy (EDS), and X-Ray Diffraction (XRD) confirmed the presence of AgNPs on the surface. Antimicrobial properties of the CoHy samples were evaluated using agar diffusion tests. The results showed distinct inhibition zones against major microorganisms found in wound infections, including Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), Staphylococcus epidermidis (S. epidermidis), Methicillin Resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa (P. aeruginosa), Candida albicans (C. albicans) and Fluconazole-resistant Candida albicans (FRCA). Cytocompatibility studies with rat bone marrow derived mesenchymal stem cells (BMSCs) in vitro showed that the adhesion density of BMScs on the CoHy loaded with 1 mg/mL was similar to the cell-only control group for the first 24 h of culture; moreover, higher cell proliferation was observed on the CoHy without AgNPs, indicating the regenerative potentials of the RSF/CMC composite hydrogels.