Developing dressing for wound dressings represents a significant challenge for the scientific community. In this study, a conductive hydrogel was synthesized to promote the wound-healing process. This hydrogel is composed of silk fibroin (SF), reduced graphene oxide (rGO), and glycerol (G). The impact of modifying the SF:rGO ratio, and the G content (%), on the physicochemical and biological properties. The hydrogels were characterized using FT-IR, SEM, XRD, TGA, swelling, mechanical resistance, and conductivity. The cytotoxicity of the materials and their wound-healing capacity in human fibroblasts were also determined. Chemical analysis revealed that the gelation of SF occurs due to the formation of beta sheet structures, which was confirmed by the shift from amide I to amide II. An Increase in the SF:rGO ratio favored swelling behavior, although increasing G reduced this effect. The swelling of the hydrogel followed a Fick diffusion mechanism. Furthermore, the increase in the SF:rGO ratio and the percentage G improved the conductivity of the materials. The hydrogels were found to be non-cytotoxic to human fibroblasts, and those containing rGO exhibited superior wound healing capacity compared to the positive control cell culture medium. Therefore, SF:rGO hydrogels could be considered promising candidates for wound dressing.