The influence of collagen as an effective substitute for gelatin was investigated on properties of chitosan/gelatin hydrogels for fibroblasts growth and attachment for wound dressing applications. We synthesized hydrogels based on chitosan associated with collagen and gelatin biopolymers (in the ratio of 1:5 and 1:1, respectively). The hydrogels properties such as morphology, swelling ratio, mechanical characteristics, water vapor loss, water vapor transmission rate (WVTR), and biodegradation were analyzed. 1 × 10 human fibroblasts were seeded per ml of hydrogels and maintained for 7 days. Cell viability was assessed by using MTT. The presence of collagen caused reduced swelling ratio, and biodegradation rate compared to chitosan/gelatin hydrogels (p < 0.05). The introduction of collagen into chitosan hydrogels improved the mechanical strength compared to gelatin. Hydrogels with collagen possessed an optimum WVTR compared to the chitosan group and hydrogels with gelatin (p < 0.05). Analyzing the morphology of hydrogels revealed that the addition of collagen leads to a homogenous and interconnected structure. Collagen impregnation promoted cell survival and attachment compared with chitosan hydrogels after 7 days (p < 0.05). Collectively, these results demonstrated the potential of the chitosan/collagen hydrogels for wound dressing applications.