Recombinant collagen (rCOL)-hyaluronic acid (HA) composite scaffolds were prepared to thoroughly investigate their biological properties.The rCOL and HA composite scaffolds were formulated via lyophilization. The scaffolds were characterized for various materials properties, including porosity, surface modification, and degradation rates. Biological properties such as in vitro cytotoxicity, cell adhesion, proliferation and migration effects were also evaluated.The water absorption, mechanical strength, degradation resistance and thermal stability of the prepared rCOL-HA composites were improved over that of the control studied. Scanning electron microscopy (SEM) revealed that the composites formed a three-dimensional network structure with uniform pore distribution. The cytotoxicity of the composites was minimal (grade I) and the material showed strong adhesion and proliferation effects when grown with mouse fibroblasts, particularly the composite material of rCOL (5% HA) group (P < 0.05).The rCOL-HA composite prepared via lyophilization after cross-linking is characterized by high porosity, high water absorption, and good interaction between the material and cells, as well as good biodegradability. Compared with rCOL materials, rCOL-HA has increased mechanical strength, water absorption and thermal stability. The biocompatibility and fibroblast proliferation of rCOL-HA have excellent biological performance, providing a new material for wound healing applications.