Abstract: The microstructure on the implants is essential for bone repairing and tissue regeneration. Especially, controllable preparation of well-defined micropattern is one of the key factors to regulate the biological fate of bone mesenchymal stem cells (BMSCs). Usually, the micropatterns are made of materials with limited degradability and poor bioactivity. In order to meet the needs for bone repairing and take the special structural and physiochemical properties of biomedical implants into account, the degradable hyaluronic acid (HA) was cross-linked with N′-(ethyliminomethylidene)-N, N-dimethyl-1,3-propylene diamine (EDCI) and adipic acid dihydrazide (ADH). Then, the HA microwell patterns were fabricated by soft-lithography technique. By turning the mass fraction of ADH and EDCI from 8.3% to 16.7% respectively, the effects of contents on the microstructure, swelling properties, degradation behavior, and compatibility of HA films were evaluated by a series of characterization methods.The experimental results showed that as the mass fraction of ADH and EDCI increased, the crosslinking degree of the film increased, and the swelling variation of gel became smaller. HA films with high mass fraction of ADH and EDCI showed a slower degradation behavior. A series of HA films with micro-array pore structure (pore diameter of 32, 96, 128, 320 μm respectively) were successfully fabricated by combining soft lithography and solvent evaporation. When co-culture with rBMSCs in vitro, the results showed that the larger micron wells (96, 320 μm) with sparse distribution presented no significant effect on the proliferation of rat bone mesenchymal stem cells (rBMSCs) compared to the smooth HA gel film, while the small micron wells (32 μm) with compact distribution can significantly promote the proliferation and osteogenic activity of rBMSCs.