Abstract: Biomimetic scaffolds with extracellular matrix (ECM)-mimicking structure have been widely investigated in wound healing applications, while insufficient mechanical strength and limited biological activity remain major challenges. Here, we present a microfluidic 3D printing biomimetic polyhydroxyalkanoates-based scaffold with excellent mechanical properties and hierarchical porous structures for enhanced wound healing. This scaffold is composed of poly(3-hydroxybutyrate-4-hydroxybutyrate) and polycaprolactone, endowing it with excellent tensile strength (2.99 MPa) and degradability (80% of weight loss within 7 d). The ECM-mimicking hierarchical porous structure allows bone marrow mesenchymal stem cells (BMSCs) and human umbilical vein endothelial cells (HUVECs) to proliferate and adhere on the scaffolds. Besides, anisotropic composite scaffolds loaded with BMSCs and HUVECs can significantly promote re-epithelization, collagen deposition and capillary formation in rat wound defects, indicating their satisfactory in vivo tissue regenerative activity. These results indicate the feasibility of polyhydroxyalkanoates-based biomimetic scaffolds for skin repair and regeneration, which also provide a promising therapeutic strategy in diverse tissue engineering applications.