The response of host blood vessels to graded distribution of macro-pores size in the process of ectopic osteogenesis.

Angiogenesis is of great importance to bone regeneration, but it remains a significant challenge to induce sufficient angiogenesis and osteogenesis within bone grafts for large bone defect healing. The aim of this study is to investigate the effects of hydroxyapatite (HA) scaffold via a novel graded pore distribution approach on vascularization and osteoinduction. Two types of graded porous scaffolds were fabricated by sugar templates-leaching techniques: (1) one with large pores of 1100-1250 μm in the center and small pores of 500-650 μm at the periphery (HALS); (2) the other with small pores of 500-650 μm in the center and large pores of 1100-1250 μm at the periphery (HASL). In vivo data showed different pore size distribution had a remarkable impact on blood vessel formation during bone formation, which led to distinct localization of new bone within the defects. After one month of implantation, the diameters of the blood vessels infiltrated on the periphery of HASL were substantially larger than those in the center though the host blood vessels were successful in infiltrating throughout the whole scaffold. In contrast, vascularization within HALS appeared to be poor with very few blood vessels formed in the center, indicating heterogeneous vascularization in the scaffolds. After 3 months of implantation, we found that HASL induced more homogeneous bone formation in the whole bone graft but new bone was only found at the periphery of HALS. This study suggests that the pores size distribution in graded scaffolds cannot only affected early stage vascularization, but also influence late stage bone formation and remodeling. The architecture of larger pores at the periphery of graded scaffold may be capable of enhancing angiogenesis and osteogenesis during large size bone defect healing.