利用鸡胚绒毛尿囊膜(CAM)中丰富的血管建立半体内模型,研究多孔磷酸钙骨修复材料的血管化.将多孔羟基磷灰石陶瓷(HA)和具有部分孔隙结构的磷酸钙骨水泥(CPCs)分别植入CAM中共培养,利用体式显微镜和扫描电子显微镜(SEM)观察CPCs表面血管的生长,并利用Image-Pro Plus和Nano Measurer定量血管密度、直径和数量;对CAM中培养的HA采用不脱钙组织学处理和观察.体式显微镜观察及定量结果显示多孔结构对血管生长影响显著,随着培养时间延长,多孔部分血管密度和数量均有明显增加,而无孔部分血管生长缓慢.CPCs表面血管以出芽方式生长,形成血管网络,直径一般小于50μm.SEM观察显示血管紧贴CPCs表面生长.苏木精-伊红(H&E)染色显示血管通过孔隙结构长入HA内部.实验结果说明三维孔隙结构利于血管的生长和长入,此实验可简便、高效、低成本地研究多孔磷酸钙骨修复材料的血管化,为快速评价生物材料血管化寻找一种新途径.
In this study,ex vivo chick chorioallantoic membrane (CAM) model with abundant blood vessels was de-veloped to investigate the vascularization of porous calcium phosphate. Porous hydroxyapatite ceramic (HA) and cal-cium phosphate cement with segmental porous structure (CPCs) were implanted into CAM, respectively. Stereomicro-scope and scanning electron microscope (SEM) were employed to observe the angiogenesis on the surface of CPCs. The vascular density, diameters and numbers were quantified by Image-Pro Plus and Nano Measurer. Furthermore, undecalcified histological staining was conducted to observe the inner angiogenesis of porous HA. The vascular density and number in the porous part of CPCs both increased significantly with the increase of culture time, while the an-giogenesis was tardily in the part of CPCs without pores. Most of the blood vessels less than 50μm on the porous part of CPCs grew in budding manner to form vascular network. SEM images showed that blood vessels grew tightly on the surface of CPCs. Histological staining demonstrated that some blood vessels grew into the inside of porous HA. These results suggest that three-dimensional porous structure may promote the angiogenesis of porous calcium phos-phate and the present CAM model may provide a convenient, efficient and with low-cost approach to evaluate vascu-larization of porous biomaterials.
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