松质骨支架与成骨细胞生物相容性的实验研究

Experimental Research of Compatibility on the Porcine Cancellous Bone Scaffold and Osteoblast.

目的:观察松质骨支架对成骨细胞的粘附、生长、增殖的影响,为骨组织工程支架的选择提供实验依据。方法:采用F icoll梯度离心法得到兔骨髓基质干细胞,经诱导培养为成骨细胞并通过相差显微镜观察、Ⅰ型胶原免疫组化法染色、四环素染色确认为成骨细胞;采用物理化学方法对猪松质骨进行处理得到猪松质骨支架,与成骨细胞体外复合培养,通过扫描电镜观察猪松骨材料对成骨细胞生长、粘附的影响。结果:骨髓基质干细胞经条件培养基诱导培养后已分化为成骨细胞,与松质骨支架联合培养1d后可见细胞附着于材料表面,但数量不多,细胞呈圆球形,7d后细胞生长密集,似葡萄状粘附于脱矿猪松质骨的孔壁上,细胞呈不规则圆球形,表面有丰富的绒毛状突起,细胞间有伪足样突起相连但分布不均。细胞相互融合,可见细胞遮盖微孔间隙。结论:松质骨材料具有良好的细胞相容性,可用作骨组织工程的支架材料。

Objective： To investigate the effect of porcine cancellous bone（PCB） on the attachment, growth and proliferation of osteoblast. Methods： The rabbit bone marrow stromal cells were gained through Ficoll centrifugal method and were cultivated in conditional culture medium. The osteoblast was identified by discrepancy microscope, collagen I immunohistochemistry and tracycline coloration. The PCB was gained through physical and chemical treatment and was cultivated with osteoblast. The effect of PCB on the attachment, growth and proliferation of osteoblast was investigated through scanning electron microscope（SEM） ld and 7d post cultivation. Results： The bone marrow stromal cell could transfer to osteoblast. After ld cultivated with PCB, the osteoblast attached on the surface of the PCB, but the number was limited, the morphoglogy of osteoblast was spherical; After 7d cultivated with PCB, the osteoblasts were denseness and became irregular spherical. There were many floss on the surface of the osteoblast and pseudopods between cells. Part of the PCB formen were enveloped. Conclusion： The PCB has good biocompatibility with osteoblast, and it can be regarded as scaffold material of bone tissue - engineering.