乳鼠成骨细胞与珊瑚-羟基磷灰石支架材料在旋转式生物反应器中培养的实验研究

目的探讨联合运用成骨细胞、珊瑚-羟基磷灰石(CHA)支架材料和自行研制的旋转式生物反应器在体外构建组织工程化骨的可行性.方法运用骨片组织培养法,分离培养Wistar乳鼠的成骨细胞,传至第3代后,形成成骨细胞-CHA复合物,然后在自行研制的生物反应器中培养.通过茜素红(ARS)染色,MTT法及其他组织化学的方法来研究在生物反应器中培养14 d时的成骨潜能.结果成骨细胞和CHA支架材料有良好的生物相容性.在生物反应器中,成骨细胞-CHA复合物培养至第14天时,CHA明显促进了细胞的增殖,ARS染色呈弱阳性反应,同时,碱性磷酸酶(ALP)的分泌减少.结论CHA材料和生物反应器可以为成骨细胞提供良好的生物学及力学环境,三者联合运用有希望成为构建组织工程骨的理想方法.

Experimental study on mice osteoblasts cocultured with coral-hydraxyapatite scaffold in rotary bioreactor

Objective To study the feasibility of construction of the tissue engineered bone with coral-hydroxyapatite(CHA) scaffold and osteoblast in the self developed rotary bioreactor. Methods Osteoblasts from the parietal bone chips of Wistar mice were isolated and cultured. The third generation cells were cultured on CHA scaffold,and then cultured in bioreactor. The osteogenic potential of the cell-CHA scaffold constructs cultured in bioreactor were studied by Alizarin Red S(ARS) staining,MTT method and histochemistry at the 14th days. Results It was showed that the osteoblasts had good biocompatibility with CHA matrix scaffold,the scaffold promoted the proliferation of osteoblast remarkably in bioreactor and weak positive reaction presented in ARS staining on the 14th days. At the same time,the secretion of alkaline phosphatase(ALP) was decreased. Conclusion The CHA material and bioreactor provides suitable biological and mechanical microenvironment for osteoblasts. The osteoblasts cocultured with CHA in bioreactor might be an ideal approach for construction of tissue engineered bone in vitro.