四环素对胶原生物衍生骨材料细胞相容性的影响

Cytocompatibility of tetracycline loaded collagen bio-derived bone with osteoblasts

目的:观察四环素-胶原生物衍生骨材料与兔成骨细胞的相容性及体外附着规律。方法:实验于2003-02/2005-03在华西医科大学组织工程实验室完成。①实验方法:取新鲜人捐献骨经脱脂、脱蛋白等理化过程,制成单纯生物衍生骨材料为载体,载体表面经I型胶原修饰得到胶原生物衍生骨材料,将四环素吸附于胶原生物衍生骨复合材料,构建出四环素-胶原生物衍生骨材料。体外培养兔骨膜成骨细胞,与胶原生物衍生骨材料、四环素-胶原生物衍生骨材料共培养。②实验评估:分别于1,3,5,7d取材,扫描电镜下观察细胞在材料表面的生长情况,测定两组材料的碱性磷酸酶活性,MTT检测细胞生长和增殖情况,流式细胞仪检测细胞周期情况。结果:①扫描电镜观察、碱性磷酸酶活性检测及MTT检测结果表明,成骨细胞在四环素-胶原生物衍生骨材料表面和孔隙内黏附和增殖良好,在胶原生物衍生骨材料较差。②流式细胞仪检测表明,四环素-胶原生物衍生骨材料无细胞毒性。结论:以胶原生物衍生骨材料作为载体,进一步吸附四环素后构建的复合材料与成骨细胞有良好的细胞相容性。

AIM： To investigate the cytocompatibility of collagen bio-derived bone loading tetracycline with osteoblast in rabbits, and analyze the adhesion law in vitro. METHODS： The experiment was carried out in the Laboratory of Tissue Engineering at West China Medical University from February 2003 to March 2005. ①Methods： The donor human bone were degreased and deproteinized to form bio-derived bone vector, in which the surface was covered with type- Ⅰ collagen for modification and tetracycline were absorbed to collagen bio-derived bone. The rabbit osteoblasts were cultured in vitro and compounded ＇to tetracycline loaded collagen bio-derived bone.②Evaluation： 1, 3, 5 and 7 days later, the growth of cells on the surface of compound materials was observed under scanning electron microscope, the activity of alkaline phosphatase （ALP） was determined, MTT method was adopted to detect the growth and proliferation of cells, and the cell cycle was tested by flow cytometry. RESULTS： ①The results of scanning electron microscope, ALP activity and MTT assay showed that, the osteoblast adhered and grew well on the surface and internal foramen of tetracycline loaded collagen bio-erived bone, while the cell proliferation was not good on collagen bio-derived bone. ②Tetracycline loaded collagen bio-derived bone materials had no toxicity to osteoblast by flow cytometry. CONCLUSION： After tetracycline is absorbed to the surface of the collagen bio-derived bone, the tetracycline loaded collagen bio-derived bone has good cytocompatibility with the osteoblasts.