摘要
目的 研究人脱细胞骨(HAB)复合经诱导的骨髓基质细胞的实验效果,观察细胞的黏附和生长情况,并对其成骨活性进行检测。方法 用15％的过氧化氢和乙醚去除人髂骨块内的结缔组织和细胞成分,消毒后制备人脱细胞骨。取材活体或新鲜尸体的骨髓行骨髓基质细胞培养,细胞纯化后加入β-甘油酸钠,地塞米松和抗坏血酸等向成骨方向诱导,并进行对照培养。通过碱性磷酸酶(ALP)和骨钙素(OCN)检测来确定骨髓基质细胞的增殖和分化情况,将诱导的骨髓基质细胞浓缩后复合到制备好的脱细胞骨块内进行复合培养。8d后通过光镜和电镜等形态学观察以及生化指标检测来确定细胞的成骨活性。结果 人髂骨块内细胞清除干净,骨基质保存良好;诱导后的骨髓基质细胞ALP和OCN水平明显高于对照组(P<0.05);复合后的人脱细胞骨培养液中ALP和OCN检测呈阳性;骨髓基质细胞在HAB支架内附着紧密,生长良好。结论 人脱细胞骨复合经诱导的骨髓基质细胞在体外具有有效的成骨功能,是一种较为理想的骨组织工程材料。
Objective To investigate the osteogenic ability of human allogenous acellular bone(HAB) loaded with bone marrow stromal cells (BMSCs) . Methods Human acellular bone was obtained from the iliac bone of fresh corpse, processed with 15% hydroperoxide and ether solution and sterilized for conservation. BMSCs from living femoral shaft bone marrow were cultured immediately in ordinary and mineralized medium composed of DMEM, fetal bovine serum, dexomethasone, β-glycerophophate and ascorbic acid. After BMSCs confirmed rapidly proliferated and differentiated toward osteoblast, which was proved by alkaline phosphatase(ALP) and osteocalcin (OCN) detect in the culture medium, they were concentrated and implanted within HAB scaffold. After this composite was soaked and cultured for another 8 days, ALP and OCN in the medium were then assayed again. The his-tologic presentation was determined under light microscope and scanning electron microscope. Results HAB was well prepared showing a complete reservation of its porous structure. ALP and OCN activities in the induced culture medium were significantly higher than in the ordinary medium( P < 0.05) . The composite bone loaded with BMSCs demonstrated a positive presentation of the osteogenic function by the biochemical assay. BMSCs were adhered and grew well in HAB scaffold. Conclusion HAB loaded with induced BMSCs has an excellent osteogenic function in vitro and presents an effective potential to be a good bone tissue engineering material.
出处
《中国骨质疏松杂志》
CAS
CSCD
2004年第1期35-38,共4页
Chinese Journal of Osteoporosis
