同种异体脱钙骨基质作为骨组织工程载体的实验研究

目的通过体外同种异体脱钙骨基质（DBM）和骨髓基质细胞（BMSC）的复合培养及体内异位成骨实验，研究DBM作为骨组织工程载体的生物相容性及成骨活性。方法参考Urist操作方法大量制备兔同种异体DBM。骨穿取兔骨髓单细胞悬液进行培养，将BMSC与同种异体DBM体外复合培养3～7d，相差显微镜、扫描电镜（SEM）和苏木精-伊红染色后光镜下观察结果。将DBM和BMSC复合培养3d的复合物植入家兔一侧骶棘肌肌袋内，分别在1、2、4周活体取材，扫描电镜和苏木精-伊红染色后光镜观察，对侧单纯植入DBM作为对照。结果体外培养发现BMSC在DBM中贴壁生长、增殖并有分泌活动。体内实验发现BMSC在DBM孔隙内均匀成骨，对照组则从DBM骨块的边缘到中心逐步成骨，成骨所需的时间长，而且成骨量要小于前者。结论DBM作为组织工程载体具有良好的生物相容性和成骨活性。

Allogenic Decalcified Bone Matrix as Carrier for Bone Tissue Engineering

Objective To study the biocompatibility and osteogenic activity of allogenic decalcified bone matrix (DBM) used as a carrier for bone tissue engineering by bone marrow stromal cells (BMSC) and DBM co-culture in vitro and ectopic bone formation in vivo.Methods Following the method described by Urist, allogenic DBM was mass-made. In vitro, DBM and BMSC from rabbits were co-cultured for 3-7 days and subjected to HE staining, and a series of histomorphological observations were performed under phase-contrast microscopy and scanning electron microscopy (SEM). In vivo the mixture of DBM/BMSC co-cultured for 3 days was planted into one side of muscles sacrospinalis of rabbits, and the DBM without BMSC was planted into other side as control. Specimens were collected at postoperative week 1, 2 and 4, and subjected to HE staining, and observed under SEM.Results During culture in vitro, the BMSCs adherent to the wall of DBM grew, proliferated and had secretive activity. The in vivo experiment revealed that BMSCs and mesenchymal cells invaded gradually and proliferated together in DBMBMSC group, and colony-forming units of chondrocytes were found. Osteoblasts, trabecular bone and medullary cavity appeared. The inflammatory reaction around muscles almost disappeared at the second week. In pure DBM group, the similar changes appeared from the surface of the DBM to center, and the volume of total regenerate bones was less than the DBMBMSC group at the same time.Conclusion DBM had good biocompatibility and ectopic induced osteogenic activity as a carrier for bone tissue engineering.