Experimental study of osteoinduction with gene transfer of bone morphogenetic protein using adenovirus vector.

骨形态发生蛋白（BMP）具有独特的骨诱导能力。BMP在软组织中诱导骨。BMP有望应用于骨重建。本研究将BMP-2植入软组织，并将携带BMP-2基因的腺病毒载体（AVV-rhBMP-2）转染肌肉。1）将rhBMP-2（2、10、50 μ g）与端肽Ⅰ型胶原混合冻干（rhBMP-2-CL）; 2）将rhBMP-2基因连接到粘粒载体中。另一方面，将腺病毒缺陷型Δ1Δ3和粘粒载体共转染到HEK 293细胞中。在细胞中扩增AVV-rhBMP-2，并纯化AVV-rhBMP-2。3）在大鼠背部肌瓣移植。植入rhBMP-2和AVV-rhBMP-2。3周后行软X线摄片，取种植体进行组织学和生化检查。4）取种植体进行HE染色，观察组织学变化，观察诱导骨形成情况。rhBMP-2-CL在10天内诱导成大量骨，包括软骨细胞，在21天内诱导成含有骨髓的骨块。AVV-rhBMP-2诱导骨颗粒分散。每一个都是骨量。与对照组相比，两组中诱导骨的量呈剂量依赖性。

Bone morphogenetic protein (BMP) has a unique ability, osteoinduction. BMP induces bone in the soft tissue. It is expected that BMP is applied in bone reconstruction. In this study, BMP is implanted in soft tissue, and adenovirus vector with BMP-2 gene (AVV-rhBMP-2) is transfected in the muscle. The induced bone is investigated.1)rhBMP-2 (2,10,50ug) and atelopeptide tipe I collagen were mixed and freezed-dried (rhBMP-2-CL).2)rhBMP-2 gene was associated into cosmid vector. On the other hand, adenovirus deficit Δ1Δ3 and cosmid vector were co-transfected to HEK293 cells. In the cells, AVV-rhBMP-2 was multiplicated and AVV-rhBMP-2 was purified.3)muscle flaps were operated on the back of rats. There rhBMP-2 and AVV-rhBMP-2 were implanted. Three weeks later the implanted areas soft X-rayed and the implants were excised and examined histologically and biochemically.4)The implanted areas were excised and HE-stained to investigate the tissues histologically.In both, induced bone were investigated. rhBMP-2-CL induced massive bone including chondrocytes in 10days, bony mass with bone marrow in 21 days. AVV-rhBMP-2 induced scattered bony particles. Each was bone mass. Comparing controls, the amount of induced bone was dose-dependent in both groups.