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

骨形态发生蛋白(BMP)具有独特的诱导成骨能力。BMP在软组织中诱导成骨。BMP有望在骨重建中得到应用。在本研究中，将BMP植入软组织，并将携带BMP-2基因的腺病毒载体(AVV-rhBMP-2)转入肌肉。1)将重组人骨形态发生蛋白-2(2，10，50ug)与多肽I型胶原混合冻干(rhBMP-2-CL)。2)将重组人骨形态发生蛋白-2基因整合到粘粒载体中。另一方面，将腺病毒缺陷型Δ-1-Δ-3与粘粒载体共转染HEK293细胞。在细胞内扩增和纯化AVV-rhBMP-2。3)大鼠背部肌瓣移植。分别植入rhBMP-2和AVV-rhBMP-2。3周后，对种植区进行软X射线检查，切除种植体，进行组织学和生化检查。4)切取种植区，HE染色观察组织学变化。重组人骨形态发生蛋白-2-CL在10d诱导大量骨形成软骨细胞，在21d诱导有骨髓的骨量。AVV-rhBMP-2诱导骨颗粒散在分布。每组均为骨量。与对照组相比，两组诱导骨量均呈剂量依赖性。

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.