【作者】 王磊；

【导师】 巢永烈； 宫苹；

【作者基本信息】 四川大学 ， 口腔临床医学， 2004， 博士

【摘要】 本研究通过使用纤维蛋白凝胶作为rhBMP-2载体,与BIO-GIDE膜,瑞诺膜两种临床效果较理想的引导再生胶原膜,构建rhBMP-2复合胶原膜,并检测了复合膜rhBMP-2的释放,并在细胞水平和临床骨形成水平探讨其生物相容性和成骨能力。为新型复合胶原膜临床应用提供更多资料。 本研究分三部分。 1、利用人纤维蛋白原和凝血酶合成纤维蛋白凝胶,将其作为rhBMP-2的载体,与胶原膜复合,获得rhBMP-2复合BIO-GIDE膜和rhBMP-2复合瑞诺膜;通过SDS-PAGE电泳和紫外分光光度法,测定复合膜rhBMP-2释放性能。 2、原代培养大鼠顶骨成骨细胞,与复合膜联合培养,通过MTT比色法检测和扫描电镜直接观察等手段,了解rhBMP-2复合胶原膜的细胞相容性。 3、建立大鼠顶骨临界性骨缺损模型,以钛膜和单纯胶原膜为对照,使用复合胶原膜结合BIO-OSS修复顶骨缺损,X线检查和组织学切片HE、新三色法染色观察2、4、8、12周后骨缺损修复情况,免疫组化染色了解BMP在缺损区的分布和数量。更多还原

【Abstract】 To investigate the utility of fibrin gel as a carrier of rhBMP-2, BIO-GIDE? and another collagen membrane RuiNuo were used as a binding bearer which were covered by rhBMP-2 combined fibrin gel layer, for the sake of BIO-GIDE? and RuiNuo all have been applied in clinical use effectively and show good outcome for guided bone regeneration. In this research, the release of rhBMP-2 from combined collagen membrane and the cellular compatibility were detected in vitro. And the osteogenetic ability of these new guided bone regeneration membranes were inspected in vivo. So the application of these combined membranes in clinic was found in stronger foundation.In this study, we have three parts: 1, in vitro, human fibrinogen and thrombase were used to make the fibrin gel, and then rhBMP-2 was added in this fibrin gel. Smear this combined gel on the rough side of BIO-GIDE? and the inner side of RuiNuo. And the combined collagen membranes were carried out. By using the SDS-PAGE tech and UV spectrophotometer, the released rhBMP-2 was quantified, and the releasing behavior of combined collagen membrane was estimated. 2, cultured primary osteoblast cells of rat were seeded on the combined collagen membrane and SEM and MTT have been given to the cultured cell at different time point. And the cellular compatibility of these combined collagen membrane was estimated. 3, the critical sized defects of rat calvarium were made and the combined collagenmembranes were used as guided bone regeneration membrane with BIO-OSS? compared with titanium and uncombined collagen membranes as control. X-ray and faistoiogical and mmutnoMstochemisfric inspection were given to inspect the new bone under the membrane and the rhBMP-2 distribution.And then, we got such results as: 1, the methods of human fibrinogen and thrombase to make fibrin gel in lab are convenient. And the outcome of SDS-PAGE and UV spectrophotometer showed a rapid and small-dosage release of rhBMP-2 in die eariy stage when the combined membrane was tested in vitro. 2, the morphologic characters of rhBMP-2 combined fibrin gel were evaluated under SEM and showed an orange-like appearance. And such kind of appearance was then approved to be comfortable for osteoblast’s attachment and proliferation m MTT. 3, fee immunohistochemistry outcomes showed that the releasing of rhBMP-2 was controlled by the fibrin and collagen membrane. Histoiogical results gave proof to osteogenetic ability of these combined collagen membranes by new bone quantity underneath the membrane in HE and Masson’s tnchrome stain.The use of fibrin gel as rhBMP-2’s carrier is feasible in lab. Moreover, a layer on the given side of collagen membrane can locate the release of rhBMP-2 for given area and occasion. Such kind of collagen membrane can accelerate the sew bone regeneration m viv?. So the technique to make fibrin gel and combine rhBMP-2 with collagen membrane may be considered for use in cine.更多还原