重组人骨形成蛋白—2／珊瑚复合人工骨的研制及其诱导成骨的？…

目的：制备珊瑚（ｃｏｒａｌ）与重组人骨形成蛋白－２（ｒｈＢＭＰ－２）复合人工骨并测试其骨诱导活性，方法：将ｒｈＢＭＰ－２与珊瑚复合后植入鼠肌袋内，６６只小鼠随机分为３组，第１组为复合骨（１：２０ｗ／ｗ）；第２组为复合骨（１：４０ｗ／ｗ）；第３组为单独珊瑚，术后１，３，６周处死动物。光镜下观察，并用图象分析法作成骨定量测定。结果：术后１周，可见复合材料表面及孔洞内有软骨形成。３周出现编织骨。６周形成     

重组人骨形成蛋白-2/珊瑚复合人工骨的动物实验研究

本研究将重组人骨形成蛋白－２（ｒｈＢＭＰ－２）和珊瑚以一定的方式复合后,植入小鼠股部肌袋和兔颅骨标准大小缺损,以单纯珊瑚植入作对照,术后不同时间取材,通过组织学方法检测其骨诱导活性和骨修复能力．结果显示：ｒｈＢＭＰ－２／珊瑚复合人工骨植入小鼠肌袋１周,诱导软骨形成,３周,形成编织骨,６周,形成含骨髓的板层骨,同时,珊瑚被部分降解吸收;复合人工骨植入兔颅骨缺损后,以引导成骨和诱导成骨双重机制完成骨修复过程,术后１２周,植入物完全被成熟的骨组织取代,其骨修复效果明显优于单纯的珊瑚．此复合人工骨具有骨传导和骨诱导活性,骨修复能力较强,是一种较理想的新型生物性植骨材料     

重组人骨形成蛋白-2-珊瑚复合人工骨修复骨缺损的生物力学研究

将珊瑚与具有骨诱导特性的重组人骨形成蛋白－２（ｒｈＢＭＰ－２）复合，制成ｒｈＢＭＰ－２－ｃｏｒａｌ复合人工骨，将其植入兔颅骨标准大小缺损，并与单纯珊瑚植入作对照。通过Ｘ线片、组织学和生物力学方法来评价此复合人工骨的骨修复能力。结果显示：ｒｈＢＭＰ－２－ｃｏｒａｌ复合人工骨具有较强的骨修复作用，植入骨缺损后，材料被逐渐降解吸收，新骨不断形成，机械强度不断增大；１２周后，植入物完全被成熟的骨组织取代，缺损区抗压强度接近正常的骨组织。该复合人工骨是一种较理想的骨移植替代材料。     

基因重组人骨形成蛋白-2与珊瑚/聚乳酸复合异位诱导成骨的实验研究

目的 :观察基因重组人骨形成蛋白 - 2 (rhBMP - 2 )和珊瑚 /聚乳酸形成复合人工骨的异位诱导成骨活性。方法 :把rhBMP - 2和珊瑚 /聚乳酸形成复合人工骨。进行小鼠肌内种植 1、3、6周后 ,组织学观察其异位诱导成骨活性。结果 :rhBMP - 2赋予珊瑚 /聚乳酸骨诱导能力 ,珊瑚 /聚乳酸则充当rhBMP - 2的载体和释放系统 ,对BMP的活性未产生不利影响。与单纯的珊瑚 /聚乳酸相比 ,这种复合人工骨以软骨内成骨的方式诱导成骨。结论 :rhBMP - 2 /珊瑚 /聚乳酸复合人工骨具有良好的生物相容性和骨诱导活性 ,是一种较理想的骨移植替代材料     

应用珊瑚作为重组人骨形成蛋白-2缓释载体的动物实验研究

将大肠杆菌表达的重组人骨形成蛋白-2（rhBMP－2）和珊瑚（coral）复合，制成rhBMP－2／coral复合人工骨，将其植入小鼠股部肌肉内，并以单纯珊瑚植入作对照，术后1、3、6周取材，组织学观察。结果显示：rhBMP－2／coral复合人工骨在植入局部诱导形成骨和软骨，表现出良好的异位诱导成骨能力；珊瑚对rhBMP－2的活性未产生不利影响，是rhBMP－2的良好缓释载体。此复合人工骨可望作为一种新型生物性植骨材料而应用于骨科和颌面外科。     

基因重组人骨形成蛋白—2与珊瑚／聚乳酸复合异位诱导成骨的实验研究

目的：观察基因重组人骨形成-2(rhBMP-2)和珊瑚／聚乳酸形成复合人工骨的异位诱导成骨活性。方法：把rhBMP-2和珊瑚／聚乳酸形成复合人工骨。进行小鼠肌内种植1，3，6周后，组织学观察其异位诱导成骨活性。结果：rhBMP-2赋予珊瑚／聚乳酸骨诱导能力，珊瑚／聚乳酸则充当rhBMP-2的载体和释放系统，对BMP的活性未不利影响。与单纯的珊瑚／聚乳酸相比，这种复合人工骨以软骨内成骨的方式诱导成骨。结论：rhBMP-2／珊瑚／聚乳酸复合人工骨具有良好的生物相容性和骨诱导活性，是一种较理想的骨移植替代材料。     

重组人骨形成蛋白-2、胶原、珊瑚复合人工骨异位诱导成骨的实验研究

目的 采用珊瑚作为载体,胶原作为缓释系统,制血出重组人骨形成蛋白－２（ｒｈＢＭＰ－２）胶原／珊瑚复合人工骨,并评价其骨诱导活性。方法 ｒｈＢＭＰ－２、胶原和珊瑚以一定的方式复合后,植入小鼠股部肌袋内,以单纯珊瑚植入用对照,术后不同取材,通过组织学方法检测骨诱导活性。结果 复合人工骨植入１周诱导软骨形成,２周形成编织骨,４周形成含骨髓的板层骨,同时珊瑚被降解吸收。结果 胶原、珊瑚是ｒｈＢＭＰ－２较理     

基因重组人骨形成蛋白-2与珊瑚/聚乳酸复合修复骨质缺损的实验研究

目的　观察生物珊瑚、聚乳酸和rhBMP-2合成人工骨(复合骨)修复兔颅骨缺损后复合骨的变化情况。方法　选择24只新西兰兔,随机分成2组,每组12只,建立兔颅骨缺损标准模型。植入复合骨,用珊瑚/聚乳酸作为对照。术后4、8、12周每组各处死4只动物,取出植入体进行扫描电镜观察和机械强度测定。结果　复合骨在植入缺损后,不仅在植入体周边部有骨组织长入,而且在整个植入体内均有新骨形成,即出现多中心成骨。复合骨在同一时间点的成骨量明显多于对照组,随时间推移,成骨量递增。在植入前,两材料间的抗压强度无明显差异;在植入后,两植入体的抗压强度则差异显著,复合骨明显高于同期的对照组。结论　生物珊瑚、聚乳酸和rhBMP-2合成人工骨在体内以传导成骨和诱导成骨双重机制完成骨修复,且有良好的机械强度,作为植骨材料具有良好的应用前景。     

重组人骨形成蛋白-2复合1%透明质酸钠骨膜内诱导成骨的实验研究

目的　观察重组人骨形成蛋白 - 2 (rhBMP 2 )复合 1%透明质酸钠骨膜内诱导成骨的组织学过程 ,探讨其诱导成骨的机制。方法　将rhBMP - 2和 1%透明质酸钠复合后植入 9只新西兰大白兔胫骨骨膜下骨皮质的表面 ,对侧植入 1%透明质酸钠作为对照 ,分别于术后第 4、8、12周处死 3只动物取标本进行大体和组织学观察。结果　rhBMP - 2复合 1%透明质酸钠组骨膜内侧有明显的新骨形成 ,新骨呈渐进性的成熟过程 ,12周时和原有骨的组织结构一致。结论　rhBMP - 2和 1%透明质酸钠复合后可以用于骨缺损的修复。     

基因重组人骨形成蛋白-2和聚乳酸复合异位诱导成骨的实验研究

目的：观察基因重组人骨形成蛋白－2（recombinant human Bone Morphofenetic protein-2,rhBMP-2)和聚乳酸（Polylactic acid,PLA)复合形成的活性涂层种植体的异位诱导成骨活性。方法：将rhBMP-2与聚乳酸复合，构建活性涂层种植体，植入兔肌内，于1，4，8周后进行X线、大体观察及组织学观察，检测异位成骨活性。结果：rh-BMP-2/PLA活性复合涂层种植体具有骨诱导能力，PLA为rhBMP-2的良好控释载体。结论：rhBMP-2/PLA涂层具有良好的生物相容性和骨诱导活性，是一种理想的种植体涂层形成。     

重组人骨形成蛋白2-珊瑚复合人工骨修复骨质缺损的实验研究

目的弥补珊瑚无骨诱导活性、成骨能力弱等缺陷。方法将珊瑚与重组人骨形成蛋白２（ｒｅｃｏｍｂｉｎａｎｔｈｕｍａｎｂｏｎｅｍｏｒｐｈｏｇｅｎｅｔｉｃｐｒｏｔｅｉｎ－２，ｒｈＢＭＰ－２）复合，制备出ｒｈＢＭＰ－２－珊瑚复合人工骨。将其植入兔颅骨直径１５ｍｍ的圆形缺损，以单纯珊瑚植入作对照。术后２、６、１２周取材，通过组织学观察和计算机图像分析，评价其骨修复能力。结果ｒｈＢＭＰ－２－珊瑚复合人工骨以传导成骨和诱导成骨双重机制完成骨修复过程，骨修复能力和骨修复效果明显优于单纯的珊瑚。结论此复合人工骨是一种较理想的骨移植替代材料。     

Effect of recombinant human bone morphogenetic protein-2 on the osseointegration of dental implants: a biomechanics study

Background Bone augmentation procedures in combination with dental implants enhance osseointegration in areas that demonstrate localized bone deficit. Clinical confirmation of a biomechanically stable interface is essential for functional implant loading.Purpose The aim of this study was to evaluate biomechanically the effect of recombinant human bone morphogenetic protein (rhBMP)-2 on implant osseointegration and correlate it with periotest and radiographic measurements.Materials and methods Hollow cylinder implants were filled with absorbable collagen sponge soaked with rhBMP-2 or left empty and implanted in dog mandibles. The animals were followed for 4, 8, and 12 weeks, periotest assessment was performed at the end of each time interval, and specimens were collected for pullout biomechanical testing and radiographic evaluation of bone-implant contact levels.Results Periotest assessment did not provide evidence of statistically significant differences between the two groups and correlated well with the radiographic bone-implant contact levels. The pullout test revealed a higher correlation between force/displacement and displacement/energy for the experimental group, suggesting that the addition of rhBMP-2 did influence the rate of osseointegration.Conclusion The results from the pullout test support the potential role of rhBMP-2 in clinical applications by promoting a biomechanically mature interface at 12 weeks. However, radiographic and periotest assessment of the bone-implant interface did not provide evidence of the differences observed with biomechanical testing.This project is part of a dissertation prepared in partial fulfillment of Dr. Sykaras requirements for the degree of Doctor of Philosophy.     

重组人血管内皮生长因子和重组人骨形态发生蛋白-7对大鼠成骨细胞增殖能力的影响

目的研究重组人血管内皮生长因子（recombinant human vascular endothelial growth factor,rhVEGF）和重组人骨形态发生蛋白-7（recombinant human bone morphogenetic protein-7,rhBMP-7）对成骨细胞增殖能力的影响。方法取大鼠颅盖骨组织块,采用改良组织块混合酶消化法培养成骨细胞,分成4组,第1组、第2组、第3组分别加3．125ng／mL不含血清的rhVEGF培养液、50．000ng／mL不含血清的rhBMP-7培养液、3．125ng／mL不含血清的rhVEGF和50．000ng／mL不含血清的rhBMP-7培养液,第4组加不含血清的培养液作为对照组,采用细胞培养技术,应用光镜、四甲基偶氮唑盐（methyl thiazolyl tetrazotium,MTT）法,分析不同浓度的rhVEGF和rhBMP．7对成骨细胞增殖的影响。结果与对照组相比,3．125ng／mL rhVEGF和50.000ng／mLrhBMP-7单独或者联合作用,在1、3、5d均有促进细胞增殖的作用,与对照组的差异均有统计学意义（P〈0.05）,各组间比较差异也有统计学意义（P〈0．05）。其中,3．125ng／mLrhVEGF和50．000ng／mL rhBMP-7单独或者两者联合作用于第3天时,成骨细胞增殖最显著。结论一定剂量的rhVEGF和rhBMP-7可明显促进成骨细胞的增殖。     

RhBMP-2-珊瑚复合修复腭板缺损的动物模型研究

目的：探讨RhBMP-2－珊瑚复合在腭板缺损修复中的可行性。方法：将珊瑚与RhBMP-2复合，制成rhBMP-2-coral复合人工骨，将其嵌入人造狗腭板骨缺损区。通过X片、扫描电镜、生物力学方法，观察复合人工骨在受植区的骨修复效果。结果：RhBMP-2－coral复合人工骨具有很强的骨修复能力，珊瑚逐渐被降解、吸收，新骨不断形成，其抗压强度不断增大，三个月后抗压强度接近正常腭板，复合人工骨完全被成熟骨组织替代。结论：RhBMP-2－coral复合人工骨是一种比较理想的腭板缺损修复材料。     

Expression of bone morphogenetic protein in the course of osteoinduction by recombinant human bone morphogenetic protein-2

To clarify the mechanism of osteoinduction by recombinant human bone morphogenetic protein-2 (rhBMP-2), we examined the time-course localization of bone morphogenetic proteins (BMPs) immunostained by an anti-BMP-2 monoclonal antibody after implantation of pellets consisting of rhBMP-2 and collagen in rat calf muscle pouch. On day 3 after implantation, BMP was detected in the entire lump, and the intensity of staining for BMP around the implant on day 7 was weaker than that on day 3. The staining for BMP decreased with time and the region of staining for BMP remained more centralized in the implant. On day 10 after implantation, BMP was observed in part of the newly induced cartilage, especially around chondrocytes. On day 14 after implantation, BMP was localized in the newly induced woven bone. On day 21, BMP staining was found in osteoblasts at the surface of the newly induced bone. Especially, the staining for BMP decreased from day 10 to day 21. These results indicate that the woven bone was replaced with mature lamellar bone from day 14 to day 21. The present findings suggest that rhBMP-2 plays an important role in osteoinduction, especially at the early stage.     

Experimental study of osteoinduction using a new material as a carrier for bone morphogenetic protein-2

We evaluated the usefulness of artificial collagen as a new carrier for recombinant human bone morphogenetic protein-2 (rhBMP-2) by comparing it with that of atelopeptide collagen, which is derived from porcine skin, and which we have previously shown to be useful for the induction of bone. rhBMP-2 5 μg with either atelopeptide collagen 3 mg or artificial collagen 3 mg was implanted into the calf muscle of 10-week-old Wistar rats (n = 3 in each group). Three rats were given artificial collagen alone and acted as controls (n = 3). Radiographic evaluation, histological analysis, and biochemical examinations were made on day 21 after implantation. Soft radiographs (wavelength 10-0.10 nm) showed opaque shadows in both groups. Histological analysis showed that new bone had formed in both experimental groups. Endochondral ossification was found at the outermost edge of the implanted collagen in the atelopeptide group. However, there was less ossification in the implanted collagen in the artificial collagen group. On biochemical examination, alkaline phosphatase activity and calcium concentrations in both experimental groups were higher than in the control group, and were higher in the atelopeptide group than in the artificial collagen group. Our results suggest that artificial collagen is useful as a carrier for rhBMP-2 designed to promote the formation of new bone.     

rhBMP-2对人牙周膜细胞骨桥蛋白表达的影响

目的　深入了解牙周膜细胞 (periodontalligamentcells,PDLC)的成骨样细胞特性 ,以及非胶原蛋白在牙周组织矿化中的作用。方法　在体外培养条件下 ,观察重组人骨形成蛋白 2(recombinanthumanbonemorphogeneticprotein 2 ,rhBMP 2 )作用前后是否对矿化相关蛋白之一的骨桥蛋白 (osteopontin ,OPN)在人PDLC中的存在及表达情况产生影响。在小玻片上培养第 5代PDLC ,分为加rhBMP 2 (5 0 μg/L)刺激的实验组和不加任何因子的空白对照组 ,以地高辛标记的OPNcDNA探针 ,采用原位杂交技术对PDLC中OPN的表达情况进行检测 ;同时做PBS替代探针和RNA酶预处理的方法学对照。结果　对照组、替代对照和RNA酶预处理的PDLC均显示为阴性反应 ,没有显示出OPN的阳性表达信号 ;实验组PDLC的胞浆中可见明显的OPN阳性反应 ,表达信号较强。结论　rhBMP 2的刺激作用可使PDLC表达出较强的OPN阳性信号 ;表明在一定条件和外界因子的诱导下 ,PDLC具有向成骨特性方向转变的潜能 ,对牙周组织再生有促进效应。     

Molded bone augmentation by a combination of barrier membrane and recombinant human bone morphogenetic protein-2

OBJECTIVES: To provide the histological background to a new method of local bone augmentation, we examined the events occurring beneath a barrier membrane applied with recombinant human bone morphogenetic protein-2 (rhBMP-2). MATERIALS AND METHODS: The effects on bone augmentation of rhBMP-2, applied with a membrane mold (BMP-Memb), over surgically-induced bone defects in rat calvaria were examined histologically, and the results compared with those from application of rhBMP-2 (BMP) alone, or of a molded membrane (Memb) alone. RESULTS: At postoperative week 2, the BMP group showed the most marked bone formation. However, the bone diminished in size by week 8. The Memb group showed slow but continuous bone formation by week 8. In the BMP-Memb group, bone filled the space in the mold at week 2, and this was maintained until week 8. Moreover, the soft tissue that had intervened between newly formed bone and the membrane in the Memb group was not evident in the BMP-Memb group, in which bone had formed directly on the membrane. CONCLUSIONS: The results suggest that the combination of rhBMP-2 and barrier membrane has advantages in producing and maintaining bone in the intended shape by inducing osteoblasts directly on the inner surface of the membrane.