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Histological and ultrastructural analysis of heterotopic osteogenesis in porous calcium phosphate ceramics
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Failure of heterotopic osteogenesis by epithelial mesenchymal cell interactions in xenogeneic transplants in the kidney
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planoepitheliale, n=2) were grafted intramuscularly into cortisone-immunosuppressed mice to test the ability of transformed transitional epithelium to induce heterotopic osteogenesis.
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The occurrence of different nerve types was analyzed at different stages of heterotopic osteogenesis, induced by allogeneic bone matrix.
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| Objective To evaluate the heterotopic osteogenenic potential of xenogeneic bone delivery of human mesenchymal stem cells transfected with bone morphogenetic protein-2 (BMP-2) gene. Methods Cell-scaffold complexes were implanted subcutaneously into the back of the nude mice in five groups respectively,ie,Ad-BMP-2 infected hBMSC plus antigen-free bovine cancellous bone (BCB) (Group A),Ad-LacZ infected hBMSC-BCB (Group B),hBMSC-BCB plus reconstructed hBMP-2 (Group C),hBMSC-BCB (Group D) and only BCB scaffolds (Group... Objective To evaluate the heterotopic osteogenenic potential of xenogeneic bone delivery of human mesenchymal stem cells transfected with bone morphogenetic protein-2 (BMP-2) gene. Methods Cell-scaffold complexes were implanted subcutaneously into the back of the nude mice in five groups respectively,ie,Ad-BMP-2 infected hBMSC plus antigen-free bovine cancellous bone (BCB) (Group A),Ad-LacZ infected hBMSC-BCB (Group B),hBMSC-BCB plus reconstructed hBMP-2 (Group C),hBMSC-BCB (Group D) and only BCB scaffolds (Group E). Bone formation and vascularization were observed by histological examination four and eight weeks after operation. Results hBMSC continuously expressed exogenous gene in vivo three days after implantation. The Group A was filled with newly formed woven bone with abundant blood capillary eight weeks after operation. Conclusions Xenogeneic bone delivery of human mesenchymal stem cells transfected by BMP-2 gene can attain heterotopic osteogenesis and induce vascularization of capillary vessels. | 目的 观察骨形态发生蛋白 - 2 (BMP - 2 )基因转染的人骨髓间质干细胞复合异种骨支架异位成骨效果。 方法 分组 :(1)BMP - 2基因转染细胞 +异种骨支架 [去抗原牛松质骨 (BCB) ];(2 )对照基因转染细胞 +BCB ;(3)未转染细胞 +重组BMP - 2 +BCB ;(4)未转染细胞+BCB ;(5 )BCB。分别将各组人工骨植入裸鼠皮下 ,于术后 4 ,8周行组织学观察。 结果 体内植入后 3d ,细胞持续表达外源基因。BMP - 2基因转染组 8周时完全由新形成的编织骨构成 ,血管丰富。 结论 BMP - 2基因转染后细胞复合BCB支架 ,可以在异位形成骨组织并诱导毛细血管长入。 | [Objective]To observe the effect of sciatic nerve and femoral nerve resection on heterotopic osteogenesis induced by recombined human bone morphogenetic protein-2(rhBMP-2) and discuss role of never innervation on bone regeneration and partial reasons for larger callus after bone fracture accompanying denervation.[Method]A total of 36 male ICR mice were divided into experimental group and control group at random.0.125mg rhBMP-2 /collagen composites were implanted into the right thigh muscle pouches after... [Objective]To observe the effect of sciatic nerve and femoral nerve resection on heterotopic osteogenesis induced by recombined human bone morphogenetic protein-2(rhBMP-2) and discuss role of never innervation on bone regeneration and partial reasons for larger callus after bone fracture accompanying denervation.[Method]A total of 36 male ICR mice were divided into experimental group and control group at random.0.125mg rhBMP-2 /collagen composites were implanted into the right thigh muscle pouches after sciatic nerve and femoral nerve section in the experimental group and after sham operation in the control group.On the 7~(th),14~(th) and 21~(st) day after implantation,roentgenographic,biochemical,histological analyses and osteoclasts TRAP staining were performed to detect the effects of sciatic nerve and femoral nerve resection on bone growth initiated by rhBMP-2.[Result]On the 7~(th),14~(th) and 21~(st) day,wet weight of new bony tissues of the experimental group was obviously greater than that of the control group.Radiography showed range of bone formation in the experimental group was larger than that in the control,but density of new bone was lower than that of the latter.Biochemical detection showed value of AKP of the experimental group was significantly higher than that of the control on the 7~(th) day,while the content of Ca of the experimental group was higher than that of the control on the 14~(th) day and the content of Ca and P of the experimental group was lower than that of the control on the 21~(st) day.Histological observation showed area of new bone of the experimental group was greater than that of the control,but trabecular bone of the former was sparser than that of the latter.Image analysis of bony tissue showed the relative number of osteoclasts in the experimental group became higher on the 21~(st) day,while volume density and width of bone trabecula in the experimental group were lower than that of the control group.TRAP staining showed osteoclasts in the new bone of the experimental group were more activated than that of the control.[Conclusion]Nerve section results in enhancement of ability of bone regeneration by BMP in the early period,but trabecular bone becomes sparse in the middle and later period because of bone resorption by osteoclasts,which indicates nerve in nervation influences bone regemeration though direct and/or indirect way. | [目的]通过rhBMP-2异位诱导成骨模型,观察坐骨神经和股神经切除对骨再生的影响,探讨神经支配在骨再生中的作用以及失神经所致骨折骨痂增大的部分机制。[方法]ICR小鼠36只随机分为实验组和对照组。行右侧股后部肌袋模型,实验组行右侧坐骨神经和股神经切除后植入含0.125 mg rhBMP-2胶原复合物。对照组仅进行神经暴露后植入等量rhBMP-2胶原复合物。于术后7、14、21 d取材,行湿重测量、放射学、生化检测、组织学观察和形态计量分析以及破骨细胞TRAP染色。[结果]湿重检测显示实验组组织块湿重明显大于对照组。X线检测实验组成骨范围较对照组明显增大,成骨组织密度不及对照组。生化检测结果显示术后第7 d实验组AKP含量明显高于对照组,术后第14 d,实验组钙含量高于对照组,术后21 d,实验组钙、磷含量均低于对照组。组织学观察显示实验组成骨范围大于对照组,成骨后期破骨细胞活跃,骨小梁稀疏。组织形态计量分析显示实验组术后21 d破骨细胞相对数增多,骨小梁体积密度、平均宽度均低于对照组。TRAP染色显示实验组破骨细胞明显较对照组活跃。[结论]在外源性BMP-2异位诱导过程中,神经切除引起骨诱导早期成骨活动的增... [目的]通过rhBMP-2异位诱导成骨模型,观察坐骨神经和股神经切除对骨再生的影响,探讨神经支配在骨再生中的作用以及失神经所致骨折骨痂增大的部分机制。[方法]ICR小鼠36只随机分为实验组和对照组。行右侧股后部肌袋模型,实验组行右侧坐骨神经和股神经切除后植入含0.125 mg rhBMP-2胶原复合物。对照组仅进行神经暴露后植入等量rhBMP-2胶原复合物。于术后7、14、21 d取材,行湿重测量、放射学、生化检测、组织学观察和形态计量分析以及破骨细胞TRAP染色。[结果]湿重检测显示实验组组织块湿重明显大于对照组。X线检测实验组成骨范围较对照组明显增大,成骨组织密度不及对照组。生化检测结果显示术后第7 d实验组AKP含量明显高于对照组,术后第14 d,实验组钙含量高于对照组,术后21 d,实验组钙、磷含量均低于对照组。组织学观察显示实验组成骨范围大于对照组,成骨后期破骨细胞活跃,骨小梁稀疏。组织形态计量分析显示实验组术后21 d破骨细胞相对数增多,骨小梁体积密度、平均宽度均低于对照组。TRAP染色显示实验组破骨细胞明显较对照组活跃。[结论]在外源性BMP-2异位诱导过程中,神经切除引起骨诱导早期成骨活动的增加,在成骨中后期失神经导致破骨细胞活动增强引起骨小梁的稀疏和骨密度的降低,提示神经支配可能通过直接或间接的方式影响骨再生活动。 | Objective To evaluate the mechanism of heterotopic osteogenesis induced by adenovirus mediated BMP-2 gene (Ad-BMP-2) transfection. Methods (1) Ad-BMP-2 was directly injected into the muscle of nude mice to induce bone formation; (2) Human mesenchymal stem cells (hBMSC), after being transfected by Ad-BMP-2, were implanted subcutaneously into the back of the nude mice in five groups respectively, ie. Ad-BMP-2 infected hBMSC plus antigen-free bovine cancellous bone (BCB)(Group A), hBMSC-BCB plus reconstructed... Objective To evaluate the mechanism of heterotopic osteogenesis induced by adenovirus mediated BMP-2 gene (Ad-BMP-2) transfection. Methods (1) Ad-BMP-2 was directly injected into the muscle of nude mice to induce bone formation; (2) Human mesenchymal stem cells (hBMSC), after being transfected by Ad-BMP-2, were implanted subcutaneously into the back of the nude mice in five groups respectively, ie. Ad-BMP-2 infected hBMSC plus antigen-free bovine cancellous bone (BCB)(Group A), hBMSC-BCB plus reconstructed hBMP-2 (Group B), Ad-LacZ infected hBMSC-BCB (Group C), hBMSC- BCB (Group D) and only BCB scaffolds (Group E). X-ray and histological examination were conducted after operation. Results (1) Heterotopic osteogenesis was found after intramuscular injection in nude mice, most of which were endochondral ossification. (2) Implanted cell fates were traced by Y chromosome fluorescence in situ hybridization and some transduced gene cells, direct involvement in bone formation was observed; host mesenchymal cells, conversion into osteoblasts induced by BMP-2 from gene expression was displayed by osteocalcin immunohistochemical staining analysis. Conclusions hBMSC infected by BMP-2 gene not only is used as the tool of growth factor delivery, but also directly involve in bone formation. However, the induction of BMP-2 is in the leading place. | 目的观察骨形态发生蛋白2基因直接及间接转染诱导异位成骨的效果和机制。方法1.将携带骨形态发生蛋白-2基因的腺病毒载体(Ad-BMP-2)直接注射到裸鼠肌内诱导成骨;2.Ad-BMP-2体外转染人骨髓间质干细胞(hBMSC)后种植异种骨支架(BCB)植入裸鼠皮下。实验分为5组:①BMP-2基因转染细胞+BCB;②对照基因转染细胞+BCB;③未转染细胞+重组BMP-2+BCB;④未转染细胞+BCB;⑤BCB。术后行X线、免疫组化及原位杂交染色观察BMP-2基因诱导下的成骨方式及成骨机制。结果1.裸鼠肌内注射后有异位成骨,其主要为软骨化骨。2.Y染色体探针荧光原位杂交法(FISH)示踪植入细胞命运,见部分转基因细胞直接参与成骨;骨钙素免疫组化检测见基因表达产生的BMP-2诱导宿主间质细胞向成骨细胞转化。结论BMP-2基因转染的hBMSC不仅作为运载生长因子的工具,而且直接参与成骨,但BMP-2的诱导趋化作用占主导地位。 |   | << 更多相关文摘 |
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