Integrin-β1、FN及F-actin在多孔钽-骨界面成骨中的表达及其意义
发布时间：2019-04-22 14:21
【摘要】：目的观察多孔钽兔股骨外侧髁植入,钽-骨结合界面成骨形态特点及成骨相关因子整合素β1(Integrinβ1)、纤维黏连蛋白(FN)、纤维状肌动蛋白(F-actin)的表达,探讨多孔钽骨内植入后钽-骨界面成骨特征及相关骨整合生物学机制。方法1材料准备及分组:将多孔钽材料制成直径为3mm、长6mm的圆柱体,异种骨制成同样大小圆柱体,经消毒,备用。选取48只6月龄新西兰兔。采取自身对照方法分为A、B两组,A组为动物左侧股骨外侧髁内多孔钽植入(实验组),B组为同一动物右侧股骨外侧髁内异种骨植入(对照组)。制备动物双侧股骨外侧髁骨缺损模型。于股骨外侧髁处钻直径3mm、深度6mm孔道,在左侧(A组)将直径为3mm、长6mm的多孔钽沿孔道植入,右侧(B组)孔道植入同等大小异种骨。术后2、4、8周3个时间点处死动物并立即于骨缺损部位取材,每个时间点16只。2术后一般状态及形态学观察:术后观察动物的日常饮食、活动及伤口愈合情况。实验动物分别于术后2、4、8周处死并取材。大体标本观察界面结合的紧密程度;石蜡切片、硬组织切片观察多孔钽与宿主骨交界处成骨形态特点;扫描电镜观察多孔钽-骨界面成骨形态特征;免疫组织化学检测成骨相关蛋白Integrin-β1、FN及F-actin表达,图像采集及Image-Pro-Plus图像分析软件进行半定量分析得出IOD值并进行统计学分析。结果1大体观察:多孔钽植入后各时间点与宿主骨结合紧密,两组均无炎症反应。2石蜡切片HE染色结果显示:早期钽-骨界面少量浆液、红细胞及纤维素渗出,晚期骨小梁形成,成骨;对照组异种骨逐渐降解,新骨形成,“爬行替代”。3硬组织切片观察:早期钽-骨界面新生骨及小血管向孔隙内生长;晚期钽-宿主骨界面新生骨逐渐增多并与宿主骨连接成片,骨小梁成熟。4扫描电镜下可见成骨细胞在多孔钽表面及孔隙内生长,骨胶原束状排列。晚期骨质成熟,骨胶原粗大并见板层骨形成。5免疫组化结果显示:Integrin-β1:多孔钽组与异种骨组术后2周、4周表达均高于术后8周,P0.05;术后2周、4周异种骨组表达高于多孔钽组,P0.05。FN:多孔钽组与异种骨组术后2周、4周表达均高于术后8周,P0.05;术后2周、4周异种骨组表达高于多孔钽组,P0.05。F-actin:多孔钽组与异种骨组术后2周、4周表达均高于术后8周,P0.05;术后2周、4周异种骨组表达高于多孔钽组,P0.05。结论多孔钽兔股骨外侧髁植入术后2周,钽-骨界面组织中Integrin-β1、FN、F-actin均有阳性表达,4周达高峰,后逐渐降低,8周时表达程度达到最低。说明钽-骨界面的相互作用初期即出现成骨细胞与多孔钽黏附作用,细胞形态改变,Integrin-β1、FN及F-actin表达增强,对早期成骨起促进作用。随着骨组织逐渐成熟,成骨细胞表面Integrin-β1与FN的结合能力减低,对成骨细胞力学刺激减弱,F-actin激活数量减少,3种成骨相关蛋白表达降低,则有利于骨整合及改建。
[Abstract]:Objective to observe the bone morphology and expression of integrin 尾 1 (integrin 尾 1) and fibronectin (FN), (F-actin) in the lateral femoral condyle implantation, tantalum-bone interface and osteogenic associated factor 尾 1 (integrin 尾 1) and fibronectin (F-actin) in porous tantalum rabbits. Objective: to investigate the osteogenic characteristics of tantalum-bone interface after porous tantalum implantation and the biological mechanism of associated bone integration. Methods (1) preparation and grouping of materials: porous tantalum materials were made into cylinder with diameter of 3 mm and long 6mm, and the xenogeneic bone was made into cylinder of the same size, which was sterilized and set aside. Forty-eight 6-month-old New Zealand rabbits were selected. Two groups were divided into two groups: group A and group B, and group A was the left lateral femoral intercondylar porous tantalum implantation (the experimental group), B was the same animal right lateral femoral intercondylar bone xenograft implantation (control group). The model of bilateral lateral femoral condylar bone defect was established. The lateral femoral condyle was drilled with 3 mm diameter and deep 6mm channel. On the left side (group A), 3 mm porous tantalum with long 6mm was implanted along the channel, and the right side (group B) was implanted with the same size xenogeneic bone. At 2, 4, 8 weeks after operation, 16 animals were killed at 3 time points after operation, and 16 animals were taken from bone defect site immediately. 2. General state and morphology observation: daily diet, activity and wound healing were observed after operation. The animals were killed at 2, 4 and 8 weeks after operation. The bone morphology at the junction of porous tantalum and host bone was observed by paraffin section and hard tissue section, the morphology of porous tantalum-bone interface was observed by scanning electron microscope (SEM), and the bone formation characteristics of porous tantalum-bone interface were observed by scanning electron microscope (SEM). The expression of osteogenic associated proteins Integrin- 尾 1, FN and F-actin was detected by immunohistochemistry. The IODs were obtained by semi-quantitative analysis with image acquisition and Image-Pro-Plus image analysis software and analyzed statistically. Results 1Gross observation: the porous tantalum was closely bound to the host bone at each time point after implantation, and there was no inflammatory reaction in both groups. 2 the results of HE staining on paraffin sections showed that a small amount of plasma, red blood cells and cellulose exudated from the tantalum-bone interface in the early stage. Bone trabecular formation and osteogenesis in late stage; In the control group, the heterogeneous bone gradually degraded, new bone formed, and "creeping substitution". 3 observation on hard tissue section: the early tantalum-bone interface new bone and small blood vessels grew into the pores; The late Ta-host bone interface was gradually increased and connected to the host bone, bone trabecula matured. 4 under scanning electron microscope, osteoblasts grew on the porous tantalum surface and pore, and bone collagenous bundle-like arrangement was observed. The results of immunohistochemistry showed that the expression of Integrin- 尾 1 in porous tantalum group and xenogeneic bone group was significantly higher than that in 8 weeks after operation (P 0.05), and the expression of TGF 尾 1 in porous tantalum group and xenogeneic bone group was significantly higher than that in 8 weeks after operation (P 0.05). The expression of P0.05.FN in the xenogeneic bone group was higher than that in the porous tantalum group at 2 weeks and 4 weeks after operation, and it was higher in the porous tantalum group and the xenogeneic bone group at 2 weeks and 4 weeks after operation than in the 8 weeks after operation, P 0.05; At 2 weeks and 4 weeks after operation, the expression of xenogeneic bone group was higher than that of porous tantalum group, and P0.05.Factin expression was higher in porous tantalum group and xenogeneic bone group at 2 weeks and 4 weeks after operation than that at 8 weeks after operation, P 0.05; At 2 weeks and 4 weeks after operation, the expression of bone xenograft was higher than that of porous tantalum group (P 0.05). Conclusion the positive expression of Integrin- 尾 1 and FN,F-actin in tantalum-bone interface was found in the lateral femoral condyle of porous tantalum rabbits 2 weeks after implantation, reached the peak at 4 weeks, then decreased gradually, and reached the lowest level at 8 weeks. These results suggest that the adhesion between osteoblasts and porous tantalum occurs at the early stage of the interaction between tantalum and bone. The morphological changes of osteoblasts and the increased expression of Integrin- 尾 1, FN and F-actin play an important role in promoting early osteogenesis. With the maturing of bone tissue, the binding ability of Integrin- 尾 1 to FN on osteoblast surface decreased, the mechanical stimulation of osteoblast decreased, the number of F-actin activation decreased, and the expression of three osteogenic related proteins decreased, which was beneficial to bone integration and remodeling.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R68
本文编号：2462913
[Abstract]:Objective to observe the bone morphology and expression of integrin 尾 1 (integrin 尾 1) and fibronectin (FN), (F-actin) in the lateral femoral condyle implantation, tantalum-bone interface and osteogenic associated factor 尾 1 (integrin 尾 1) and fibronectin (F-actin) in porous tantalum rabbits. Objective: to investigate the osteogenic characteristics of tantalum-bone interface after porous tantalum implantation and the biological mechanism of associated bone integration. Methods (1) preparation and grouping of materials: porous tantalum materials were made into cylinder with diameter of 3 mm and long 6mm, and the xenogeneic bone was made into cylinder of the same size, which was sterilized and set aside. Forty-eight 6-month-old New Zealand rabbits were selected. Two groups were divided into two groups: group A and group B, and group A was the left lateral femoral intercondylar porous tantalum implantation (the experimental group), B was the same animal right lateral femoral intercondylar bone xenograft implantation (control group). The model of bilateral lateral femoral condylar bone defect was established. The lateral femoral condyle was drilled with 3 mm diameter and deep 6mm channel. On the left side (group A), 3 mm porous tantalum with long 6mm was implanted along the channel, and the right side (group B) was implanted with the same size xenogeneic bone. At 2, 4, 8 weeks after operation, 16 animals were killed at 3 time points after operation, and 16 animals were taken from bone defect site immediately. 2. General state and morphology observation: daily diet, activity and wound healing were observed after operation. The animals were killed at 2, 4 and 8 weeks after operation. The bone morphology at the junction of porous tantalum and host bone was observed by paraffin section and hard tissue section, the morphology of porous tantalum-bone interface was observed by scanning electron microscope (SEM), and the bone formation characteristics of porous tantalum-bone interface were observed by scanning electron microscope (SEM). The expression of osteogenic associated proteins Integrin- 尾 1, FN and F-actin was detected by immunohistochemistry. The IODs were obtained by semi-quantitative analysis with image acquisition and Image-Pro-Plus image analysis software and analyzed statistically. Results 1Gross observation: the porous tantalum was closely bound to the host bone at each time point after implantation, and there was no inflammatory reaction in both groups. 2 the results of HE staining on paraffin sections showed that a small amount of plasma, red blood cells and cellulose exudated from the tantalum-bone interface in the early stage. Bone trabecular formation and osteogenesis in late stage; In the control group, the heterogeneous bone gradually degraded, new bone formed, and "creeping substitution". 3 observation on hard tissue section: the early tantalum-bone interface new bone and small blood vessels grew into the pores; The late Ta-host bone interface was gradually increased and connected to the host bone, bone trabecula matured. 4 under scanning electron microscope, osteoblasts grew on the porous tantalum surface and pore, and bone collagenous bundle-like arrangement was observed. The results of immunohistochemistry showed that the expression of Integrin- 尾 1 in porous tantalum group and xenogeneic bone group was significantly higher than that in 8 weeks after operation (P 0.05), and the expression of TGF 尾 1 in porous tantalum group and xenogeneic bone group was significantly higher than that in 8 weeks after operation (P 0.05). The expression of P0.05.FN in the xenogeneic bone group was higher than that in the porous tantalum group at 2 weeks and 4 weeks after operation, and it was higher in the porous tantalum group and the xenogeneic bone group at 2 weeks and 4 weeks after operation than in the 8 weeks after operation, P 0.05; At 2 weeks and 4 weeks after operation, the expression of xenogeneic bone group was higher than that of porous tantalum group, and P0.05.Factin expression was higher in porous tantalum group and xenogeneic bone group at 2 weeks and 4 weeks after operation than that at 8 weeks after operation, P 0.05; At 2 weeks and 4 weeks after operation, the expression of bone xenograft was higher than that of porous tantalum group (P 0.05). Conclusion the positive expression of Integrin- 尾 1 and FN,F-actin in tantalum-bone interface was found in the lateral femoral condyle of porous tantalum rabbits 2 weeks after implantation, reached the peak at 4 weeks, then decreased gradually, and reached the lowest level at 8 weeks. These results suggest that the adhesion between osteoblasts and porous tantalum occurs at the early stage of the interaction between tantalum and bone. The morphological changes of osteoblasts and the increased expression of Integrin- 尾 1, FN and F-actin play an important role in promoting early osteogenesis. With the maturing of bone tissue, the binding ability of Integrin- 尾 1 to FN on osteoblast surface decreased, the mechanical stimulation of osteoblast decreased, the number of F-actin activation decreased, and the expression of three osteogenic related proteins decreased, which was beneficial to bone integration and remodeling.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R68
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