發(fā)布時(shí)間：2019-04-22 14:21

【摘要】：目的觀察多孔鉭兔股骨外側(cè)髁植入,鉭-骨結(jié)合界面成骨形態(tài)特點(diǎn)及成骨相關(guān)因子整合素β1(Integrinβ1)、纖維黏連蛋白(FN)、纖維狀肌動(dòng)蛋白(F-actin)的表達(dá),探討多孔鉭骨內(nèi)植入后鉭-骨界面成骨特征及相關(guān)骨整合生物學(xué)機(jī)制。方法1材料準(zhǔn)備及分組:將多孔鉭材料制成直徑為3mm、長6mm的圓柱體,異種骨制成同樣大小圓柱體,經(jīng)消毒,備用。選取48只6月齡新西蘭兔。采取自身對照方法分為A、B兩組,A組為動(dòng)物左側(cè)股骨外側(cè)髁內(nèi)多孔鉭植入(實(shí)驗(yàn)組),B組為同一動(dòng)物右側(cè)股骨外側(cè)髁內(nèi)異種骨植入(對照組)。制備動(dòng)物雙側(cè)股骨外側(cè)髁骨缺損模型。于股骨外側(cè)髁處鉆直徑3mm、深度6mm孔道,在左側(cè)(A組)將直徑為3mm、長6mm的多孔鉭沿孔道植入,右側(cè)(B組)孔道植入同等大小異種骨。術(shù)后2、4、8周3個(gè)時(shí)間點(diǎn)處死動(dòng)物并立即于骨缺損部位取材,每個(gè)時(shí)間點(diǎn)16只。2術(shù)后一般狀態(tài)及形態(tài)學(xué)觀察:術(shù)后觀察動(dòng)物的日常飲食、活動(dòng)及傷口愈合情況。實(shí)驗(yàn)動(dòng)物分別于術(shù)后2、4、8周處死并取材。大體標(biāo)本觀察界面結(jié)合的緊密程度;石蠟切片、硬組織切片觀察多孔鉭與宿主骨交界處成骨形態(tài)特點(diǎn);掃描電鏡觀察多孔鉭-骨界面成骨形態(tài)特征;免疫組織化學(xué)檢測成骨相關(guān)蛋白Integrin-β1、FN及F-actin表達(dá),圖像采集及Image-Pro-Plus圖像分析軟件進(jìn)行半定量分析得出IOD值并進(jìn)行統(tǒng)計(jì)學(xué)分析。結(jié)果1大體觀察:多孔鉭植入后各時(shí)間點(diǎn)與宿主骨結(jié)合緊密,兩組均無炎癥反應(yīng)。2石蠟切片HE染色結(jié)果顯示:早期鉭-骨界面少量漿液、紅細(xì)胞及纖維素滲出,晚期骨小梁形成,成骨;對照組異種骨逐漸降解,新骨形成,“爬行替代”。3硬組織切片觀察:早期鉭-骨界面新生骨及小血管向孔隙內(nèi)生長;晚期鉭-宿主骨界面新生骨逐漸增多并與宿主骨連接成片,骨小梁成熟。4掃描電鏡下可見成骨細(xì)胞在多孔鉭表面及孔隙內(nèi)生長,骨膠原束狀排列。晚期骨質(zhì)成熟,骨膠原粗大并見板層骨形成。5免疫組化結(jié)果顯示:Integrin-β1:多孔鉭組與異種骨組術(shù)后2周、4周表達(dá)均高于術(shù)后8周,P0.05;術(shù)后2周、4周異種骨組表達(dá)高于多孔鉭組,P0.05。FN:多孔鉭組與異種骨組術(shù)后2周、4周表達(dá)均高于術(shù)后8周,P0.05;術(shù)后2周、4周異種骨組表達(dá)高于多孔鉭組,P0.05。F-actin:多孔鉭組與異種骨組術(shù)后2周、4周表達(dá)均高于術(shù)后8周,P0.05;術(shù)后2周、4周異種骨組表達(dá)高于多孔鉭組,P0.05。結(jié)論多孔鉭兔股骨外側(cè)髁植入術(shù)后2周,鉭-骨界面組織中Integrin-β1、FN、F-actin均有陽性表達(dá),4周達(dá)高峰,后逐漸降低,8周時(shí)表達(dá)程度達(dá)到最低。說明鉭-骨界面的相互作用初期即出現(xiàn)成骨細(xì)胞與多孔鉭黏附作用,細(xì)胞形態(tài)改變,Integrin-β1、FN及F-actin表達(dá)增強(qiáng),對早期成骨起促進(jìn)作用。隨著骨組織逐漸成熟,成骨細(xì)胞表面Integrin-β1與FN的結(jié)合能力減低,對成骨細(xì)胞力學(xué)刺激減弱,F-actin激活數(shù)量減少,3種成骨相關(guān)蛋白表達(dá)降低,則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.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R68

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