發(fā)布時(shí)間：2018-05-13 21:07

  本文選題：FG + bFGF　； 參考：《甘肅農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：目前,骨折愈合研究的熱點(diǎn)已經(jīng)轉(zhuǎn)向?yàn)閯?chuàng)口創(chuàng)造良好的生物學(xué)環(huán)境和有效控制感染,這是骨外科迫切需要解決的問(wèn)題,也是成功治療骨折的關(guān)鍵。BMP能誘導(dǎo)未分化的間充質(zhì)細(xì)胞分化形成軟骨和新生骨,對(duì)骨量、骨的發(fā)生和重建具有多向調(diào)節(jié)作用;bFGF能顯著促進(jìn)細(xì)胞的有絲分裂和血管形成,加速軟骨的成熟和骨化,可在BMP啟動(dòng)骨重建后協(xié)同骨修復(fù);骨外傷感染中的幾種主要致病菌對(duì)妥布霉素的耐藥性低,敏感性較高;并且妥布霉素對(duì)BMP的促骨愈合效果沒(méi)有抑制作用,反而可促進(jìn)骨折部位的修復(fù)。FG作為一種低抗原性的生物大分子材料,符合BMP和bFGF載體的理想條件,因此,本實(shí)驗(yàn)將纖維蛋白膠作為一種支架材料和抗生素藥物緩釋載體,復(fù)合可在骨重建過(guò)程中發(fā)揮重要作用的兩種因子rhBMP-2和bFGF以及妥布霉素,制備成一種具有抗生素緩釋系統(tǒng)和細(xì)胞因子載體作用的復(fù)合物,以評(píng)估該復(fù)合物對(duì)骨折愈合的影響,并從病理組織學(xué)及分子生物學(xué)角度初步探討該復(fù)合物在骨折愈合過(guò)程中的作用機(jī)理,為骨組織工程提供實(shí)驗(yàn)依據(jù)和理論基礎(chǔ)。本實(shí)驗(yàn)選取12只犬作為實(shí)驗(yàn)動(dòng)物,在無(wú)菌條件下用骨科擺鋸建立標(biāo)準(zhǔn)犬脛骨骨折模型。每只犬以右側(cè)后肢為實(shí)驗(yàn)肢,左側(cè)后肢為對(duì)照肢。實(shí)驗(yàn)肢采用常規(guī)內(nèi)固定方法+FG/rh BMP-2/bFGF/妥布霉素復(fù)合物進(jìn)行骨折整復(fù)固定,對(duì)照肢僅采用常規(guī)內(nèi)固定方法進(jìn)行骨折整復(fù)固定。于術(shù)后4、8、12、16周隨機(jī)選取三只取骨折處組織樣品,制備石蠟切片,HE染色、MASSON三色染色法與甲苯胺藍(lán)染色觀察骨折處組織學(xué)變化,并采用免疫組織化學(xué)方法檢測(cè)中4種因子的表達(dá)情況,實(shí)驗(yàn)結(jié)果如下:1.組織學(xué)變化:實(shí)驗(yàn)組骨折愈合早期成骨細(xì)胞、血管內(nèi)皮細(xì)胞和成軟骨細(xì)胞數(shù)目增多、增殖分化快,炎性清除期短,較對(duì)照組膠原纖維和骨基質(zhì)的沉積量多,骨小梁形成和改建快,提前進(jìn)入骨痂改造期,表明FG/rhBMP-2/bFGF/妥布霉素復(fù)合物在骨修復(fù)期可增強(qiáng)細(xì)胞、黏附增殖作用,為骨折愈合創(chuàng)造良好的細(xì)胞微環(huán)境,可增加細(xì)胞與細(xì)胞因子相互作用,促進(jìn)組織生長(zhǎng),提高骨修復(fù)及改建質(zhì)量。2.VEGF陽(yáng)性率的表達(dá):實(shí)驗(yàn)組和對(duì)照組VEGF陽(yáng)性率均呈先升高后降低趨勢(shì),第8周最高,實(shí)驗(yàn)組第8周與12周差異顯著(P0.01),其余時(shí)間點(diǎn)差異不顯著;對(duì)照組第4、8、12周差異均顯著(P0.01),術(shù)后第4周和第8周,實(shí)驗(yàn)組VEGF陽(yáng)性率顯著高于對(duì)照組(P0.01),第12周和16周差異性不顯著。表明FG/rhBMP-2/bFGF/妥布霉素復(fù)合物在骨折愈合早期促進(jìn)內(nèi)皮細(xì)胞遷移、增殖,毛細(xì)血管新生,提高骨折局部的細(xì)胞代謝,優(yōu)化骨折局部細(xì)胞微環(huán)境。3.PDGF陽(yáng)性率的表達(dá):術(shù)后各時(shí)間點(diǎn),實(shí)驗(yàn)組和對(duì)照組PDGF陽(yáng)性率均呈先增長(zhǎng)后下降趨勢(shì),實(shí)驗(yàn)組第8周最高,各相鄰時(shí)間點(diǎn)間差異均顯著(P0.01);對(duì)照組第12周最高,第4周和8周、第12周和第16周相比差異顯著,其余相鄰時(shí)間點(diǎn)間差異不顯著(P0.01)。術(shù)后第4周和第8周實(shí)驗(yàn)組PDGF陽(yáng)性率均顯著高于對(duì)照組(P0.01),第12周兩組差異不顯著,第16w對(duì)照組陽(yáng)性率高于實(shí)驗(yàn)組,兩組差異不顯著。表明FG/rhBMP-2/bFGF/妥布霉素復(fù)合物提高堿性成纖維細(xì)胞的增殖、分化,可提前增強(qiáng)成骨細(xì)胞和成纖維細(xì)胞的分泌能力,促進(jìn)膠原纖維生成與沉積。加快骨折愈合。4.IGF、TGF-β1陽(yáng)性率的表達(dá):IGF、TGF-β1陽(yáng)性率均呈先增長(zhǎng)后下降趨勢(shì),均為第12周最高,IGF實(shí)驗(yàn)組陽(yáng)性率各時(shí)間點(diǎn)間差異均顯著(P0.01),對(duì)照組第4周、8周和12周差異均顯著(P0.01),實(shí)驗(yàn)組與對(duì)照組相比,第4周、8周和12周差異均顯著(P0.01);TGF-β實(shí)驗(yàn)組陽(yáng)性率各時(shí)間點(diǎn)間差異均顯著(P0.01),對(duì)照組第4周、8周和12周差異均顯著(P0.01),實(shí)驗(yàn)組與對(duì)照組相比,第4周、8周和12周差異均顯著(P0.01)。表明FG/rhBMP-2/bFGF/妥布霉素能促進(jìn)成骨細(xì)胞以自分泌的方式增殖分化和增加膠原合成。刺激細(xì)胞募集和增殖,啟動(dòng)修復(fù)過(guò)程。加速骨基質(zhì)的鈣化,同時(shí)改建骨小梁,加快愈合進(jìn)程。結(jié)論:FG/rhBMP-2/bFGF/妥布霉素復(fù)合物在骨折早期為細(xì)胞和細(xì)胞因子創(chuàng)建良好微環(huán)境,促進(jìn)多種細(xì)胞趨化、黏附、分化、增殖以及骨折端血管的形成和長(zhǎng)入,維持藥物和細(xì)胞因子較高濃度,骨修復(fù)作用增加,能協(xié)調(diào)骨折愈合改建期破骨細(xì)胞和成骨細(xì)胞相互作用,加快骨改建進(jìn)程。
[Abstract]:At present, the hot spot of fracture healing research has turned to create a good biological environment and effective control of infection for the wound, which is an urgent problem in bone surgery. It is also the key.BMP for the successful treatment of fracture. It can induce undifferentiated mesenchymal cells to differentiate into cartilage and new bone, and have multi direction for bone mass, bone formation and reconstruction. BFGF can significantly promote mitosis and angiogenesis of cells, accelerate the maturation and ossification of cartilage, and can be combined with bone repair after the BMP start of bone reconstruction; several major pathogens in bone trauma infection have low resistance to tobramycin and high sensitivity; and tobramycin does not inhibit the effect of BMP on bone healing. The repair of.FG as a low antigenicity biological macromolecular material conforms to the ideal conditions for BMP and bFGF carriers. Therefore, this experiment uses fibrin glue as a scaffold material and antibiotic drug sustained-release carrier, combined with two factors, rhBMP-2 and bFGF, which can play an important role in the process of bone reconstruction. In order to evaluate the effect of the compound on fracture healing, a complex of antibiotic sustained-release system and cytokine carrier was prepared to evaluate the effect of the compound on fracture healing. The mechanism of the compound in the process of fracture healing was preliminarily discussed from the histopathological and molecular biological angles, which provided experimental basis and theoretical basis for bone tissue engineering. In this experiment, 12 dogs were selected as experimental animals. In the aseptic condition, a standard canine tibial fracture model was set up in the Department of orthopedics. The right hind limbs were the experimental limbs and the left hind limbs were the control limbs. The experimental limbs were fixed by the routine internal fixation method +FG/rh BMP-2/bFGF/ tobramycin complex, and the control limbs were only used in routine internal fixation. A fixed method was used to fix the fracture and fix. Three tissue samples were selected at random in 4,8,12,16 weeks after the operation. The paraffin section, HE staining, MASSON staining and toluidine blue staining were used to observe the histological changes of the fracture, and the expression of the 4 factors was detected by immunohistochemistry. The experimental results were as follows: 1. tissues Study changes: the early osteoblast of fracture healing in the experimental group increased, the number of vascular endothelial cells and chondrocytes increased, the proliferation and differentiation were fast, the inflammatory clearance period was short, the deposition of collagen fibers and bone matrix in the control group was more, the formation and reconstruction of bone trabeculae were fast, and the FG/rhBMP-2/bFGF/ tobramycin complex was repaired in the bone repair. It can enhance cell and adhesion and proliferate, create a good cell microenvironment for fracture healing, increase the interaction of cell and cytokine, promote tissue growth, improve the expression of.2.VEGF positive rate of bone repair and remodeling: the positive rate of VEGF in the experimental group and the control group all increased first and then decreased, the highest in eighth weeks and eighth weeks in the experimental group. The difference between the 12 weeks was significant (P0.01) and the difference of other time points was not significant. The difference of the control group at week 4,8,12 was significant (P0.01). The positive rate of VEGF in the experimental group was significantly higher than that of the control group (P0.01) at the fourth and eighth weeks postoperatively, and the difference between the twelfth and the 16 weeks was not significant. It showed that the FG/rhBMP-2/ bFGF/ tobramycin complex promoted endothelial cell migration early in the fracture healing. Shift, proliferation, capillary neovascularization, improve the cell metabolism in the part of the fracture, and optimize the expression of.3.PDGF positive rate in the microenvironment of the fracture. The positive rates of PDGF in the experimental group and the control group all increased first after the operation, the highest in the experimental group eighth weeks, and the difference between the adjacent time points (P0.01), and the highest in the control group for Twelfth weeks, Fourth weeks and 8 weeks, compared with twelfth weeks and sixteenth weeks, the difference was not significant (P0.01). The positive rate of PDGF in the experimental group was significantly higher than that of the control group (P0.01) at fourth weeks and eighth weeks after the operation, and the difference of two groups in twelfth weeks was not significant. The positive rate of the 16W control group was higher than that of the experimental group, and the difference of the two groups was not significant. It showed that the FG/rhBMP-2/bFGF/ was duly distributed. Mycomycin complex enhanced the proliferation of basic fibroblasts, enhanced the secretion of osteoblasts and fibroblasts in advance, promoted the formation and deposition of collagen fibers, accelerated the.4.IGF and TGF- beta 1 positive rate of fracture healing: the positive rates of IGF, TGF- beta 1 were all first increased and then decreased, both were the highest in twelfth weeks, and the positive rate of IGF experimental group. The difference in each time point was significant (P0.01), the difference was significant (P0.01) in the control group for fourth weeks and at the 8 and 12 weeks. Compared with the control group, the difference was significant (P0.01) at fourth weeks, 8 and 12 weeks in the experimental group, and the difference in the positive rate of the TGF- beta test group was significant (P0.01), the control group was fourth weeks, and the difference between the 8 weeks and the 12 weeks was significant (P0.01). The experimental group and the control group were all significantly different (P0.01). The difference between fourth weeks, 8 and 12 weeks was significant (P0.01). It showed that FG/rhBMP-2/bFGF/ tobramycin could promote the proliferation and differentiation of osteoblasts in the autocrine way and increase collagen synthesis, stimulate cell recruitment and proliferation, start the repair process, accelerate calcification of bone matrix, reconstruct trabecular bone and accelerate the healing process. Conclusion: FG/rhBMP-2/bFGF/ tob. Mycin complex creates a good microenvironment for cells and cytokines at the early stage of fracture to promote the chemotaxis, adhesion, differentiation, proliferation, and formation and growth of the blood vessels at the end of the fracture, maintaining a high concentration of drugs and cytokines, increasing the effect of bone repair, and coordinating the interaction of osteoclasts and osteoblasts in the healing period of fracture healing and accelerating the interaction of bone cells and osteoblasts. Bone remodeling process.

【學(xué)位授予單位】：甘肅農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S858.292

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 周正楠;張巖;邱旭升;施鴻飛;張子韜;陳一心;;骨折愈合過(guò)程中VEGF的表達(dá)與調(diào)控及其作用的研究進(jìn)展[J];中國(guó)矯形外科雜志;2015年24期

2 張虎;;骨折X線攝片不當(dāng)誤診2例分析[J];中國(guó)骨與關(guān)節(jié)損傷雜志;2015年S1期

3 李棋;李箭;;堿性成纖維細(xì)胞生長(zhǎng)因子及纖維蛋白膠在骨科的運(yùn)用[J];中國(guó)組織工程研究;2015年12期

4 李芳;;X線平片與CR在診斷隱匿性骨折診斷中的應(yīng)用比較[J];大家健康(學(xué)術(shù)版);2014年21期

5 王玉婷;陳加源;蔣瑾;李麗佳;;非骨化性纖維瘤的X射線及CT表現(xiàn)特征分析[J];實(shí)用醫(yī)院臨床雜志;2014年04期

6 馮家義;;230例脊柱創(chuàng)傷X線平片與螺旋CT的對(duì)照分析[J];中國(guó)醫(yī)學(xué)工程;2013年11期

7 李曉;郭衛(wèi);楊毅;尉然;杜志業(yè);;四肢長(zhǎng)骨骨巨細(xì)胞瘤伴病理性骨折的外科治療[J];北京大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2013年05期

8 肖裕華;李紹琴;潘文峰;李建飛;陳宗和;;纖維蛋白膠復(fù)合骨形態(tài)發(fā)生蛋白與VEGF修復(fù)兔橈骨缺損的實(shí)驗(yàn)研究[J];實(shí)用臨床醫(yī)學(xué);2013年06期

9 周長(zhǎng)臣;張建設(shè);李焰;;生長(zhǎng)因子在骨折愈合過(guò)程中促血管生成作用的研究進(jìn)展[J];西南國(guó)防醫(yī)藥;2013年04期

10 劉爽;李善昌;劉占領(lǐng);魏巍;;骨折愈合過(guò)程中骨形態(tài)發(fā)生蛋白與細(xì)胞外基質(zhì)磷酸糖蛋白表達(dá)的相關(guān)性[J];中國(guó)老年學(xué)雜志;2013年02期

相關(guān)博士學(xué)位論文 前1條

1 尹德馨;新劑型神經(jīng)生長(zhǎng)因子聯(lián)合丙戊酸促進(jìn)大鼠坐骨神經(jīng)損傷再生協(xié)同作用的實(shí)驗(yàn)研究[D];吉林大學(xué);2015年

相關(guān)碩士學(xué)位論文 前3條

1 修金濤;多孔自凝固磷酸鈣/纖維蛋白血管化的分析研究[D];第四軍醫(yī)大學(xué);2013年

2 崔林江;骨折合并腦外傷時(shí)IGF-Ⅱ?qū)钦塾系挠绊慬D];青島大學(xué);2007年

3 王帥;生物蛋白膠—加替沙星混合劑預(yù)防肌腱粘連和局部感染的臨床研究[D];青島大學(xué);2007年

，

本文編號(hào)：1884799