本文選題：脫細(xì)胞真皮基質(zhì) + 堿性成纖維細(xì)胞生長因子��； 參考：《山東大學(xué)》2017年碩士論文

【摘要】：背景與目的:引導(dǎo)組織再生(GTR)或引導(dǎo)骨再生(GBR)是牙周/骨組織再生醫(yī)學(xué)領(lǐng)域的重要技術(shù),當(dāng)今醫(yī)學(xué)的發(fā)展不僅要求GTR膜作為屏障,而且希望其能作為載體釋放藥物,如抗炎藥,生長因子,粘附因子等促進(jìn)缺損區(qū)域的愈合。堿性成纖維細(xì)胞生長因子(bFGF)對干細(xì)胞有明顯促增殖作用,并有促血管形成及神經(jīng)再生等功能。骨形態(tài)發(fā)生蛋白-2(BMP-2)以促成骨分化為特色。我們認(rèn)為BMP-2和bFGF均可作為GTR和GBR屏障膜材料的備選負(fù)載因子。另外,脫細(xì)胞真皮基質(zhì)(ADM)來自于天然皮膚,是一種被去除細(xì)胞結(jié)構(gòu)的細(xì)胞外基質(zhì),并且其本身具有一定的促進(jìn)組織再生的生物活性,目前商品化的ADM膜已在臨床廣泛應(yīng)用但是,是否負(fù)載BMP-2或bFGF的ADM膜材料更能有效加速骨缺損愈合有待探索,本實(shí)驗(yàn)的目的是比較負(fù)載bFGF、BMP-2的ADM屏障膜材料在骨缺損修復(fù)中的作用,并探討其可能的不同機(jī)制。材料與方法:(1)膜材料負(fù)載因子。無菌生理鹽水分別配置200ng/ml的bFGF和800ng/ml的BMP-2,取大小相等的1cm2的膜材料,0.5ml溶液浸泡過夜,真空凍干機(jī)凍干膜材料備用。采用ELISA和體外緩釋系統(tǒng)檢測載藥膜材料的釋藥情況。(2)實(shí)驗(yàn)分為四組,分別為空白對照組(control),單純膜材料組(ADM),負(fù)載BMP-2的膜材料組(ADM+BMP-2),負(fù)載bFGF的膜材料組(ADM+bFGF)。在8w齡的雌性Wistar大鼠的顱骨左側(cè)頂骨上制備8mm×7mm橢圓全厚骨缺損,分別將上述膜材料覆蓋于缺損表面,并設(shè)置空白對照組。于術(shù)后1周,2周和8周處死實(shí)驗(yàn)動物、取材。(3)術(shù)后1周和2周,采用CD34-/CD90+免疫熒光雙染觀察骨缺損愈合過程中材料對間充質(zhì)干細(xì)胞的募集情況。(4)術(shù)后8周,通過HE染色,Micro CT掃描分析比較四個(gè)實(shí)驗(yàn)組骨缺損的整體愈合情況。(5)術(shù)后8周,免疫組化檢測骨橋蛋白(OPN)的表達(dá)以檢測干細(xì)胞成骨分化的情況。所得數(shù)據(jù)全部采用均數(shù)±標(biāo)準(zhǔn)差(Mean±SD)表示。應(yīng)用SPSS 17.0軟件包,對相關(guān)數(shù)據(jù)進(jìn)行單因素方差分析,α=0.05。結(jié)果:(1)負(fù)載因子并凍干的膜材料在形態(tài)、結(jié)構(gòu)上未發(fā)生明顯改變。(2)術(shù)后實(shí)驗(yàn)動物術(shù)后成活率約90%,缺損處無明顯炎癥,并無明顯免疫排斥反應(yīng)發(fā)生。(3)CD34-/CD90+免疫熒光雙染結(jié)果顯示ADM+bFGF在1周和2周時(shí)對干細(xì)胞的募集作用很強(qiáng),明顯高于其他三組(p0.01)。(4)組織學(xué)HE染色觀察發(fā)現(xiàn),ADM組、ADM+BMP-2組及ADM+bFGF組缺損處修復(fù)組織量明顯多于空白對照組;8w時(shí)MicroCT掃描分析結(jié)果顯示,所有植有ADM膜的實(shí)驗(yàn)組骨愈合情況明顯好于空白對照組,ADM+BMP-2組、ADM+bFGF組骨缺損基本完全愈合,新生骨體積明顯多于其他兩組。(5)骨橋蛋白(OPN)免疫組化染色顯示8周時(shí)ADM+BMP-2組、ADM+bFGF組的平均光密度(IOD)無明顯差異,但顯著高于空白對照組和單純ADM組(p0.05)。結(jié)論:與傳統(tǒng)生長因子BMP-2相比,bFGF對干細(xì)胞有早期募集和促增殖的作用,同時(shí)保留了其成骨向分化能力,其與GTR膜材料的結(jié)合不僅發(fā)揮了屏障膜的空間保持功能,而且可有效加速骨缺損的愈合。
[Abstract]:Background & objective: guided tissue Regeneration (GTRR) or guided Bone Regeneration (GBR) is an important technology in periodontal / bone regeneration medicine. The development of modern medicine not only requires GTR membrane to act as a barrier, but also hopes that it can be used as a carrier to release drugs, such as anti-inflammatory drugs. Growth factor, adhesion factor and so on promote the healing of defect area. Basic fibroblast growth factor (bFGF) can promote the proliferation of stem cells and promote angiogenesis and nerve regeneration. Bone morphogenetic protein-2 (BMP-2) is characterized by bone differentiation. We consider that BMP-2 and bFGF can be used as alternative load factors for GTR and GBR barrier membrane materials. In addition, acellular dermal matrix (ADM) comes from natural skin and is an extracellular matrix with removed cellular structure, and it has certain biological activity to promote tissue regeneration. At present, commercial ADM membrane has been widely used in clinical practice. Whether ADM membrane material loaded with BMP-2 or bFGF can accelerate bone defect healing more effectively remains to be explored. The purpose of this study was to compare the role of ADM barrier membrane loaded with bFGF- BMP-2 in bone defect repair and to explore its different mechanisms. Materials and methods 1) membrane material loading factor. 200ng/ml bFGF and 800ng/ml BMP-2 were prepared with aseptic saline respectively. The membrane material of 1cm2 of equal size was soaked in 0.5ml solution for the night. The vacuum freeze-drying machine was used for freeze-drying membrane material. Elisa and in vitro slow-release system were used to detect the drug release of drug-loaded membrane material. The experiment was divided into four groups: blank control group, simple membrane material group, ADM BMP-2 group, and bFGF loaded membrane material group. 8mm 脳 7mm elliptical full-thickness bone defects were prepared on the left parietal bone of female Wistar rats aged 8 weeks. The above membrane materials were covered on the surface of the defect respectively, and a blank control group was set up. The experimental animals were killed at 1 week, 2 weeks and 8 weeks after operation. After 1 and 2 weeks of operation, the recruitment of mesenchymal stem cells (MSCs) was observed by CD34 / CD90 immunofluorescence double staining. Eight weeks after operation, osteopontin (OPN) expression was detected by immunohistochemistry to detect osteogenic differentiation of stem cells. All the data were expressed as mean 鹵standard deviation (mean 鹵SD). Using SPSS 17.0 software package, the single factor ANOVA of related data was carried out, 偽 0.05. Results (1) the morphology and structure of the lyophilized membrane material were not changed significantly. The survival rate of the experimental animals after operation was about 90%, and there was no obvious inflammation in the defect. The results of immunofluorescence double staining of CD34 / CD90 showed that ADM bFGF had a strong recruitment effect on stem cells at week 1 and week 2. The results of HE staining showed that the defects of ADM BMP-2 and ADM bFGF in ADM group were significantly higher than those in control group at 8 weeks. The bone healing of all experimental groups with ADM membrane was better than that of ADM BMP-2 group, and the bone defect of ADM bFGF group was almost completely healed. The mean optical density (IOD) of ADM BMP-2 / ADM bFGF group was significantly higher than that of blank control group and ADM group at 8 weeks, but significantly higher than that of blank control group and ADM group. Conclusion: compared with the traditional growth factor BMP-2, bFGF has the function of early recruitment and proliferation of stem cells, while preserving its ability of osteogenic differentiation. The combination of bFGF and GTR membrane material not only plays a role in maintaining the space of barrier membrane. And it can accelerate the healing of bone defect effectively.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R781

【參考文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 路煒;異種脫細(xì)胞真皮基質(zhì)膜對大鼠顱骨缺損的修復(fù)作用[D];山東大學(xué);2014年

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本文編號：2015892