【摘要】：目的探討利用大網(wǎng)膜包裹構(gòu)建血管化組織工程骨修復(fù)兔橈骨大段骨缺損能力,評(píng)估大網(wǎng)膜包裹構(gòu)建大段血管化組織工程骨的可行性及其對(duì)骨缺損的修復(fù)效果,為組織工程骨用于臨床提供及補(bǔ)充實(shí)驗(yàn)依據(jù)。 方法采用全骨髓貼壁培養(yǎng)法結(jié)合換液傳代分離培養(yǎng)、純化兔骨髓間充質(zhì)干細(xì)胞(rBMSCs),通過(guò)觀察細(xì)胞形態(tài)、多向分化誘導(dǎo)及電鏡下觀察成骨誘導(dǎo)前后細(xì)胞的結(jié)構(gòu)改變對(duì)其進(jìn)行鑒定,MTT法測(cè)定第三代細(xì)胞的增殖情況,并繪制出生長(zhǎng)曲線。取成骨誘導(dǎo)后生長(zhǎng)狀態(tài)良好第三代細(xì)胞兔骨髓間充質(zhì)干細(xì)胞接種到自制的生物衍生骨支架材料上構(gòu)建組織工程骨,將體外構(gòu)建的組織工程骨(TEB)植入體內(nèi),分為大網(wǎng)膜包裹組織工程骨組和單純組織工程骨組。術(shù)后不同時(shí)間點(diǎn)觀察兩組移植物的表面變化及周圍組織反應(yīng),術(shù)后4周,8周,12周行X線攝片檢查移植物修復(fù)骨缺損情況,行標(biāo)本大體觀察及組織學(xué)檢測(cè),觀察成骨及血管化情況。 結(jié)果兩組移植物植入兔橈骨缺損處后,術(shù)口生長(zhǎng)良好,未發(fā)現(xiàn)明顯的全身及局部的毒性和炎癥反應(yīng),每只動(dòng)物的手術(shù)切口均達(dá)到一期愈合。大網(wǎng)膜包裹組在術(shù)后12周時(shí)X線表明植入物與宿主骨連接處基本愈合,而單純組術(shù)后12周與宿主骨愈合不甚滿意。同時(shí)大網(wǎng)膜包裹組骨修復(fù)區(qū)的骨灰度值都高于單純組織工程骨組,且差異有統(tǒng)計(jì)學(xué)意義(P.05)；術(shù)后4周,8周,12周通過(guò)分析骨組織學(xué)切片,表明不同時(shí)間點(diǎn)大網(wǎng)膜包裹組新生骨小梁面積都明顯高于單純組織工程骨組,且差異有統(tǒng)計(jì)學(xué)意義(P0.01)；同時(shí)從骨組織學(xué)分析材料的血管化面積,表明在同一時(shí)間點(diǎn)大網(wǎng)膜包裹組新生血管面積均高于單純組織工程骨組(P0.05),差異有統(tǒng)計(jì)學(xué)意義。 結(jié)論利用具有豐富血管網(wǎng)的大網(wǎng)膜包裹組織工程骨植入兔橈骨缺損處,促進(jìn)組織工程骨的血管化及成骨能力,提高了組織工程骨修復(fù)骨缺損的質(zhì)量。
[Abstract]:Objective to evaluate the feasibility of vascularized tissue engineering bone encapsulated with greater omentum to repair the defect of rabbit radius, and to evaluate the feasibility of constructing vascularized tissue engineered bone with greater omentum and its effect on repairing bone defect. To provide clinical and supplementary experimental basis for tissue engineering bone. Methods the (rBMSCs), of rabbit bone marrow mesenchymal stem cells was purified by whole bone marrow adherent culture combined with the culture of fluid exchange, and the morphology of the cells was observed. The structural changes of osteoblasts before and after osteoblast induction were observed by multidirectional differentiation and electron microscopy. The proliferation of the third generation cells was determined by MTT assay and the growth curve was drawn. After osteogenic induction, bone marrow mesenchymal stem cells (BMSCs) of the third generation of rabbits in good growth state were inoculated into self-made scaffold of bioderived bone to construct tissue engineered bone, and the tissue engineered bone (TEB) was implanted in vivo. It was divided into two groups: omentum wrapped tissue-engineered bone group and simple tissue-engineered bone group. The surface changes and tissue responses of the two groups were observed at different time points after operation. X ray films were performed at 4 weeks and 8 weeks to 12 weeks after operation, and the bone defects were repaired by the grafts. The gross and histological changes of the grafts were observed. Osteogenesis and vascularization were observed. Results after the two groups of grafts were implanted into the radius defect of rabbits, the operation mouth grew well, no obvious systemic and local toxicity and inflammatory reaction were found, and the surgical incision of each animal was healed in one stage. In the omentum encapsulated group, X-ray showed that the junction between the implant and the host bone was basically healed at 12 weeks after operation, but the healing of the host bone in the simple group was not satisfactory at 12 weeks after operation. At the same time, the degree of bone ashes in the greater omentum encapsulated group was higher than that in the simple tissue engineered bone group (P.05), and the histological sections of bone were analyzed at 4 weeks and 8 weeks to 12 weeks after operation. The results showed that the trabecular area of the new bone in the greater omentum encapsulated group was significantly higher than that in the simple tissue engineered bone group at different time points (P0.01), and the vascularized area of the material was analyzed from the bone histology. The results showed that the area of neovascularization in omentum encapsulated group was higher than that in simple tissue engineered bone group at the same time point (P0.05), and the difference was statistically significant. Conclusion the tissue engineering bone encapsulated by greater omentum with abundant vascular network was implanted into the defect of rabbit radius, which promoted the vascularization and osteogenesis of tissue engineering bone, and improved the quality of tissue engineering bone in repairing bone defect.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R329

【參考文獻(xiàn)】

中國(guó)期刊全文數(shù)據(jù)庫(kù) 前10條

1 陸云濤,原林,焦培峰,余磊,邱小忠,張黎聲,鐘世鎮(zhèn);甲醛固定源同種異體骨移植材料的體外實(shí)驗(yàn)研究[J];第一軍醫(yī)大學(xué)學(xué)報(bào);2004年01期

2 黃濤;孟志斌;賈丙申;李俊;王挺銳;譚海濤;王晟;;全骨髓貼壁法分離培養(yǎng)rBMSCs及成骨誘導(dǎo)探討[J];廣東醫(yī)學(xué);2010年09期

3 孔勁松;陳振光;鄭曉暉;陶圣祥;楊玉華;林海濤;;帶血管蒂組織瓣預(yù)構(gòu)組織工程骨血管化的研究[J];武漢大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2008年01期

4 黨洪勝;趙猛;嚴(yán)永祥;劉家國(guó);;胎兔成骨細(xì)胞與復(fù)合肝細(xì)胞生長(zhǎng)因子/異種脫蛋白松質(zhì)骨的體外培養(yǎng)[J];中南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2008年04期

5 陳濱 ,裴國(guó)獻(xiàn) ,王珂 ,金丹 ,魏寬海 ,任高宏;筋膜瓣促組織工程骨體內(nèi)再血管化的實(shí)驗(yàn)研究[J];解放軍醫(yī)學(xué)雜志;2002年06期

6 雷榮昌,翦新春;松質(zhì)骨支架與成骨細(xì)胞生物相容性的實(shí)驗(yàn)研究[J];口腔醫(yī)學(xué)研究;2005年05期

7 黃艷;李瑩輝;;影響骨髓間質(zhì)干細(xì)胞向成骨細(xì)胞分化的調(diào)控因素[J];中國(guó)生物工程雜志;2006年05期

8 江華;肖增明;;骨髓間充質(zhì)干細(xì)胞在骨科疾病修復(fù)中的應(yīng)用[J];中國(guó)臨床康復(fù);2006年45期

9 李強(qiáng);孫正義;嚴(yán)冬雪;劉文忠;姚新德;;脫鈣骨基質(zhì)材料的生物學(xué)表現(xiàn):降解性能、孔隙率及其黏附性能特征[J];中國(guó)組織工程研究與臨床康復(fù);2007年31期

10 李濤;臧洪敏;唐天駟;;1,25-(OH)_2VitD_3促進(jìn)組織工程骨的血管化和骨化[J];中國(guó)組織工程研究與臨床康復(fù);2008年11期

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