本文關(guān)鍵詞：兔自體外周血間充質(zhì)干細(xì)胞對腱—骨界面愈合影響的實(shí)驗(yàn)研究　出處：《安徽醫(yī)科大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 外周血間充質(zhì)干細(xì)胞 腱—骨界面愈合 前交叉韌帶 交叉韌帶重建

【摘要】：目的探討兔自體外周血間充質(zhì)干細(xì)胞（MSCs）對前交叉韌帶（ACL）重建后腱-骨界面愈合的影響。 方法以粒細(xì)胞集落刺激因子作為外周血MSCs動員劑，體外分離、培養(yǎng)、擴(kuò)增獲得外周血MSCs。48只4~6月齡新西蘭白兔按1-48編號，隨機(jī)均分為A、B、C三組，以后肢右膝關(guān)節(jié)作為研究對象，采用趾長屈肌腱作為ACL重建移植物。A組兔將外周血間充質(zhì)干細(xì)胞-生物蛋白膠（FG-MSCs）混合注入兔右膝ACL重建模型股骨道頂端間隙內(nèi)；B組兔僅注射生物蛋白膠；C組兔交叉韌帶重建后不做其他處理。分別在2周、4周、8周、12周各時間點(diǎn)每組各處死4只兔子進(jìn)行解剖學(xué)觀察，取材制成石蠟切片行蘇木素-伊紅染色(Hematoxylin-Eosin Staining,HE)、甲苯胺藍(lán)染色(Toluidine Blue Staining,TB)、Masson三色染色（Masson trichrome staining）進(jìn)行組織形態(tài)學(xué)觀察，并進(jìn)行腱-骨界面各時間點(diǎn)成纖維細(xì)胞計數(shù)及組織形態(tài)學(xué)Buark評分，以了解腱-骨界面愈合情況。 結(jié)果（1）①A組：術(shù)后2周可見腱-骨界面間隙由疏松結(jié)締組織填充，可見少量成纖維細(xì)胞，移植物與骨道之間分離；術(shù)后4周可見大量疏松結(jié)締組織、成纖維細(xì)胞和少量類軟骨細(xì)胞及膠原纖維，新生骨形成，腱-骨間隙已不可見；術(shù)后8周在腱-骨界面可見大量類軟骨細(xì)胞，出現(xiàn)Sharpey纖維集結(jié)成束，膠原纖維合成增多,排列較規(guī)則；術(shù)后12周腱-骨間形成致密結(jié)締組織，可見大量排列有序的Sharpey纖維以及纖維軟骨、鈣化軟骨形成，未出現(xiàn)潮線。②B、C組：術(shù)后2周腱-骨界面僅見少量疏松結(jié)締組織，少量成纖維細(xì)胞增生,未見膠原纖維，同時腱-骨界面明顯分離；術(shù)后4周腱-骨間可見少量疏松結(jié)締組織及少量新生骨細(xì)胞長入，散在分布的類軟骨細(xì)胞和膠原纖維，且未見與骨道垂直連接；術(shù)后8周腱-骨界面為較多類軟骨細(xì)胞及膠原纖維，Sharpey纖維散在分布；術(shù)后12周有較多排列無序的類軟骨細(xì)胞出現(xiàn)，，膠原纖維密集排列不規(guī)則，Sharpey纖維排列紊亂。（2）術(shù)后各時間點(diǎn)三組腱-骨界面高倍鏡下（HE染色）成纖維細(xì)胞計數(shù)資料采用單向方差分析及SNK法檢驗(yàn)：A組*與B組**和A組*與C組***在各時間點(diǎn)比較均有統(tǒng)計學(xué)意義（P＜0.05），而B組**與C組***在各時間點(diǎn)比較無統(tǒng)計學(xué)意義。（3）基于高倍鏡下腱-骨界面的纖維血管組織和Sharpey纖維評定的組織形態(tài)學(xué)Buark評分統(tǒng)計學(xué)資料采用Wilcoxon秩和檢驗(yàn)：12周內(nèi)A組與B組組織形態(tài)學(xué)Buark評分中位數(shù)間的差異有統(tǒng)計學(xué)意義（P＜0.01），即A組組織形態(tài)學(xué)Buark評分較B組高。 結(jié)論兔自體外周血MSCs通過頂端注射的方式能夠促進(jìn)腱-骨界面的愈合。
[Abstract]:Objective to investigate the effect of autologous peripheral blood mesenchymal stem cells (MSCs) on the healing of tendon bone interface after the reconstruction of anterior cruciate ligament (ACL) in rabbits.
Methods with granulocyte colony-stimulating factor for peripheral blood MSCs mobilization agent, in vitro isolation, culture, amplification of peripheral blood MSCs.48 4~6 month old New Zealand white rabbits by 1-48 number, were randomly divided into A, B, C three groups, after limb knee joint as the research object, as ACL reconstruction graft in group.A the peripheral blood mesenchymal stem cells and fibrin glue with flexor digitorum longus tendon (FG-MSCs) mixed into ACL model of rabbit femur reconstruction of right knee top clearance; B group were only injected with fibrin glue; no other treatment in group C after anterior cruciate ligament reconstruction. In 2 weeks, 4 weeks, 8 week, 12 weeks each time point in each group were sacrificed 4 rabbits were observed in paraffin section, hematoxylin eosin staining (Hematoxylin-Eosin, Staining, HE), toluidine blue staining (Toluidine Blue Staining, TB), Masson trichrome staining (Masson trichrome staining) organization form In order to understand the healing of the tendon bone interface, the fibrous cell count and the histomorphology Buark score at the time point of the tendon bone interface were observed.
Results (1) A group: 2 weeks after surgery, the tendon bone interface by loose connective tissue filled the gap, a small amount of fibroblasts, separation between graft and bone; 4 weeks after operation showed a large amount of loose connective tissue, fibroblasts and a small amount of cartilage cells and collagen fibers, new bone formation that tendon bone gap is not visible; 8 weeks after operation in the tendon bone interface showed a large number of chondrocytes, Sharpey fiber bundles of collagen synthesis increased, a regular arrangement; 12 weeks after operation, the tendon bone formation of dense connective tissue, showing a large number of ordered Sharpey fiber and fiber cartilage. Calcified cartilage formation, without tidal line. The B, C group: after 2 weeks of tendon bone interface but only a small amount of loose connective tissue, a small amount of fibroblast proliferation, collagen fibers were at the same time, the tendon bone interface were separated; after 4 weeks of tendon bone can be seen between the small amount of loose connective tissue And a small amount of new bone cells, scattered cartilage cells and collagen fibers, and no bone and vertical connection; 8 weeks after operation, the tendon bone interface into chondrocytes and collagen fibers, Sharpey fibers were distributed; 12 weeks after surgery more cartilage cell disorder arrangement, dense collagen fibers arranged in irregular, disordered arrangement of Sharpey fiber. (2) at each time point after operation three groups of tendon bone interface at high magnification (HE staining) fibroblast count data using one-way analysis of variance and SNK test: A group and B group and A * * * * * * * in the group and C group at each time point were statistically significant (P < 0.05), B group and C group * * * * * in each time point is not significant. (3) fibrovascular tissue and Sharpey fibers evaluated at high magnification, the tendon bone interface morphology Buark Score statistical data using the Wilcoxon rank test based on: 12 weeks There was significant difference in the median of Buark score between group A and group B (P < 0.01), that is, the histomorphological Buark score of group A was higher than that of the B group.
Conclusion the autologous peripheral blood MSCs can promote the healing of the tendon bone interface by injection of the apical injection.

【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R681;R-332

【參考文獻(xiàn)】

中國期刊全文數(shù)據(jù)庫 前5條

1 陳崗;吳海山;;前交叉韌帶重建術(shù)后腱-骨界面愈合研究[J];國際骨科學(xué)雜志;2010年01期

2 沈光思;徐又佳;周海斌;郭霞;牛文鑫;丁祖泉;董啟榕;;前交叉韌帶重建術(shù)股骨隧道角度與隧道壁承受應(yīng)力關(guān)系的研究[J];臨床骨科雜志;2008年05期

3 薛海濱,敖英芳,于長隆;應(yīng)用半腱肌腱重建前交叉韌帶末端形成的特點(diǎn)[J];中國運(yùn)動醫(yī)學(xué)雜志;2002年02期

4 王永健;敖英芳;;自體半腱肌腱重建兔前交叉韌帶腱骨愈合和止點(diǎn)形成實(shí)驗(yàn)研究[J];中國運(yùn)動醫(yī)學(xué)雜志;2007年01期

5 陳百成;孫然;王曉峰;邵德成;陸搏;陳競清;;前交叉韌帶重建術(shù)后骨隧道擴(kuò)大的產(chǎn)生及轉(zhuǎn)歸的 MRI 隨訪結(jié)果[J];中華外科雜志;2007年02期

本文編號：1357757