本文選題：組織工程 + 骨髓間充質(zhì)干細(xì)胞　； 參考：《中南大學(xué)學(xué)報(醫(yī)學(xué)版)》2017年08期

【摘要】：目的:探討骨髓間充質(zhì)干細(xì)胞(bone marrow mesenchymal stem cells,BMSCs)和肋軟骨細(xì)胞(costal chondrocytes,CCs)共培養(yǎng)構(gòu)建組織工程軟骨及共培養(yǎng)細(xì)胞修復(fù)五指山小型豬膝關(guān)節(jié)軟骨缺損的可行性,為臨床修復(fù)關(guān)節(jié)軟骨缺損提供理論依據(jù)和奠定實(shí)驗(yàn)基礎(chǔ)。方法:密度梯度離心法分離五指山小型豬BMSCs,雙酶消化法分離CCs。取第3代BMSCs和第2代CCs,隨機(jī)分為3組:BMSCs:CCs為1:2的共培養(yǎng)組為A組,單純CCs為B組,單純BMSCs為C組。繪制細(xì)胞生長曲線圖,測定軟骨細(xì)胞外分泌基質(zhì)糖胺多糖(glycosaminoglycan,GAG)的能力。將12只五指山小型豬隨機(jī)分為共培養(yǎng)細(xì)胞/膠原膜實(shí)驗(yàn)組、膠原膜對照組和空白組。共培養(yǎng)細(xì)胞/膠原膜實(shí)驗(yàn)組髁間窩軟骨缺損處植入共培養(yǎng)細(xì)胞/膠原膜,膠原膜對照組單純植入膠原膜,空白組不做任何植入,于術(shù)后第8和16周分別處死6只動物,每組2只。取材行大體觀察、軟骨組織學(xué)評分及病理組織學(xué)檢測。結(jié)果:密度梯度離心法分離的BMSCs生長良好,雙酶消化法分離的CCs生物活性良好,共培養(yǎng)細(xì)胞生長良好。術(shù)后16周共培養(yǎng)細(xì)胞/膠原膜實(shí)驗(yàn)組修復(fù)組織呈透明軟骨樣,表面光滑平坦,周圍軟骨及軟骨下骨整合良好,而膠原膜對照組和空白組為纖維性修復(fù)和無修復(fù)。共培養(yǎng)細(xì)胞/膠原膜實(shí)驗(yàn)組修復(fù)組織大體評分明顯優(yōu)于膠原膜對照組和空白組(均P0.05),膠原膜對照組和空白組之間差異無統(tǒng)計學(xué)意義(P0.05)。結(jié)論:BMSCs,CCs和共培養(yǎng)細(xì)胞均適合作為軟骨組織工程的種子細(xì)胞,其中共培養(yǎng)細(xì)胞(BMSCs:CCs為1:2)更具優(yōu)勢;共培養(yǎng)細(xì)胞復(fù)合膠原膜修復(fù)軟骨缺損近期效果滿意。
[Abstract]:Objective: to investigate the feasibility of co-culture of bone marrow mesenchymal stem cells (bone marrow mesenchymal stem cells) and costal chondrocytes (costal chondrocytes) to construct tissue engineered cartilage and co-culture cells to repair articular cartilage defects in Wuzhishan miniature pig. To provide theoretical basis and experimental basis for clinical repair of articular cartilage defects. Methods: BMSCs were isolated from Wuzhishan miniature pig by density gradient centrifugation and CCs were isolated by double enzyme digestion. The third generation of BMSCs and the second generation of CCS were randomly divided into three groups: group A, group B, group C, group B: BMSCs: 1. Cell growth curves were plotted to determine the ability of chondrocytes to secrete glycosaminoglycan polysaccharide (GAG). Twelve Wuzhishan miniature pigs were randomly divided into co-cultured cells / collagen membrane experimental group, collagen membrane control group and blank group. The co-cultured cells / collagen membrane was implanted into the cartilage defect of the intercondylar fossa in the experimental group, the collagen membrane was implanted only in the control group, and the blank group was not implanted at all. Six animals were killed at the 8th and 16th week after operation. 2 rats in each group. Gross observation, cartilage histological score and histopathological examination were performed. Results: BMSCs isolated by density gradient centrifugation grew well, CCs isolated by double enzyme digestion had good bioactivity and co-cultured cells grew well. 16 weeks after operation, the cultured cells / collagen membrane in the experimental group showed hyaline cartilage, smooth surface and good integration of the surrounding cartilage and subchondral bone, while the collagen membrane control group and the blank group were fibrous repair and no repair. The gross score of repair tissue in co-cultured cells / collagen membrane group was significantly better than that in collagen membrane control group and blank group (P0.05), but there was no significant difference between collagen membrane control group and blank group (P0.05). Conclusion both the CCs and the co-cultured cells are suitable for the seed cells of cartilage tissue engineering, among which the co-cultured cells (BMSCs: CCs is 1:2) have more advantages, and the short-term effect of the co-cultured cells combined with collagen membrane on the repair of cartilage defects is satisfactory.
【作者單位】： 中南大學(xué)湘雅二醫(yī)院骨科;中南大學(xué)湘雅醫(yī)學(xué)院附屬�？卺t(yī)院骨科中心;
【基金】：海南省自然科學(xué)基金(814379)~~
【分類號】：R684

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