论文部分内容阅读
目的 探讨不同浓度软骨细胞提供的软骨微环境诱导骨髓基质干细胞(BMSC)体外构建软骨的可行性。方法 将体外分别培养扩增的猪BMSC与耳软骨细胞按不同比例混合(9:1,8:2),均以5.0×107/mL的细胞终浓度接种于聚羟基乙酸/聚乳酸(PGA/PLA)支架作为共培养组,以相同终浓度的单纯软骨细胞和单纯BMSC分别接种作为阳性及阴性对照,以20％上述浓度的单纯软骨细胞接种作为低软骨细胞浓度对照。各标本于体外培养4周时取材,通过大体观察、组织学以及免疫组化等方法对新生软骨进行初步评价。结果 各组细胞均与材料粘附良好。8:2共培养组及阳性对照组在体外培养4周时,外观已类似软骨组织并基本保持了材料的大小和形状,组织学显示有较连续的成熟软骨形成,免疫组化也均显示有大量Ⅱ型胶原分泌。9:1共培养组在培养过程中稍有缩小和变形,组织学上仅在培养物的边缘可见到连续的软骨样组织。阴性对照组明显皱缩变形,组织学未见成熟软骨陷窝。低软骨细胞浓度组复合物明显变薄,只在局部形成了不连续的软骨组织,新生软骨量明显少于共培养各组及阳性对照组。结论软骨细胞能够提供软骨微环境诱导BMSC成软骨分化并形成软骨,20％浓度的软骨细胞已能够达到良好的诱导效果。
Objective To investigate the feasibility of in vitro cartilage induction by bone marrow stromal stem cells (BMSC) induced by different concentrations of chondrocytes. Methods Porcine BMSCs and auricular cartilage cells cultured in vitro were mixed in different ratios (9: 1, 8: 2) and all were inoculated into PGA / PLA at a final cell concentration of 5.0 × 107 / mL. PLA) scaffolds as co-culture group were inoculated with pure chondrocytes and pure BMSCs at the same final concentration respectively as positive and negative controls, and chondrocytes inoculated at a concentration of 20% were used as low chondrocyte concentration control. The specimens were harvested in vitro for 4 weeks, and primary cartilage was evaluated by gross observation, histology and immunohistochemistry. Results All cells adhered well to the material. 8: 2 co-culture group and positive control group cultured in vitro for 4 weeks, the appearance has been similar to the cartilage and the basic material to maintain the size and shape, histology showed more continuous mature cartilage, immunohistochemistry also showed that there A large number of type Ⅱ collagen secretion. The 9: 1 co-culture group was slightly reduced and deformed during culture, with histologically only continuous cartilage-like tissue visible at the edges of the culture. Negative control group was significantly shrinkage deformation, histology no mature cartilage lacuna. Low chondrocyte concentration of the composite group was significantly thinner, only in the local formation of discontinuous cartilage tissue, new cartilage was significantly less than the co-culture group and the positive control group. Conclusion Chondrocytes can provide cartilage microenvironment to induce cartilage differentiation and cartilage formation in BMSCs. Chondrocytes in 20% concentration have been able to achieve good induction effect.