Repairing articular cartilage defect of the knee joint by synovial mesenchymal stem cells and chondrocytes

doi:10.3969/j.issn.2095-4344.2017.09.018

Abstract

Abstract:

BACKGROUND: As is known to all, it is difficult to repair cartilage defect of the knee joint. Fortunately, it is reported that a chitosan scaffold with the mixture of synovial mesenchymal stem cells and chondrocytes enables the formation of cartilage matrix.
OBJECTIVE: To observe the chondrogenic differentiation of synovial mesenchymal stem cells co-cultured with chondrocytes on a chitosan/type I collagen composite scaffold after being transplanted into an animal model.
METHODS: The synovial mesenchymal stem cells and chondrocytes harvested from the knee joint of Sprague-Dawley rats were co-cultured at a ratio of 1:2 on the chitosan/type I collagen composite scaffolds and transplanted into the defect site of the knee joint in rat models of cartilage defect of the knee joint.
RESULTS AND CONCLUSION: After 4, 8, 12 weeks, the hematoxylin-eosin staining, the toluidine blue staining of aggrecan, and safranine O staining showed that there were increasing chondrocytes in the rat knee joint. The immunohistochemical staining of type II collagen showed the positive expression of type II collagen in the cartilage tissues and extracellular matrix of the knee joint. To conclude, the combination of two kinds of cells cultured in the chitosan/type II collagen composite scaffold can form cartilage-like tissues in vivo, and improve the quality of cartilage repair.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Tissue Engineering, Chondrocytes, Synovial Membrane

CLC Number:

R394.2

Hu Xin, Yin Xiao-peng, Gong Zhong-cheng, Shao Bo, Ling Bin, Liu Hui, Rekemu Abasi, Wang Bing, Hu Lu-lu, Wang Yue, Lin Zhao-quan. Repairing articular cartilage defect of the knee joint by synovial mesenchymal stem cells and chondrocytes[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(9): 1408-1413.

Figures/Tables 2

2.1 实验动物数量分析全部大鼠均进入结果分析。 2.2 一般情况观察大鼠膝关节软骨缺损植入后手术切口均为Ⅰ期愈合，植入后大鼠膝关节活动佳，手术对大鼠步态未见明显影响，未出现术区感染迹象，术区缝线均故位良好，植入后3周可见手术区域缝线呈自然性脱落，部分缝线残存于术区。植入后大鼠觅食未见明显异常。见图2。 2.3 软骨缺损修复情况植入后4周，软骨缺损修复处可见局部粗糙状，修复区域可见微黄，可见明显边界；4组缺损修复处与周围软骨组织平齐，可见壳聚糖支架材料与周围组织局部融合。修复8周，缺损处修复缺损组织表面不光滑且不平整，仍可见凹陷缺损区域，整合一般；4组缺损修复处与周围软骨组织可见平齐，壳聚糖支架材料与周围组织融合尚好，未见周围软组织局部明显坏死感染迹象发生。植入后12周，4组缺损处修复组织面较4，8周时改善明显增大，边界模糊不清；4组缺损修复处与周围软骨组织相似未见明显边界，边界不清晰，可见局部残留支架成分，软骨缺损处表面光滑，见图2。 2.4 细胞支架复合物植入后组织形态苏木精-伊红染色：植入后4周，4组细胞支架复合物中可见少许细胞，支架材料最外层多分布细胞及细胞外基质，支架材料中心未见明显细胞分布，植入后8周，可见支架材料细胞及细胞外基质分布较植入后4周时明显，植入后12周可见细胞及细胞外基质比较4，8周时广泛分布。植入后4周，各组未见软骨样细胞表型，植入后8周可见软骨细胞样表型出现于细胞支架复合物周围，深红色为支架材料，植入后12周，可见各组软骨细胞表型较4，8周明显，见图3。甲苯胺蓝染色：植入后4周，单纯软骨细胞组、滑膜间充质干细胞︰软骨细胞1︰2组可见细胞核染色为深蓝色，细胞质和细胞外基质均为蓝色，滑膜间充质干细胞︰软骨细胞1︰2组可见散在类软骨样组织。植入后8周，滑膜间充质干细胞︰软骨细胞1︰2组类软骨样组织增多比较植入后4周明显增多，细胞质和细胞外基质均呈蓝染阳性，单纯滑膜间充质干细胞组未见明显改变，植入后12周，滑膜间充质干细胞：软骨细胞1︰2组中软骨组织增多较植入后4，8周明显，细胞及细胞外基质呈蓝染强阳性，见图4。番红O染色：植入后4周，单纯软骨细胞组、滑膜间充质干细胞：软骨细胞1︰2组可见少许软骨基质，软骨细胞核为蓝染，分布于支架材料外层。植入后8周，单纯软骨细胞组、滑膜间充质干细胞：软骨细胞1︰2组可见细胞及软骨基质向材料中心分布趋势。植入后12周，可见细胞及软骨细胞基质较植入后4，8周表达明显增强，深蓝色为支架材料，见图5。免疫组织化学染色：植入后4周，单纯软骨细胞组、滑膜间充质干细胞︰软骨细胞1︰2组可见新生软骨基质部分出现不同程度的Ⅱ型胶原阳性反应，部分细胞及细胞外基质可见少许棕黄色颗粒分布，基底部着色较深。植入后8周，可见滑膜间充质干细胞组：软骨细胞1︰2组中，新生软骨基质染色显示为阳性表达，部分棕黄色颗粒比较4周深染色。植入后12周，Ⅱ型胶原免疫组织化学染色染色呈强阳性表达，软骨细胞中的基质染色呈现棕褐色且基底部着色比较深，见图6。"

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