Effect of poly(hydroxybutyrate-co-hydroxyoctanoate)/collagen composite osteochondral scaffold combined with bone marrow mesenchymal stem cells in the repair of articular osteochondral defects

doi:10.3969/j.issn.2095-4344.0403

Abstract

Abstract:

BACKGROUND: Preliminary experiments have confirmed that poly(hydroxybutyrate-co-hydroxyoctanoate) (PHBHOx)/collagen composite osteochondral scaffold exhibits desirable pore structure and biocompatibility.
OBJECTIVE: To investigate the effect of PHBHOx/collagen composite osteochondral scaffold carrying bone marrow mesenchymal stem cells in the repair of articular osteochondral defects in the weight-bearing area of rabbits.
METHODS: Cone-shaped osteochondral defects were created in the femoral medial condyle of the right knee of 30 New Zealand white rabbits. Then, the rabbit models were randomized into three groups and underwent implantation of PHBHOx/collagen composite osteochondral scaffold carrying bone marrow mesenchymal stem cells in the scaffold-cell group, PHBHOx/collagen composite osteochondral scaffold in the scaffold group and no intervention in the control group. At 4 and 12 weeks after surgery, animals were sacrificed for gross, Micro-CT, histological and immunohistochemical collagen II observations.
RESULTS AND CONCLUSION: At 12 weeks after surgery, Micro-CT scanning results suggested that osteochondral defects were not repaired in the control group, repaired incompletely in the scaffold group with many new bone trabeculae, and repaired completely in the scaffold-cell group. Histological results showed that at 4 weeks after surgery, the defects in the control group were filled with amorphous tissues, subchondral bone formation just occurred in the scaffold group but increased in the scaffold-cell group. At 12 weeks after surgery, trabecular bone structure with no cartilage lacuna was observed in the control group; incomplete subchondral bone formation was observed in the scaffold group, and the cartilage layer was repaired by the fibrous tissues; in the scaffold-cell group, osteochondral defect repair was complete, with the emergence of tidal line, and the newborn cartilage was completely integrated with the surrounding normal tissue in addition to a similar thickness. At 12 weeks after surgery, collagen II basically did not express in the control group, weakly expressed in the scaffold group and highly expressed in the scaffold-cell group. In short, the PHBHOx/collagen composite osteochondral scaffold promotes the repair of articular osteochondral defects in the weight-bearing area.

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: Mesenchymal Stem Cells, Cartilage, Tissue Engineering

CLC Number:

R394.2

Jian Xiao-dong, Zhang Yong-hong, Zhang Yu, Liu Lai-you. Effect of poly(hydroxybutyrate-co-hydroxyoctanoate)/collagen composite osteochondral scaffold combined with bone marrow mesenchymal stem cells in the repair of articular osteochondral defects[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(1): 7-12.

Figures/Tables 1

2.1 实验动物数量分析 30只造模新西兰大白兔全部进入结果分析。 2.2 各组修复组织大体形态观察结果植入后4，12周，所有动物关节液正常，膝关节内未观察到滑膜炎症、骨赘形成。植入后4，12周时，空白对照组骨软骨缺损未修复，缺损区被纤维样组织覆盖；植入后4周时，对照组可见软骨缺损轮廓清楚，12周时缺损区中央区域凹陷，修复组织与周围正常软骨边界清晰；植入后4周时，实验组缺损区边界轮廓尚存，12周时缺损区被类透明软骨样组织覆盖，表面光滑，颜色与周围的正常软骨相似，新生软骨与周围正常的软骨的边界模糊，见图3，4。 2.3 骨缺损修复的检测结果 Micro-CT扫描结果表明，软骨下骨的修复是从周围开始向中心进行的，缺损区的范围逐渐减小。直到植入后12周时，空白对照组软骨下骨缺损未修复，留有较大的凹陷；对照组软骨下骨缺损虽然仍未完全修复，但凹陷较空白对照组小，再生骨小梁结构较多；实验组植入后4周缺损周围出现较多的骨重建，12周时软骨下骨缺损区得到完全修复，见图3，4。 2.4 各组修复处苏木精-伊红、番红O组织学染色结果植入后4周，空白对照组缺损区残留大的凹陷，缺损区被无定形的组织填充，12周时缺损区无骨小梁结构出现，细胞分布杂乱无章，无软骨陷窝结构，番红O染色显示糖胺多糖表达很少；对照组植入后4周时软骨下骨区域开始出现骨组织的重建，修复软骨番红O染色显示糖胺多糖基本上不表达，植入12周时软骨下骨的重建不充分，软骨层的修复是由纤维组织构成，细胞呈簇分布，软骨层与软骨下骨层界限不明显，新生软骨层的番红O染色较淡，颜色不均一，糖胺多糖的表达较少；实验组植入后4周时软骨下骨区域出现较多的骨组织重建，12周时骨软骨缺损区修复完全，软骨层与软骨下骨层界限清楚，潮线出现，新生软骨细胞形态为圆形，位于软骨陷窝内，呈柱状分布，软骨层厚度与周围正常软骨相似，表面光滑，与周围正常组织完全融合，番红O染色颜色更深，表明糖胺多糖的表达接近于正常软骨，见图3，4。 2.5 各组修复处Ⅱ型胶原免疫组织化学染色结果植入后12周时，空白对照组Ⅱ型胶原基本上不表达，表明修复的软骨为纤维组织；对照组软骨层Ⅱ型胶原表达较弱；实验组软骨基质Ⅱ型胶原表达多，与周围的正常软骨相似，可清楚看到潮线，这与番红O染色结果一致，见图3，4。"

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