Micro-fracture enhanced by autologous bone marrow mesenchymal stem cells extracellular matrix scaffold to treat articular cartilage defects in the knee of pigs

doi:10.3969/j.issn.2095-4344.2014.43.004

Abstract

Abstract:

BACKGROUND: Micro-fracture surgery method is simple, easy to operate, which is an effective way to treat articular cartilage defects, but there are still some problems such as regenerated fibrocartilage and regenerated cartilage degradation. Scholars have focused on the use of various methods to improve the micro-fracture effect on repairing cartilage defects.
OBJECTIVE: To explore the effects of micro-fracture enhanced by autologous bone marrow mesenchymal stem cells extracellular matrix (aBMSC-dECM) scaffold for treating cartilage defects in minipig models.
METHODS: Bone marrow was extracted from the minipigs and bone marrow mesenchymal stem cells were obtained. aBMSC-dECM membranes were collected. Cross-linking and freeze-drying technology were used to make the three-dimensional porous aBMSC-dECM scaffold. Full thickness cartilage defects, 2 mm in depth and 6 mm in diameter, were created on the femoral condyles and trochlea grooves of the two knees of the minipigs. The right knees were treated with micro-fracture as control and the left were treated with micro-fracture enhanced by aBMSC-dECM scaffold. Six months later, histological examination and Wakitani score were used to evaluate the cartilage regeneration, and glycosaminoglycans and DNA contents in the regenerative tissue were determined.
RESULTS AND CONCLUSION: After 6 months, the tissue treated by micro-fracture enhanced by aBMSC-dECM scaffold got better surface and integrated with the surrounding cartilage. Safranin O and fast green staining and Masson staining showed that the regenerated cartilage of the left knee, with abundant matrix and dense bone trabeculae, was better than that of the right. Wakitani score of the left knee was higher than that of the right. Glycosaminoglycans content of the left knee was much more than that of the right, while the DNA content was lower in the left knee than the right knee. Better results were observed in the left knee undergoing micro-fracture enhanced by aBMSC-dECM scaffold, and improvements in the femoral condyles and trochlea grooves showed no differences.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: bone marrow, mesenchymal stem cells, extracellular matrix, cartilage, articular

CLC Number:

R318

Li Xiang-quan, Tang Cheng, Song Ke-rong, Jin Cheng-zhe. Micro-fracture enhanced by autologous bone marrow mesenchymal stem cells extracellular matrix scaffold to treat articular cartilage defects in the knee of pigs [J]. Chinese Journal of Tissue Engineering Research, 2014, 18(43): 6907-6913.

Figures/Tables 4

2.1 大体观察结果术后6个月，手术伤口愈合良好，无化脓感染现象，6只实验动物均存活。打开关节腔可见，实验组和对照组膝关节均无明显关节炎表现，关节液成淡黄色，透明。实验组股骨滑车和股骨髁均可见良好的软骨修复，对照组股骨滑车软骨修复较好，股骨髁未见明显再生组织填充。实验组股骨滑车和股骨髁填充组织有光泽，且在外观上与周围软骨组织类似，而对照组股骨滑车再生组织表面毛糙，与周围软骨相差较大(图2)。 2.2 组织学染色结果术后6个月，番红固绿染色可见实验组股骨滑车和股骨髁表面平整，软骨区域着红色，软骨下骨成绿色，染色深，滑车区域和股骨髁区域染色无明显差异。对照组股骨滑车表面平整，修复组织着色浅，软骨下骨修复欠佳；对照组股骨髁表面缺损较大，未见软骨修复(图3)。Masson染色结果类似番红固绿染色，实验组软骨下骨修复良好，对照组修复欠佳(图4)。天狼星红染色可见实验组胶原排列情况明显优于对照组(图5)。 2.3 组织学评分术后6个月，实验组的股骨滑车Wakitani评分为13.38±0.98，股骨髁为14.00±0.89，对照组股骨滑车评分为9.67±1.21，股骨髁为2.00±0.89。实验组股骨髁和股骨滑车评分高于对照组，差异有显著性意义(P < 0.05)；实验组股骨髁和股骨滑车Wakitani评分差异无显著性意义(P > 0.05)；对照组股骨滑车评分与股骨髁比较差异有显著性意义(P < 0.05)，见图6。 2.4 生化成分分析实验组和对照组在术后6个月内均可测到糖胺聚糖的表达。实验组股骨髁糖胺聚糖的含量为(92.938 3±3.362 8) mg/g，股骨滑车为(93.533 3± 3.840 9) mg/g，对照组股骨髁糖胺聚糖的含量为(9.896 7± 4.370 4) mg/g，股骨滑车为(44.773 3±3.916 4) mg/g，正常组织糖胺聚糖含量为(93.756 7±4.315 5) mg/g。实验组股骨髁和股骨滑车的糖胺聚糖含量差异无显著性意义(P > 0.05)；对照组股骨滑车糖胺聚糖含量高于股骨髁，差异有显著性意义(P < 0.05)。实验组股骨滑车和股骨髁糖胺聚糖含量相应高于对照组，差异有显著性意义(P < 0.05)，见图7。术后6个月，实验组股骨髁和股骨滑车DNA含量分别为(0.518 3±0.134 5) mg/g，(0.526 7±0.161 5) mg/g，对照组股骨髁和股骨滑车DNA含量分别为(1.566 7± 0.206 6) mg/g和(2.318 3±0.200 1) mg/g，正常组织为(0.931 7±0.089 5) mg/g；实验组股骨髁和股骨滑车DNA含量差异无显著性意义(P > 0.05)，但均相应低于对照组，差异有显著性意义(P < 0.05)。对照组股骨滑车DNA含量低于股骨髁，差异有显著性意义(P < 0.05)。实验组的DNA含量和正常软骨差异有显著性意义(P < 0.05)，见图7。实验组股骨髁和股骨滑车糖胺聚糖/DNA含量明显高于对照组，差异有显著性意义(P < 0.05)；对照组股骨髁数值低于股骨滑车，但差异无显著性意义(P > 0.05)，见图7。"

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