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过度表达骨髓间充质干细胞的 Runx2 可改善膝关节软骨缺损的修复。

Overexpressing Runx2 of BMSCs Improves the Repairment of knee Cartilage Defects.

机构信息

Wuhan Sports University, Wuhan 430079, China.

Department of Orthopedics, Wuhan Third Hospital, Tongren Hospital of Wuhan University, Wuhan 430060, China.

出版信息

Curr Gene Ther. 2020;20(5):395-404. doi: 10.2174/1566523220666201005110339.

Abstract

BACKGROUND

Recruitment of gene modifying bone marrow mesenchymal stem cells (BMSCs) has been considered an alternative to single-cell injection in articular cartilage repair.

PURPOSE

This study aimed to investigate whether the effect of runt-related transcription factor 2(Runx2) overexpression bone marrow mesenchymal stem cells in vivo could improve the quality of repaired tissue of a knee cartilage defect in a rabbit model.

METHODS

Thirty-two New Zealand rabbits were randomly divided into four groups. The blank group (Con) did not receive anything, the model group (Mo) was administered saline, the simple stem cell group (MSCs) received MSCs injection, and the Runx2 transfection group (R-MSCs) received Runx2 overexpression MSCs injection. After adapting to the environment for a week, a 5 mm diameter cylindrical osteochondral defect was created in the center of the medial femoral condyle. Cell and saline injections were performed in the first and third weeks after surgery. The cartilage repair was evaluated by macroscopically and microscopically at 4 and 8 weeks.

RESULTS

Macroscopically, defects were filled and surfaces were smoother in the MSCs groups than in the Mo group at 4th week. Microscopically, the R-MSCs group showed coloration similar to surrounding normal articular cartilage tissue at 8 weeks in masson trichrome staining. The COL-II, SOX9, and Aggrecan mRNA expressions of MSCs were enhanced at 4 weeks compared with R-MSCs, then the expression reduced at 8 weeks, but was still higher than Mo group level (P<0.05). The western blot examination revealed that the COL-IIand SOX9 expression of MSCs was higher than R-MSCs at 4 weeks, then the expression reduced at 8 weeks, but was still higher than the Mo level (P<0.05). The IL-1β content in the joint fluid also revealed that cartilage repair with R-MSCs was better than that with MSCs at 8 weeks (P<0.05).

CONCLUSION

The R-MSCs group showed cellular morphology and arrangement similar to surrounding normal articular cartilage tissue, and Runx2 overexpression of MSCs resulted in overall superior cartilage repair as compared with MSCs at 8 weeks.

摘要

背景

招募基因修饰的骨髓间充质干细胞(BMSCs)已被认为是关节软骨修复中单细胞注射的替代方法。

目的

本研究旨在探讨体内过表达 runt 相关转录因子 2(Runx2)的骨髓间充质干细胞对兔模型膝关节软骨缺损修复组织质量的影响。

方法

32 只新西兰兔随机分为四组。空白组(Con)不接受任何治疗,模型组(Mo)给予生理盐水,单纯干细胞组(MSCs)给予干细胞注射,Runx2 转染组(R-MSCs)给予过表达 Runx2 的干细胞注射。术后第 1 周和第 3 周分别进行细胞和生理盐水注射。术后 4 周和 8 周通过大体和显微镜评估软骨修复情况。

结果

大体上,第 4 周时 MSCs 组的缺损填充和表面较 Mo 组更光滑。Masson 三色染色显示,R-MSCs 组在第 8 周时的染色与周围正常关节软骨组织相似。第 4 周时,MSCs 的 COL-II、SOX9 和 Aggrecan mRNA 表达增强,8 周时表达降低,但仍高于 Mo 组水平(P<0.05)。Western blot 检测显示,第 4 周时 MSCs 的 COL-II 和 SOX9 表达高于 R-MSCs,8 周时表达降低,但仍高于 Mo 组水平(P<0.05)。关节液中 IL-1β 含量也表明,8 周时 R-MSCs 修复的软骨优于 MSCs(P<0.05)。

结论

R-MSCs 组的细胞形态和排列与周围正常关节软骨组织相似,与 MSCs 相比,8 周时 Runx2 过表达的 MSCs 可获得整体更好的软骨修复效果。

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