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软骨素酶 ABC 对兔关节软骨部分厚度缺损模型中滑膜来源间充质干细胞黏附及行为的影响。

Effect of chondroitinase ABC on adhesion and behavior of synovial membrane-derived mesenchymal stem cells in rabbit partial-thickness chondral defects.

机构信息

Department of Orthopedic Surgery, Seoul National University College of Medicine, #28 Yongondong, Chongnogu, Seoul, 110-744, Republic of Korea.

出版信息

J Orthop Res. 2013 Aug;31(8):1293-301. doi: 10.1002/jor.22353. Epub 2013 Apr 29.

Abstract

Transplanted cells may have difficulty attaching to the surface of partial-thickness chondral lesions because of the anti-adhesive properties of the proteoglycan rich matrix. Therefore, the current study attempts to evaluate the effect of chondroitinase ABC (chABC) on the adhesion and behavior of transplanted synovial membrane-derived mesenchymal stem cells (SDSCs) in rabbit partial-thickness chondral defects. In ex vivo adhesion experiments, chABC treatment (0.1 U/ml) was increased in SDSC attachment to the cartilage explants, and significantly diminished by pretreatment with neutralizing antibody against fibronectin. In the in vivo experiments, 1 day and 4 weeks after the chABC treatment (0.1 and 1 U/ml), the immunoreactivity (IR) against CS-56 (intact chondroitin sulfate antibody) was markedly decreased; however, the IR of 2B6 (stub of the chondroitin 4-sulfate chain), 3B3 (stub of the chondroitin 6-sulfate chain), and fibronectin was increased. At 12 weeks, this IR returned to normal except in the high-dose chABC-treated group (1 U/ml). Furthermore, the attachment of SDSCs to the chondral defects after chABC treatment was increased at 7 days compared with that in the chondral defects pretreated with saline. However, the tissue repaired by SDSCs was negatively stained for type II collagen at 12 weeks. In conclusion, these results showed that the exposure to fibronectin by chABC treatment enhances the attachment of SDSCs to partial-thickness chondral defects. However, the tissue regenerated by SDSCs showed lack of hyaline cartilage regeneration. Thus, to understand the fate of transplanted MSCs in cartilage defect is very important for successful cell therapies.

摘要

移植细胞可能难以附着在部分厚度软骨病变的表面,因为富含蛋白聚糖的基质具有抗附着特性。因此,本研究试图评估软骨素酶 ABC(chABC)对兔部分厚度软骨缺损中滑膜衍生间充质干细胞(SDSC)移植后的黏附和行为的影响。在体外黏附实验中,chABC 处理(0.1 U/ml)增加了 SDSC 与软骨外植体的附着,而用纤连蛋白中和抗体预处理则显著减少了 SDSC 的附着。在体内实验中,chABC 处理(0.1 和 1 U/ml)后 1 天和 4 周,CS-56(完整软骨素硫酸抗体)的免疫反应性(IR)明显降低;然而,2B6(软骨素 4-硫酸盐链的残基)和 3B3(软骨素 6-硫酸盐链的残基)以及纤连蛋白的 IR 增加。12 周时,除高剂量 chABC 处理组(1 U/ml)外,IR 恢复正常。此外,chABC 处理后 SDSC 对软骨缺损的附着在 7 天内较盐水预处理的软骨缺损增加。然而,12 周时,由 SDSC 修复的组织对 II 型胶原呈阴性染色。总之,这些结果表明,chABC 处理暴露纤连蛋白增强了 SDSC 对部分厚度软骨缺损的附着。然而,由 SDSC 再生的组织显示缺乏透明软骨再生。因此,了解移植 MSC 在软骨缺损中的命运对于成功的细胞治疗非常重要。

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