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用于骨关节炎修复的人软骨细胞的体外分离与培养。

In vitro isolation and cultivation of human chondrocytes for osteoarthritis renovation.

作者信息

Xu Jiaming, Zhang Changqing

机构信息

Department of Orthopedics, The Sixth People's Hospital Affiliated to Shanghai Jiao Tong University, 600 Yishan Road, 200233, Shanghai Province, China.

出版信息

In Vitro Cell Dev Biol Anim. 2014 Aug;50(7):623-9. doi: 10.1007/s11626-014-9742-5. Epub 2014 Jun 11.

Abstract

The purpose of this study was to evaluate the repair effects of chondrocytes that were cultured in vitro on osteoarthritis (OA). Chondrocytes were isolated from fetal rabbits and cultured in Biosilon microcarriers. Sixty rabbits were randomly divided into three groups equally (blank group, model group, treatment group). The rabbit knee OA model was established by inducing papain. Rabbits in the treatment group were injected with the chondrocytes that were cultured in vitro. Hematoxylin-eosin (HE) staining and gross morphologic observation were conducted. Expression level of cytokines such as IL-1bβ, IL-6, and TNF-α in cartilage synovial cells was also analyzed by an ELISA assay. The cultured chondrocyte was validated by a positive stain of type II collagen and vimentin by immunofluorescence. Compared to the model group, the articular cartilage of the rabbit knee in the treatment group showed a normal color, smooth surface, and none of malacia and coloboma. HE staining indicated that the articular surface of the treatment group tended to be smooth and flat; the matrix stained tinge and the cartilage destruction and fiber hyperplasia of the synovia were lightened. The expression levels of IL-1bβ, IL-6, and TNF-α also declined in the treatment group. OA symptoms were improved by treating with chondrocytes. In summary, the animal experiment in the present study indicated that chondrocyte injection played an active effect on renovation of OA.

摘要

本研究的目的是评估体外培养的软骨细胞对骨关节炎(OA)的修复效果。从胎兔中分离软骨细胞,并在Biosilon微载体中培养。60只兔子随机均分为三组(空白组、模型组、治疗组)。通过木瓜蛋白酶诱导建立兔膝骨关节炎模型。治疗组兔子注射体外培养的软骨细胞。进行苏木精-伊红(HE)染色和大体形态学观察。还通过酶联免疫吸附测定法分析软骨滑膜细胞中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)等细胞因子的表达水平。通过免疫荧光法对II型胶原蛋白和波形蛋白进行阳性染色,验证培养的软骨细胞。与模型组相比,治疗组兔膝的关节软骨颜色正常,表面光滑,无软化和缺损。HE染色表明,治疗组的关节表面趋于光滑平整;基质染色变淡,滑膜的软骨破坏和纤维增生减轻。治疗组中IL-1β、IL-6和TNF-α的表达水平也下降。通过软骨细胞治疗改善了OA症状。总之,本研究中的动物实验表明,注射软骨细胞对OA的修复起到了积极作用。

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