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体内 RNAi 软骨细胞组织工程化软骨同种异体移植物的实验研究。

Experimental study of tissue-engineered cartilage allograft with RNAi chondrocytes in vivo.

机构信息

Department of Otolaryngology - Head and Neck Surgery, The Second Hospital, Xi'an Jiaotong University, Xi'an, People's Republic of China.

Department of Dermatology, The Second Hospital, Xi'an Jiaotong University, Xi'an, People's Republic of China.

出版信息

Ther Clin Risk Manag. 2014 May 8;10:335-40. doi: 10.2147/TCRM.S51518. eCollection 2014.

Abstract

PURPOSE

To determine the effects of RNA interference (RNAi) on chondrocyte proliferation, function, and immunological rejection after allogenic tissue-engineered cartilage transplantation within bone matrix gelatin scaffolds.

METHODS

Seven million rat normal and RNAi chondrocytes were harvested and separately composited with fibrin glue to make the cell suspension, and then transplanted subcutaneously into the back of Sprague Dawley rats after being cultured for 10 days in vitro. Untransplanted animals served as the control group. The allograft and immunological response were examined at 1, 2, 4, 8, and 12 months postoperatively with hematoxylin and eosin histochemical staining, immunohistochemical staining (aggrecan, type II collagen, class I and II major histocompatibility complex), and flow cytometry for peripheral blood cluster of differentiation 4(+) (CD4(+)) and CD8(+) T-cells.

RESULTS

There was no infection or death in the rats except one, which died in the first week. Compared to the control group, the RNAi group had fewer eukomonocytes infiltrated, which were only distributed around the graft. The ratio of CD4(+)/CD8(+) T-cells in the RNAi group was significantly lower than the normal one (P<0.05). There were many more positively stained chondrocytes and positively stained areas around the cells in the RNAi group, which were not found in the control group.

CONCLUSION

The aggrecanase-1 and aggrecanase-2 RNAi for chondrocytes decreased the immunological rejection effect.

摘要

目的

在骨基质明胶支架内进行同种异体组织工程软骨移植后,确定 RNA 干扰 (RNAi) 对软骨细胞增殖、功能和免疫排斥的影响。

方法

收获 700 万只正常和 RNAi 大鼠软骨细胞,并分别与纤维蛋白胶混合制成细胞悬液,体外培养 10 天后皮下移植到 Sprague Dawley 大鼠背部。未移植的动物作为对照组。术后 1、2、4、8 和 12 个月,采用苏木精-伊红组织化学染色、免疫组织化学染色(聚集蛋白聚糖、Ⅱ型胶原、Ⅰ类和Ⅱ类主要组织相容性复合体)和外周血 CD4+(cluster of differentiation 4+)和 CD8+T 细胞流式细胞术检测同种异体移植物和免疫反应。

结果

除一只大鼠在第一周死亡外,其余大鼠均无感染或死亡。与对照组相比,RNAi 组的白细胞浸润较少,仅分布在移植物周围。RNAi 组 CD4+/CD8+T 细胞的比值明显低于正常组(P<0.05)。RNAi 组细胞周围有更多的阳性染色软骨细胞和阳性染色区域,而对照组则没有。

结论

软骨细胞的基质金属蛋白酶-1 和基质金属蛋白酶-2 RNAi 降低了免疫排斥反应的效果。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/58da/4020888/4feb78973e25/tcrm-10-335Fig1.jpg

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