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通过体内电穿孔将基因直接导入大鼠关节软骨

Direct gene transfer into rat articular cartilage by in vivo electroporation.

作者信息

Grossin Laurent, Cournil-Henrionnet Christel, Mir Lluis M, Liagre Bertrand, Dumas Dominique, Etienne Stéphanie, Guingamp Corinne, Netter Patrick, Gillet Pierre

机构信息

Unité Mixte de Recherches 7561, Centre National de la Recherche Scientifique-Université Henri Poincaré Nancy 1, Faculté de Médecine, Avenue de la Forêt de Haye, BP184, F-54505 Vandoeuvre lès Nancy, France.

出版信息

FASEB J. 2003 May;17(8):829-35. doi: 10.1096/fj.02-0518com.

Abstract

To establish a system for efficient direct in vivo gene targeting into rat joint, we have evaluated a strategy of gene transfer by means of the delivery of external electric pulses (EP) to the knee after intra-articular injection of a reporter gene (GFP). Rats were killed at various times after the electro gene-therapy to analyze GFP gene expression by immunohistochemistry. GFP staining was detected in the superficial, middle, and deep zones of the patellar cartilage at days 2 and 9, and thereafter only in the deep zone (months 1 and 2). The average percentage of GFP-positive cells was estimated at 30% both one and 2 months after the gene transfer. Moreover, no pathologic change caused by the EP was detected in the cartilage. The level and stability of the long-term GFP expression found in this study demonstrate the feasibility of a treatment of joint disorders (inflammatory or degenerative, focal or diffuse) using electric gene transfer.

摘要

为建立一种高效的将基因直接体内靶向大鼠关节的系统,我们评估了一种基因转移策略,即在关节腔内注射报告基因(绿色荧光蛋白,GFP)后,通过向膝关节施加外部电脉冲(EP)来进行基因传递。在电基因治疗后的不同时间点处死大鼠,通过免疫组织化学分析GFP基因表达。在第2天和第9天,在髌软骨的浅、中、深区均检测到GFP染色,此后仅在深区(第1个月和第2个月)检测到。基因转移后1个月和2个月,GFP阳性细胞的平均百分比估计均为30%。此外,在软骨中未检测到由电脉冲引起的病理变化。本研究中发现的长期GFP表达的水平和稳定性证明了使用电基因转移治疗关节疾病(炎症性或退行性、局灶性或弥漫性)的可行性。

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