Cad-Ⅱ基因转染的兔骨髓间充质干细胞自体移植术后的表达研究

Expression of Cad-Ⅱ gene in bone marrow mesenchymal stem cells transfected with Cad-ⅡcDNA after autografted into the bone defects

[目的]研究Cad-Ⅱ基因转染的骨髓间充质干细胞在骨缺损局部移植后Cad-Ⅱ基因的表达变化。[方法]20只日本长耳白兔制作双侧髂骨缺损模型,将体外培养扩增的自体骨髓干细胞经脂质体介导转染Cad-Ⅱ基因,再与胶原海绵复合移植于髂骨缺损处,术后4周取材,RT-PCR和免疫组化方法检测Cad-Ⅱ基因的表达。[结果]术后4周各组骨缺损区已有新骨生成,Cad-Ⅱ基因转染移植组和对照组均检测到Cad-ⅡmRNA的表达,但转染组Cad-ⅡmRNA的表达显著高于对照组(P<0.01)。免疫组织化学染色结果显示,与对照组比较转染移植各组Cad-Ⅱ阳性细胞数量也显著增多(P<0.01)。[结论]转染了Cad-Ⅱ基因的骨髓间充质干细胞使骨缺损区Cad-Ⅱ基因的表达增高。

[ Objective] To study the expression of Cad - Ⅱ gene in bone marrow mesenchymal stem cells （MSCs） transfected with Cad - Ⅱ cDNA after autografted into the bone defects. [ Method ] The experimental model of ilium segment defect was established in 20 Japanese white rabbits. The rabbit MSCs were isolated, cultured and expanded in vitro, and then the MSCs, transfeeted with Cad -Ⅱ and compounded with collagen sponge were autografted into the ilium segment defect. At 4 weeks of operation, the MSCs/collagen sponge were excised, and the expression of Cad - Ⅱ was evaluated with RT - PCR and immunohistochemical methods. [ Result] All of the bone defects treated with implants exhibited new bone formation at 4 weeks postoperatively. In the transfection group, Cad - Ⅱ gene mRNA expression was higher than that in the control group （ P 〈 0.01 ） . The results of immunohistochemical staining showed there was a significant increase of the positive cells in the transfection group as compared with the control group （P 〈 0. 01 ） . [ Conclusion ] MSCs transfected with Cad - 1/ is highly expressed after autografted into the bone defects.