不锈钢血管支架蛋白涂层和携带质粒DNA的研究

Methodological Studies of Protein Coating and Plasmid DNA Delivery Based on Stainless Steel Coronary Stents

目的:探讨在不锈钢冠状动脉支架上携带质粒基因,为心血管再狭窄基因治疗的临床应用提供试验依据。方法:支架表面用交联蛋白涂层;采用双官能团偶联剂将抗DNA抗体以化学键结合在蛋白涂层上,再与表达绿色荧光蛋白的质粒DNA免疫偶联;用同位素标记的抗体评价抗体与支架结合的稳定性,用体外细胞培养试验验证支架携带基因的转染效果。结果:交联蛋白涂层上化学键结合抗DNA抗体的量比单纯物理吸附高约15倍,支架上通过抗DNA抗体免疫偶联质粒DNA的细胞转染效率明显高于单纯物理吸附携带基因的支架。结论:本研究成功地在金属冠状动脉支架上用化学和免疫偶联携带质粒基因,实现了细胞培养中局部高效的基因表达。

Objective; To investigate stainless steel coronary stent-based plasmid gene delivery system and evaluate its feasibility and effectiveness to gene therapy of cardiovascular restenosis. Methods： 316L stainless steel stents were formulated with protein coatings. Anti-DNA antibodies were covalently bound to the protein surface by using a bi-functional cross-linking agent SPDP. Report plasmid of green fluorescent protein （pEGFP） was bound on the anti-DNA immobilized stents by immune affinity. The feasibility and stability of anti-DNA antibody covalently bound to the stent were evaluated by means of isotope labeling antibody. Cell culture in vitro was employed to assess transduction effects of the gene tethered on the stents. Results： The amount of chemically coupled antibody on the protein coating stents was fifteen times higher than the physically absorbed control stents, and the transfection efficiency of anti-DNA antibody chemically coupled with plasmid DNA was significantly better than that of the control stents. Conclusion： The anti-DNA antibody chemically coupled with plasmid DNA is successfully used to stainless stents and obtain a local high efficient gene expression in the cell culture.