中文摘要：

目的评价胶原膜上通过抗体结合腺病毒或质粒DNA作为基因投递载体的可行性及效果。方法采用交联剂N-琥珀酰亚胺基3-（2-吡啶二硫基）丙酸酯（SPDP）将抗腺病毒或抗DNA抗体共价键结合在胶原膜上,通过这些抗体将腺病毒载体（Ad-GFP）和质粒DNA（pEGFP-C1）结合在膜上。采用同位素标记的质粒DNA（pDNA）在体外测定基因的释放,用细胞转染试验评价这种新型基因递送体系的功能。结果通过SPDP交联剂成功地将抗体共价键连接在胶原膜上,并结合了表达绿色荧光蛋白（GFP）的腺病毒和质粒DNA。细胞实验发现胶原膜上有大量表达GFP的阳性细胞,而膜以外的区域则没有GFP阳性细胞,表明具有局部递送特征。偶联Ad-GFP的胶原膜最大转染效率达到92.8%,而且在10^7-10^10病毒粒子/mL范围内,转染效率呈正剂量相关性。偶联pEGFP-C1的胶原膜诱导的细胞转染效率约为21.8%,结合32P标记的pDNA的胶原膜体外基因释放持续13天以上。用胶原涂层的不锈钢血管支架进行了同样的试验,在细胞培养中支架表面有大量GFP阳性细胞,其他区域没有GFP阳性细胞。结论胶原膜携带抗体偶联基因是一种新型的基因投递体系,可实现高效和局部定位的基因转染。

英文摘要：

Objective To evaluate the possibility and efficiency of collagen films as gene delivery system using antibody-tethered adenovirus or pDNA.Methods Anti-adenoviral or anti-DNA antibodies were covalently bound to the collagen surface by a cross linking reagent of N-Succinimidyl3-（2-pyridyldithio） propionate （SPDP）. These antibodies were used to tether adenovirus vectors （Ad-GFP） and pDNA （pEGFP-C1）. The DNA released from collagen films was measured in vitro by isotope label pDNA and the function of this novel gene delivery system was evaluated in cell culture. Results Anti-adenoviral monoclonal antibodies and anti-DNA antibodies were covalently attached to collagen films through the bi-functional cross-linker SPDP. In cell culture studies, GFP positive cells were detected only on the surface of collagen films but not on the portion out of collagen films, which demonstrated highly localized gene delivery. In the experiments of collage films tethered Ad-GFP, the terminal cell transduction was 92.8 % . Viral loading and transduction efficiency were positive correlation when viral Ad-GFP ranging from 8.5 × 10^7 to 8.5 × 10^10 viral particles/mL. In the experiments of collagen films tethered pEGFP-C1, the transduction efficiency was about 21.8%, the slow release of pDNA lasted more than 13 days. Metal vascular stents, that have been coated with collagen and treated in the same way, showed localized gene transduction on the surface of stents in cell culture too. Conclusions A novel gene delivery system from collagen film carrying antibody-tethered DNA was developed. The system showed its highly efficient and localized gene tranduction in cell culture.