摘要
目的 考察N-亚甲基磷酸化壳聚糖（ NMPCS）基因纳米粒子的体外细胞毒性及基因转染效率.方法 采用均相反应法制备了NMPCS,用复凝聚法制备了NMPCS/DNA纳米粒子；通过MTT实验考察了NMPCS及其与DNA复合物对HeLa细胞的细胞毒性,以荧光索酶质粒为报告基因考察了NMPCS及NMPCS-CaZ＋载体介导的体外基因转染效率.结果 NMPCS及其与DNA的复合物在体外表现出很小的细胞毒性,远远低于同等浓度时聚乙烯亚胺（PEI）的毒性.通过对壳聚糖进行亚甲基磷酸化修饰后,可大幅提高载体的基因转染效率.结论 N-亚甲基磷酸化壳聚糖有望成为一种新型、安全、高效的非病毒基因载体.
Objective To investigate the cytotoxicity and gene transfection mediated by NMPCS/DNA nanoparticles. Methods N-methylene phosphonic chitosan （NMPCS） was synthesized using one-step reaction under homogeneous conditions. The NMPCS/DNA nanoparticles were prepared using complex coacervation method. The cytotoxicity of NMPCS alone and its complexes with plasmid DNA were determined by MTF assay on HeLa cells. The gene transfection mediated by NMPCS/DNA nanoparticles were investigated using pGL3-control vector as reporter gene. Results The MTF results suggested that the NMPCS and NMPCS/DNA complexes showed significantly lower cytotoxicity than PEI and PEI/DNA complexes, respectively. The gene transfection mediated by NMPCS/DNA nanoparticles were greatly improved compared with unmodified chitosan. Conclusion NMPCS would demonstrate great potential as a novel, safe, efficient non-viral vector for gene delivery.
出处
《国际生物医学工程杂志》
CAS
2012年第3期165-168,F0003,共5页
International Journal of Biomedical Engineering
基金
国家自然科学基金资助项目（81100100）
天津市自然科学基金面上项目（10JCYBJC10600）
教育部新教师基金（20091106120048）
协和青年科研基金资助项目
