摘要
目的探讨不同修饰度氯化壳聚糖季铵盐纳米粒对小鼠骨髓来源树突状细胞转染效率的影响,为以壳聚糖纳米粒为佐剂的树突状细胞生物免疫疫苗研究提供理论依据。方法采用卤代法修饰合成不同修饰度的氯化壳聚糖季铵盐;离子交联法制备不同修饰度的氯化壳聚糖季铵盐/p EGFP-C1纳米粒;激光粒度仪测粒径和Zate电位,酶保护实验评价纳米粒质量。用改良Insba法,体外分离培养树突状细胞,用不同修饰度的氯化壳聚糖季铵盐纳米粒做体外转染实验,验证其细胞毒性和转染效率。结果纳米粒粒径:(220±15)nm,Zeta电位:(29±5)m V,包封率99.8%,氯化壳聚糖季铵盐纳米粒对树突状细胞体外转染效率比壳聚糖纳米粒提高了10%。结论氯化壳聚糖季铵盐纳米粒能有效提高p EGFP-C1质粒在树突状细胞中的转染效率。
OBJECTIVE To investigate the effect of chlorinated N-trimethyl chitosan （TMC·Cl） nanoparticle with different modified degrees on the transfection efficiency of mouse marrow-derived dendritic cells （DC） to provide a theoretical basis for the study of DC biological immune vaccine with chitosan （CS） nanoparticle adjuvant.METHODS TMC·Cl/pEGFP-C1 nanoparticles with different modified degrees were prepared by ion-crosslinking method. Particle size and Zate potential were measured by laser particle size analyzer. Nanoparticle quality was evaluated by enzyme protection experiment. DC were cultured in vitro by a modified Insba method, and TMC·Cl nanoparticles with different degrees of modification were analyzed by in vitro transfection experiments to verify the cytotoxicity and transfection efficiency.RESULTS The nanoparticle size was （220±15） nm, Zeta potential was （29±5） mV, encapsulation efficiency was 99.8%, and the transfection efficiency of TMC·Cl nanoparticles on DC in vitro increased by 10% than that of CS nanoparticles.CONCLUSION TMC·Cl nanoparticles can effectively improve the transfection efficiency of pEGFP-C1 plasmid in DC.
作者
冯胜
贡长慧
沈文静
李军
FENG Sheng, GONG Chang-hui, SHEN Wen-jing, LI Jun(Key Laboratory of Tropical Biological Resources of Ministry of Education, Department of Pharmacy of Ocean College, Hainan University, Haikou 570228, Chin)
出处
《中国药学杂志》
CAS
CSCD
北大核心
2018年第9期707-712,共6页
Chinese Pharmaceutical Journal
基金
国家自然科学基金项目资助(31560261)
海南省自然科学基金项目资助(818MS039)
关键词
氯化壳聚糖季铵盐
纳米粒
树突状细胞
细胞转染
chlorinated N-trimethyl chitosan
nanoparticle
dendritic cell
cell transfection
