氯化壳聚糖季铵盐纳米粒对树突状细胞的转染效率研究

冯胜,贡长慧,沈文静,李军

中国药学杂志 ›› 2018, Vol. 53 ›› Issue (9) : 707-712.

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中国药学杂志 ›› 2018, Vol. 53 ›› Issue (9) : 707-712. DOI: 10.11669/cpj.2018.09.010
论著

氯化壳聚糖季铵盐纳米粒对树突状细胞的转染效率研究

  • 冯胜,贡长慧,沈文静,李军*
作者信息 +

Transfection Efficiency of Chlorinated N-trimethyl Chitosan Nanoparticle on Dendritic Cells

  • FENG Sheng, GONG Chang-hui, SHEN Wen-jing, LI Jun*
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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/pEGFP-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)mV,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pEGFP-C1质粒在树突状细胞中的转染效率。

Abstract

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.

关键词

氯化壳聚糖季铵盐 / 纳米粒 / 树突状细胞 / 细胞转染

Key words

chlorinated N-trimethyl chitosan / nanoparticle / dendritic cell / cell transfection

引用本文

导出引用
冯胜,贡长慧,沈文静,李军. 氯化壳聚糖季铵盐纳米粒对树突状细胞的转染效率研究[J]. 中国药学杂志, 2018, 53(9): 707-712 https://doi.org/10.11669/cpj.2018.09.010
FENG Sheng, GONG Chang-hui, SHEN Wen-jing, LI Jun. Transfection Efficiency of Chlorinated N-trimethyl Chitosan Nanoparticle on Dendritic Cells[J]. Chinese Pharmaceutical Journal, 2018, 53(9): 707-712 https://doi.org/10.11669/cpj.2018.09.010
中图分类号: R917   

参考文献

[1] LAURA D L, JENNIFER C R, LONNIE D S. Design of modular non-viral gene therapy vectors[J] . Biomaterials, 2006, 27(7):947-954.
[2] BALDRICK P. The safety of chitosan as a pharmaceutical ex-cipient [J] . Regul Toxicol Pharmacol, 2010, 56(3):290-299.
[3] DU Y X, LI L, HUANG F, et al. Preparation and performance of chitosan/gene transfer system[J] . Polym Mater Sci Eng(高分子材料科学与工程), 2012, 28(7):108-111,115.
[4] National Health and Family Planning Commission. 2013 A Summary of China′s Health and Family Planning Statistics(2013年中国卫生和计划生育统计摘要)[M] . Beijing:Peking Union Medical College Press, 2013.
[5] RADFORD K J, TULLETT K M, LAHOUD M H. Dendritic cells and cancer immunotherapy[J] . Curr Opin Immunol, 2014, 27:26-32.
[6] APETOH L, LOCHER C, GHIRINGHELLI F, et al. Harnessing dendritic cells in cancer[J] . Semin Immunol, 2011, 23(1):42-49.
[7] DING F, SHI X, LI X, et al. Homogeneous synthesis and characterization of quaternized chitin in NaOH/urea aqueous solution[J] . Carbohydr Polym, 2012, 87(1):422-426.
[8] JI J H. Determination of the degree of deacetylation of chitosan by different methods[J] . Chem Eng(化学工程师), 2016, 30(11):11,23-25.
[9] ZHU N, WANG H T, LI C H, et al. Isolation,induced culture and identification of immature dendritic cells in vitro[J] . Chin J Immunol(中国免疫学杂志), 2017, 33(7):1043-1047.
[10] SHI X W, DONG H B, DU Y M. Chitin/Chitosan Materials and Applications(甲壳素/壳聚糖材料及应用)[M] . Beijing:Chemical Industry Press, 2015,9:59-62.
[11] MAO S R, SUN W, KISSEL T. Chitosan-based formulations for delivery of DNA and si RNA[J] . Adv Drug Deliv Rev, 2010, 62(1):12-27.
[12] MA Q, FAN J. Research progress of chitosan nanoparticles as a carrier[J] . Northwest Pharm J(西北药学杂志), 2015, 30(2):213-215.
[13] ROLINSKI J, HUS I. Dendritic-cell tumor vaccines [J] . Transplant Proc, 2010, 42(8):3306-3308.
[14] ISHIGAMI S, NATSUGOE S, TOKUDA K, et al. Clinical impact of intratumoral natural killer cell and dendritic cell infiltration in gastric cancer[J] . Cancer Lett, 2000, 159(1):103-108.
[15] LIECHTY W B, CALDORERA-MOORE M, PHILLIPS M A, et al. Advanced molecular design of biopolymers for transmucosaland intracellular deliveryof chemotherapeutic agents andbiological therapeutics[J] . J Controlled Release, 2011, 155(2):119-127.

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)
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