中国组织工程研究

中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (30): 4489-4495.doi: 10.3969/j.issn.2095-4344.2016.30.012

• 药物控释材料 drug delivery materials • 上一篇    下一篇

甲氨喋呤长循环脂质体对人骨肉瘤细胞的体外抗瘤活性

秦迎州，陈 汉，张 洋，张其清，刘玲蓉   

  1. 中国医学科学院北京协和医学院，生物医学工程研究所，天津市生物医学材料重点实验室，天津市  300192
  • 出版日期:2016-07-15 发布日期:2016-07-15
  • 通讯作者: 刘玲蓉，副研究员，中国医学科学院北京协和医学院，生物医学工程研究所，天津市生物医学材料重点实验室，天津市 300192
  • 作者简介:秦迎州，女，1990年生，山东省枣庄市人，汉族，中国医学科学院北京协和医学院在读硕士，主要从事生物医学材料研究。
  • 基金资助:

    国家自然科学基金面上项目(31170917)，负责人刘玲蓉：分级模拟胶原特征结构的胶原模拟多肽及其对间充质干细胞的分化调控作用

Long-circulating methotrexate-loaded liposomes exhibit an antitumor effect on human osteosarcoma in vitro

Qin Ying-zhou, Chen Han, Zhang Yang, Zhang Qi-qing, Liu Ling-rong   

  1. Institute of Biomedical Engineering, Chinese Academy of Medical Science & Peking Union Medical College, Tianjin 300192, China
  • Online:2016-07-15 Published:2016-07-15
  • Contact: Liu Ling-rong, Associate researcher, Institute of Biomedical Engineering, Chinese Academy of Medical Science & Peking Union Medical College, Tianjin 300192, China
  • About author:Qin Ying-zhou, Studying for master’s degree, Institute of Biomedical Engineering, Chinese Academy of Medical Science & Peking Union Medical College, Tianjin 300192, China
  • Supported by:

    the National Natural Science Foundation of China, No. 31170917

PDF

273

被引次数

1

摘要：

 

文章快速阅读：

文题释义：
甲氨喋呤：为叶酸类似物，进入机体后可与叶酸还原酶和二氢叶酸还原酶结合，从而与叶酸产生竞争拮抗作用，进而影响DNA和RNA合成，抑制细胞增殖。甲氨喋呤是治疗儿童白血病和淋巴瘤的一种重要的化学药物。
脂质体：作为一种新型给药系统，具有不良反应小、无免疫原性和可生物降解等优点。将甲氨喋呤包载入脂质体中可显著降低药物毒性，改善药物抗肿瘤效果。

 

背景：脂质体作为一种新型给药系统，具有不良反应小、无免疫原性和可生物降解等优点，将甲氨喋呤包载入脂质体中可显著降低药物毒性，改善药物抗肿瘤效果。
目的：制备甲氨喋呤长循环脂质体，考察其对人骨肉瘤细胞MG-63的抗瘤活性。
方法：采用薄膜分散法制备传统甲氨喋呤脂质体，后插入法制备甲氨喋呤长循环脂质体(甲氨喋呤初始质量浓度分别为9.1，1.82，0.364 g/L)，分别采用超速离心法、葡聚糖凝胶G-10微柱离心法和葡聚糖凝胶G-50微柱离心法去除甲氨喋呤长循环脂质体未包载的游离药物，对3种方法纯化后脂质体的回收率、粒径、形态、包封率及药脂比等理化性质进行表征。分别以0，1，5，25 mg/L 甲氨喋呤长循环脂质体(选择超速离心法与葡聚糖凝胶G-50微柱离心法纯化的脂质体)和游离甲氨喋呤稀释液培养人骨肉瘤细胞MG-63，24，48 h后采用MTS法检测细胞毒性。
结果与结论：①甲氨喋呤长循环脂质体的粒径在200 nm左右，为均匀的球形或近球形；②回收率实验结果显示，葡聚糖凝胶G-50微柱离心法是最合适的甲氨喋呤长循环脂质体纯化方法，具有回收完全和快速的优点；③甲氨喋呤初始质量浓度相同时，超速离心法、葡聚糖凝胶G-10微柱离心法纯化的脂质体包封率与药脂比明显低于葡聚糖凝胶G-50微柱离心法纯化的脂质体；④体外细胞毒性实验表明，在同质量浓度条件下，甲氨喋呤长循环脂质体的细胞毒性远高于游离甲氨喋呤，葡聚糖凝胶G-50微柱离心法纯化的甲氨喋呤长循环脂质体细胞活性普遍比超速离心法纯化的脂质体细胞毒性高；⑤结果表明，甲氨喋呤长循环脂质体对于人骨肉瘤细胞具有较强的体外抗瘤活性。

ORCID: 0000-0003-3203-2034(秦迎州)

 

关键词: 生物材料, 材料相容性, 甲氨喋呤, 长循环, 脂质体, 人骨肉瘤细胞MG-63, 国家自然科学基金

Abstract:

BACKGROUND: Liposomes as a new drug delivery system are characterized by few adverse reactions, no immunogenicity and biodegradation. Furthermore, methotrexate-loaded liposomes can significantly reduce drug toxicity and improve anti-tumor effect.
OBJECTIVE: To prepare long-circulating methotrexate-loaded liposomes and to evaluate its antitumor activity in MG-63 in vitro.
METHODS: The methotrexate-loaded liposomes were prepared using the film dispersion method, and the long-circulating ones were prepared using the post-insertion method. The initial concentrations of methotrexate were 9.1, 1.82, 0.364 g/L. The ultracentrifugation method and spin column method with Sephadex G-10 or G-50 as packing were employed to separate free drugs from the methotrexate-loaded liposomes. Their recovery, size, morphology, encapsulation efficiency and drug-to-lipid ratio were evaluated. The cytotoxity of the long-circulating methotrexate-loaded liposomes purified with ultracentrifugation method and spin column G-50 method under three dose levels (0, 1, 5, 25 mg/L) were determined by MTS method.
RESULTS AND CONCLUSION: According to the recovery rates of three methods, the spin column G-50 method was considered as optimal for the long-circulating methotrexate-loaded liposomes. The long-circulating liposomes were spherical or ellipsoidal under transmission electron microscope, about  200 nm in size. At the certain initial concentration of methotrexate, the encapsulation efficiency and drug-to-lipid ratio of the liposomes purified using the spin column G-50 method were remarkably higher than those purified using the other two methods. At the same mass concentration, the cytotoxity of the liposomes purified with ultracentrifugation or spin column G-50 was significantly higher than that of free methotrexate, and furthermore, the cytotoxity of the liposomes purified with spin column G-50 was higher than that of the liposomes purified with ultracentrifugation method. To conclude, the long-circulating methotrexate-loaded liposomes show a higher antitumor activity than free methotrexate in MG-63 cells in vitro, providing the basis for further investigation of its antitumor effect on human osteosarcoma in vivo.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Liposomes, Methotrexate, Osteosarcoma, Tissue Engineering

中图分类号: