阿霉素壳寡糖纳米粒的制备及体外抗肿瘤研究

Preparation of doxorubicin-loaded chitosan oligosaccharide nanoparticles and its anti-tumor effect in vitro

目的制备负载阿霉素的壳寡糖纳米粒,并研究其理化性质和体外抗肿瘤细胞毒性。方法采用离子凝胶法制备负载阿霉素的壳寡糖纳米粒;透射电镜观察纳米粒形态,激光粒度仪测定粒径和表面电位,紫外分光光度法测量包封率、载药量,考察载药纳米粒的体外释药特性;采用MTT法对载药壳寡糖纳米粒在体外乳腺癌细胞株MCF-7的细胞毒作用进行评价。结果制得的阿霉素壳寡糖纳米粒呈球形或类球形,形态较为完整,平均粒径为(136.77±1.21)nm,表面电位为(20.53±0.31)m V,包封率为(56.99±1.40)%,载药量为(15.49±0.38)%,168 h的累积释放率为72.15%;阿霉素和载药纳米粒对MCF-7细胞增殖的抑制作用存在明显的浓度和时间依赖性,且载药纳米粒对MCF-7细胞增殖的抑制作用随时间增加而逐渐强于游离阿霉素。结论此方法制备的阿霉素壳寡糖纳米粒粒径较小,药物释放具有明显的缓释作用,并具有较好的抗肿瘤作用。

Objective To prepare doxorubicin-loaded chitosan oligosaccharide nanoparticles（ DOX-CSO-NPs）, and investigate the physiochemical properties and anti-tumor activity in vitro. Methods DOX-CSO-NPs were prepared by the method of ionic cross-linking; the shape of nanoparticles were observed by transmission electron microscope（ TEM）,the paticle size and surface potential were determined by laser particle size analyzer,encapsulation efficiency and drug-loading rate were measured by ultraviolet spectrometry. The drug release characteristics of nanoparticles in vitro were preliminarily evaluated. The cytotoxicity of drug-loaded nanoparticles to MCF-7 cells in vitro were also investigated by MTT assay.Results DOX-CSO-NPs were spherical and complete in shape as shown under the transmission election microscope,the average size,Zeta potential,encapsulation efficiency and drug-loading rate were（ 136. 77 ± 1. 21） nm,（ 20. 53 ± 0. 31） m V,（ 56. 99 ± 1. 40） % and（ 15. 49 ± 0. 38） %,respectively. The total amount of accumulative release was 72. 15% at the 168 th hour. Free doxorubicin and DOX-CSO-NPs could inhibit the proliferation of MCF-7 cells in dose and time dependent manners in vitro obviously. Furthermore,the inhibition of the DOX-CSO-NPs to the MCF-7 cells was increasing with time and stronger than free doxorubicin. Conclusion The particle size of DOX-CSO-NPs prepared by this method is smal and features a good quality of slow releasing,and it has better anti-tumor effect.