负载柳氮磺吡啶缓释纳米粒的制备及体外评价  

Preparation and in vitro evaluation of sustained-release nanoparticles loaded with sulfasalazine

作　　者：钟文静 周子涵 王浩宇 李尚勇 夏玉军[1] Zhong Wenjing;Zhou Zihan;Wang Haoyu;Li Shangyong;Xia Yujun(Department of Special Medicine,Basic Medical College,Qingdao University,Qingdao 266000,Shandong Province,China;Basic Medical College,Qingdao University,Qingdao 266000,Shandong Province,China)

机构地区：[1]青岛大学基础医学院特种医学系,山东省青岛市266000 [2]青岛大学基础医学院,山东省青岛市266000

出　　处：《中国组织工程研究》2023年第16期2461-2466,共6页Chinese Journal of Tissue Engineering Research

基　　金：山东省自然科学基金(ZR2019BD027),项目负责人:李尚勇。

摘　　要：背景:柳氮磺吡啶是治疗溃疡性结肠炎的常用药,传统给药方式吸收差、利用度低,且常伴随严重的毒副反应。与传统给药方式相比,基于结肠靶向纳米给药能够保护药物免受不良环境的影响,改善药物的局部吸收和生物利用度。目的:制备柳氮磺吡啶@聚乳酸-羟基乙酸共聚物-壳聚糖-果胶-壳聚糖纳米粒,对其进行表征和体外评价,并在细胞水平验证纳米颗粒用作药物载体的安全性。方法:采用O/W乳液法和静电逐层自组装技术制备柳氮磺吡啶@聚乳酸-羟基乙酸共聚物-壳聚糖-果胶-壳聚糖纳米粒,利用透射电镜观察了纳米粒的形态,激光粒度仪检测了纳米粒的电位与粒径,紫外分光光度计法检测其载药率和包封率。将纳米粒溶解于不同pH值(1.2,6.8,7.4)PBS中,检测纳米粒对柳氮磺吡啶的释放情况。将不同质量浓度(10,20,50,100,200 mg/L)的纳米粒溶液与巨噬细胞共培养,48 h后,采用CCK8法检测细胞活性;将20 mg/L的纳米粒溶液与巨噬细胞共培养,24 h后,罗丹明荧光染色细胞摄取纳米粒情况。结果与结论:①透射电镜下可见,纳米粒呈椭球形,粒径大小均一;纳米粒粒径为290.9 nm,电位为19.8 mV,分散指数为0.295,单个纳米颗粒具有良好的分散性;纳米粒的包封率为75.68%,载药量为22.24%;②体外药物释放实验显示,在36 h内,随着PBS pH值的升高,纳米粒的释放速率加快,纳米粒在pH=1.2的PBS中几乎不释放柳氮磺吡啶,在pH=7.4的PBS中可以缓慢持续释放柳氮磺吡啶,表明纳米粒具有pH值依赖性和良好的缓释特征;③不同质量浓度的米粒浓度对巨噬细胞的活性未造成明显影响,未表现细胞毒性作用;④细胞摄取实验显示,培养6 h后,细胞开始摄取纳米粒,12-24 h纳米颗粒进入了巨噬细胞;⑤柳氮磺吡啶@聚乳酸-羟基乙酸共聚物-壳聚糖-果胶-壳聚糖纳米粒具有良好的的缓释特性与细胞相容性,能被巨噬细胞摄取且摄取效率�BACKGROUND:Sulfasalazine has been commonly used in the treatment of ulcerative colitis.Traditional administration methods have poor absorption,low availability,and are often accompanied by serious toxic side effects.Compared with traditional drug delivery methods,colon-targeted nano-delivery can protect the drug from adverse environment and improve the local absorption and bioavailability of the drug.OBJECTIVE:Sulfasalazine@polylactic acid-glycolic acid copolymer-chitosan-pectin-chitosan nanoparticles were prepared for characterization and vitro evaluation,and the safety of nanoparticles as drug carriers was verified at the cellular level.METHODS:The O/W emulsion method and electrostatic layer-by-layer self-assembly technique were used to prepare sulfasalazine@polylactic acid-glycolic acid copolymer-chitosan-pectin-chitosan nanoparticles.The morphology of the nanoparticles was observed using transmission electron microscopy.Laser particle sizer was used to detect nanoparticle potential and particle size.Drug loading and encapsulation rates of nanoparticles were determined by ultraviolet spectrophotometry.The nanoparticles were dissolved in PBS with different pH values(1.2,6.8,7.4)to detect the release of sulfasalazine by the nanoparticles.The nanoparticle solutions of different mass concentrations(10,20,50,100,200 mg/L)were co-cultured with macrophages,and after 48 hours,the cell viability was detected by CCK8 assay.The 20 mg/L nanoparticle solution was co-cultured with macrophages,and after 24 hours,rhodamine fluorescent staining was performed for the uptake of nanoparticles by cells.RESULTS AND CONCLUSION:(1)Transmission electron microscopy images showed that the nanoparticles were ellipsoidal with uniform particle size;with a particle size of 290.9 nm,a potential of 19.8 mV and a dispersion index of 0.295,with good dispersion of the individual nanoparticles.The encapsulation rate of the nanoparticles was 75.68%and the drug loading was 22.24%.(2)In vitro release experiments showed that within 36 hours,the relea

关 键 词：柳氮磺吡啶 聚乳酸-羟基乙酸共聚物 壳聚糖 果胶 纳米粒 体外评价 溃疡性结肠炎 

分 类 号：R516.3[医药卫生—内科学] R318[医药卫生—临床医学] R8