中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (16): 2461-2466.doi: 10.12307/2023.134

• 纳米生物材料 nanobiomaterials •    下一篇

负载柳氮磺吡啶缓释纳米粒的制备及体外评价

钟文静1，周子涵1，王浩宇2，李尚勇2，夏玉军1   

  1. 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
  • 收稿日期:2022-03-11 接受日期:2022-05-14 出版日期:2023-06-08 发布日期:2022-11-10
  • 通讯作者: 夏玉军，教授，青岛大学基础医学院特种医学系，山东省青岛市 266000
  • 作者简介:钟文静，女，1997年生，山东省菏泽市人，汉族，青岛大学基础医学院在读硕士，主要从事纳米材料载药的制备研究。
  • 基金资助:
    山东省自然科学基金(ZR2019BD027)，项目负责人：李尚勇

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

Zhong Wenjing1, Zhou Zihan1, Wang Haoyu2, Li Shangyong2, Xia Yujun1   

  1. 1Department of Special Medicine, Basic Medical College, Qingdao University, Qingdao 266000, Shandong Province, China; 2Basic Medical College, Qingdao University, Qingdao 266000, Shandong Province, China
  • Received:2022-03-11 Accepted:2022-05-14 Online:2023-06-08 Published:2022-11-10
  • Contact: Xia Yujun, Professor, Department of Special Medicine, Basic Medical College, Qingdao University, Qingdao 266000, Shandong Province, China
  • About author:Zhong Wenjing, Master candidate, Department of Special Medicine, Basic Medical College, Qingdao University, Qingdao 266000, Shandong Province, China
  • Supported by:
    Natural Science Foundation of Shandong Province, No. ZR2019BD027 (to LSY)

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摘要：


文题释义：

柳氮磺吡啶：是水杨酸与磺胺吡啶的偶氮化合物，为磺胺类抗菌药，具有抗菌和免疫抑制作用，在肠道内被该处细菌分解为磺胺吡啶与5-氨基水杨酸。
缓释系统：是指通过适宜的方法延缓药物在体内的释放、吸收、代谢以及排泄的过程，从而延长药物作用时间或者减轻其毒副作用的给药系统。

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高。
https://orcid.org/0000-0002-7169-8261(钟文静)

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

Abstract: 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 release rate of nanoparticles increased with the increase of PBS pH. The prepared nanoparticles barely released sulfasalazine at low pH=1.2 and released slowly and continuously up to pH 7.4, indicating that the nanoparticles had pH-dependent and good slow release characteristics. (3) Different concentrations of nanoparticles had no obvious effects on the activity of macrophages and did not show cytotoxic effects. (4) Cellular uptake experiments showed that nanoparticles could be taken up by cells after 6 hours of culture. The nanoparticles entered the macrophages within 12-24 hours. (5) The sulfasalazine@polylactic acid-glycolic acid copolymer-chitosan-pectin-chitosan nanoparticles have good slow-release properties and cytocompatibility, can be taken up by macrophages with high uptake efficiency. 

Key words: sulfasalazine, polylactic acid-glycolic acid copolymer, chitosan, pectin, nanoparticles, in vitro evaluation, ulcerative colitis

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