曲安奈德和霉酚酸酯纳米粒子的制备及其生物学特性评价

doi:10.13481/j.1671-587X.20230430

摘要/Abstract

摘要：

目的探讨治疗甲状腺相关眼病（TAO）药物曲安奈德（TA）和霉酚酸酯（MMF）聚乙二醇-聚乳酸羟基乙酸（PEG-PLGA）纳米粒子的最佳制备工艺及体外释放性能，评价其经眶周注射治疗TAO的安全性。方法以PEG-PLGA共聚物为原料，采用乳化法分别制备TA纳米粒子（TA NPs组）和MMF纳米粒子（MMF NPs组），以包封率为评价指标进行工艺优化。采用透射电子显微镜观察纳米粒子形态表现，采用Zetasizer粒径电位分析仪检测各组纳米粒子的粒径和电位。在体外眶周组织模拟液中，采用紫外分光光度法检测各组纳米粒子的释放性质，计算药物释放率，结合临床用药规则对药物的整体释放性质进行分析。人视网膜色素上皮hRPE-19细胞分为空白对照组、不同浓度TA组、不同浓度MMF组、不同浓度TA NPs组和不同浓度MMF NPs组，采用MTT法检测各组细胞活性，评价制剂的安全性。结果制备载有TA和MMF的纳米粒子（TA NPs和MMF NPs），包封率分别为47.66%和16.52%，平均粒径约为600 nm，其电位均符合眶周注射的基本要求。透射电子显微镜下观察，TA NPs和MMF NPs表观圆整，均一化程度较高。体外释放体系检测TA NPs和MMF NPs的释放特性均符合临床上TAO治疗药物的给药特性，均可持续释放3周以上，其初始释放率不高，体外释放曲线较为平稳。MTT法检测，不同浓度TA组、MMF组、TA NPs组和MMF NPs组在较低浓度下无明显的细胞抑制；较高浓度下，与相同浓度TA组比较，40、80和160 nmol·L^－1 TA NPs组细胞活性明显升高（P<0.01）；与相同浓度MMF组比较，50、100和200 nmol·L^－1 MMF NPs组细胞活性明显升高（P<0.01）。结论 TA NPs和MMF NPs的物化表征参数符合眶周注射制剂的基本要求，且制备工艺简单，安全性良好，具有优良的载药和缓释效果，与TAO的临床用药治疗需求一致。

Abstract:

Objective To discuss the optimal preparation process and the in vitro release properties of the poly （lactic-co-glycolic acid）poly（ethylene glycol）（PEG-PLGA） nanoparticles of triamcinolone acetonide（TA） and mycophenolate mofetil（MMF） in the treatment of thyroid associated ophthalmopathy（TAO）， and to evaluate its safety in the treatment of TAO by periorbital injection. Methods The TA nanoparticles （TA NPs group） and MMF nanoparticles （MMF NPs group） were prepared with PEG-PLGA as the raw material by emulsification method. The process optimization was carried out with the encapsulation efficiency as the evaluation indicator. The morphology of nanoparticles in various groups was observed under transmission electron microscope；the sizes of nanoparticles in various groups were detected by zetasizer particle size and potential analyzer. The release properties of nanoparticles in various groups were detected by ultraviolet spectrophotometry in vitro and the release rates of the drug were calculated，and then the release properties of the drugs were analyzed on the basis of clinical medication rules. The human retinal pigment epithelial hRPF-19 cells were divided into blank control group，different concentrations of TA groups，different concentrations of MMF groups，different concentrations of TA NPs group， and different concentrations of MMF NPs groups.The viabilities of the cells were evaluated by MTT test and the safty of preparation was evaluated. Results The TA-loaded and MMF-loaded nanoparticles were prepared （TA NPs and MMF NPs）. The encapsulation efficiencies of TA NPs and MMF NPs were 47.66% and 16.52%， with an average particle size of 600 nm. The potential of the nanoparticles met the basic requirements of periorbital injection. Under the microscope， the TA NPs and MMF NPs showed round appearance and a high degree of uniformity. The detection results of drug release system in vitro showed that the release characteristics of TA-NPs and MMF-NPs met the administration characteristics of TAO treatment drugs and the sustained release was able to last more than 3 weeks. The initial release rate was low and the release curve was stable. The MTT results showed that there were no significant cell inhibitions in different concentrations of TA groups， MMF groups， TA NPs groups，and MMF NPs groups at the lower concentration. At the higher concentration，compared with same concentration of TA group， the viabilities of the cells in 40，80，and 160 nmol·L^－1 TA NPs groups were increased （P<0.01）； compared with same concentration of MMF group，the viabilities of the cells in 50，100，and 200 nmol·L^－1 MMF NPs groups were increased （P<0.01）. Conclusion The physicochemical parameters of TA NPs and MMF NPs meet the basic requirements for the periorbital injection with simple preparation process and good safety and have better drug-loaded and sustained release effects， which can be consistent with the requirements for clinical drug therapy in the treatment of TAO.

Key words: Thyroid-associated ophthalmopathy, Triamcinolone acetonide, Mycophenolate mofetil, Nanoparticles, In vitro release

中图分类号:

R777.5

丛云毅,李光宇. 曲安奈德和霉酚酸酯纳米粒子的制备及其生物学特性评价[J]. 吉林大学学报(医学版), 2023, 49(4): 1060-1066.

Yunyi CONG,Guangyu LI. Preparations of triamcinolone acetonide and mycophenolate mofetil nanoparticles and their biological evaluations[J]. Journal of Jilin University(Medicine Edition), 2023, 49(4): 1060-1066.

图/表 3

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