Subconjunctival nano- and microparticles sustain retinal delivery of budesonide, a corticosteroid capable of inhibiting VEGF expression
Subconjunctival nano- and microparticles sustain retinal delivery of budesonide, a corticosteroid capable of inhibiting VEGF expression
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DOI：
10.1167/iovs.02-0791
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发表时间：
2003-03-01
影响因子：
4.4
通讯作者：
Ayalasomayajula, SP
中科院分区：
文献类型：
--
作者：
Kompella, UB;Bandi, N;Ayalasomayajula, SP
目的.本研究的目的是确定布地奈德是否抑制视网膜色素上皮细胞系（ARPE-19）中血管内皮生长因子（VEGF）的表达，并确定结膜下施用的布地奈德纳米颗粒和微粒是否维持视网膜药物水平。分别通过ELISA、RT-PCR和细胞活力测定法测定布地奈德（100 pM至10 μ M）对ARPE-19细胞中VEGF分泌、VEGF mRNA表达和细胞毒性的影响。为了确定糖皮质激素受体参与观察到的布地奈德的作用，在糖皮质激素受体拮抗剂（RU 486）存在下还进行了分泌和mRNA表达研究。采用溶剂挥发法制备了含有布地奈德的DL-聚乳酸（PLA）纳米粒和微米粒，并对颗粒的大小、形态、包封率和体外释放进行了表征。将布地奈德-PLA纳米颗粒和微粒结膜下给予Sprague-Dawley大鼠的一只眼睛，并在第1、7和14天结束时测定两只眼睛的视网膜、玻璃体、透镜和角膜中的药物水平。在没有细胞毒性的浓度下，布地奈德以剂量依赖性方式抑制ARPE-19细胞中VEGF分泌以及mRNA表达。RU 486治疗阻止了布地奈德介导的VEGF分泌和VEGF mRNA表达的抑制。布地奈德-聚乳酸纳米（345轮辋）和微粒（3.6微米），与65%和99%的包封率，分别持续布地奈德在体外释放。结膜下给药后，布地奈德-PLA纳米粒和微粒在视网膜和其他眼组织中产生持续的布地奈德水平。布地奈德能够通过糖皮质激素抑制VEGF表达。受体活性结膜下给药的布地奈德-PLA纳米和微粒维持视网膜药物递送。
PURPOSE. The purpose of this study was to determine whether budesonide inhibits expression of vascular endothelial growth factor (VEGF) in a retinal pigment epithelial cell line (ARPE-19) and to determine whether subconjunctivally administered budesonide nano- and microparticles sustain retinal drug levels.METHODS. The effect of budesonide (100 pM to 10 muM) on VEGF secretion, expression of VEGF mRNA, and cytotoxicity were determined in ARPE-19 cells by ELISA, RT-PCR, and a cell-viability assay, respectively. To determine the involvement of glucocorticoid receptor in the observed effects of budesonide, secretion and mRNA expression studies were also performed in the presence of a glucocorticoid receptor antagonist (RU486). DL-Polylactide (PLA) nano- and microparticles containing budesonide were prepared by a solvent evaporation technique, and the particles were characterized for size, morphology, encapsulation efficiency, and in vitro release. Budesonide-PLA nano- and microparticles were administered subconjunctivally to one eye of Sprague-Dawley rats and drug levels in the retina, vitreous, lens, and cornea of both eyes were determined at the end of 1, 7, and 14 days.RESULTS. At concentrations devoid of cytotoxicity, budesonide inhibited VEGF secretion as well as mRNA expression in ARPE-19 cells in a dose-dependent manner. RU486 treatment prevented budesonide-mediated inhibition of VEGF secretion and VEGF mRNA expression. Budesonide-PLA nano- (345 rim) and microparticles (3.6 mum), with an encapsulation efficiency of 65% and 99%, respectively, sustained budesonide release in vitro. After subconjunctival administration, both budesonide-PLA nano- and microparticles produced sustained budesonide levels in the retina and other ocular tissues.CONCLUSIONS. Budesonide is capable of inhibiting VEGF expression through glucocorticoid. receptor activity. Subconjunctivally administered budesonide-PLA nano- and microparticles sustain retinal drug delivery.