Systemic and Intravitreal Delivery of Dendrimers to Activated Microglia/Macrophage in Ischemia/Reperfusion Mouse Retina
Systemic and Intravitreal Delivery of Dendrimers to Activated Microglia/Macrophage in Ischemia/Reperfusion Mouse Retina
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DOI：
10.1167/iovs.14-16250
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发表时间：
2015-07-01
影响因子：
4.4
通讯作者：
Lutty, Gerard
中科院分区：
文献类型：
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作者：
Kambhampati, Siva P.;Clunies-Ross, Alexander J. M.;Lutty, Gerard
目的。小胶质细胞激活和相关的神经炎症在许多视网膜疾病的发病机制中起着关键作用，包括病毒感染、糖尿病和视网膜变性。针对激活的小胶质细胞和巨噬细胞并减轻炎症的策略在治疗这些疾病方面可能是有价值的。我们寻求开发以树枝状大分子为基础的制剂，以病理依赖的方式靶向视网膜小胶质细胞和巨噬细胞，并通过静脉或玻璃体内递送药物。方法：在正常和缺血/再灌注(I/R)小鼠眼睛中评估了花菁染料(Cy5)-偶联树突状大分子(D-Cy5)的视网膜摄取。使用兔Iba-1抗体和Cy3标记的二抗(小胶质细胞/巨噬细胞)，用免疫荧光法检测树枝状大分子对小胶质细胞/巨噬细胞的摄取。用荧光分光光度法定量测定视网膜和其他器官的摄取。结果：D-Cy5在玻璃体腔注射72小时和静脉注射24小时后从正常眼中几乎完全清除。在I/R损伤后激活的小胶质细胞中，D-Cy5被激活的小胶质细胞/巨噬细胞(IBA1(+)细胞)保留到玻璃体内和静脉注射后21d。在I/R眼中，如果静脉注射剂量增加30倍，D-Cy5在玻璃体内和静脉内的相对滞留率是相当的。结论：玻璃体内和全身树突状分子靶向激活的小胶质细胞，无论哪种途径给药，在质量上都显示出相似的视网膜生物分布。这些结果为开发树枝状大分子药物制剂作为治疗与小胶质细胞或巨噬细胞激活相关的视网膜疾病，如老年性黄斑变性、糖尿病视网膜病变和视网膜退化提供了概念验证见解。
PURPOSE. Microglial activation and associated neuroinflammation play a key role in the pathogenesis of many diseases of the retina, including viral infection, diabetes, and retinal degeneration. Strategies to target activated microglia and macrophages and attenuate inflammation may be valuable in treating these diseases. We seek to develop dendrimer-based formulations that target retinal microglia and macrophages in a pathology-dependent manner, and deliver drugs, either intravenously or intravitreally.METHODS. Retinal uptake of cyanine dye (Cy5)-conjugated dendrimer (D-Cy5) was assessed in normal and ischemia/reperfusion (I/R) mouse eyes. Microglia/macrophage uptake of the dendrimer was assessed with immunofluorescence using rabbit Iba-1 antibody with Cy3-tagged secondary antibody (microglia/macrophage). Uptake in retina and other organs was quantified using fluorescence spectroscopy.RESULTS. Clearance of D-Cy5 from normal eyes was almost complete by 72 hours after intravitreal injection and 24 hours after intravenous delivery. In eyes with activated microglia after I/R injury, D-Cy5 was retained by activated microglia/macrophage (Iba1(+) cells) up to 21 days after intravitreal and intravenous administration. In I/R eyes, the relative retention of intravitreal and intravenous D-Cy5 was comparable, if a 30-fold higher intravenous dose was used.CONCLUSIONS. Intravitreal and systemic dendrimers target activated microglia and show qualitatively similar retinal biodistribution when administered by either route. Results provide proof-of-concept insights for developing dendrimer drug formulations as treatment options for retinal diseases associated with microglia or macrophage activation such as age-related macular degeneration, diabetic retinopathy, and retinal degenerations.