Preferential and Increased Uptake of Hydroxyl-Terminated PAMAM Dendrimers by Activated Microglia in Rabbit Brain Mixed Glial Culture.
Preferential and Increased Uptake of Hydroxyl-Terminated PAMAM Dendrimers by Activated Microglia in Rabbit Brain Mixed Glial Culture.
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DOI：
10.3390/molecules23051025
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发表时间：
2018-04-27
期刊：
影响因子：
--
通讯作者：
Kannan S
中科院分区：
文献类型：
--
作者：
Alnasser Y;Kambhampati SP;Nance E;Rajbhandari L;Shrestha S;Venkatesan A;Kannan RM;Kannan S
聚酰胺-胺（PAMAM）树枝状聚合物是具有可调物理化学特征的多功能纳米颗粒，使其成为中枢神经系统（CNS）靶向药物递送的有希望的候选者。全身给药的树枝状聚合物已被证明定位于活化的神经胶质细胞中，其介导CNS中的神经炎症。这些树枝状聚合物将药物特异性地递送到活化的小胶质细胞，在多种脑损伤模型中产生显著的神经改善，包括在新生兔脑瘫模型中。为了进一步了解树枝状聚合物细胞摄取的机制，我们利用从新生兔分离的原代胶质细胞的体外模型来评估活化和非活化胶质细胞对羟基封端的第4代PAMAM树枝状聚合物（D4-OH）摄取的差异。我们使用荧光标记的D4-OH（D-Cy 5）作为研究树枝状聚合物摄取机制的工具。使用共聚焦显微镜和流式细胞术通过荧光定量测定D4-OH PAMAM树枝状聚合物的摄取。我们的结果表明，尽管混合细胞群中的小胶质细胞在该体外系统中表现出对树枝状聚合物的早期吸收，但与静息小胶质细胞相比，激活的小胶质细胞吸收更多的树枝状聚合物。星形胶质细胞表现出延迟和有限的摄取。我们还说明了不同的途径抑制剂之间的休息和激活的小胶质细胞的摄取机制的差异。静息和活化的小胶质细胞主要采用内吞途径，这在活化的小胶质细胞中得到增强。此外，我们证明了羟基终止的树枝状聚合物被初级小胶质细胞使用其他机制，包括胞饮，小窝，和水通道的树枝状聚合物摄取。
Polyamidoamine (PAMAM) dendrimers are multifunctional nanoparticles with tunable physicochemical features, making them promising candidates for targeted drug delivery in the central nervous system (CNS). Systemically administered dendrimers have been shown to localize in activated glial cells, which mediate neuroinflammation in the CNS. These dendrimers delivered drugs specifically to activated microglia, producing significant neurological improvements in multiple brain injury models, including in a neonatal rabbit model of cerebral palsy. To gain further insight into the mechanism of dendrimer cell uptake, we utilized an in vitro model of primary glial cells isolated from newborn rabbits to assess the differences in hydroxyl-terminated generation 4 PAMAM dendrimer (D4-OH) uptake by activated and non-activated glial cells. We used fluorescently-labelled D4-OH (D-Cy5) as a tool for investigating the mechanism of dendrimer uptake. D4-OH PAMAM dendrimer uptake was determined by fluorescence quantification using confocal microscopy and flow cytometry. Our results indicate that although microglial cells in the mixed cell population demonstrate early uptake of dendrimers in this in vitro system, activated microglia take up more dendrimer compared to resting microglia. Astrocytes showed delayed and limited uptake. We also illustrated the differences in mechanism of uptake between resting and activated microglia using different pathway inhibitors. Both resting and activated microglia primarily employed endocytotic pathways, which are enhanced in activated microglial cells. Additionally, we demonstrated that hydroxyl terminated dendrimers are taken up by primary microglia using other mechanisms including pinocytosis, caveolae, and aquaporin channels for dendrimer uptake.
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影响因子：
4.4
作者：
Kambhampati, Siva P.;Clunies-Ross, Alexander J. M.;Lutty, Gerard
通讯作者：
Lutty, Gerard
DOI：
10.3390/molecules21040428
发表时间：
2016-03-29
期刊：
Molecules (Basel, Switzerland)
影响因子：
--
作者：
Enciso AE;Neun B;Rodriguez J;Ranjan AP;Dobrovolskaia MA;Simanek EE
通讯作者：
Simanek EE
影响因子：
--
作者：
Labieniec, Magdalena;Watala, Cezary
通讯作者：
Watala, Cezary
影响因子：
17.1
作者：
Kannan S;Dai H;Navath RS;Balakrishnan B;Jyoti A;Janisse J;Romero R;Kannan RM
通讯作者：
Kannan RM
影响因子：
4.9
作者：
Albertazzi, Lorenzo;Serresi, Michela;Beltram, Fabio
通讯作者：
Beltram, Fabio