Accumulation and cellular localization of nanoparticles in an ex vivo model of acute lung injury.
Accumulation and cellular localization of nanoparticles in an ex vivo model of acute lung injury.
复制标题
急性肺损伤离体模型中纳米粒子的积累和细胞定位。
DOI：
10.1016/j.jss.2016.11.007
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Shah,AshishS
中科院分区：
文献类型：
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作者：
Grimm,JoshuaC;Zhang,Fan;Magruder,JonathanT;Crawford,ToddC;Mishra,Manoj;Rangaramanujam,KannanM;Shah,AshishS
背景纳米药物在减轻急性肺损伤（ALI）方面的益处目前尚不清楚。因此，我们引入了第四代聚酰胺胺树枝状聚合物与中性表面性质（树枝状聚合物）到我们establishedex vivoanimal模型，并试图确定其生物分布，以确定其细胞的摄取概况，并评估其作为药物输送候选人的潜力，用于治疗缺血-再灌注诱导的ALI.MethodsEight兔心肺块收获和暴露于18小时的冷缺血。然后用兔供体血再灌注心肺块。将树枝状聚合物与异硫氰酸荧光素（D-FITC）缀合用于定位和定量研究。将D-FITC（30 mg或150 mg）注射到旁路回路中，并获得基线、1小时和2小时组织样本以确定摄取百分比。低（10×）和高（40×）放大倍数的图像，获得使用共聚焦显微镜，以确认的积累，并确定细胞的dendrimer.ResultsFour目标心肺块暴露于30毫克和4至150毫克的D-FITC。在调整干重后，再灌注2小时后30和150 mg样品的平均摄取分别为灌注剂量的0.79 ± 0.16%和0.39 ± 0.22%。共聚焦成像表明dendrimer摄取上皮细胞和macrophages. Conclusionscently标记的dendrimer表现出损伤依赖性的组织积累在各种不同的细胞类型。这种独特的方法将允许缀合和递送具有减轻ALI损伤同时避免全身毒性的潜力的多种药剂。
BackgroundThe benefit of nanomedicine in mitigating acute lung injury (ALI) is currently unknown. Therefore, we introduced the generation IV polyamidoamine dendrimers with neutral surface property (dendrimer) into our establishedex vivoanimal model and sought to determine their biodistribution to define their cellular uptake profile and to evaluate their potential as a drug delivery candidate for the treatment of ischemia–reperfusion–induced ALI.MethodsEight rabbit heart–lung blocks were harvested and exposed to 18 h of cold ischemia. The heart–lung blocks were then reperfused with rabbit donor blood. Dendrimer was conjugated to fluorescein isothiocyanate (D-FITC) for localization and quantification studies. D-FITC (30 mg or 150 mg) was injected into the bypass circuit and baseline, 1- and 2-h tissue samples were obtained to determine percent uptake. Low (10×) and high (40×) magnification images were obtained using confocal microscopy to confirm the accumulation and to determine the cellular targets of the dendrimer.ResultsFour heart–lung blocks were exposed to 30 mg and four to 150 mg of D-FITC. After adjusting for dry weight, the mean uptake in the 30 and 150 mg samples after 2 h of reperfusion were 0.79 ± 0.16% and 0.39 ± 0.22% of perfused doses, respectively. Confocal imaging demonstrated dendrimer uptake in epithelial cells and macrophages.ConclusionsFluorescently tagged dendrimers demonstrated injury-dependent tissue accumulation in a variety of different cell types. This unique approach will allow conjugation to and delivery of multiple agents with the potential of mitigating ALI injury while avoiding systemic toxicity.