Integrated drug-in-cell therapy of in-stent restenosis

 描述（由申请人提供）：由于缺乏有效性和安全性相结合的治疗方法，通过血管成形术治疗以缓解动脉粥样硬化性阻塞的动脉损伤触发的再狭窄（再狭窄）仍然是一个挑战。旨在加速血管愈合和内皮再生的策略有可能预防再狭窄，同时避免目前使用的治疗的副作用。然而，为了实现这一潜力，需要显著提高其效率。该提案涉及基于内皮细胞与载药磁性纳米载体的双重功能化的整合的细胞内药物抗再狭窄策略，以向它们提供支架靶向递送的能力和增强的抗再狭窄有效性。后者是通过诱导和激活内皮一氧化氮合酶与雌二醇配制在磁性纳米粒子（MNP）的形式，其前体，雌二醇苯甲酸酯（E2 Bz）。目前的研究将建立可行性，使用这种方法来防止支架内再狭窄，并实现促进再内皮化的动脉粥样硬化大鼠模型再现人类diseases.The的关键特征，将解决以下具体目标：目的1：药物在细胞的方法评估和优化的文化和共培养研究。E2 Bz负载的生物可降解的MNP将被配制，并且它们的大小、组成、磁性和药物释放曲线将被表征。MNP的细胞相容性、内化和降解动力学以及MNP功能化内皮细胞的E2 Bz诱导的一氧化氮合成及其对共培养平滑肌细胞的生长抑制作用将是Aim 1实验的主要终点。目的2：靶向、再内皮化和疗效研究。将在大鼠动脉粥样硬化/颈动脉支架植入模型中研究磁靶向递送功能化细胞后的动脉定位、组织分布和内皮恢复。程度 在递送后两周通过计算机化形态测定法测定再狭窄的发生率，并且使用用负载有空白MNP的细胞处理的动物作为对照来描绘细胞功能化的药理学作用。

 DESCRIPTION (provided by applicant): Injury-triggered renarrowing (restenosis) of arteries treated with angioplasty to relieve atherosclerotic obstruction remains a challenge due to a lack of therapies combining adequate efficacy and safety. Strategies aimed at accelerated vascular healing and endothelium regeneration have potential to prevent restenosis while avoiding the side effects of the currently used therapies. However, in order to realize this potential a significant improvement in their efficiency is required. This proposal is concerned with an integrated drug-in-cell antirestenotic strategy based on dual functionalization of endothelial cell with drug-loaded magnetic nanocarriers to provide them with capacity for stent-targeted delivery and enhanced antirestenotic effectiveness. The latter is achieved by inducing and activating endothelial nitric oxide synthase with estradiol formulated in magnetic nanoparticles (MNP) in the form of its precursor, estradiol benzoate (E2Bz). The present studies will establish feasibilit of using this approach for preventing in-stent restenosis and achieving facilitated reendothelialization in a validated rat model of atherosclerosis reproducing key features of human disease .The following specific aims will be addressed: AIM 1: Drug-in-cell approach evaluation and optimization in culture and co-culture studies. E2Bz-loaded biodegradable MNP will be formulated, and their size, composition, magnetic properties and drug release profile will be characterized. Cell compatibility, internalization and degradation kinetics of MNP, as well as E2Bz-induced nitric oxide synthesis by MNP-functionalized endothelial cells and its growth inhibitory effect on co-cultured smooth muscle cells will be the main endpoints of the Aim 1 experiments. AIM 2: Targeting, reendothelialization and efficacy studies. Arterial localization, tissue distribution and endothelium recovery after magnetically targeted delivery of functionalized cells will be studied in the rat atherosclerosis/ carotid stenting model. The extent of restenosis will be determined two weeks post-delivery by computerized morphometry, and the pharmacological effect of cell functionalization will be delineated using animals treated with cells loaded with blank MNP as controls.