Uniform field controlled magnetic cell targeting to stents

均匀场控制的磁性细胞靶向支架

• 批准号: 8625329
• 负责人: Michael Chorny
• 金额: $ 41.04万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-16 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8625329
• 关键词: Address; Adverse effects; Aftercare; Angioplasty; Animals; Arterial Injury; Arteries; Autologous; Biological Assay; Bioluminescence; Blood Vessels; Cell Line; Cell Proliferation; Cell Survival; Cells; Characteristics; Cultured Cells; Dose; Drug Formulations; Drug or chemical Tissue Distribution; Endothelial Cells; Energy Transfer; Family suidae; Firefly Luciferases; Generations; Growth; Healed; Homing; Image; In Vitro; Kinetics; Label; Luciferases; Magnetism; Measurement; Measures; Mediating; Methodology; Methods; Modeling; Modification; Patients; Polymers; Precipitation; Procedures; Process; Property; Publications; Rattus; Relative (related person); Reporter; Research; Scheme; Site; Stenosis; Stents; Techniques; Therapeutic; Therapeutic Effect; Toxic effect; Vascular Diseases; base; biomaterial compatibility; computerized; healing; implantation; in vivo; in vivo Model; injured; innovation; luminescence; morphometry; nanoparticle; novel; particle; physical property; prevent; programs; repaired; research study; restenosis; targeted delivery; uptake

DESCRIPTION (provided by applicant): The proposed research will explore the hypothesis that magnetically guided targeting of endothelial cells (EC) to stented arteries can reduce vessel reocclusion (restenosis) following stent angioplasty. Re-endothelialization oriented strategies have potential to prevent restenosis while avoiding the side effects of the currently used therapies. However, in order to fully realize this potential a significant improvement in cell delivery efficiency is required. This study will investigate two novel ideas: 1) the application of magnetic nanoparticles (MNP) for providing EC with magnetic responsiveness adequate for their targeted delivery to stented arteries; and 2) using a uniform field-controlled magnetic targeting of EC to reversibly magnetizable stents for enhanced reendothelialization and inhibition of restenosis post stenting. The present studies will address the following specific aims: AIM 1: MNP characterization with respect to their physical properties, cell compatibility, uptake and degradation kinetics. Biodegradable fluorescent-labeled MNP will be formulated using a modification of the polymer precipitation approach. The size, composition and magnetic properties of MNP will be characterized, and their cell compatibility, internalization and degradation kinetics will be the main endpoints of the Aim 1 experiments. The magnetic responsiveness of MNP and EC treated with MNP will be measured by alternating gradient magnetometer. The kinetic studies will be performed in cultured cells using fluorimetry and global FQrster Resonance Energy Transfer measurements. Cell compatibility of MNP will be determined using fluorimetric cell toxicity assays. AIM 2: EC targeting and reendothelialization studies. Arterial localization and tissue distribution after magnetic vs. non- magnetic delivery of MNP-impregnated EC stably expressing firefly luciferase will be studied by in vivo bioluminescence and luminometry, respectively, in the rat carotid stenting model. The local kinetic profiles of stent-targeted cells will be compared 1, 7 and 28 days post treatment with endothelialization determined using a quantitative immunohistochemical strategy. AIM 3: antirestenotic efficacy of targeted EC. The extent of restenosis in animals treated under magnetic conditions with MNP-loaded EC will be determined by computerized morphometry and compared that in non-magnetic delivery and 'stenting only' controls four weeks post delivery.

描述（由申请人提供）：拟议的研究将探讨以下假设：磁引导内皮细胞（EC）靶向支架动脉可以减少支架血管成形术后血管再闭塞（再狭窄）。以再内皮化为导向的策略有可能预防再狭窄，同时避免当前使用的疗法的副作用。然而，为了充分实现这一潜力，需要显着提高细胞递送效率。本研究将研究两个新颖的想法：1）应用 磁性纳米粒子（MNP），用于为 EC 提供足够的磁响应能力，以将其定向输送到支架动脉； 2) 使用均匀场控制的 EC 磁性靶向可逆磁化支架，以增强再内皮化并抑制支架置入后的再狭窄。目前的研究将解决以下具体目标： 目标 1：MNP 的物理特性、细胞相容性、摄取和降解动力学表征。可生物降解的荧光标记 MNP 将使用聚合物沉淀方法的改进来配制。 MNP 的尺寸、组成和磁性将被表征，其细胞相容性、内化和降解动力学将成为 Aim 1 实验的主要终点。将通过交替梯度磁力计测量MNP和用MNP处理的EC的磁响应性。动力学研究将使用荧光测定法和全局 FQrster 共振能量转移测量在培养细胞中进行。 MNP 的细胞相容性将通过荧光细胞毒性测定来确定。目标 2：EC 靶向和再内皮化研究。磁性与非磁性输送后的动脉定位和组织分布 将在大鼠颈动脉支架模型中分别通过体内生物发光和发光测定法研究稳定表达萤火虫荧光素酶的 MNP 浸渍 EC。支架靶向细胞的局部动力学特征将在治疗后 1、7 和 28 天与使用定量免疫组织化学策略测定的内皮化进行比较。目标 3：靶向 EC 的抗再狭窄功效。在磁性条件下用负载 MNP 的 EC 治疗的动物的再狭窄程度将通过计算机形态测定法确定，并在分娩后 4 周比较非磁性输送和“仅支架”对照中的再狭窄程度。