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: 8459382
• 负责人: Michael Chorny
• 金额: $ 39.87万
• 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/8459382
• 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：在支架血管成形术后，磁性引导的内皮细胞（EC）靶向支架动脉可以减少血管重纹（再狭窄）。重新皮层化的策略具有预防再狭窄的潜力，同时避免了当前使用的疗法的副作用。但是，为了充分意识到这一潜力，需要对细胞输送效率进行重大提高。这项研究将研究两个新颖的想法：1） 磁性纳米颗粒（MNP），用于为EC提供足够的磁反应能力，足以将其靶向递送到支架动脉中； 2）使用EC的均匀场控制磁靶，以可逆地磁性支架，以增强支架后的再狭窄的再狭窄和抑制。本研究将解决以下特定目的：目标1：MNP表征其物理特性，细胞兼容性，摄取和降解动力学。可生物降解的荧光标记的MNP将使用聚合物沉淀方法的修饰配制。将表征MNP的大小，组成和磁性，其细胞兼容性，内在化和降解动力学将是AIM 1实验的主要终点。 MNP和EC用MNP处理的磁反应能力将通过交替的梯度磁力计测量。动力学研究将在培养的细胞中使用荧光和全局FQRSTER共振能量传递测量进行。 MNP的细胞兼容性将使用氟化细胞毒性测定确定。 AIM 2：EC靶向和再去性皮化研究。磁性与非磁性输送后的动脉定位和组织分布 在大鼠颈动脉支架支架模型中，分别研究了MNP浸渍的EC稳定表达的萤火虫荧光素酶。在治疗后1、7和28天将使用定量免疫组织化学策略确定的内皮化1、7和28天，将比较靶向支架的细胞的局部动力学特征。 AIM 3：靶向EC的抗雷神疗效。在磁性条件下以MNP负载的EC处理的动物的再狭窄程度将由计算机化的形态计算，并比较了在非磁性输送和“仅支架”控制后四周输送后进行的。