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.

Public Health Relevance

Reopening of narrowed blood vessels is currently achieved using invasive techniques, such as stent implantation, that may disrupt the protective inner cell lining of the artery, thereby causing severe side effects. We propose a method for repairing the protective layer using cells impregnated with biodegradable magnetic particles. This treatment is expected to enable magnetic cell targeting to the stent and promote the vessel healing process.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL111118-03
Application #
8625329
Study Section
Gene and Drug Delivery Systems Study Section (GDD)
Program Officer
Lee, Albert
Project Start
2012-04-16
Project End
2017-02-28
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
3
Fiscal Year
2014
Total Cost
$787,250
Indirect Cost
$317,250
Name
Children's Hospital of Philadelphia
Department
Type
DUNS #
073757627
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Tengood, Jillian E; Alferiev, Ivan S; Zhang, Kehan et al. (2014) Real-time analysis of composite magnetic nanoparticle disassembly in vascular cells and biomimetic media. Proc Natl Acad Sci U S A 111:4245-50