title: Fibrinolytic PLGA nanoparticles for slow clot lysis within abdominal aortic aneurysms attenuate proteolytic loss of vascular elastic matrix.
abstract: Abdominal aortic aneurysms (AAAs) involve chronic overexpression of proteases in the aortic wall that result in disruption of elastic fibers and consequent loss of vessel elasticity. Nearly 75% of AAAs contain flow-obstructing, fibrin-rich intraluminal thrombi (ILT), which act as a) a bioinert shield, protecting the underlying AAA wall from high hemodynamic stresses, and b) a reservoir of inflammatory cells and proteases that cause matrix breakdown. For these reasons, restoring flow through the aorta lumen and facilitating transmural diffusion of therapeutics from circulation to the AAA wall must be achieved by slow thrombolysis of the ILT to render it porous without rapid breakdown. Intravenously dosed tissue plasminogen activator (tPA) has been shown to rapidly lyse ILTs in acute stroke and myocardial infarctions. For future use in opening up AAA segments, in this study, we investigated the ability of tPA released from poly(lactic-co-glycolic acid) nanoparticles (PLGA NPs) to slowly lyse fibrin clots without inducing proteolytic injury and matrix synthesis-inhibitory effects on cultured rat aneurysmal smooth muscle cells (EaRASMCs). Fibrin clot lysis time was greatly extended over that in presence of exogenous tPA. Surface functionalization of NPs with a cationic amphiphile allowed them to bind to anionic fibrin clot, release tPA at a slower rate and to lyse the clot as a front proceeding outwards in unlike the more rapid and homogenous lysis that occurred due to anionic PLGA NPs. Elastic matrix content was decreased in EaRASMC cultures exposed to byproducts of clot lysis with exogenous tPA, but not tPA-NPs, and was likely due to increased proteolytic activity (MMPs, plasmin) in EaRASMC cultures exposed to exogenous tPA-lysed clots. Our results suggest that gradual ILT lysis via slow release of tPA from NPs will be likely beneficial over exogenous tPA delivery in preserving elastic matrix content and attenuating matrilysis in the adjoining AAA wall, in vivo, while rendering the ILT porous to facilitate transmural delivery of endoluminally delivered AAA therapeutics.
authors.affiliation: Department of Biomedical Engineering, The Cleveland Clinic Foundation, 9500 Euclid Avenue, ND 20, Cleveland, OH 44195, USA.
authors.forename: Balakrishnan
authors.initials: B
authors.lastname: Sivaraman
authors.affiliation: Department of Biomedical Engineering, The Cleveland Clinic Foundation, 9500 Euclid Avenue, ND 20, Cleveland, OH 44195, USA; Department of Biomedical Engineering, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Sylvester
authors.affiliation: Department of Biomedical Engineering, The Cleveland Clinic Foundation, 9500 Euclid Avenue, ND 20, Cleveland, OH 44195, USA; Department of Biomedical Engineering, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, USA. Electronic address: ramamua@ccf.org.
authors.forename: Anand
authors.initials: A
authors.lastname: Ramamurthi
mesh_terms: D000818:Animals; D017544:Aortic Aneurysm, Abdominal / Q000188:drug therapy* / Q000378:metabolism; D003692:Delayed-Action Preparations / Q000737:chemistry / Q000494:pharmacology; D004195:Disease Models, Animal; D004548:Elasticity / Q000187:drug effects; D005109:Extracellular Matrix / Q000378:metabolism*; D005343:Fibrinolytic Agents / Q000737:chemistry / Q000494:pharmacology; D019344:Lactic Acid / Q000737:chemistry / Q000494:pharmacology; D008297:Male; D011100:Polyglycolic Acid / Q000737:chemistry / Q000494:pharmacology; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D051381:Rats; D017207:Rats, Sprague-Dawley; D015912:Thrombolytic Therapy / Q000379:methods*
chemical_list: D003692:Delayed-Action Preparations; D005343:Fibrinolytic Agents; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid
keywords: Aortic aneurysms; Elastic matrix; Fibrinolysis; Nanoparticles; Regenerative matrix repair; Smooth muscle cells