title: Magnetically-responsive, multifunctional drug delivery nanoparticles for elastic matrix regenerative repair.
abstract: Arresting or regressing growth of abdominal aortic aneurysms (AAAs), localized expansions of the abdominal aorta are contingent on inhibiting chronically overexpressed matrix metalloproteases (MMPs)-2 and -9 that disrupt elastic matrix within the aortic wall, concurrent with providing a stimulus to augmenting inherently poor auto-regeneration of these matrix structures. In a recent study we demonstrated that localized, controlled and sustained delivery of doxycycline (DOX; a tetracycline-based antibiotic) from poly(lactic-co-glycolic acid) nanoparticles (PLGA NPs), enhances elastic matrix deposition and MMP-inhibition at a fraction of the therapeutically effective oral dose. The surface functionalization of these NPs with cationic amphiphiles, which enhances their arterial uptake, was also shown to have pro-matrix regenerative and anti-MMP effects independent of the DOX. Based on the hypothesis that the incorporation of superparamagnetic iron oxide NPs (SPIONs) within these PLGA NPs would enhance their targetability to the AAA site under an applied external magnetic field, we sought to evaluate the functional effects of NPs co-encapsulating DOX and SPIONs (DOX-SPION NPs) on elastic matrix regeneration and MMP synthesis/activity in vitro within aneurysmal smooth muscle cell (EaRASMC) cultures. The DOX-SPION NPs were mobile under an applied external magnetic field, while enhancing elastic matrix deposition 1.5-2-fold and significantly inhibiting MMP-2 synthesis and MMP-2 and -9 activities, compared to NP-untreated control cultures. These results illustrate that the multifunctional benefits of NPs are maintained following SPION co-incorporation. Additionally, preliminary studies carried out demonstrated enhanced targetability of SPION-loaded NPs within proteolytically-disrupted porcine carotid arteries ex vivo, under the influence of an applied external magnetic field. Thus, this dual-agent loaded NP system proffers a potential non-surgical option for treating small growing AAAs, via controlled and sustained drug release from multifunctional, targetable nanocarriers.
  
  
  
  Proactive screening of high risk elderly patients now enables early detection of abdominal aortic aneurysms (AAAs). There are no established drug-based therapeutic alternatives to surgery for AAAs, which is unsuitable for many elderly patients, and none which can achieve restore disrupted and lost elastic matrix in the AAA wall, which is essential to achieve growth arrest or regression. We have developed a first generation design of polymer nanoparticles (NPs) for AAA tissue localized delivery of doxycycline, a modified tetracycline drug at low micromolar doses at which it provides both pro-elastogenic and anti-proteolytic benefits that can augment elastic matrix regenerative repair. The nanocarriers themselves are also uniquely chemically functionalized on their surface to also provide them pro-elastin-regenerative & anti-matrix degradative properties. To provide an active driving force for efficient uptake of intra-lumenally infused NPs to the AAA wall, in this work, we have rendered our polymer NPs mobile in an applied magnetic field via co-incorporation of super-paramagnetic iron oxide NPs. We demonstrate that such modifications significantly improve wall uptake of the NPs with no significant changes to their physical properties and regenerative benefits. Such NPs can potentially stimulate structural repair in the AAA wall following one time infusion to delay or prevent AAA growth to rupture. The therapy can provide a non-surgical treatment option for high risk AAA patients.
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic, 9500 Euclid Avenue, ND 20, Cleveland, OH 44195, USA.
authors.forename: Balakrishnan
authors.initials: B
authors.lastname: Sivaraman
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic, 9500 Euclid Avenue, ND 20, Cleveland, OH 44195, USA; Department of Biology, University of Akron, Akron, OH, USA.
authors.forename: Ganesh
authors.initials: G
authors.lastname: Swaminathan
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic, 9500 Euclid Avenue, ND 20, Cleveland, OH 44195, USA.
authors.forename: Lee
authors.initials: L
authors.lastname: Moore
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic, 9500 Euclid Avenue, ND 20, Cleveland, OH 44195, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Fox
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic, 9500 Euclid Avenue, ND 20, Cleveland, OH 44195, USA.
authors.forename: Dhruv
authors.initials: D
authors.lastname: Seshadri
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic, 9500 Euclid Avenue, ND 20, Cleveland, OH 44195, USA.
authors.forename: Shataakshi
authors.initials: S
authors.lastname: Dahal
authors.affiliation: Department of Cell Biology and Physiology, Washington University, St. Louis, MO, USA; Department of Molecular Medicine, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, OH, USA.
authors.forename: Ivan
authors.initials: I
authors.lastname: Stoilov
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic, 9500 Euclid Avenue, ND 20, Cleveland, OH 44195, USA.
authors.forename: Maciej
authors.initials: M
authors.lastname: Zborowski
authors.affiliation: Department of Cell Biology and Physiology, Washington University, St. Louis, MO, USA; Department of Molecular Medicine, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, OH, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Mecham
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic, 9500 Euclid Avenue, ND 20, Cleveland, OH 44195, USA; Department of Cell Biology and Physiology, Washington University, St. Louis, MO, USA; Department of Molecular Medicine, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, OH, USA. Electronic address: ramamua@ccf.org.
authors.forename: Anand
authors.initials: A
authors.lastname: Ramamurthi
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003692:Delayed-Action Preparations / Q000008:administration & dosage* / Q000737:chemistry / Q000528:radiation effects; D003911:Dextrans / Q000008:administration & dosage* / Q000528:radiation effects; D004318:Doxycycline / Q000008:administration & dosage* / Q000737:chemistry; D005109:Extracellular Matrix / Q000187:drug effects / Q000528:radiation effects; D060526:Magnetic Fields; D058185:Magnetite Nanoparticles / Q000008:administration & dosage* / Q000528:radiation effects; D008297:Male; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000528:radiation effects*; D053769:Nanocapsules / Q000008:administration & dosage / Q000737:chemistry / Q000528:radiation effects; D011829:Radiation Dosage; D051381:Rats; D017207:Rats, Sprague-Dawley; D012038:Regeneration / Q000187:drug effects* / Q000528:radiation effects
chemical_list: D003692:Delayed-Action Preparations; D003911:Dextrans; D058185:Magnetite Nanoparticles; D053769:Nanocapsules; C097921:ferumoxtran-10; D004318:Doxycycline
keywords: Aortic aneurysms; Drug delivery; Elastic matrix; Magnetic targeting; Nanoparticles; Regenerative matrix repair; Smooth muscle cells