Drug Delivery and Translational Research

, Volume 8, Issue 4, pp 954–963 | Cite as

Optimizing endothelial cell functionalization for cell therapy of vascular proliferative disease using a direct contact co-culture system

  • Mark R. Battig
  • Ilia Fishbein
  • Robert J. Levy
  • Ivan S. Alferiev
  • David Guerrero
  • Michael ChornyEmail author
Original Article


Increased susceptibility to thrombosis, neoatherosclerosis, and restenosis due to incomplete regrowth of the protective endothelial layer remains a critical limitation of the interventional strategies currently used clinically to relieve atherosclerotic obstruction. Rapid recovery of endothelium holds promise for both preventing the thrombotic events and reducing post-angioplasty restenosis, providing the rationale for developing cell delivery strategies for accelerating arterial reendothelialization. The successful translation of experimental cell therapies into clinically viable treatment modalities for restoring vascular endothelium critically depends on identifying strategies for enhancing the functionality of endothelial cells (EC) derived from high cardiovascular risk patients, the target group for the majority of angioplasty procedures. Enhancing EC-associated nitric oxide (NO) synthesis by inducing overexpression of NO synthase (NOS) has shown promise as a way of increasing paracrine activity and restoring function of EC. In the present study, we developed a direct contact co-culture approach compatible with highly labile effectors, such as NO, and applied it for determining the effect of EC functionalization via NOS gene transfer on the growth of co-cultured arterial smooth muscle cells (A10 cell line) exhibiting the defining characteristics of neointimal cells. Bovine aortic endothelial cells magnetically transduced with inducible NOS-encoding adenovirus (Ad) formulated in zinc oleate-based magnetic nanoparticles (MNP[iNOSAd]) strongly suppressed growth of proliferating A10 and attenuated the stimulatory effect of a potent mitogen, platelet-derived growth factor (PDGF-BB), whereas EC functionalization with free iNOSAd or MNP formulated with a different isoform of the enzyme, endothelial NOS, was associated with lower levels of NO synthesis and less pronounced antiproliferative activity toward co-cultured A10 cells. These results show feasibility of applying magnetically facilitated gene transfer to potentiate therapeutically relevant effects of EC for targeted cell therapy of restenosis. The direct contact co-culture methodology provides a sensitive and reliable tool with potential utility for a variety of biomedical applications.


Direct contact co-culture system Stent angioplasty Nitric oxide Endothelial cell Smooth muscle cell Magnetic nanoparticle 



This research was supported by US National Heart, Lung, and Blood Institute grants T32-HL007915 (MRB), R01-HL111118, and R21-HL131016 (MC); a grant from The W.W. Smith Charitable Trust; and The Children’s Hospital of Philadelphia Research Funds including the William J. Rashkind Endowment, Erin’s Fund, and The Kibel Foundation (RJL).

Compliance with ethical standards

Competing interests

The authors declare that they have no conflict of interest.


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Copyright information

© Controlled Release Society 2017

Authors and Affiliations

  1. 1.Division of Cardiology, The Children’s Hospital of Philadelphia, and Department of PediatricsPerelman School of MedicinePhiladelphiaUSA

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