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(Circulation. 2007;116:I-55 – I-63.)
© 2007 American Heart Association, Inc.

Cell Transplantation and Tissue Regeneration

Protein Precoating of Elastomeric Tissue-Engineering Scaffolds Increased Cellularity, Enhanced Extracellular Matrix Protein Production, and Differentially Regulated the Phenotypes of Circulating Endothelial Progenitor Cells

Virna L. Sales, MD; George C. Engelmayr, Jr, PhD; John A. Johnson, Jr, BS; Jin Gao, PhD; Yadong Wang, PhD; Michael S. Sacks, PhD; John E. Mayer, Jr, MD

From the Department of Cardiac Surgery (V.L.S., J.A.J., J.E.M.), Children’s Hospital Boston, Harvard Medical School, Boston, Mass; Children’s Hospital Boston, Department of Bioengineering (G.C.E., M.S.S.), Engineered Tissue Mechanics Lab, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pa; Coulter Department of Biomedical Engineering (J.G., Y.W.), Georgia Institute of Technology, Emory University, Atlanta, Ga; Harvard-MIT Division of Health Sciences and Technology (G.C.E.), Massachusetts Institute of Technology, Cambridge, Mass.

Correspondence to Virna L. Sales, MD, Department of Cardiac Surgery, Children’s Hospital Boston, 300 Longwood Ave, Boston, MA 02115. E-mail virna.sales{at}cardio.chboston.org

Background— Optimal cell sources and scaffold-cell interactions remain unanswered questions for tissue engineering of heart valves. We assessed the effect of different protein precoatings on a single scaffold type (elastomeric poly (glycerol sebacate)) with a single cell source (endothelial progenitor cells).

Methods and Results— Elastomeric poly (glycerol sebacate) scaffolds were precoated with laminin, fibronectin, fibrin, collagen types I/III, or elastin. Characterized ovine peripheral blood endothelial progenitor cells were seeded onto scaffolds for 3 days followed by 14 days incubation. Endothelial progenitor cells were CD31⁺, vWF⁺, and -SMA^– before seeding confirmed by immunohistochemistry and immunoblotting. Both precoated and uncoated scaffolds demonstrated surface expression of CD31⁺ and vWF⁺, -SMA⁺ cells and were found in the "interstitium" of the scaffold. Protein precoating of elastomeric poly (glycerol sebacate) scaffolds revealed significantly increased cellularity and altered the phenotypes of endothelial progenitor cells, which resulted in changes in cellular behavior and extracellular matrix production. Moreover, mechanical flexure testing demonstrated decreased effective stiffness of the seeded scaffolds compared with unseeded controls.

Conclusions— Scaffold precoating with extracellular matrix proteins can allow more precise "engineering" of cellular behavior in the development of tissue engineering of heart valves constructs by altering extracellular matrix production and cell phenotype.

Key Words: extracelllular matrix • pulmonary valve • stem cells • transplantation