title: Smooth muscle cell seeding of decellularized scaffolds: the importance of bioreactor preconditioning to development of a more native architecture for tissue-engineered blood vessels.
abstract: Vascular smooth muscle cells (VSMCs) impart important functional characteristics in the native artery and, therefore, should logically be incorporated in the development of tissue-engineered blood vessels. However, the native architecture and low porosity of naturally derived biomaterials (i.e., decellularized vessels) have impeded efforts to seed and incorporate a VSMC layer in tissue-engineered blood vessels. To this end, the goal of this study was to develop improved methods for seeding, proliferation, and maturation of VSMCs on decellularized porcine carotid arteries. Decellularized vessels were prepared in the absence and presence of the adventitial layer, and statically seeded with a pipette containing a suspension of rat aortic VSMCs. After cell seeding, recellularized engineered vessels were placed in a custom bioreactor system for 1-2 weeks to enhance cellular proliferation, alignment, and maturation. Initial attachment of VSMCs was dramatically enhanced by removing the adventitial layer of the decellularized porcine artery. Moreover, cyclic bioreactor conditioning (i.e., flow and pressure) resulted in increased VSMC proliferation and accelerated formation of a muscularized medial layer in the absence of the adventitial layer during the first week of preconditioning. Fura-2-based digital imaging microscopy revealed marked and reproducible depolarization-induced calcium mobilization after bioreactor preconditioning in the absence, but not in the presence, of the adventitia. The major finding of this investigation is that bioreactor preconditioning accelerates the formation of a significant muscular layer on decellularized scaffolds, in particular on adventitia-denuded scaffolds. Further, the VSMC layer of bioreactor-preconditioned vessels was capable of mobilizing calcium in response to cellular depolarization. These findings represent an important first step toward the development of tissue-engineered vascular grafts that more closely mimic native vasculature.
authors.affiliation: Wake Forest Institute for Regenerative Medicine, Wake Forest University School of Medicine , Winston-Salem, North Carolina, USA.
authors.forename: Saami K
authors.initials: SK
authors.lastname: Yazdani
authors.forename: Benjamin
authors.initials: B
authors.lastname: Watts
authors.forename: Masood
authors.initials: M
authors.lastname: Machingal
authors.forename: Yagna P R
authors.initials: YP
authors.lastname: Jarajapu
authors.forename: Mark E
authors.initials: ME
authors.lastname: Van Dyke
authors.forename: George J
authors.initials: GJ
authors.lastname: Christ
mesh_terms: D000818:Animals; D019149:Bioreactors; D001808:Blood Vessels; D002478:Cells, Cultured; D032389:Myocytes, Smooth Muscle / Q000166:cytology*; D051381:Rats; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods*; D054457:Tissue Scaffolds