Smooth muscle cell seeding of decellularized scaffolds: the importance of bioreactor preconditioning to development of a more native architecture for tissue-engineered blood vessels.

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.

[1]  Z. Naito,et al.  Effects of extracellular matrix on phenotype modulation and MAPK transduction of rat aortic smooth muscle cells in vitro. , 2000, Experimental and molecular pathology.

[2]  J. Pickering,et al.  Evidence for a role of collagen synthesis in arterial smooth muscle cell migration. , 1998, The Journal of clinical investigation.

[3]  Young Ha Kim,et al.  Elastic biodegradable poly(glycolide-co-caprolactone) scaffold for tissue engineering. , 2003, Journal of biomedical materials research. Part A.

[4]  L A Geddes,et al.  Small intestinal submucosa as a large diameter vascular graft in the dog. , 1989, The Journal of surgical research.

[5]  E. Chaikof,et al.  Regulation of syndecan-4 expression with mechanical stress during the development of angioplasty-induced intimal thickening. , 2002, Journal of vascular surgery.

[6]  Chrysanthi Williams,et al.  Small-diameter artificial arteries engineered in vitro. , 2005, Circulation research.

[7]  A. Ratcliffe.  Tissue engineering of vascular grafts. , 2000, Matrix biology : journal of the International Society for Matrix Biology.

[8]  R. Nerem,et al.  Altered response of vascular smooth muscle cells to exogenous biochemical stimulation in two- and three-dimensional culture. , 2003, Experimental cell research.

[9]  Jan Feijen,et al.  Biological characterisation of vascular grafts cultured in a bioreactor. , 2006, Biomaterials.

[10]  I. Nishio,et al.  Effect of centrifugal force and catecholamines on glycosaminoglycans synthesis of vascular smooth muscle cells in culture. , 1990, Atherosclerosis.

[11]  Masood A. Machingal,et al.  Engineering of Large Diameter Vessels , 2011 .

[12]  Julian B. Chaudhuri,et al.  Vascular Tissue Engineering: Bioreactor Design Considerations for Extended Culture of Primary Human Vascular Smooth Muscle Cells , 2007, ASAIO journal.

[13]  Laura E Niklason,et al.  Blood vessels engineered from human cells. , 2006, Trends in cardiovascular medicine.

[14]  N. L'Heureux,et al.  Human tissue-engineered blood vessels for adult arterial revascularization , 2007, Nature Medicine.

[15]  A. Goldberg,et al.  TNF‐α increases ubiquitin‐conjugating activity in skeletal muscle by up‐regulating UbcH2/E220k , 2003 .

[16]  R. Virmani,et al.  Pathological Correlates of Late Drug-Eluting Stent Thrombosis: Strut Coverage as a Marker of Endothelialization , 2007, Circulation.

[17]  M. Yamamoto,et al.  Type I collagen promotes modulation of cultured rabbit arterial smooth muscle cells from a contractile to a synthetic phenotype. , 1993, Experimental cell research.

[18]  R. Nerem,et al.  Genetic modification of smooth muscle cells to control phenotype and function in vascular tissue engineering. , 2004, Tissue engineering.

[19]  Laura E. Niklason,et al.  An Ultrastructural Analysis of Collagen in Tissue Engineered Arteries , 2007, Annals of Biomedical Engineering.

[20]  R. Guignard,et al.  Mechanical loading modulates the differentiation state of vascular smooth muscle cells. , 2006, Tissue engineering.

[21]  Christopher J. O’Callaghan,et al.  Mechanical Strain–Induced Extracellular Matrix Production by Human Vascular Smooth Muscle Cells: Role of TGF-&bgr;1 , 2000, Hypertension.

[22]  G. Di Luozzo,et al.  Vascular smooth muscle cell effect on endothelial cell endothelin-1 production. , 2000, Journal of vascular surgery.

[23]  U. A. Stock,et al.  Tissue Engineering of Ovine Aortic Blood Vessel Substitutes Using Applied Shear Stress and Enzymatically Derived Vascular Smooth Muscle Cells , 2004, Annals of Biomedical Engineering.

[24]  Dual cell seeding and the use of zymogen tissue plasminogen activator to improve cell retention on polytetrafluoroethylene grafts. , 2001, Journal of vascular surgery.

[25]  D. Vorp,et al.  Characterization of the response of bone marrow-derived progenitor cells to cyclic strain: implications for vascular tissue-engineering applications. , 2004, Tissue engineering.

[26]  J. Wood,et al.  Combined Vascular Endothelial Growth Factor and Platelet-Derived Growth Factor Inhibition in Rat Cardiac Allografts: Beneficial Effects on Inflammation and Smooth Muscle Cell Proliferation , 2005, Transplantation.

[27]  Byung-Soo Kim,et al.  Mechano-active tissue engineering of vascular smooth muscle using pulsatile perfusion bioreactors and elastic PLCL scaffolds. , 2005, Biomaterials.

[28]  K. Peck,et al.  Genomic analysis of smooth muscle cells in three‐dimensional collagen matrix , 2003, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[29]  K. Dora.  Cell-cell communication in the vessel wall , 2001, Vascular medicine.

[30]  A. Lumsden,et al.  Roles of Hemodynamic Forces in Vascular Cell Differentiation , 2005, Annals of Biomedical Engineering.

[31]  L. Germain,et al.  A human tissue‐engineered vascular media: a new model for pharmacological studies of contractile responses , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[32]  B. Sumpio,et al.  Cyclic stretch induces the expression of vascular endothelial growth factor in vascular smooth muscle cells. , 2001, Endothelium : journal of endothelial cell research.

[33]  Robert M Nerem,et al.  Equibiaxial strain stimulates fibroblastic phenotype shift in smooth muscle cells in an engineered tissue model of the aortic wall. , 2006, Biomaterials.

[34]  R T Tranquillo,et al.  A fibrin-based arterial media equivalent. , 2003, Journal of biomedical materials research. Part A.

[35]  Cornelius Borst,et al.  Mechanical properties of porcine and human arteries: implications for coronary anastomotic connectors. , 2003, The Annals of thoracic surgery.

[36]  K. J. Grande-Allen,et al.  Effects of static and cyclic loading in regulating extracellular matrix synthesis by cardiovascular cells. , 2006, Cardiovascular research.

[37]  Laura E. Niklason,et al.  Mechanical Properties and Compositions of Tissue Engineered and Native Arteries , 2007, Annals of Biomedical Engineering.

[38]  J. Cha,et al.  Time-dependent modulation of alignment and differentiation of smooth muscle cells seeded on a porous substrate undergoing cyclic mechanical strain. , 2006, Artificial organs.

[39]  E Bell,et al.  A blood vessel model constructed from collagen and cultured vascular cells. , 1986, Science.

[40]  H. Ives,et al.  Mechanical strain increases PDGF-B and PDGF beta receptor expression in vascular smooth muscle cells. , 1999, Biochemical and biophysical research communications.

[41]  M G Walker,et al.  Tissue engineering of blood vessels , 2006, The British journal of surgery.

[42]  R Langer,et al.  Functional arteries grown in vitro. , 1999, Science.

[43]  Anthony Atala,et al.  Functional small-diameter neovessels created using endothelial progenitor cells expanded ex vivo , 2001, Nature Medicine.

[44]  Axel Haverich,et al.  Tissue engineering of small diameter vascular grafts. , 2002, European journal of vascular and endovascular surgery : the official journal of the European Society for Vascular Surgery.

[45]  W. Zeng,et al.  Development of anti-atherosclerotic tissue-engineered blood vessel by A20-regulated endothelial progenitor cells seeding decellularized vascular matrix. , 2008, Biomaterials.

[46]  M. Merrilees,et al.  The effect of centrifugal force on glycosaminoglycan production by aortic smooth muscle cells in culture. , 1977, Atherosclerosis.

[47]  Jennifer S. Park,et al.  Regulation of vascular smooth muscle cells and mesenchymal stem cells by mechanical strain. , 2006, Molecular & cellular biomechanics : MCB.

[48]  D. Mooney,et al.  Cyclic strain inhibits switching of smooth muscle cells to an osteoblast‐like phenotype , 2003, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[49]  Robert M. Nerem,et al.  Dynamic Mechanical Conditioning of Collagen-Gel Blood Vessel Constructs Induces Remodeling In Vitro , 2000, Annals of Biomedical Engineering.

[50]  Michael Horrocks,et al.  Endothelial and Smooth Muscle Cell Seeding onto Processed Ex Vivo Arterial Scaffolds Using 3D Vascular Bioreactors , 2004, ASAIO journal.

[51]  M. Jacob,et al.  Effect of elastin peptides on ion fluxes in mononuclear cells, fibroblasts, and smooth muscle cells. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[52]  Cheng Li,et al.  In vitro and in vivo studies of ePTFE vascular grafts treated with P15 peptide , 2005, Journal of biomaterials science. Polymer edition.

[53]  Jennifer S. Park,et al.  Differential effects of equiaxial and uniaxial strain on mesenchymal stem cells , 2004, Biotechnology and bioengineering.

[54]  Jan P Stegemann,et al.  Biomechanics and Mechanotransduction in Cells and Tissues Mechanical , biochemical , and extracellular matrix effects on vascular smooth muscle cell phenotype , 2005 .