SAM-based cell transfer to photopatterned hydrogels for microengineering vascular-like structures.

A major challenge in tissue engineering is to reproduce the native 3D microvascular architecture fundamental for in vivo functions. Current approaches still lack a network of perfusable vessels with native 3D structural organization. Here we present a new method combining self-assembled monolayer (SAM)-based cell transfer and gelatin methacrylate hydrogel photopatterning techniques for microengineering vascular structures. Human umbilical vein cell (HUVEC) transfer from oligopeptide SAM-coated surfaces to the hydrogel revealed two SAM desorption mechanisms: photoinduced and electrochemically triggered. The former, occurs concomitantly to hydrogel photocrosslinking, and resulted in efficient (>97%) monolayer transfer. The latter, prompted by additional potential application, preserved cell morphology and maintained high transfer efficiency of VE-cadherin positive monolayers over longer culture periods. This approach was also applied to transfer HUVECs to 3D geometrically defined vascular-like structures in hydrogels, which were then maintained in perfusion culture for 15 days. As a step toward more complex constructs, a cell-laden hydrogel layer was photopatterned around the endothelialized channel to mimic the vascular smooth muscle structure of distal arterioles. This study shows that the coupling of the SAM-based cell transfer and hydrogel photocrosslinking could potentially open up new avenues in engineering more complex, vascularized tissue constructs for regenerative medicine and tissue engineering applications.

[1]  C. van Nostrum,et al.  Novel crosslinking methods to design hydrogels. , 2002, Advanced drug delivery reviews.

[2]  Bruce K Milthorpe,et al.  Engineering thick tissues--the vascularisation problem. , 2007, European cells & materials.

[3]  A. Khademhosseini,et al.  Cell-laden microengineered gelatin methacrylate hydrogels. , 2010, Biomaterials.

[4]  H. Kaji,et al.  Transfer of two-dimensional patterns of human umbilical vein endothelial cells into fibrin gels to facilitate vessel formation. , 2010, Chemical communications.

[5]  Micah Dembo,et al.  The dynamics and mechanics of endothelial cell spreading. , 2005, Biophysical journal.

[6]  Peter T C So,et al.  Three-dimensional cellular deformation analysis with a two-photon magnetic manipulator workstation. , 2002, Biophysical journal.

[7]  M. Tarlov,et al.  UV photopatterning of alkanethiolate monolayers self-assembled on gold and silver , 1993 .

[8]  Narutoshi Hibino,et al.  Cell-seeding techniques in vascular tissue engineering. , 2010, Tissue engineering. Part B, Reviews.

[9]  Stefan J. Janusz,et al.  Photooxidation of self-assembled monolayers by exposure to light of wavelength 254 nm: a static SIMS study. , 2005, The journal of physical chemistry. B.

[10]  T. Okano,et al.  Cell Attachment–Detachment Control on Temperature-Responsive Thin Surfaces for Novel Tissue Engineering , 2010, Annals of Biomedical Engineering.

[11]  T. Okano,et al.  Mechanism of cell detachment from temperature-modulated, hydrophilic-hydrophobic polymer surfaces. , 1995, Biomaterials.

[12]  D. Laird Connexin phosphorylation as a regulatory event linked to gap junction internalization and degradation. , 2005, Biochimica et biophysica acta.

[13]  J. Vörös,et al.  The quantification of single cell adhesion on functionalized surfaces for cell sheet engineering. , 2010, Biomaterials.

[14]  H. Hattori,et al.  Encouraging effect of cadherin-mediated cell-cell junctions on transfer printing of micropatterned vascular endothelial cells. , 2009, Langmuir : the ACS journal of surfaces and colloids.

[15]  Kyongbum Lee,et al.  Vascularization strategies for tissue engineering. , 2009, Tissue engineering. Part B, Reviews.

[16]  J. Kohn,et al.  Tissue spreading on implantable substrates is a competitive outcome of cell–cell vs. cell–substratum adhesivity , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[17]  G. Whitesides,et al.  Self-assembled monolayers of thiolates on metals as a form of nanotechnology. , 2005, Chemical reviews.

[18]  L. Niklason,et al.  Scaffold-free vascular tissue engineering using bioprinting. , 2009, Biomaterials.

[19]  Joe Tien,et al.  Formation of perfused, functional microvascular tubes in vitro. , 2006, Microvascular research.

[20]  S. Chien,et al.  Effects of disturbed flow on vascular endothelium: pathophysiological basis and clinical perspectives. , 2011, Physiological reviews.

[21]  A. Leturque,et al.  Ultraviolet A radiation transiently disrupts gap junctional communication in human keratinocytes. , 2003, American journal of physiology. Cell physiology.

[22]  Satoru Takeda,et al.  In vitro formation of capillary networks using optical lithographic techniques. , 2007, Biochemical and biophysical research communications.

[23]  Alison P McGuigan,et al.  Vascularized Organoid Engineered by Modular Assembly Enables Blood Perfusion , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[24]  J. Hemminger,et al.  Photooxidation of thiols in self-assembled monolayers on gold , 1993 .

[25]  K. Rowlen,et al.  OZONE-INDUCED OXIDATION OF SELF-ASSEMBLED DECANETHIOL : CONTRIBUTING MECHANISM FOR PHOTOOXIDATION ? , 1998 .

[26]  A. Khademhosseini,et al.  Microscale technologies for tissue engineering and biology. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[27]  Elisabetta Dejana,et al.  The control of vascular integrity by endothelial cell junctions: molecular basis and pathological implications. , 2009, Developmental cell.

[28]  A. Goldstein,et al.  Examination of membrane rupture as a mechanism for mammalian cell detachment from fibronectin-coated biomaterials. , 2003, Journal of biomedical materials research. Part A.

[29]  Kytai Truong Nguyen,et al.  Photopolymerizable hydrogels for tissue engineering applications. , 2002, Biomaterials.

[30]  Masayuki Yamato,et al.  Cell sheet technology and cell patterning for biofabrication , 2009, Biofabrication.

[31]  D. Wirtz,et al.  Programmed subcellular release for studying the dynamics of cell detachment , 2009, Nature Methods.

[32]  J. Fukuda,et al.  Spatio-temporal detachment of single cells using microarrayed transparent electrodes. , 2011, Biomaterials.

[33]  Shengyuan Yang,et al.  Micromachined force sensors for the study of cell mechanics , 2005 .

[34]  A. Khademhosseini,et al.  A cell-laden microfluidic hydrogel. , 2007, Lab on a chip.

[35]  Claus Duschl,et al.  Control of cell detachment in a microfluidic device using a thermo-responsive copolymer on a gold substrate. , 2007, Lab on a chip.

[36]  Robert Langer,et al.  Long-Term Stability of Self-Assembled Monolayers in Biological Media , 2003 .

[37]  James G Truslow,et al.  The role of cyclic AMP in normalizing the function of engineered human blood microvessels in microfluidic collagen gels. , 2010, Biomaterials.

[38]  Kristi S Anseth,et al.  Photocrosslinking of gelatin macromers to synthesize porous hydrogels that promote valvular interstitial cell function. , 2009, Tissue engineering. Part A.

[39]  G. Leggett,et al.  Oxidation of self-assembled monolayers by UV light with a wavelength of 254 nm. , 2001, Journal of the American Chemical Society.

[40]  B. Isakson,et al.  Biological and biophysical properties of vascular connexin channels. , 2009, International review of cell and molecular biology.

[41]  Shaoyi Jiang,et al.  Ultra-low fouling peptide surfaces derived from natural amino acids. , 2009, Biomaterials.

[42]  M Cornelissen,et al.  Structural and rheological properties of methacrylamide modified gelatin hydrogels. , 2000, Biomacromolecules.

[43]  James G Truslow,et al.  Effect of mechanical factors on the function of engineered human blood microvessels in microfluidic collagen gels. , 2010, Biomaterials.

[44]  Ali Khademhosseini,et al.  Directed assembly of cell-laden hydrogels for engineering functional tissues , 2010, Organogenesis.

[45]  A. Khademhosseini,et al.  Electrochemical desorption of self-assembled monolayers for engineering cellular tissues. , 2009, Biomaterials.

[46]  Hiroaki Suzuki,et al.  Engineering of capillary-like structures in tissue constructs by electrochemical detachment of cells. , 2010, Biomaterials.