Structural and functional characterisation of poly(vinyl alcohol) and heparin hydrogels.

Synthetic scaffolds show great promise for use in tissue engineering due to their ability to mimic some aspects of the extracellular matrix, however, their use has been hindered by the lack of inherent recognition sites that are required for protein and cell interactions. Heparan sulfate (HS), a glycosaminoglycan polysaccharide present in the basement membrane and on the cell surface, binds growth factors and cytokines and enhances the signalling of these ligands by forming complexes with their receptors. This study focuses on the formation of photopolymerised hydrogels derived from methacrylated macromers of poly(vinyl alcohol) (PVA) and heparin, with the aim of imparting the growth factor activation property of heparin to the synthetic scaffolds. It was shown that the methacrylate group attachment on heparin did not result in the fragmentation of heparin molecules, and that the biological activity of the methacrylated heparin was preserved as determined by tests on its anticoagulation properties and ability to signal fibroblast growth factor-2 (FGF-2). The addition of heparin into the PVA hydrogels resulted in an increase in mass swelling ratio from 5.8 for pure PVA to 6.5 and 6.6 for PVA/heparin co-hydrogels of 19/1 and 17.5/2.5 (w/w) compositions, respectively. It is believed that heparin molecules can be added into a synthetic PVA scaffold without adversely affecting the structural and mechanical stability of the PVA scaffold. The tensile moduli of the co-hydrogels remained close to that of PVA hydrogels (61 kPa), even up to 2.5% heparin composition (PVA/hep 17.5/2.5). Finally, the co-hydrogels were found to retain the growth factor signalling activity of heparin at equilibrium.

[1]  Renato V Iozzo,et al.  Heparan sulfate: a complex polymer charged with biological activity. , 2005, Chemical reviews.

[2]  V. Fuster,et al.  Guide to Anticoagulant Therapy: Heparin: A Statement for Healthcare Professionals From the American Heart Association , 2001, Circulation.

[3]  J. Veerkamp,et al.  Development of tailor-made collagen-glycosaminoglycan matrices: EDC/NHS crosslinking, and ultrastructural aspects. , 2000, Biomaterials.

[4]  D. Papy-Garcia,et al.  Improved and simple micro assay for sulfated glycosaminoglycans quantification in biological extracts and its use in skin and muscle tissue studies. , 2003, Glycobiology.

[5]  Zhengliang L. Wu,et al.  A new strategy for defining critical functional groups on heparan sulfate , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[6]  K. Anseth,et al.  Heparin functionalized PEG gels that modulate protein adsorption for hMSC adhesion and differentiation. , 2005, Acta biomaterialia.

[7]  A. Muhlebach,et al.  New water-soluble photo crosslinkable polymers based on modified poly(vinyl alcohol) , 1997 .

[8]  Peter Carmeliet,et al.  Manipulating angiogenesis in medicine , 2004, Journal of internal medicine.

[9]  Zhengliang L. Wu,et al.  The Involvement of Heparan Sulfate (HS) in FGF1/HS/FGFR1 Signaling Complex* , 2003, The Journal of Biological Chemistry.

[10]  J. Hubbell,et al.  Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering , 2005, Nature Biotechnology.

[11]  K. Anseth,et al.  Multifunctional Hydrogels that Promote Osteogenic Human Mesenchymal Stem Cell Differentiation Through Stimulation and Sequestering of Bone Morphogenic Protein 2 , 2007 .

[12]  A. Hoffman,et al.  PEG-cross-linked heparin is an affinity hydrogel for sustained release of vascular endothelial growth factor , 2006, Journal of biomaterials science. Polymer edition.

[13]  A. Khademhosseini,et al.  Hydrogels in Biology and Medicine: From Molecular Principles to Bionanotechnology , 2006 .

[14]  P. Martens,et al.  Synthesis and characterization of degradable hydrogels formed from acrylate modified poly(vinyl alcohol) macromers , 2002 .

[15]  Laura A. Poole-Warren,et al.  A photo-crosslinked poly(vinyl alcohol) hydrogel growth factor release vehicle for wound healing applications , 2003, AAPS PharmSci.

[16]  F. Cavalieri,et al.  Study of gelling behavior of poly(vinyl alcohol)-methacrylate for potential utilizations in tissue replacement and drug delivery. , 2004, Biomacromolecules.

[17]  Glenn D Prestwich,et al.  Injectable glycosaminoglycan hydrogels for controlled release of human basic fibroblast growth factor. , 2005, Biomaterials.

[18]  K. Anseth,et al.  The effect of heparin-functionalized PEG hydrogels on three-dimensional human mesenchymal stem cell osteogenic differentiation. , 2007, Biomaterials.

[19]  Jennifer L West,et al.  Effects of nitric oxide releasing poly(vinyl alcohol) hydrogel dressings on dermal wound healing in diabetic mice , 2002, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[20]  Stephanie J Bryant,et al.  In situ forming degradable networks and their application in tissue engineering and drug delivery. , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[21]  J G Flanagan,et al.  Heparin is required for cell-free binding of basic fibroblast growth factor to a soluble receptor and for mitogenesis in whole cells , 1992, Molecular and cellular biology.

[22]  Kristi S Anseth,et al.  Synthesis and characterization of photocrosslinkable, degradable poly(vinyl alcohol)-based tissue engineering scaffolds. , 2002, Biomaterials.

[23]  Li-Fang Wang,et al.  Synthesis and characterization of chondroitin sulfate-methacrylate hydrogels , 2003 .

[24]  N Pallua,et al.  Modulation of angiogenic potential of collagen matrices by covalent incorporation of heparin and loading with vascular endothelial growth factor. , 2004, Tissue engineering.

[25]  D. Mooney,et al.  Hydrogels for tissue engineering. , 2001, Chemical reviews.

[26]  M. Sefton,et al.  In vitro platelet interactions with a heparin-polyvinyl alcohol hydrogel. , 1989, Journal of biomedical materials research.

[27]  P. Martens,et al.  Tailoring the degradation of hydrogels formed from multivinyl poly(ethylene glycol) and poly(vinyl alcohol) macromers for cartilage tissue engineering. , 2003, Biomacromolecules.

[28]  Kristi S Anseth,et al.  Material-based regulation of the myofibroblast phenotype. , 2007, Biomaterials.

[29]  Glenn D Prestwich,et al.  In situ crosslinkable hyaluronan hydrogels for tissue engineering. , 2004, Biomaterials.

[30]  Kristi S. Anseth,et al.  Synthesis and Characterization of Photopolymerized Multifunctional Hydrogels: Water-Soluble Poly(Vinyl Alcohol) and Chondroitin Sulfate Macromers for Chondrocyte Encapsulation , 2004 .

[31]  J. Melrose,et al.  Not All Perlecans Are Created Equal , 2002, The Journal of Biological Chemistry.

[32]  M. Petitou,et al.  1976-1983, a critical period in the history of heparin: the discovery of the antithrombin binding site. , 2003, Biochimie.

[33]  R. Sasisekharan,et al.  FGF‐2/fibroblast growth factor receptor/heparin‐like glycosaminoglycan interactions: a compensation model for FGF‐2 signaling , 1999, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[34]  Kristi S. Anseth,et al.  Characterization of hydrogels formed from acrylate modified poly(vinyl alcohol) macromers , 2000 .

[35]  M. Steinmetz,et al.  IL3-dependent mouse clones that express B-220 surface antigen, contain ig genes in germ-line configuration, and generate B lymphocytes in vivo , 1985, Cell.

[36]  J. Melrose,et al.  Determination of the average molecular size of glycosaminoglycans by fast protein liquid chromatography , 1993 .

[37]  R. Linhardt,et al.  Chemical microdetermination of heparin in plasma. , 1997, Analytical biochemistry.

[38]  Dong-An Wang,et al.  Heterogeneous-Phase Reaction of Glycidyl Methacrylate and Chondroitin Sulfate: Mechanism of Ring-Opening−Transesterification Competition , 2003 .

[39]  Jennifer T. Blundo,et al.  Effect of Poly(vinyl alcohol) Macromer Chemistry and Chain Interactions on Hydrogel Mechanical Properties , 2007 .