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Biomimetic Macroporous Hydrogels: Protein Ligand Distribution and Cell Response to the Ligand Architecture in the Scaffold

Savina, Irina N.; Dainiak, Maria LU ; Jungvid, Hans; Mikhalovsky, Sergey V. and Galaev, Igor LU (2009) In Journal of Biomaterials Science. Polymer Edition 20(12). p.1781-1795
Abstract
Macroporous hydrogels (MHs), cryogels, are a new type of biomaterials for tissue engineering that can be produced from any natural or synthetic polymer that forms a gel. Synthetic MHs are rendered bioactive by surface or bulk modifications with extracellular matrix components. In this study, cell response to the architecture of protein ligands, bovine type-I collagen (CG) and human fibrinogen (Fg), immobilised using different methods on poly(2-hydroxyethyl methacrylate) (pHEMA) macroporous hydrogels (MHs) was analysed. Bulk modification was performed by cross-linking cryo-co-polymerisation of HEMA and poly(ethylene glycol) diacrylate (PEGA) in the presence of proteins (CG/ pHEMA and Fg/pHEMA MHs). The polymer surface was modified by... (More)
Macroporous hydrogels (MHs), cryogels, are a new type of biomaterials for tissue engineering that can be produced from any natural or synthetic polymer that forms a gel. Synthetic MHs are rendered bioactive by surface or bulk modifications with extracellular matrix components. In this study, cell response to the architecture of protein ligands, bovine type-I collagen (CG) and human fibrinogen (Fg), immobilised using different methods on poly(2-hydroxyethyl methacrylate) (pHEMA) macroporous hydrogels (MHs) was analysed. Bulk modification was performed by cross-linking cryo-co-polymerisation of HEMA and poly(ethylene glycol) diacrylate (PEGA) in the presence of proteins (CG/ pHEMA and Fg/pHEMA MHs). The polymer surface was modified by covalent immobilisation of the proteins to the active epoxy (ep) groups present on pHEMA after hydrogel fabrication (CG-epHEMA and Fg-epHEMA MHs). The concentration of proteins in protein/pHEMA and protein-epHEMA MHs was 80-85 and 130-140 mu g/ml hydrogel, respectively. It was demonstrated by immunostaining and confocal laser scanning microscopy that bulk modification resulted in spreading of CG in the polymer matrix and spot-like distribution of Fg. On the contrary, surface modification resulted in spot-like distribution of CG and uniform spreading of Fg, which evenly coated the surface. Proliferation rate of fibroblasts was higher on MHs with even distribution of the ligands, i.e., on Fg-epHEMA and CG/ pHEMA. After 30 days of growth, fibroblasts formed several monolayers and deposited extracellular matrix filling the pores of these MHs. The best result in terms of cell proliferation was obtained on Fg-epHEMA. The ligands displayed on surface of these scaffolds were in native conformation, while in bulk-modified CG/ pHEMA MHs most of the proteins were buried inside the polymer matrix and were less accessible for interactions with specific antibodies and cells. The method used for MH modification with bioligands strongly affects spatial distribution, density and conformation of the ligand on the scaffold surface, which, in turn, influence cell-surface interactions. The optimal type of modification varies depending on intrinsic properties of proteins and MHs. (C) Koninklijke Brill NV, Leiden, 2009 (Less)
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Contribution to journal
publication status
published
subject
keywords
ligand, Biomimetic macroporous hydrogel, protein ligand distribution, conformation, cell response
in
Journal of Biomaterials Science. Polymer Edition
volume
20
issue
12
pages
1781 - 1795
publisher
VSP BV
external identifiers
  • wos:000271021100008
  • scopus:70350341131
ISSN
0920-5063
DOI
10.1163/156856208X386390
language
English
LU publication?
yes
id
d7ce6329-d785-46ae-8fd7-c783fc1a4f06 (old id 1506320)
date added to LUP
2009-11-23 15:39:17
date last changed
2017-10-29 03:45:55
@article{d7ce6329-d785-46ae-8fd7-c783fc1a4f06,
  abstract     = {Macroporous hydrogels (MHs), cryogels, are a new type of biomaterials for tissue engineering that can be produced from any natural or synthetic polymer that forms a gel. Synthetic MHs are rendered bioactive by surface or bulk modifications with extracellular matrix components. In this study, cell response to the architecture of protein ligands, bovine type-I collagen (CG) and human fibrinogen (Fg), immobilised using different methods on poly(2-hydroxyethyl methacrylate) (pHEMA) macroporous hydrogels (MHs) was analysed. Bulk modification was performed by cross-linking cryo-co-polymerisation of HEMA and poly(ethylene glycol) diacrylate (PEGA) in the presence of proteins (CG/ pHEMA and Fg/pHEMA MHs). The polymer surface was modified by covalent immobilisation of the proteins to the active epoxy (ep) groups present on pHEMA after hydrogel fabrication (CG-epHEMA and Fg-epHEMA MHs). The concentration of proteins in protein/pHEMA and protein-epHEMA MHs was 80-85 and 130-140 mu g/ml hydrogel, respectively. It was demonstrated by immunostaining and confocal laser scanning microscopy that bulk modification resulted in spreading of CG in the polymer matrix and spot-like distribution of Fg. On the contrary, surface modification resulted in spot-like distribution of CG and uniform spreading of Fg, which evenly coated the surface. Proliferation rate of fibroblasts was higher on MHs with even distribution of the ligands, i.e., on Fg-epHEMA and CG/ pHEMA. After 30 days of growth, fibroblasts formed several monolayers and deposited extracellular matrix filling the pores of these MHs. The best result in terms of cell proliferation was obtained on Fg-epHEMA. The ligands displayed on surface of these scaffolds were in native conformation, while in bulk-modified CG/ pHEMA MHs most of the proteins were buried inside the polymer matrix and were less accessible for interactions with specific antibodies and cells. The method used for MH modification with bioligands strongly affects spatial distribution, density and conformation of the ligand on the scaffold surface, which, in turn, influence cell-surface interactions. The optimal type of modification varies depending on intrinsic properties of proteins and MHs. (C) Koninklijke Brill NV, Leiden, 2009},
  author       = {Savina, Irina N. and Dainiak, Maria and Jungvid, Hans and Mikhalovsky, Sergey V. and Galaev, Igor},
  issn         = {0920-5063},
  keyword      = {ligand,Biomimetic macroporous hydrogel,protein ligand distribution,conformation,cell response},
  language     = {eng},
  number       = {12},
  pages        = {1781--1795},
  publisher    = {VSP BV},
  series       = {Journal of Biomaterials Science. Polymer Edition},
  title        = {Biomimetic Macroporous Hydrogels: Protein Ligand Distribution and Cell Response to the Ligand Architecture in the Scaffold},
  url          = {http://dx.doi.org/10.1163/156856208X386390},
  volume       = {20},
  year         = {2009},
}