title: The modulation of corneal keratocyte and epithelial cell responses to poly(2-hydroxyethyl methacrylate) hydrogel surfaces: phosphorylation decreases collagenase production in vitro.
abstract: We examined the regulation of collagenase production by rabbit keratocyte, epithelial and mixed keratocyte/epithelial cell cultures which were exposed to poly(2-hydroxyethyl methacrylate) (PHEMA) hydrogel surfaces with different chemistries and morphologies (sponge and homogeneous gels). Tissue culture modified polystyrene (TCP), used as a control surface, induced the maximum collagenase response with all cell culture types. Copolymer homogeneous gels containing 2-ethoxyethyl methacrylate (EEMA) or methyl methacrylate (MMA) induced a high response in keratocyte cultures, whilst PHEMA hydrogels induced a moderate response and the phosphorylated PHEMA (phos-PHEMA) hydrogel induced no response. Epithelial cells cultured on PHEMA, copolymer and phos-PHEMA hydrogels produced less collagenase activity than the keratocyte cells. The profile of collagenases produced by epithelial cells in response to phos-PHEMA was different to that for the other hydrogels. Co-cultured cells produced higher levels of collagenase (relative to the TCP) in response to hydrogels than did either the keratocytes or epithelial cells alone, but the response of phos-PHEMA was still the lowest. The overall enzyme response to the sponge hydrogels was lower than that to the homogeneous hydrogels, although this effect was less prominent in the keratocyte cultures. The markedly reduced and alternative collagenase responses to phosphorylated surfaces was not a consequence of cell death, and may be a phenomenon related to changes in cell surface charge and morphology.
authors.affiliation: Lions Eye Institute, Centre for Ophthalmology and Visual Science, Nedlands, Western Australia, Australia.
authors.forename: B W
authors.initials: BW
authors.lastname: Ziegelaar
authors.forename: J H
authors.initials: JH
authors.lastname: Fitton
authors.forename: A B
authors.initials: AB
authors.lastname: Clayton
authors.forename: S T
authors.initials: ST
authors.lastname: Platten
authors.forename: M A
authors.initials: MA
authors.lastname: Maley
authors.forename: T V
authors.initials: TV
authors.lastname: Chirila
mesh_terms: D000818:Animals; D001672:Biocompatible Materials; D002364:Caseins / Q000378:metabolism; D018929:Cell Culture Techniques; D002470:Cell Survival; D018920:Coculture Techniques; D017364:Collagenases / Q000096:biosynthesis*; D003315:Cornea / Q000166:cytology / Q000201:enzymology*; D004847:Epithelial Cells / Q000166:cytology / Q000201:enzymology*; D020100:Hydrogels; D020007:Implants, Experimental; D015603:Keratinocytes / Q000166:cytology / Q000201:enzymology*; D061965:Matrix Metalloproteinase Inhibitors; D010766:Phosphorylation; D011102:Polyhydroxyethyl Methacrylate; D011817:Rabbits; D013499:Surface Properties
chemical_list: D001672:Biocompatible Materials; D002364:Caseins; D020100:Hydrogels; D061965:Matrix Metalloproteinase Inhibitors; D011102:Polyhydroxyethyl Methacrylate; D017364:Collagenases