Corneal epithelial cell adhesion and growth on EGF-modified aminated PDMS.
Growth factor tethering has significant potential to mediate cellular responses in biomaterials and tissue engineering. We have previously demonstrated that epidermal growth factor (EGF) can be tethered to polydimethylsiloxane (PDMS) substrates and that these surfaces promoted interactions with human corneal epithelial cells in vitro. The goal of the current work was to better understand the specific effects of the tethered growth factor on the cells. The EGF was reacted with a homobifunctional N-hydroxysuccinimide (NHS) polyethylene glycol (PEG) derivative, and then bound to allyamine plasma-modified PDMS. Human corneal epithelial cells were seeded on the surfaces and cultured in serum-free medium for periods of up to 5 days. Cell growth was monitored and quantified by trypsinization and counting with a Coulter counter. Expression of matrix proteins and alpha(6)-integrins was assessed by immunostaining and confocal microscopy. A centrifugation assay was used to determine cell adhesion under an applied detachment force. Binding of EGF was found to significantly increase cell numbers and coverage across the surfaces at 5 days of culture in vitro. Immunofluorescence experiments indicate increased expression of fibronectin, laminin, and alpha(6)-integrins on the EGF-modified surfaces, and expression is localized at the cell-material interface as observed by confocal microscopy. In accordance with these results, the highest quantity of adherent cells is found on the EGF-modified subtrates at 5 days of culture. The results provide initial evidence that binding of EGF may be used to improve the epithelialization of and the adhesion of the cells on a polymeric artificial cornea device.
Klenkler
Bettina J
BJ
Department of Chemical Engineering, McMaster University, Hamilton, Ontario, Canada L8S 4L7.
Dwivedi
Dhruva
D
West-Mays
Judith A
JA
Sheardown
Heather
H
D000586:Amination / Q000187:drug effects; D002448:Cell Adhesion / Q000187:drug effects; D002452:Cell Count; D049109:Cell Proliferation / Q000187:drug effects; D004129:Dimethylpolysiloxanes / Q000494:pharmacology*; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D019573:Epithelium, Corneal / Q000166:cytology*; D005353:Fibronectins / Q000378:metabolism; D005452:Fluoresceins / Q000378:metabolism; D005453:Fluorescence; D006801:Humans; D039503:Integrin alpha6 / Q000378:metabolism; D007797:Laminin / Q000378:metabolism; D018613:Microscopy, Confocal; D011092:Polyethylene Glycols / Q000494:pharmacology; D013499:Surface Properties / Q000187:drug effects
D004129:Dimethylpolysiloxanes; D005353:Fibronectins; D005452:Fluoresceins; D039503:Integrin alpha6; D007797:Laminin; D011092:Polyethylene Glycols; D004815:Epidermal Growth Factor; C007740:fluorexon