A high-density PEG interfacial layer alters the response to an EGF tethered polydimethylsiloxane surface.

Previously, epidermal growth factor (EGF)-modified surfaces have shown promise in supporting cellular growth and adhesion on synthetic polymeric substrates. Surfaces prepared using a novel modification technique were investigated in the current work for their ability to support corneal epithelialization, important to the integration of a synthetic artificial cornea. EGF could be tethered to PDMS surfaces via a high-density, hetero-bifunctional PEG-NSC linking layer with a tunable surface concentration of up to 300 ng/cm(2). Only a small fraction of the EGF on these surfaces could be removed with SDS rinsing, indicative of covalent tethering. Studies with human corneal epithelial cells suggest a relatively linear increase in the number of corneal epithelial cells with increasing EGF concentration at all times. However, confluence was not achieved at any time point. It is believed that the presence of the non-adsorbent PEG layer, useful for preventing non-specific adsorption of proteins, may limit the cellular response by minimizing the adsorption of adhesion molecules. The effects of the EGF alone are clearly not sufficient to result in epithelialization of an artificial cornea surface. Altering both the adhesion and growth of corneal epithelial cells in a controlled manner may be necessary for epithelialization of an artificial cornea.

EGF-grafted PDMS surfaces in artificial cornea applications.

Lack of epithelial cell coverage has remained a persistent problem in the design of an artificial cornea. In this work, polydimethylsiloxane (PDMS) surfaces were modified with epidermal growth factor (EGF) to improve the growth of corneal epithelial cells. The EGF was covalently tethered to PDMS substrates aminated by plasma polymerization of allylamine via a homobifunctional polyethylene glycol (PEG) spacer. Surface modification was confirmed by contact angle and X-ray photoelectron spectroscopy measurements. By varying the ratio of EGF to PEG from 1:50 to 1:5, EGF amounts from 40 to 90 ng/cm2 could be bound, as determined by surface plasmon resonance (SPR) and 125I radiolabelling. Human corneal epithelial cells on the various modified surfaces were cultured both in the presence and absence of EGF in the culture medium to determine the effect of covalently bound EGF on the cells. The results demonstrated that covalently bound EGF on the surfaces is active with respect to promoting epithelial cell coverage. This was significant when compared to unmodified controls.