P 140 Automated technology of clinical study of eye biomechanics and diagnosis according to cornea photoelasticity

SERUM DRAMATICALLY ENHANCES ORIENTATION AND MIGRATION OF CULTURED BOVINE CORNEALEPITHELIAL CELISINANAPPLIEDELECTRICFIELD(RF) ZHAOM’, AGIUS-FERNANDEZ A~,FORRESTERP,McCAIGC’ ‘Department of Biomedical Sciences, Marischal College, ‘Deparz%uvrt of Ophthalmology, University ofAberdeen, Aberdeen AB9 IAS, Scotkmd(X) d Wounded epithelia generate small dc electric fields, whilst many cell types, including rabbit corneal epithelial cells reorient and migrate directionally when cultured in a dc field. We have studied the effects of an externally applied dc field on cultured bovine corneal epithelial cells. M&&& Primary culture of bovine corneal epithelial cells were exposed to a dc field for Sh. Media were exchanged into the cultures immediately prior to field application. Serial photographs were taken and analyzed. RM In media containing 10% FCS, cells showed marked galvanotropic response. At lOOmV/mm, cells reoriented perpendicularly after about 20-30 min in EF, whilst at ISOmVimm migration towards the cathode at a mean rate of 13.3&6ur& (n=40) was evident. Cell sheets (about 3-30 cells together) also migrated cathodally at a slightly higher rate of 17.0~1.9uti for the leading edge and 17.4+1.9uti for the trailing edge (n=l5). Much larger sheets also migrated cathodally. For both the dissociated cells and cell sheets, ruffled membranes and lamellipodia were abundant at the leading edges, and cell processes underwent retraction at the trailing edges. The directional migration of the cell and cell sheets showed no marked increase beyond 15OmV/mm to 250mVimm. Addition of 25ng/ml EGF in the serum containing media had no obvious influence on the cell response to EF. By contrast, in serum free medium, we found cell reorientation at 200 mVimm, but no obvious directional movement at field strengths up to 25OmV/mm. Conclusions The results demonstrate a much lower threshold for galvanotropic and galvanotaxic response of cultured corneal epithelial cells when serum is present in the medium. Thus element(s) within serum promote electric field directed migration of cornea1 epithelial cells. In combination, electric fields and serum components may enhance cornea1 wound healing. (supported by the Wellcome Trust)