Abstract
Purpose :
At physiological conditions, wounded cornea normally generates wound current, which directly correlates with its wound healing rate. Dynamic changes in wound-generated electric currents show that electrical signaling is an active response to injury. The present study is aimed to study the effect of an external electric field (EF) application triggered by alternate current (AC) in stromal wound healing.
Methods :
Primary human corneal fibroblasts (CF) from the human cornea were cultured by using an explant culture method. The purity of CFs in culture was confirmed by vimentin antibody immune staining whereas the transformation of CFs into “repair-phenotype” was analyzed by staining with alpha-smooth muscle actin (α-SMA). Using a dedicated cell chamber, electrical stimulation of CFs were performed by applying an electric field of 5-20 V/m at the chamber bottom with frequency of 20 Hz continuously for 24 hours and 48 hours respectively in the presence or absence of trophic factor substance P (SP).
Results :
CFs morphology was normal and no cell death was observed after an EF application for 24 or 48 hours when compared to control CFs either in the presence or absence of SP after microscopic observation. External EF application to CFs without SP or application of SP, without EF, for 48 hours could not induce any “repair-phenotype” as observed by α-SMA immunocytochemistry. However, after 24 hours EF stimulation in the presence of SP, CFs shows increased expression of α-SMA and differences in Akt signaling pathway.
Conclusions :
Application of alternate electric fields may contribute to augment the effect of trophic factors during corneal wound healing response, which may have therapeutic consequences in stromal wound healing.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.