The indirect effect of AM substrates on hiCEC re-epithelialisation following injury.

Re-epithelialisation of hiCEC in response to the substrates was assessed using a scratch assay and the wound areas were measured from micrographs taken at day 0, 2, 4, 6 and 10 days (A). Wound areas at days 2, 6 and 8 were compared to wound areas at day 0 using ImageJ software and percentage wound closure rates were calculated (B). Cell migration and re-epithelialisation was evident in all of the wounds over an 8 day period. Denuded and cryopreserved AM substrates demonstrated no additive effect on re-epithelialisation compared to hiCEC cultured without AM. Dried, trehalose and raffinose treated AM significantly promoted re-epithelialisation and wound healing compared to hiCEC cultured with cryopreserved AM. Data are expressed as mean ± SEM based on three separate experiments. *p<0.05 increase in re-epithelialisation compared to corresponding time points in cells grown with cryopreserved AM.