Suppression of interleukin 1 alpha and interleukin 1 beta in human limbal epithelial cells cultured on the amniotic membrane stromal matrix

Aims-Amniotic membrane (AM) transplantation reduces inflammation in a varie ty of ocular surface disorders. The aim of this study was to determine if A M stroma suppresses the expression of the IL-1 gene family in cultured huma n corneal limbal epithelial cells. Methods-Human corneal limbal epithelial cells were cultured from limbocorne al explants of donor eyes on plastic or on the AM stroma. Transcript expres sion of IL-1 alpha, IL-1 beta, IL-1 receptor antagonist (RA), and GAPDH was compared with or without addition of lipopolysaccharide to their serum-fre e media for 24 hours using RNAse protection assay (RPA). Their protein prod uction in the supernatant was analysed by ELISA. Results-Expression of IL-1 alpha and IL-1 beta transcripts and proteins was significantly reduced by cells cultured on the AM stromal matrix compared with plastic cultures whether lipopolysaccharide was added or not. Moreover , expression of IL-1 RA by cells cultured in the lipopolysaccharide-free me dium was upregulated by AM stromal matrix. The ratio between IL-1 RA and IL -1 alpha protein levels in AM cultures was higher than in plastic cultures. Conclusions-AM stromal matrix markedly suppresses lipopolysaccharide induce d upregulation of both IL-1 alpha and IL-1 beta. These data may explain in part the effect of AM. transplantation in reducing ocular surface inflammat ion, underscoring the unique feature of the AM as a substrate for tissue en gineering.