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Br J Ophthalmol 2001;85:444-449 ( April )

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

Abraham Solomona, Mark Rosenblatta, Dagoberto Monroya, Zhonghua Jia, Stephen C Pflugfeldera, Scheffer C G Tsenga b

a Ocular Surface and Tear Center, Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, Florida, USA, b Department of Cell Biology and Anatomy, University of Miami School of Medicine, Miami, Florida, USA

Correspondence to: Scheffer C G Tseng, MD, PhD, Bascom Palmer Eye Institute, William L McKnight Vision Research Center, 1638 NW 10th Avenue Miami, FL 33136, USA stseng{at}bpei.med.miami.edu

Accepted for publication 1 November 2000

AIMS---Amniotic membrane (AM) transplantation reduces inflammation in a variety of ocular surface disorders. The aim of this study was to determine if AM stroma suppresses the expression of the IL-1 gene family in cultured human corneal limbal epithelial cells.
METHODS---Human corneal limbal epithelial cells were cultured from limbocorneal explants of donor eyes on plastic or on the AM stroma. Transcript expression of IL-1alpha , IL-1beta , IL-1 receptor antagonist (RA), and GAPDH was compared with or without addition of lipopolysaccharide to their serum-free media for 24 hours using RNAse protection assay (RPA). Their protein production in the supernatant was analysed by ELISA.
RESULTS---Expression of IL-1alpha and IL-1beta 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 medium was upregulated by AM stromal matrix. The ratio between IL-1 RA and IL-1alpha protein levels in AM cultures was higher than in plastic cultures.
CONCLUSIONS---AM stromal matrix markedly suppresses lipopolysaccharide induced upregulation of both IL-1alpha and IL-1beta . These data may explain in part the effect of AM transplantation in reducing ocular surface inflammation, underscoring the unique feature of the AM as a substrate for tissue engineering.

© 2001 by British Journal of Ophthalmology


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