Purpose. In previous studies, we have found that the bovine defensin BNP-1 [RLCRIVVIRVCR] stimulated the growth of corneal epithelial cells and is an effective broad range antimicrobial. This study examined the effects of BNP-1 on the growth of human corneal endothelial cells to determine whether it was safe to use as an antibiotic in corneal preservative media. Methods. A cell line of human corneal endothelial cells which contained the large T antigen from SV40 (HEN-LTag) [Wilson et al., IOVS 36:32(1995)] was used to test the ability of BNP-1 to stimulate cellular growth. The dose response of BNP-1 was determined in Eagles' minimal essential culture medium in the presence and absence of both 50% Optisol and calf serum (0 to 10%). The cells were tested in the presence of BNP-1 for various periods of time up to six days. Results. The addition of BNP-1 to the HEN-LTag cells growing in tissue culture media plus 50% Optisol stimulated cellular growth, even at concentrations as high as 100 ug/ml for six days, with optima stimulation found between 10 to 30 ug/ml. This is also the concentration range which is found to be best for antibiotic activity. Cells growing in 10% calf serum plus 50% Optisol for six days showed a slight inhibition of growth at the higher concentrations of BNP-1, but at 5 ug/ml, there was a slight stimulation in growth. However, if no Optisol was present, BNP-1 was slightly stimulatory for cell growth for the entire concentration range in 10% calf serum. Optisol alone inhibited the growth stimulatory effects of calf serum, but BNP-1 seemed to moderate this effect. Conclusion. Bovine neutrophil antibiotic peptide BNP-1 stimulated the growth of human corneal endothelial cells at a wide range of concentrations. Since the BNP-1 was benificial for the growth of the human corneal endothelial cells during a six-day period, it is likely that it would be a safe antibiotic to use as a corneal preservative.
|Original language||English (US)|
|Journal||Investigative Ophthalmology and Visual Science|
|State||Published - Feb 15 1996|
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