Effect of the bovine neutrophil antibiotic dodeca-peptide (BNP-1) on the growth of human corneal endothelial cells in optisol and tissue culture media

S. A. Griffith, D. L. McCartney, Christopher J Murphy, Mark J Mannis, T. W. Reid

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Abstract

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 languageEnglish (US)
JournalInvestigative Ophthalmology and Visual Science
Volume37
Issue number3
StatePublished - Feb 15 1996

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Culture Media
Neutrophils
Endothelial Cells
Anti-Bacterial Agents
Peptides
Growth
Serum
Optisol
Defensins
Eagles
Viral Tumor Antigens
Epithelial Cells
Cell Line

ASJC Scopus subject areas

  • Ophthalmology

Cite this

@article{d83f84f07d954d559f1110af73cc8d26,
title = "Effect of the bovine neutrophil antibiotic dodeca-peptide (BNP-1) on the growth of human corneal endothelial cells in optisol and tissue culture media",
abstract = "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.",
author = "Griffith, {S. A.} and McCartney, {D. L.} and Murphy, {Christopher J} and Mannis, {Mark J} and Reid, {T. W.}",
year = "1996",
month = "2",
day = "15",
language = "English (US)",
volume = "37",
journal = "Investigative Ophthalmology and Visual Science",
issn = "0146-0404",
publisher = "Association for Research in Vision and Ophthalmology Inc.",
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T1 - Effect of the bovine neutrophil antibiotic dodeca-peptide (BNP-1) on the growth of human corneal endothelial cells in optisol and tissue culture media

AU - Griffith, S. A.

AU - McCartney, D. L.

AU - Murphy, Christopher J

AU - Mannis, Mark J

AU - Reid, T. W.

PY - 1996/2/15

Y1 - 1996/2/15

N2 - 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.

AB - 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.

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