Evaluation of various compounds to inhibit activity of matrix metalloproteinases in the tear film of horses with ulcerative keratitis

Franck J. Ollivier, Dennis E. Brooks, Maria E. Kallberg, Andras M. Komaromy, Mary Utter, Stacy E. Andrew, Kirk N. Gelatt, Gary R. Stevens, Timothy D. Blalock, Gysbert Botho van Setten, Gregory S. Schultz

Research output: Contribution to journalReview article

65 Citations (Scopus)

Abstract

Objective - To examine in vitro effects of various antiproteolytic compounds on activity of matrix metalloproteinase (MMP)-2 and -9 in the tear film of horses with active corneal ulcers. Sample Population - Samples of tear film obtained from the eyes of 34 horses with active ulcerative keratitis. Procedure - Horses were sedated, and tear samples were collected from the lower fornix of 34 ulcerated eyes by use of capillary tubes. The protease inhibitors 0.2% EDTA, 0.1% doxycycline, 10% N-acetylcysteine (NAC), 0.1% solution of a modified dipeptide that contains hydroxamic acid (ie, ilomostat), 0.1% α1-proteinase inhibitor (PI), 0.5% α1-PI, and 100% fresh equine serum (ES) were used to treat pooled samples. Amount of latent and active MMP-2 and -9 was measured by optical density scanning of gelatin zymograms of treated and untreated tear samples. Results - Pooled tear samples obtained from ulcerated eyes contained the latent and active forms of MMP-2 and -9. Compared with MMP activity in untreated samples, total MMP activity (sum of all bands detected) observed on the gelatin zymogram gels was reduced by 99.4% by EDTA, 96.3% by doxycycline, 98.8% by NAC, 98.9% by ilomostat, 52.4% by 0.1% α1-PI, 93.6% by 0.5% α1-PI, and 90.0% by ES. Conclusions and Clinical Relevance - We documented that EDTA, doxycycline, NAC, ilomostat, α1-PI, and ES inhibited MMP activity in vitro. Because these compounds use different mechanisms to inhibit various families of proteases in the tear film of horses, a combination of these protease inhibitors may be beneficial for treatment of corneal ulcers in horses.

Original languageEnglish (US)
Pages (from-to)1081-1087
Number of pages7
JournalAmerican Journal of Veterinary Research
Volume64
Issue number9
DOIs
StatePublished - Sep 1 2003
Externally publishedYes

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Corneal Ulcer
keratitis
metalloproteinases
Matrix Metalloproteinases
Tears
films (materials)
Horses
proteinase inhibitors
horses
Peptide Hydrolases
acetylcysteine
gelatinase A
doxycycline
Doxycycline
Matrix Metalloproteinase 2
Matrix Metalloproteinase 9
Acetylcysteine
Edetic Acid
eyes
sampling

ASJC Scopus subject areas

  • veterinary(all)

Cite this

Evaluation of various compounds to inhibit activity of matrix metalloproteinases in the tear film of horses with ulcerative keratitis. / Ollivier, Franck J.; Brooks, Dennis E.; Kallberg, Maria E.; Komaromy, Andras M.; Utter, Mary; Andrew, Stacy E.; Gelatt, Kirk N.; Stevens, Gary R.; Blalock, Timothy D.; van Setten, Gysbert Botho; Schultz, Gregory S.

In: American Journal of Veterinary Research, Vol. 64, No. 9, 01.09.2003, p. 1081-1087.

Research output: Contribution to journalReview article

Ollivier, FJ, Brooks, DE, Kallberg, ME, Komaromy, AM, Utter, M, Andrew, SE, Gelatt, KN, Stevens, GR, Blalock, TD, van Setten, GB & Schultz, GS 2003, 'Evaluation of various compounds to inhibit activity of matrix metalloproteinases in the tear film of horses with ulcerative keratitis', American Journal of Veterinary Research, vol. 64, no. 9, pp. 1081-1087. https://doi.org/10.2460/ajvr.2003.64.1081
Ollivier, Franck J. ; Brooks, Dennis E. ; Kallberg, Maria E. ; Komaromy, Andras M. ; Utter, Mary ; Andrew, Stacy E. ; Gelatt, Kirk N. ; Stevens, Gary R. ; Blalock, Timothy D. ; van Setten, Gysbert Botho ; Schultz, Gregory S. / Evaluation of various compounds to inhibit activity of matrix metalloproteinases in the tear film of horses with ulcerative keratitis. In: American Journal of Veterinary Research. 2003 ; Vol. 64, No. 9. pp. 1081-1087.
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abstract = "Objective - To examine in vitro effects of various antiproteolytic compounds on activity of matrix metalloproteinase (MMP)-2 and -9 in the tear film of horses with active corneal ulcers. Sample Population - Samples of tear film obtained from the eyes of 34 horses with active ulcerative keratitis. Procedure - Horses were sedated, and tear samples were collected from the lower fornix of 34 ulcerated eyes by use of capillary tubes. The protease inhibitors 0.2{\%} EDTA, 0.1{\%} doxycycline, 10{\%} N-acetylcysteine (NAC), 0.1{\%} solution of a modified dipeptide that contains hydroxamic acid (ie, ilomostat), 0.1{\%} α1-proteinase inhibitor (PI), 0.5{\%} α1-PI, and 100{\%} fresh equine serum (ES) were used to treat pooled samples. Amount of latent and active MMP-2 and -9 was measured by optical density scanning of gelatin zymograms of treated and untreated tear samples. Results - Pooled tear samples obtained from ulcerated eyes contained the latent and active forms of MMP-2 and -9. Compared with MMP activity in untreated samples, total MMP activity (sum of all bands detected) observed on the gelatin zymogram gels was reduced by 99.4{\%} by EDTA, 96.3{\%} by doxycycline, 98.8{\%} by NAC, 98.9{\%} by ilomostat, 52.4{\%} by 0.1{\%} α1-PI, 93.6{\%} by 0.5{\%} α1-PI, and 90.0{\%} by ES. Conclusions and Clinical Relevance - We documented that EDTA, doxycycline, NAC, ilomostat, α1-PI, and ES inhibited MMP activity in vitro. Because these compounds use different mechanisms to inhibit various families of proteases in the tear film of horses, a combination of these protease inhibitors may be beneficial for treatment of corneal ulcers in horses.",
author = "Ollivier, {Franck J.} and Brooks, {Dennis E.} and Kallberg, {Maria E.} and Komaromy, {Andras M.} and Mary Utter and Andrew, {Stacy E.} and Gelatt, {Kirk N.} and Stevens, {Gary R.} and Blalock, {Timothy D.} and {van Setten}, {Gysbert Botho} and Schultz, {Gregory S.}",
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AU - Ollivier, Franck J.

AU - Brooks, Dennis E.

AU - Kallberg, Maria E.

AU - Komaromy, Andras M.

AU - Utter, Mary

AU - Andrew, Stacy E.

AU - Gelatt, Kirk N.

AU - Stevens, Gary R.

AU - Blalock, Timothy D.

AU - van Setten, Gysbert Botho

AU - Schultz, Gregory S.

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N2 - Objective - To examine in vitro effects of various antiproteolytic compounds on activity of matrix metalloproteinase (MMP)-2 and -9 in the tear film of horses with active corneal ulcers. Sample Population - Samples of tear film obtained from the eyes of 34 horses with active ulcerative keratitis. Procedure - Horses were sedated, and tear samples were collected from the lower fornix of 34 ulcerated eyes by use of capillary tubes. The protease inhibitors 0.2% EDTA, 0.1% doxycycline, 10% N-acetylcysteine (NAC), 0.1% solution of a modified dipeptide that contains hydroxamic acid (ie, ilomostat), 0.1% α1-proteinase inhibitor (PI), 0.5% α1-PI, and 100% fresh equine serum (ES) were used to treat pooled samples. Amount of latent and active MMP-2 and -9 was measured by optical density scanning of gelatin zymograms of treated and untreated tear samples. Results - Pooled tear samples obtained from ulcerated eyes contained the latent and active forms of MMP-2 and -9. Compared with MMP activity in untreated samples, total MMP activity (sum of all bands detected) observed on the gelatin zymogram gels was reduced by 99.4% by EDTA, 96.3% by doxycycline, 98.8% by NAC, 98.9% by ilomostat, 52.4% by 0.1% α1-PI, 93.6% by 0.5% α1-PI, and 90.0% by ES. Conclusions and Clinical Relevance - We documented that EDTA, doxycycline, NAC, ilomostat, α1-PI, and ES inhibited MMP activity in vitro. Because these compounds use different mechanisms to inhibit various families of proteases in the tear film of horses, a combination of these protease inhibitors may be beneficial for treatment of corneal ulcers in horses.

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