Effect of matrix metalloproteinases activity on outflow in perfused human organ culture

John M B Bradley, Janice Vranka, Christine M. Colvis, Dean M. Conger, J. Preston Alexander, Aurelie S. Fisk, John R. Samples, Ted Acott

Research output: Contribution to journalArticle

263 Citations (Scopus)

Abstract

PURPOSE. To test the hypothesis that extracellular matrix turnover, mediated by the matrix metalloproteinases, modulates aqueous humor outflow facility in a human outflow model. METHODS. Matrix metalloproteinase activity was manipulated and outflow facility evaluated using perfused human anterior segment organ culture. Purified matrix metalloproteinases, tissue inhibitors of metalloproteinases (TIMPs), and several families of synthetic inhibitors of matrix metalloproteinases were added to the perfusion medium. Matrix metalloproteinase expression was increased by adding recombinant interleukin (IL)-1α. Kinetic inhibition analysis was conducted for stromelysin, gelatinase A, and gelatinase B with the various inhibitors. Live- dead staining was used to evaluate culture viability. RESULTS. Increasing metalloproteinase activity, by adding purified metalloproteinases or by inducing their expression by IL-1α treatment, increased outflow facility. Inhibition of endogenous trabecular metalloproteinase activity using TIMP or several families of synthetic metalloproteinase inhibitors reduced outflow rates. The elevation and the reduction of outflow rates were reversible, with changes requiring 1 to 3 days. Kinetic enzyme inhibition analysis produced 50% inhibitory concentration values for these inhibitors that were compatible with the concentration ranges for outflow inhibition. CONCLUSIONS. The ability of several specific matrix metalloproteinase inhibitors to reduce outflow facility implies that endogenous extracellular matrix turnover by these enzymes was required for the maintenance of trabecular outflow resistance, at least in this human culture model. These observations provide support for the hypothesis that controlled extracellular matrix turnover is important in the regulation of aqueous humor outflow facility.

Original languageEnglish (US)
Pages (from-to)2649-2658
Number of pages10
JournalInvestigative Ophthalmology and Visual Science
Volume39
Issue number13
StatePublished - Dec 1998

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Organ Culture Techniques
Metalloproteases
Matrix Metalloproteinases
Extracellular Matrix
Tissue Inhibitor of Metalloproteinases
Matrix Metalloproteinase Inhibitors
Aqueous Humor
Interleukin-1
Matrix Metalloproteinase 3
Matrix Metalloproteinase 2
Matrix Metalloproteinase 9
Enzymes
Inhibitory Concentration 50
Perfusion
Maintenance
Staining and Labeling
HSCH2CH(CH2CH(CH3)2)CO-Phe-Ala-NH2

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Bradley, J. M. B., Vranka, J., Colvis, C. M., Conger, D. M., Alexander, J. P., Fisk, A. S., ... Acott, T. (1998). Effect of matrix metalloproteinases activity on outflow in perfused human organ culture. Investigative Ophthalmology and Visual Science, 39(13), 2649-2658.

Effect of matrix metalloproteinases activity on outflow in perfused human organ culture. / Bradley, John M B; Vranka, Janice; Colvis, Christine M.; Conger, Dean M.; Alexander, J. Preston; Fisk, Aurelie S.; Samples, John R.; Acott, Ted.

In: Investigative Ophthalmology and Visual Science, Vol. 39, No. 13, 12.1998, p. 2649-2658.

Research output: Contribution to journalArticle

Bradley, JMB, Vranka, J, Colvis, CM, Conger, DM, Alexander, JP, Fisk, AS, Samples, JR & Acott, T 1998, 'Effect of matrix metalloproteinases activity on outflow in perfused human organ culture', Investigative Ophthalmology and Visual Science, vol. 39, no. 13, pp. 2649-2658.
Bradley, John M B ; Vranka, Janice ; Colvis, Christine M. ; Conger, Dean M. ; Alexander, J. Preston ; Fisk, Aurelie S. ; Samples, John R. ; Acott, Ted. / Effect of matrix metalloproteinases activity on outflow in perfused human organ culture. In: Investigative Ophthalmology and Visual Science. 1998 ; Vol. 39, No. 13. pp. 2649-2658.
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abstract = "PURPOSE. To test the hypothesis that extracellular matrix turnover, mediated by the matrix metalloproteinases, modulates aqueous humor outflow facility in a human outflow model. METHODS. Matrix metalloproteinase activity was manipulated and outflow facility evaluated using perfused human anterior segment organ culture. Purified matrix metalloproteinases, tissue inhibitors of metalloproteinases (TIMPs), and several families of synthetic inhibitors of matrix metalloproteinases were added to the perfusion medium. Matrix metalloproteinase expression was increased by adding recombinant interleukin (IL)-1α. Kinetic inhibition analysis was conducted for stromelysin, gelatinase A, and gelatinase B with the various inhibitors. Live- dead staining was used to evaluate culture viability. RESULTS. Increasing metalloproteinase activity, by adding purified metalloproteinases or by inducing their expression by IL-1α treatment, increased outflow facility. Inhibition of endogenous trabecular metalloproteinase activity using TIMP or several families of synthetic metalloproteinase inhibitors reduced outflow rates. The elevation and the reduction of outflow rates were reversible, with changes requiring 1 to 3 days. Kinetic enzyme inhibition analysis produced 50{\%} inhibitory concentration values for these inhibitors that were compatible with the concentration ranges for outflow inhibition. CONCLUSIONS. The ability of several specific matrix metalloproteinase inhibitors to reduce outflow facility implies that endogenous extracellular matrix turnover by these enzymes was required for the maintenance of trabecular outflow resistance, at least in this human culture model. These observations provide support for the hypothesis that controlled extracellular matrix turnover is important in the regulation of aqueous humor outflow facility.",
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N2 - PURPOSE. To test the hypothesis that extracellular matrix turnover, mediated by the matrix metalloproteinases, modulates aqueous humor outflow facility in a human outflow model. METHODS. Matrix metalloproteinase activity was manipulated and outflow facility evaluated using perfused human anterior segment organ culture. Purified matrix metalloproteinases, tissue inhibitors of metalloproteinases (TIMPs), and several families of synthetic inhibitors of matrix metalloproteinases were added to the perfusion medium. Matrix metalloproteinase expression was increased by adding recombinant interleukin (IL)-1α. Kinetic inhibition analysis was conducted for stromelysin, gelatinase A, and gelatinase B with the various inhibitors. Live- dead staining was used to evaluate culture viability. RESULTS. Increasing metalloproteinase activity, by adding purified metalloproteinases or by inducing their expression by IL-1α treatment, increased outflow facility. Inhibition of endogenous trabecular metalloproteinase activity using TIMP or several families of synthetic metalloproteinase inhibitors reduced outflow rates. The elevation and the reduction of outflow rates were reversible, with changes requiring 1 to 3 days. Kinetic enzyme inhibition analysis produced 50% inhibitory concentration values for these inhibitors that were compatible with the concentration ranges for outflow inhibition. CONCLUSIONS. The ability of several specific matrix metalloproteinase inhibitors to reduce outflow facility implies that endogenous extracellular matrix turnover by these enzymes was required for the maintenance of trabecular outflow resistance, at least in this human culture model. These observations provide support for the hypothesis that controlled extracellular matrix turnover is important in the regulation of aqueous humor outflow facility.

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