Characterization of the major matrix degrading metalloproteinase of human corneal stroma. Evidence for an enzyme/inhibitor complex

Brown, Donald J.; Chwa, Marilyn; Escobar, Ma. del Carmen; Kenney, M. Cristina

doi:10.1016/0014-4835(91)90123-v

Cited by 52 publications

(28 citation statements)

References 36 publications

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“…They express aldehyde dehydrogenase and keratocan, 1,2 exhibit a dendritic=stellate morphology, interconnect using multiple long cellular processes, 2,3 and help maintain the extracellular matrix (ECM) by synthesizing its components. [4][5][6][7][8] After injury to the corneal stroma, keratocytes transform via a wound healing response into the more active phenotype of fibroblasts, 9 and from there sometimes into myofibroblasts. 10,11 These repair cells migrate to the wound area and secrete a repair ECM (collagen and proteoglycans) 2 that is often fibrotic, fails to restore normal function, and may cause scarring and contraction of the cornea.…”

Section: Introductionmentioning

confidence: 99%

Effect of Serum and Insulin Modulation on the Organization and Morphology of Matrix Synthesized by Bovine Corneal Stromal Cells

Bueno

Saeidi

Melotti

et al. 2009

Tissue Engineering Part A

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The in vitro production of highly organized collagen fibrils by corneal keratocytes in a three-dimensional scaffold-free culture system presents a unique opportunity for the direct observation of organized matrix formation. The objective of this investigation was to develop such a culture system in a glass substrate (for optical accessibility) and to directly examine the effect of reducing serum and=or increasing insulin on the stratification and secretion of aligned matrix by fourth-to fifth-passage bovine corneal stromal keratocytes. Medium concentrations of 0%, 1%, or 10% fetal bovine serum and 0% or 1% insulin-transferrin-selenium were investigated. Highresolution differential interference contrast microscopy, quick-freeze=deep-etch, and conventional transmission electron microscopy were used to monitor the evolution, morphology, and ultrastructure of the cell-matrix constructs. In a medium containing 1% each of serum and insulin-transferrin-selenium, stromal cells stratified and secreted abundant and locally aligned matrix, generating the thickest cell-matrix constructs (allowing handling with forceps). The results of this study have the potential to significantly advance the field of developmental functional engineering of load-bearing tissues by (i) elucidating cues that modulate in vitro cell secretion of organized matrix and (ii) establishing an optically accessible cell culture system for investigating the mechanism of cell secretion of aligned collagen fibrils.

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Section: Introductionmentioning

confidence: 99%

Effect of Serum and Insulin Modulation on the Organization and Morphology of Matrix Synthesized by Bovine Corneal Stromal Cells

Bueno

Saeidi

Melotti

et al. 2009

Tissue Engineering Part A

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“…Hydrolysis of this substrate was catalyed only by the soluble protein preparations that also contained the M r 43,000 zymographic activity and were generally those that had been maintained in culture medium for periods exceeding 30 days. In addition to supporting the hypothesis that MMP-2 can become activated in organ cultured corneas, these observations also suggested that the M r 43,000 form of MMP-2 is the catalytically active species present, perhaps because it cannot effectively bind the tissue inhibitors of metalloproteinases 2 and 1 that are co-secreted by keratocytes [9,11,21]. With respect to the activity status of the M r 62,000 form of corneal MMP-2, this has been questioned previously [10,11,20].…”

Section: Discussionmentioning

confidence: 68%

“…The major protease found in corneal stromal tissue is inactive matrix metalloproteinase 2 or proMMP-2 [9,10]. It is known to become activated in the culture medium of metabolically stressed corneal keratocyte cultures [11], and, when activated, will cleave type IV collagen, the major component of basement membranes and type VI collagen, a component of the anchoring fibrils [12].…”

Section: Introductionmentioning

confidence: 99%

Matrix Metalloproteinase 2 Activation in Cultured Corneas

Smith¹,

El-Rakhawy²,

Easty³

2000

Ophthalmic Res

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Purpose: Corneas that are maintained in tissue culture medium shed their epithelial cells and repopulation following graft surgery is an essential facet of the healing process. Failure to do so may be a result of structural damage to the epithelial basement membrane of a donor cornea. The purpose of the present investigation was to ascertain whether MMP-2, the matrix metalloproteinase produced by corneal keratocytes, may be activated during storage and hence cleave the type IV collagen component of the epithelial cell basement membrane. Methods: Fresh and transplant rejected corneas that had been stored in culture medium for varying time periods and of known donor age were collected. The soluble protein fractions of these corneas were obtained. Their MMP-2 proteins were visualised by zymography on SDS gelatin polyacrylamide gels and assayed for activity against nitrophenyl acetate and denatured [³H]type I collagen. Results: The stromal tissue of fresh, normal corneas produced inactive MMP-2 of M_r 66,000. Although the cultured corneas did not up-regulate MMP-2 production, they contained additional MMP-2 activities of M_r 62,000 and M_r 43,000. The appearance of these additional MMP-2 activities correlated with corneal culture time but not donor age. The ability to cleave denatured [³H]type I collagen correlated with the appearance of the M_r 43,000 activity but not the M_r 62,000 activity. Conclusion: Activated MMP-2 is produced in cultured corneas. For this reason the corneas donated for all graft procedures should not be held in culture medium for periods exceeding 4 weeks.

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“…MMPs are a group of closely related enzymes, and are responsible for connective tissue degradation (Brown et al, 1991). Of these, MMP-2 and MMP-9 were known initially as gelatinases because they degrade gelatin.…”

Section: Discussionmentioning

confidence: 99%

Effects of Synthetic Inhibitor of Metalloproteinase and Cyclosporin A on Corneal Haze after Excimer Laser Photorefractive Keratectomy in Rabbits

Chang

Kook

Lee

et al. 1998

Experimental Eye Research

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Characterization of the major matrix degrading metalloproteinase of human corneal stroma. Evidence for an enzyme/inhibitor complex

Cited by 52 publications

References 36 publications

Effect of Serum and Insulin Modulation on the Organization and Morphology of Matrix Synthesized by Bovine Corneal Stromal Cells

Effect of Serum and Insulin Modulation on the Organization and Morphology of Matrix Synthesized by Bovine Corneal Stromal Cells

Matrix Metalloproteinase 2 Activation in Cultured Corneas

Effects of Synthetic Inhibitor of Metalloproteinase and Cyclosporin A on Corneal Haze after Excimer Laser Photorefractive Keratectomy in Rabbits

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