Abstract
Aim
To compare matrix metalloproteinase (MMP) localisation in anterior keratectomy (AK) and lamellar keratectomy (LK) wounds.
Methods
Wounds were produced in one eye of 24 rabbits. The AK wounds were made to approximately 120 μm in depth and then allowed to re-epithelialise. The LK wounds were of similar depth, but the anterior stroma and epithelium were replaced after a second deeper keratectomy had been performed. Immunohistochemistry was used to localise the MMP-1, -2, -3, and -9 at intervals from 4 h to 14 days following surgery. The contralateral eyes acted as controls.
Results
After an AK wound MMP-1 was present at the leading edge of migrating epithelium after 18 h, while MMP-2 and -9 were localised behind the advancing epithelial edge. The presence of these enzymes rapidly fell to low levels after epithelial closure. There was only faint MMP-3 localisation between days 3 and 7. After an LK wound, MMP-1, -3, and -9 were not detected in the stromal interface, but MMP-2 was present at all time points.
Conclusions
This study suggests that after an AK wound, MMP-1 is a key mediator of epithelial migration, while MMP-2 and -9, and to a lesser extent MMP-3, may participate in the remodelling of corneal stroma and the reformation of epithelial basement membrane. In contrast, an LK wound results in a much lower stimulus for MMP activation. The action of MMP-2 in stromal repair is thus partly independent of epithelial injury.
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References
Sivak JM, Fini ME . MMPs in the eye: emerging roles for matrix metalloproteinases in ocular physiology. Prog Retinal Eye Res 2002; 21: 1–14.
Wong TTL, Sethi C, Daniels JT, Limb GA, Murphy G, Khaw PT . Matrix metalloproteinases in disease and repair processes in the anterior segment. Surv Ophthalmol 2002; 47: 239–256.
Daniels JT, Cambrey AD, Occleston NL, Garrett Q, Tarnuzzer RW, Schultz GS et al. Matrix metalloproteinase inhibition modulates fibroblast-mediated matrix contraction and collagen production in vitro. Invest Ophthalmol Vis Sci 2003; 44: 1104–1110.
Fini ME, Girard MT . Expression of collagenolytic/gelatinolytic metalloproteinases by normal cornea. Invest Ophthalmol Vis Sci 1990; 31: 1779–1788 (published erratum appears in Invest Ophthalmol Vis Sci 1990; 31: 2229).
Matsubara M, Girard MT, Kublin CL, Cintron C, Fini ME . Differential roles for two gelatinolytic enzymes of the matrix metalloproteinase family in the remodelling cornea. Dev Biol 1991; 147: 425–439.
Kenney MC, Chwa M, Alba A, Saghizadeh M, Huang ZS, Brown DJ . Localisation of TIMP-1, TIMP-2, TIMP-3, gelatinase A and gelatinase B in pathological human corneas. Curr Eye Res 1998; 1998: 238–246.
Ottino P, Taheri F, Bazen HE . Platelet-activating factor induces the gene expression of TIMP-1, -2, and PAI-1: imbalance between the gene expression of MMP-9 and TIMP-1 and -2. Exp Eye Res 2002; 74: 393–402.
Kjeldsen L, Sengelov H, Lollike K, Nielsen MH, Borregaard N . Isolation and characterization of gelatinase granules from human neutrophils. Blood 1994; 83: 1640–1649.
Girard MT, Matsubara M, Kublin C, Tessier MJ, Cintron C, Fini ME . Stromal fibroblasts synthesize collagenase and stromelysin during long-term tissue remodeling. J Cell Sci 1993; 104: 1001–1011.
Daniels JT, Limb GA, Saarialho-Kere U, Murphy G, Khaw PT . Human corneal epithelial cells require MMP-1 for HGF-mediated migration on collagen I. Invest Ophthalmol Vis Sci 2003; 44: 1048–1055.
Matsubara M, Zieske JD, Fini ME . Mechanism of basement membrane dissolution preceding corneal ulceration. Invest Ophthalmol Vis Sci 1991; 32: 3221–3237.
Fini ME, Parks WC, Rinehart WB, Girard MT, Matsubara M, Cook JR et al. Role of matrix metalloproteinases in failure to re-epithelialize after corneal injury. Am J Pathol 1996; 149: 1287–1302.
Mohan R, Chintala SK, Jung JC, Villar WV, McCabe F, Russo LA et al. Matrix metalloproteinase gelatinase B (MMP-9) coordinates and effects epithelial regeneration. J Biol Chem 2002; 18(277): 2065–2072.
Jain S, Khoury JM, Chamon W, Azar DT . Corneal light scattering after laser in situ keratomileusis and photorefractive keratectomy. Am J Ophthalmol 1995; 120: 532–534.
Hersh PS, Brint SF, Maloney RK, Durrie DS, Gordon M, Michelson MA et al. Photorefractive keratectomy versus laser in situ keratomileusis for moderate to high myopia. A randomized prospective study. Ophthalmology 1998; 105: 1512–1522.
Park CK, Kim JH . Comparison of wound healing after photorefractive keratectomy and laser in situ keratomileusis in rabbits. J Cat Refract Surg 1999; 25: 842–850.
Tuft SJ, Zabel RW, Marshall J, Corneal repair following keratectomy . A comparison between conventional surgery and laser photoablation. Invest Ophthalmol Vis Sci 1989; 30: 1769–1777.
Nishida T, Tanaka T . Extracellular matrix and growth factors in corneal wound healing. Curr Opin Ophthalmol 1996; 7: 2–11.
Fini ME . Keratocyte and fibroblast phenotypes in the repairing cornea. Prog Retinal Eye Res 1999; 18: 529–551.
Tuft SJ, Gartry DS, Rawe IM, Meek KM . Photorefractive keratectomy: implications of corneal wound healing. Br J Ophthalmol 1993; 77: 243–247.
Seiler T, Holschbach A, Derse M, Jean B, Genth U . Complications of myopic photorefractive keratectomy with the excimer laser. Ophthalmology 1994; 101: 153–160.
Corbett MC, Prydal JI, Verma S, Oliver KM, Pande M, Marshall J . An in vivo investigation of the structures responsible for corneal haze after photorefractive keratectomy and their effect on visual function. Ophthalmology 1996; 103: 1366–1380.
Brown RA, McFarland CD . Regulation of growth plate cartilage degradation in vitro: effects of calcification and a low molecular weight angiogenic factor (ESAF). Bone Miner 1992; 17: 49–57.
Maddox PH, Jenkins D . 3-Aminopropyltriethoxysilane (APES): a new advance in section adhesion. J Clin Pathol 1987; 40: 1256–1257.
Hembry RM, Murphy G, Cawston TE, Dingle JT, Reynolds JJ . Characterisation of a specific antiserum for mammalian collagenase from several species: Immunolocalisation of collagenase in rabbit chondrocytes and uterus. J Cell Sci 1986; 81: 105–123.
Hipps DS, Hembry RM, Docherty AJ, Reynolds JJ, Murphy G . Purification and characterisation of human 72-kDa gelatinase (type IV collagenase) Use of immunolocalisation to demonstrate the non-coordinate regulation of the 72-kDa and 95-kDa gelatinases by human fibroblasts. Biol Chem Hoppe Seyler 1991; 372: 287–296.
Murphy G, Hembry RM, Reynolds JJ . Characterisation of a specific antiserum to rabbit stromelysin and demonstration of the synthesis of collagenase and stromelysin by stimulated rabbit articular chondrocytes. Coll Relat Res 1986; 6: 351–363.
Murphy G, Ward R, Hembry RM, Reynolds JJ, Kuhn K, Tryggvason K . Characterisation of gelatinase from pig polymorphonuclear leucocytes. A metalloproteinase resembling tumour type IV collagenase. Biochem J 1989; 258: 463–472.
Gallegos NC, Smales C, Savage FJ, Hembry RM, Boulos PB . The distribution of matrix metalloproteinases and tissue inhibitor of metalloproteinases in colorectal cancer. Surg Oncol 1995; 4: 111–119.
Vaalamo M, Weckroth M, Puolakkainen P, Kere J, Saarinen P, Lauharanta J et al. Patterns of matrix metalloproteinase and TIMP-1 expression in chronic and normally healing human cutaneous wounds. Br J Dermatol 1996; 135: 52–59.
Pilcher BK, Dumin JA, Sudbeck BD, Krane SM, Welgus HG, Parks WC et al. The activity of Collagenase-1 is required for keratinocyte migration on a type I collagen Matrix. J Cell Biol 1997; 137: 1445–1457.
Madlener M, Parks WC, Werner S . Matrix metalloproteinases (MMPs) and their physiological inhibitors (TIMPs) are differentially expressed during excisional skin wound repair. Exp Cell Res 1998; 242: 201–210.
Azar DT, Pluznik D, Jain S, Khoury JM . Gelatinase B and A expression after laser in situ keratomileusis and photorefractive keratectomy. Arch Ophthalmol 1998; 116: 1206–1208.
Yu Q, Stamenkovic I . Cell surface-localized matrix metalloproteinase-9 proteolytically activates TGF-beta and promotes tumour invasion and angiogenesis. Genes Dev 2000; 14: 163–176.
Lu PC, Ye H, Maeda M, Azar DT . Immunolocalisation and gene expression of matrilysin during corneal wound healing. Invest Ophthalmol Vis Sci 1999; 40: 20–27.
Kure T, Chang JH, Kato T, Hernandez-Quintela E, Ye H, Lu PC et al. Corneal neovascularisation after excimer keratectomy wounds in matrilysin-deficient mice. Invest Ophthalmol Vis Sci 2003; 44: 137–144.
Ye HQ, Maeda M, Yu FS, Azar DT . Differential expression of MT1-MMP (MMP-14) and collagenase III (MMP-13) genes in normal and wounded rat corneas. Invest Ophthalmol Vis Sci 2000; 41: 2894–2899.
Acknowledgements
This work was supported in part by a BUPA scholarship, Moorfields Eye Hospital locally organized research and the Peel Research Trust. Professor G Murphy generously provided the MMP antibodies. Dr F Savage provided technical advice.
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Mulholland, B., Tuft, S. & Khaw, P. Matrix metalloproteinase distribution during early corneal wound healing. Eye 19, 584–588 (2005). https://doi.org/10.1038/sj.eye.6701557
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DOI: https://doi.org/10.1038/sj.eye.6701557
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