Abstract
Purpose
To examine the alteration in structure and matrix composition of epithelial basement membrane (BM) during the healing of alkali-burned rabbit cornea, and the roles of matrix metalloproteinases (MMPs) in these alterations.
Methods
The central cornea of one eye of 78 albino rabbits was exposed to 1 N NaOH for 180 s under general and topical anesthesia and allowed to heal with or without subconjunctival injection of GM6001 (an MMP inhibitor). Cryosections of affected corneas were observed by H&E staining, immunohistochemistry for type IV collagen subtypes, or in situ zymography for detection of localization of MMP activity.
Results
Uninjured corneal epithelial BM exhibited α5 (IV)-immunoreactivity, but lacked the α1/α2-immunoreactivity of collagen IV. Epithelial BM in healing burned cornea transiently exhibited α1/α2-immunoreactivity. Examination by in situ zymography showed an upregulation of MMP activity in the regenerated central epithelium and anterior stroma of the burned corneas at days 7 and 14. GM6001 suppressed degradation of α5-containing epithelial BM in vivo and also in organ culture.
Conclusions
Epithelial BM was degraded by endogenous MMPs during healing following an alkali burn in rabbit cornea. GM6001 had an inhibitory effect on the degradation of the epithelial basement membrane in burned cornea in vivo. Jpn J Ophthalmol 2006;50:90–95 © Japanese Ophthalmological Society 2006
Similar content being viewed by others
References
HE Kaufman MB McDonald BA Barron SR Waltman (1998) The cornea EditionNumber1st Ed. Churchill Livingston New York 610–617
S Saika S Kobata N Hashizume Y Okada O Yamanaka (1993) ArticleTitleEpithelial basement membrane in alkali-burned corneas in rats. Immunohistochemical study Cornea 12 383–390 Occurrence Handle1:STN:280:ByuC3sbjtFQ%3D Occurrence Handle8306658
S Saika S Kobata N Hashizume O Yamanaka Y Okada A Ooshima (1995) ArticleTitleType IV collagen in the basement membrane of the corneal epithelium after alkali burns in guinea pigs Ophthalmic Res 27 129–135 Occurrence Handle1:CAS:528:DyaK2MXos1Cksbw%3D Occurrence Handle8538989
H Matsuda GK Smelser (1973) ArticleTitleEpithelium and stroma in alkali-burned corneas Arch Ophthalmol 89 396–401 Occurrence Handle1:STN:280:CSyC2Mngs1c%3D Occurrence Handle4697217
RR Pfister (1983) ArticleTitleThe effects of chemical injury on the ocular surface Ophthalmology 90 601–609 Occurrence Handle1:STN:280:BiyB1Mfnt1w%3D Occurrence Handle6888853
S Saika K Uenoyama K Hiroi H Tanioka K Takase M Hikita (1993) ArticleTitleAscorbic acid phosphate ester and wound healing in rabbit corneal alkali burns: epithelial basement membrane and stroma Graefes Arch Clin Exp Ophthalmol 231 221–227 Occurrence Handle1:CAS:528:DyaK3sXksVSrtbw%3D Occurrence Handle8486304
M Matsubara MT Girard CL Kublin C Cintron ME Fini (1991) ArticleTitleDifferential roles for two gelatinolytic enzymes of the matrix metalloproteinase family in the remodelling cornea Dev Biol 147 425–439 Occurrence Handle10.1016/0012-1606(91)90300-R Occurrence Handle1:CAS:528:DyaK38XotFCgsg%3D%3D Occurrence Handle1655541
M Matsubara JD Zieske ME Fini (1991) ArticleTitleMechanism of basement membrane dissolution preceding corneal ulceration Invest Ophthalmol Vis Sci 32 3221–3237 Occurrence Handle1:STN:280:By2D1cvkvFA%3D Occurrence Handle1660857
VM Kahari U Saarialho-Kere (1997) ArticleTitleMatrix metalloproteinases in skin Exp Dermatol 6 199–213 Occurrence Handle1:CAS:528:DyaK2sXmslCgtrs%3D Occurrence Handle9450622
WC Parks (1999) ArticleTitleMatrix metalloproteinases in repair Wound Repair Regen 7 423–432 Occurrence Handle10.1046/j.1524-475X.1999.00423.x Occurrence Handle1:STN:280:DC%2BD3c%2FpvVyrtQ%3D%3D Occurrence Handle10633001
L Ravanti VM Kahari (2000) ArticleTitleMatrix metalloproteinases in wound repair Int J Mol Med 6 391–407 Occurrence Handle1:CAS:528:DC%2BD3cXntFOnsrw%3D Occurrence Handle10998429
Y Herouy (2001) ArticleTitleMatrix metalloproteinases in skin pathology Int J Mol Med 7 3–12 Occurrence Handle1:CAS:528:DC%2BD3MXjtFykuw%3D%3D Occurrence Handle11115601
EA O'Toole (2001) ArticleTitleExtracellular matrix and keratinocyte migration Clin Exp Dermatol 26 525–530 Occurrence Handle10.1046/j.1365-2230.2001.00891.x Occurrence Handle11678882
M Shimoda M Ishizaki T Saiga N Yamanaka (1997) ArticleTitleExpression of matrix metalloproteinases and tissue inhibitor of metalloproteinase by myofibroblasts—morphological study on corneal wound healing Nippon Ganka Gakkai Zasshi (J Jpn Ophthalmol Soc) 101 371–379 Occurrence Handle1:STN:280:ByiA3Mvmtlw%3D
HQ Ye M Maeda FS Yu DT Azar (2000) ArticleTitleDifferential expression of MT1-MMP (MMP-14) and collagenase III (MMP-13) genes in normal and wounded rat corneas Invest Ophthalmol Vis Sci 41 2894–2899 Occurrence Handle1:STN:280:DC%2BD3cvivVWrsw%3D%3D Occurrence Handle10967042
T Nishida S Nakagawa T Awata Y Ohashi K Watanabe R Manabe (1983) ArticleTitleFibronectin promotes epithelial migration of cultured rabbit cornea in situ J Cell Biol 97 1653–1657 Occurrence Handle10.1083/jcb.97.5.1653 Occurrence Handle1:STN:280:BiuD3s3mvV0%3D Occurrence Handle6415071
S Saika Y Kawashima Y Okada et al. (1999) ArticleTitleImmunohistochemical and ultrastructural analysis of dysplastic epithelium of human ocular surface: basement membrane and intermediate filament Cornea 18 343–352 Occurrence Handle1:STN:280:DyaK1M3mslSrsw%3D%3D Occurrence Handle10336039
ME Fini WC Parks WB Rinehart et al. (1996) ArticleTitleRole of matrix metalloproteinases in failure to re-epithelialize after corneal injury Am J Pathol 149 1287–1302 Occurrence Handle1:CAS:528:DyaK28XmsFOmtbg%3D Occurrence Handle8863676
BG Hudson ST Reeders K Tryggvason (1993) ArticleTitleType IV collagen: structure, gene organization, and role in human disease J Biol Chem 268 26033–26036 Occurrence Handle1:CAS:528:DyaK3sXmsVKrtLw%3D Occurrence Handle8253711
M Ishizaki A Westerhausen-Larson J Kino T Hayashi WWY Kao (1993) ArticleTitleDistribution of collagen IV in human ocular tissues Invest Ophthalmol Vis Sci 34 2680–2689 Occurrence Handle1:STN:280:ByyA2cnhslQ%3D Occurrence Handle7688359
Y Ninomiya M Kagawa K Iyama et al. (1995) ArticleTitleDifferential expression of two basement membrane collagen genes, COL4A6 and COL4A5, demonstrated by immunofluorescence staining using peptide-specific monoclonal antibodies J Cell Biol 130 1219–1229 Occurrence Handle10.1083/jcb.130.5.1219 Occurrence Handle1:CAS:528:DyaK2MXnvVWht70%3D Occurrence Handle7657706
M Sugimoto T Oohashi Y Ninomiya (1994) ArticleTitleThe genes COL4A5 and COL4A6, coding for basement membrane collagen chains alpha 5 (IV) and alpha 6 (IV), are located head-to-head in close proximity on human chromosome Xq22, and COL4A6 is transcribed from two alternative promoters Proc Natl Acad Sci USA 91 11679–11683 Occurrence Handle1:CAS:528:DyaK2MXitlyrtLc%3D Occurrence Handle7972123
Y Sado M Kagawa Y Kishiro et al. (1995) ArticleTitleEstablishment by the rat lymph node method of epitope-defined monoclonal antibodies recognizing the six different alpha chains of human type IV collagen Histochem Cell Biol 104 267–275 Occurrence Handle10.1007/BF01464322 Occurrence Handle1:CAS:528:DyaK2MXosVWhurg%3D Occurrence Handle8548560
AV Ljubimov RE Burgeson RJ Butkowski et al. (1996) ArticleTitleExtracellular matrix alterations in human corneas with bullous keratopathy Invest Ophthalmol Vis Sci 37 997–1007 Occurrence Handle1:STN:280:BymB3Mfgt1Y%3D Occurrence Handle8631643
ML Corcoran DE Kleiner WG Stetler-Stevenson (1995) Regulation of matrix metalloproteinases during extracelluar matrix turnover S Zderic (Eds) Muscle matrix and bladder function Plenum Press New York 151–159
AFC Richard (1998) The molecular and cellular biology of wound repair EditionNumbersecond ed. Plenum Press New York 442–461
T Sakimoto J Shoji M Sawa (2003) ArticleTitleActive form of gelatinases in tear fluid in patients with corneal ulcer or ocular burn Jpn J Ophthalmol 47 423–426 Occurrence Handle1:CAS:528:DC%2BD3sXntVCmsb8%3D Occurrence Handle12967855
KP Xu Y Ding J Ling Z Dong FS Yu (2004) ArticleTitleWound-induced HB-EGF ectodomain shedding and EGFR activation in corneal epithelial cells Invest Ophthalmol Vis Sci 45 813–820 Occurrence Handle14985295
Author information
Authors and Affiliations
Corresponding author
Additional information
The abstract of this paper was presented by Dr. Kato at the annual meeting of the Association for Research in Vision and Ophthalmology (Fort Lauderdale, FL, USA, in 1998 and 2003) as an oral and a poster presentation, respectively.
About this article
Cite this article
Kato, T., Saika, S. & Ohnishi, Y. Effects of the Matrix Metalloproteinase Inhibitor GM6001 on the Destruction and Alteration of Epithelial Basement Membrane During the Healing of Post-alkali Burn in Rabbit Cornea. Jpn J Ophthalmol 50, 90–95 (2006). https://doi.org/10.1007/s10384-005-0287-8
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10384-005-0287-8