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.
OBJECTIVE
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 alpha5 (IV)-immunoreactivity, but lacked the alpha1/alpha2-immunoreactivity of collagen IV. Epithelial BM in healing burned cornea transiently exhibited alpha1/alpha2-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 alpha5-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.
Kato
Tadashi
T
Department of Ophthalmology, Wakayama Medical University School of Medicine, 811-1 Kimiidera, Wakayama 641-0012, Japan. alioth@pb3.so-net.ne.jp
Saika
Shizuya
S
Ohnishi
Yoshitaka
Y
D000818:Animals; D001485:Basement Membrane / Q000293:injuries / Q000378:metabolism / Q000473:pathology; D002057:Burns, Chemical / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D024141:Collagen Type IV / Q000096:biosynthesis; D004151:Dipeptides / Q000627:therapeutic use*; D004195:Disease Models, Animal; D019573:Epithelium, Corneal / Q000293:injuries / Q000378:metabolism / Q000473:pathology*; D005126:Eye Burns / Q000139:chemically induced / Q000188:drug therapy / Q000473:pathology*; D005260:Female; D008297:Male; D061965:Matrix Metalloproteinase Inhibitors; D009924:Organ Culture Techniques; D011480:Protease Inhibitors / Q000627:therapeutic use*; D011817:Rabbits; D012972:Sodium Hydroxide / Q000633:toxicity; D016896:Treatment Outcome; D014945:Wound Healing / Q000187:drug effects*
D024141:Collagen Type IV; D004151:Dipeptides; D061965:Matrix Metalloproteinase Inhibitors; C078131:N-(2(R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl)-L-tryptophan methylamide; D011480:Protease Inhibitors; D012972:Sodium Hydroxide