Effect of topical fluoroquinolones on the expression of matrix metalloproteinases in the cornea

Victor E Reviglio, Melinda A Hakim, Jae K Song, Terrence P O'Brien, Victor E Reviglio, Melinda A Hakim, Jae K Song, Terrence P O'Brien

Abstract

Background: Matrix metalloproteinases play an important role in extracellular matrix deposition and degradation. Based on previous clinical observations of corneal perforations during topical fluoroquinolone treatment, we decided to evaluate the comparative effects of various fluoroquinolone eye drops on the expression of matrix metalloproteinases (MMPs) in cornea.

Methods: Eighty female Lewis rats were divided into two experimental groups: intact and wounded corneal epithelium. Uniform corneal epithelial defects were created in the right eye with application of 75% alcohol in the center of the tissue for 6 seconds. The treatment groups were tested as follows: 1) Tear drops: carboxymethylcellulose sodium 0.5 % (Refresh, Allergan); 2) Ciprofloxacin 0.3% (Ciloxan, Alcon); 3) Ofloxacin 0.3%(Ocuflox, Allergan); 4) Levofloxacin 0.5%(Quixin, Santen). Eye drops were administered 6 times a day for 48 hours. Rats were sacrificed at 48 hours. Immunohistochemical analysis and zymography were conducted using antibodies specific to MMPs-1, 2, 8 and 9.

Results: MMP-1, MMP-2, MMP-8 and MMP-9 expression were detected at 48 hrs in undebrided corneal epithelium groups treated with the topical fluoroquinolones. No statistical difference was observed in quantitative expression of MMPs among ciprofloxacin 0.3%, ofloxacin 0.3%, levofloxacin 0.5%. When the artificial tear group and the fluoroquinolone groups with corneal epithelial defect were compared, increased expression of MMPs was observed as a result of the wound healing process. However, the fluoroquinolone treated group exhibited high statistically significantly levels of MMPs expression.

Conclusions: Our study provides preliminary evidence that topical application of fluoroquinolone drugs can induce the expression of MMP-1, MMP-2, MMP-8 and MMP-9 in the undebrided corneal epithelium compared to artificial tear eye drops.

Figures

Figure 1
Figure 1
Immunolocalization of MMP-1 in intact corneal epithelium at 48 hours is shown for each treatment group. Panel a: artificial tears; panel b: Ciprofloxacin 0.3%; panel c: Ofloxacin 0.3%; panel d: Levofloxacin 0.5%. A photomicrograph of the artificial tear group shows negative corneal staining for MMP-1. However, intense corneal epithelial and superficial stromal expression of MMP-1 was identified in intact corneal epithelium groups treated with fluoroquinolone eye drops (panel's b-d). The presence of MMP-1 was detected mostly in the corneal epithelial cells (arrows). (Immunohistochemistry, ×400 magnification).
Figure 2
Figure 2
A representative western blot of MMP-1 and MMP-8 expression of supernatant samples from control (artificial tear) and the fluoroquinolone test groups. Corneas with debrided (OD) and intact (OS) epithelium were analyzed. 1. MW; 2. Tears OS; 3. Tears OD; 4. Ciprofloxacin OS; 5. Ciprofloxacin OD; 6. Ofloxacin OS; 7. Ofloxacin OD; 8. Levofloxacin OS; 9. Levofloxacin OD The molecular sizes of both MMPs were calculated based on a molecular weight standard and recorded in kilodaltons (kDa). The fluoroquinolone eye drops increased the expression of both metalloproteinases, MMP-1 (42 kDa) and of MMP-8 (64 kDa) in intact corneas; whereas, the expression of these MMPs was negative in the controls treated with artificial tears.
Figure 3
Figure 3
Typical gelatin zymography analyses of gelatinase A (MMP-2) and gelatinase B (MMP-9) expression in artificial tear group rat and in the test groups were performed. Corneal conditioned medium samples from debrided (OD) and intact (OS) corneal epithelium of each group were analyzed. 1. Tears OD; 2. Tears OS; 3. Levofloxacin 0.5% OD; 4. Levofloxacin 0.5% OS; 5. Ciprofloxacin OD; 6. Ciprofloxacin OS; 7. Ofloxacin OD; 8. Ofloxacin OS. MMP-9 (92 kDa) and MMP-2 (72 kDa) expression were up regulated in fluoroquinolone treated corneas and were present at basal level in unwounded corneas receiving artificial tears. Corneal epithelium debridement up regulated the expression of MMP-9 and MMP-2 in all test groups.
Figure 4
Figure 4
Effects of fluoroquinolone eye drops on the corneal rat's metalloproteinases expression. The bands densities of western immunoblots for MMP-1 (A) and MMP-8 (B) and zymograms for MMP-2 (C), MMP-9 (D) were subjected to densitometry analysis, the statistical significance was determined by one-way ANOVA test (values are means, error bars). P < 0.05 was considered statistically significant when compared fluoroquinolones groups vs. artificial tear group. Note the highest values of MMPs levels from topical fluoroquinolone groups compared to artificial tear group in unwounded corneas.

References

    1. Graves A, Henry M, O'Brien TP, Hwang DG, Van Buskirk A, Trousdale MD. In Vitro Susceptibilities of Bacterial Ocular Isolates to Fluoroquinolones. CORNEA. 2001;20:301–305. doi: 10.1097/00003226-200104000-00012.
    1. O'Brien TP, Maguire MG, Fink NE, Alfonso E, McDonnell P. Efficacy of ofloxacin vs cefazolin and tobramycin in the therapy of bacterial keratitis. Arch Ophthalmol. 1995;113:1257–1265.
    1. Harrell RM. Fluoroquinolone-induced tendinopathy. What do we know? South Med J. 1999;92:622–625.
    1. Huston KA. Achilles tendonitis and tendon rupture due to fluoroquinolone antibiotics (Letter) N Eng J Med. 1994;331:748. doi: 10.1056/NEJM199409153311116.
    1. Redmond AO. Risk-benefit experience of ciprofloxacin use in pediatric patients in the United Kingdom. Pediatr Infect Dis J. 1997;16:147–9. doi: 10.1097/00006454-199701000-00040. discussion 160–2.
    1. Burstein NL. Corneal cytotoxicity of topically applied drugs, vehicles and preservatives. Surv Ophthalmol. 1980;25:15–30.
    1. Cutarelli PE, Lass JH, Lazarus HM, Putman SC, Jacobs R. Topical fluoroquinolones: antimicrobial activity and in vitro corneal epithelial toxicity. Curr Eye Res. 1991;10:557–563.
    1. Seitz B, Hayashi S, Wee WR, LaBree L, McDonnell PJ. In vitro effects of aminoglycosides and fluoroquinolones on keratocytes. Invest Ophthal Vis Sci. 1996;37:656–665.
    1. Morlet N, Minassian D, Butcher J. Ofloxacin Study Groups. Risk factors for treatment of suspected microbial keratitis. Br J Ophthalmol. 1999;83:1027–1031.
    1. Bekoe NA, Li Q, Ashraf F, O'Brien TP. Effects of ciprofloxacin, ofloxacin and gentamicin on corneal cells and wound healing. Invest Ophthalmol Vis Sci. 1999;40:S548.
    1. Mallari PLT, McCarty DJ, Daniell M, Taylor H. Increased incidence of corneal perforation after topical fluoroquinolone treatment for microbial keratitis. Am J Ophthalmol. 2001;131:131–133. doi: 10.1016/S0002-9394(00)00642-5.
    1. Massova I, Kotra LP, Fridman R, et al. Matrix metalloproteinases: structures, evolution and diversification. FASEB J. 1998;12:1075–1095.
    1. Fini ME, Parks WC, Rinehart WB, Girard MT, et al. Role of matrix metalloproteinases in failure to re-epithelialize after corneal injury. Am J Pathol. 1996;149:1287–302.
    1. Ye HQ, Azar DT. Statement of gelatinases A and B, and TIMPs 1 and 2 during wound healing. Invest Ophthalmol Vis Sci. 1998;39:913–21.
    1. Leibowitz HM. Antibacterial effectiveness of ciprofloxacin 3% ophthalmic solution in the treatment of bacterial conjunctivitis. Am J Ophthalmol. 1991;112:29S–33S.
    1. Moreira LB, Lee RF, Oliveira C, LaBree L, McDonnell PJ. Effect of topical fluoroquinolones on corneal re-epithelialization after excimer laser keratectomy. J Cataract Refract Surg. 1997;23:845–848.
    1. Essepian JP, Rajpal R, O'Brien TP. Tandem scanning confocal microscopic analysis of ciprofloxacin corneal deposits in vivo. Cornea. 1995;14:402–407.
    1. Kanellopoulos AJ, Miller F, Wittpenn JR. Deposition of topical ciprofloxacin to prevent re-epithelialization of corneal defect (Letter) Am J Ophthalmol. 1994;117:258–259.
    1. Fini ME, Yue YJT, Sugar J. Collagenolytic / gelatinolytic metalloproteinases in normal and keratoconus corneas. Curr Eye Res. 1992;11:849–62.
    1. Birkedal-Hansen H, Moore WG, Bodden MK, et al. Matrix metalloproteinases: a review. Crit Rev Oral Biol Med. 1993;4:197–250.
    1. O'Brien TP, Li QJ, Sauerburger F, Reviglio VE, et al. The role of matrix metalloproteinases in ulcerative keratolysis associated with perioperative diclofenac use. Ophthalmology. 2001;108:656–659. doi: 10.1016/S0161-6420(00)00590-X.
    1. Riesbeck K, Sigvardsson M, Leanderson T, Forsgren A. Superinduction of cytokine gene transcription by ciprofloxacin. J Immunol. pp. 343–52. 1994 Jul 1.
    1. Williams RJ, III, Attia E, Wickiewicz TL, Hannafin JA. The effect of ciprofloxacin on tendon, paratenon, and capsular fibroblast metabolism. Am J Sports Med. 2000;28:364–9.
    1. Shakibaei M, Stahlmann R. Ultrastructure of Achilles tendon from rats after treatment with fleroxacin. Arch Toxicol. 2001;75:97–102. doi: 10.1007/s002040000203.
    1. Egerbacher M, Seiberl G, Wolfesberger B, Walter I. Ciprofloxacin causes Cytoskeletal changes and detachment of human and rat chondrocytes in vitro. Arch Toxicol. 2000;73:557–63. doi: 10.1007/s002040050008.
    1. Shakibaei M, Stahlmann R. Ultrastructural changes induced by the des-F(6)-quinolone garenoxacin (BMS-284756) and two fluoroquinolones in Achilles tendon from immature rats. Arch Toxicol. 2003.

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