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

Reviglio VE, Hakim MA, Song JK, O'Brien TP - BMC Ophthalmol (2003)

Bottom Line: 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.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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Ocular Microbiology and Immunology Laboratory, Refractive Surgery Research Laboratory, Wilmer Eye Institute, Johns Hopkins University, Baltimore, Maryland, USA. victorwilmer@aol.com

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.

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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).
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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).

Mentions: We also examined the MMP expression after the application of commercially available fluoroquinolone eye drops. Analysis of immunohistochemical staining disclosed a positive staining for metalloproteinase at corneal epithelium and superficial stroma. Interestingly, the percentage of eyes positively staining for MMP-1 and MMP-8 was not found to be significantly different between unwounded groups treated with each of the commercially available fluoroquinolone eye drops. This result indicates that the fluoroquinolone ophthalmic solutions are the major trigger for the increased expression of these MMPs in the normal corneal epithelium. In previous studies, we also observed MMP-8 expression at the level of the corneal epithelium associated with ulcerative keratolysis. These findings identify a novel source of MMP-8 expression at the corneal epithelial cells as a non-PMN cellular source of this collagenase. Using hematoxylin-eosin stain, the polymorpho nuclear neutrophils (PMNs) were identified on the basis of their characteristic multilobed nucleus and staining pattern. In the current study, the fluoroquinolone treatment causes a non significant increase of PMNs in unwounded corneas compared to the artificial tear group at 48 hours. Figure 1 illustrates MMP-1 immunostaining primarily in the epithelial cells and occasionally in the superficial stromal keratocytes of intact corneas of the fluoroquinolone treated groups.


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

Reviglio VE, Hakim MA, Song JK, O'Brien TP - BMC Ophthalmol (2003)

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).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC239861&req=5

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).
Mentions: We also examined the MMP expression after the application of commercially available fluoroquinolone eye drops. Analysis of immunohistochemical staining disclosed a positive staining for metalloproteinase at corneal epithelium and superficial stroma. Interestingly, the percentage of eyes positively staining for MMP-1 and MMP-8 was not found to be significantly different between unwounded groups treated with each of the commercially available fluoroquinolone eye drops. This result indicates that the fluoroquinolone ophthalmic solutions are the major trigger for the increased expression of these MMPs in the normal corneal epithelium. In previous studies, we also observed MMP-8 expression at the level of the corneal epithelium associated with ulcerative keratolysis. These findings identify a novel source of MMP-8 expression at the corneal epithelial cells as a non-PMN cellular source of this collagenase. Using hematoxylin-eosin stain, the polymorpho nuclear neutrophils (PMNs) were identified on the basis of their characteristic multilobed nucleus and staining pattern. In the current study, the fluoroquinolone treatment causes a non significant increase of PMNs in unwounded corneas compared to the artificial tear group at 48 hours. Figure 1 illustrates MMP-1 immunostaining primarily in the epithelial cells and occasionally in the superficial stromal keratocytes of intact corneas of the fluoroquinolone treated groups.

Bottom Line: 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.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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Ocular Microbiology and Immunology Laboratory, Refractive Surgery Research Laboratory, Wilmer Eye Institute, Johns Hopkins University, Baltimore, Maryland, USA. victorwilmer@aol.com

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.

Show MeSH
Related in: MedlinePlus