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1 339 Vol. 32, pp , 2004 : in vitro : in vitro In vitro 6 3 b in vitro in vitro in vitro in vitro

2 340 evidence-based medicine: EBM 1 b a 1 2 b 3 b in vitro : in vitro: 1 : b a 1b 10 b XE 0.5 TG a 1 b

3 a FL BSS PLUS ml 10 BSS PLUS 20 ml FL superficial punctuate keratopathy;spk AD 12 A area, D density SPK 4 tear film break up time BUT Schirmer I BUT 3 BUT 5.0 Schirmer I mm mm 5 paired t-test Mann- Whitney U-test 5 in vitro: Epilife HCGS KC-6450 Cell Titer 96 One Solution Cell Proliferation Assay cm ml 96 well 50 ml ml ml 37 C nm

4 342 Fig. 1. Time course of corneal fluorescein uptake after topical administration of antiglaucoma eyedrop. Timolol group n 10 Unoprostone group n 10 Pilocarpine group n 10 paired t test p 0.05 Values are mean SD. ANOVA Bonferroni Dunnett[s test Fig ng ml ng ml p ng ml ng ml 1 1 b XE 10 TG Fig ng ml ng ml p ng ml ng ml p ng ml ng ml ng ml

5 343 Fig. 2. Time course of corneal fluorescein uptake after addition of antiglaucoma eyedrop. Latanoprost adition group n 22 Unoprostone adition group n 21 Dorzolamide adition group n 12 paired t test p 0.05 Values are mean SD Fig ng ml ng ml ng ml p ng ml ng ml n 5 6 n 5 1 Fig. 4, Table ng ml ng ml ng ml ng ml ng ml b ng ml 0.5 TG ng ml 0.5 XE ng ml ng ml 125

6 344 Fig. 3. Corneal fluorescein uptake versus number of antiglaucoma eyedrop. Values are mean SD ng ml ng ml SPK TG XE SPK Fig. 5 BUT TG XE BUT Schirmer I mm mm 0.5 TG mm mm mm mm mm 0.5 XE mm mm mm 2 Fig. 6, Table n 4 n 4 n 3 5 b 120 ng ml 0.5 XE TG ng ml ng ml ng ml ng ml ng ml ng ml b 126

7 345 Fig. 4. Fluorescein uptake after asingle antiglaucoma eyedrop. Pilocarpine: PL Unoprostone: UP Latanoprost: LP Timolol: TM Timoptol XE: XE Rysmon TG: TG Betaxolol: BX Carteolol: CT Nipradilol: NP Values are mean SD. Table 1. Background of Patients in the Group Receiving Eye Drops for a Long Period 130 ng ml ng ml ng ml ng ml b ng ml b ng ml 81.3 ng ml 89.5 ng ml b p Fig. 7, 127

8 346 Fig. 5. Fluorescein staining score after asingle antiglaucoma eyedrop. Pilocarpine: PL Unoprostone: UP Latanoprost: LP Timolol: TM Timoptol XE: XE Rysmon TG: TG Betaxolol: BX Carteolol: CT Nipradilol: NP Values are mean SD. Table TG n 2 n 4 n 4 n ng ml 218.5ng ml n 8 XE ng ml n 6 4 Fig. 8, Table n 4 TG n 2 n ng ml 0.5 XE ng ml n ng ml Table ng ml 4 Fig. 9 : n

9 347 Fig. 6. Fluorescein uptake after varying combination of two antiglaucoma eyedrops. Unoprostone: UP Latanoprost: LP Timolol: TM Timoptol XE: XE Rysmon TG: TG Betaxolol: BX Carteolol: CT Nipradilol: NP Values are mean SD. Table 2. Background of Patients in the Group Receiving Eye Drops for a Long Period b

10 348 Fig. 7. Fluorescein uptake after varying combination of three antiglaucoma eyedrops. Pilocarpine: PL Unoprostone: UP Latanoprost: LP Timolol: TM Timoptol XE : XE Rysmon TG: TG Betaxolol: BX Carteolol: CT Nipradilol: NP Dorzolamide: DO Bunazosin: BU Values are mean SD. Table 3. Background of Patients in the Group Receiving Eye Drops for a Long Period b

11 349 Fig. 8. Corneal fluorescein uptake after varying combination of four antiglaucoma eyedrops. Pilocarpine: PL Unoprostone: UP Latanoprost: LP Timolol: TM Timoptol XE: XE Rysmon TG: TG Betaxolol: BX Carteolol: CT Nipradilol: NP Dorzolamide: DO Bunazosin: BU Dipivefrin: DI Values are mean SD. Table 4. Background of patients in the group receiving eye drops for a long period

12 350 Fig. 9. Inhibitory e#ects of topical antiglaucoma eyedrops on corneal epithelial proliferation. : Unoprostone : Latanoprost : Carteolol : Timolol : Betaxolol : Nipradilol : Dorzolamide Values are mean SD SPK delayed staining ml

13 b 6 b BUT b 29 Hoh 30 b BUT Schirmer XE TG 32 SPK b 0.5 XE TG 10 C C TG 33 BUT BUT 6.7 Schirmer test 8.6 mm 133

14 352 2 b 140 ng ml b 150 ng ml 4 b 120 ng ml 3 b a 1 b b 83.7 ng ml XE ng ml ng ml 3 b b b b ng ml XE TG ng ml 3 b tight junction

15 in vitro b b b 37 reflex loop in vitro in vitro in vitro SPK ; 107: ; 15: ; 15: TGTM ; 53: b 2001; 105:

16 ; 17: ; 104: ; 55: ; 10: ; 10: ; 98: ; 48: ; 37: ; 13: ; 108: ; 57: : : Wolosin, J. M. Regeneration of resistance and ion transport in rabbit corneal epithelium after induced surface cell exfoliation. J. Membr. Biol 1988; 104: Wang, Y. ZO-1 in corneal epithelium; Stratal distribution and synthesis induction by outer cell removal. Exp. Eye Res 1993; 57: Huang, AJW. Paracellular permeability of corneal and conjunctival epithelia. Invest. Ophthalmol. Vis. Sci 1989; 30: Yokoi N, Kinoshita S. Clinical evaluation of corneal epithelial barrier function with the slit lamp fluorophotometer. Cornea 1995; 14: ; 58: ; 8: ; 48: F2a 1998; 102: ; 105: : b 1995; 49: ; 49: Hoh H & Nastainczyk W. b-blocker und Hornhautsensibilitat. Fortschr Ophthalmol 1991; 88: Timolol GS ; 12: ; 18: T TG 2003; 20:

17 KT-210 III ; 13: : Iwata, T.: In vitro studies on the permeability of the corneal epithelium with horseradish peroxidase Jpn J. ophthalmol. 1975; 19: Van Buskirk EM. Corneal Anesthesia after timolol maleate therapy. Am J Ophthalmol 1979; 88: Yokoi N, Komuro A, Takehisa Y, Kinoshita S: Clinical assessment of conjunctival damage and tear film stability in drug-induced epithelial keratopathy. In Lass JH Eds : Advances in corneal research: Selected transaction of the world cornea congress. Plenum Press, New York, 1998;

18 356 Abstract E#ects of Antiglaucoma Eye Drops on the Cornea: In clinic in vitro Naoto Tokuda, Yumiko Aoyama, Jun Inoue and Satoki Ueno Purpose: To elucidate the e#ects of antiglaucoma eyedrops on the cornea in clinic and in vitro. Patients and methods: In the clinic corneal epithelial barrier function was examined by fluorophotometry in 538 eyes of 828 glaucoma patients treated with various antiglaucoma eyedrops. In vitro, to investigate the e#ects of antiglaucoma eyedrops on the proliferation of human corneal epitherial cells. Results: Fluorescein uptake in the cornea increased following administration of antiglaucoma eyedrops and reached a plateau by 6 months. Among eyes concurrently treated with more than 1 medication, the barrier function of eyes treated with 2 or 3 medications was significantly lower than that of eyes treated with 1 medication. As to eyes treated with a single medication, parasymapathetic nerve stimulating agents and prostaglandin analogues did not reveal marked change in the barrier function, while beta-blockers reduced the barrier function. In the case of multidrug use, a combined regimen of prostaglandin analogues and beta-blockers, specifically betaxolol and timolol, a#ected the corneal epithelial barrier. These results suggest that topical administration of prostaglandin analogues inhibits the proliferation of human corneal epithelial cells more strongly than topical administration of beta-blockers. Although timolol has a low toxicity on corneal epithelial cells in vitro, it may alter the corneal epithelial barrier function when applied clinically and may a#ect corneal sensitivity and tear fluid dynamics. Therefore, caution should be taken when administering timolol in combination with other antiglaucoma eyedrops. Prostaglandin analogues has a high toxicity on corneal epithelial cells in vitro, however prostaglandin analogues did not reveal marked change in the barrier function. Conclusions: This study demonstrates that clinical as well as in vitro evaluations are important for assessing the e#ects of antiglaucoma eyedrops on the corneal epithelium. Department of Ophthalmology, St Marianna University School of Medicine 138

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