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. 2008 Sep;92(9):1275-82.
doi: 10.1136/bjo.2008.138768.

Conjunctival and corneal reactions in rabbits following short- and repeated exposure to preservative-free tafluprost, commercially available latanoprost and 0.02% benzalkonium chloride

H Liang  1 , C Baudouin, A Pauly, F Brignole-Baudouin
Affiliations

Conjunctival and corneal reactions in rabbits following short- and repeated exposure to preservative-free tafluprost, commercially available latanoprost and 0.02% benzalkonium chloride

H Liang et al. Br J Ophthalmol. 2008 Sep.

Abstract

Aim: To compare the conjunctival and corneal reactions of commercially available solution of latanoprost (Xalatan) and preservative-free (PF) tafluprost in rabbits.

Methods: The rabbits received 50 microl of phosphate-buffered saline (PBS), PF-tafluprost 0.0015%, latanoprost 0.005% or benzalkonium chloride (BAK) 0.02%; all solutions were applied at 5 min intervals for a total of 15 times. The ocular surface toxicity was investigated using slit-lamp biomicroscopy examination, flow cytometry (FCM) and on imprints for CD45 and tumour necrosis factor-receptor 1 (TNFR1) conjunctival impression cytology (CIC) and corneal in vivo confocal microscopy (IVCM). Standard immunohistology also assessed inflammatory/apoptotic cells.

Results: Clinical observation and IVCM images showed the highest ocular surface toxicity with latanoprost and BAK, while PF-tafluprost and PBS eyes presented almost normal corneoconjunctival aspects. FCM showed a higher expression of CD45+ and TNFR1+ in latanoprost- or BAK-instilled groups, compared with PF-tafluprost and PBS groups. Latanoprost induced fewer positive cells for inflammatory marker expressions in CIC specimens compared with BAK-alone, both of which were higher than with PF-tafluprost or PBS. Immunohistology showed the same tendency of toxic ranking.

Conclusion: The authors confirm that rabbit corneoconjunctival surfaces presented a better tolerance when treated with PF-tafluprost compared with commercially available latanoprost or BAK solution.

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Conflict of interest statement

Competing interests: None.

Figures

Figure 1
Figure 1. Photographs of rabbit eyes 4 h after the 15 instillations of phosphate-buffered saline (A: PBS), preservative-free tafluprost (B: PF tafluprost), latanoprost (C) or 0.02% benzalkonium chloride (D: BAK). (E) Draize test scores at H4 and D1 after treatment instillation. *p<0.005 compared with PBS; #p<0.01 compared with PF-tafluprost; §p<0.05 compared with latanoprost.
Figure 2
Figure 2. In vivo confocal images of rabbit corneal superficial epithelium (line 1), basal epithelium (line 2), anterior stroma (line 3), limbus (line 4) and conjunctival stroma (line 5) after instillations of: phosphate-buffered saline (A: PBS), preservative-free tafluprost (B: PF-tafluprost), latanoprost (C) or benzalkonium chloride (D: BAK), at D1. The scale bar indicates 100 μm. (E) In vivo confocal microscopy (IVCM) score evaluation at H4 and D1 after treatment instillation. *p<0.0005 compared with PBS; #p<0.0005 compared with PF-tafluprost; §p<0.005 compared with latanoprost.
Figure 3
Figure 3. Cresyl violet-stained conjunctival impression cytology from rabbits instilled with phosphate-buffered saline (A: PBS), preservative-free tafluprost (B: PF-tafluprost), latanoprost (C) or 0.02% benzalkonium chloride (D: 0.02% BAK) (original size × 40). Conjunctival impression cytology expressions of (E) CD45+ and (F) tumour necrosis factor-receptor 1 (TNFR1) evaluated by flow cytometry. *p<0.05 compared with PBS; #p<0.05 compared with PF-tafluprost; §p<0.05 compared with latanoprost.
Figure 4
Figure 4. Immunohistology of CD45+ and TUNEL+ cells in rabbit cryosections at D1 after instillations of: phosphate-buffered saline (A: PBS), preservative-free tafluprost (B: PF-tafluprost), latanoprost (C) or benzalkonium chloride (D: BAK). The scale bars indicate 100 μm. (E) CD45+ and (F) TUNEL+ cell counts in limbus and conjunctiva of rabbit cryosections at D1 after eye-drop instillations. *p<0.0001 compared with PBS; #p<0.0001 compared with PF-tafluprost.
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