PDF《JOURNAL OF OCULAR PHARMACOLOGY AND》

Evaluation of a Novel Arti?cial Tear in the Prevention and Treatment of Dry Eye in an Animal Model Yujing She,1 Jinyang Li,1 Bing Xiao,1 Huihui Lu,

1 Haixia Liu,2 Peter A.

Simmons,2 Joseph G. Vehige,2 and Wei Chen1 Abstract Purpose: To evaluate effects of a novel multi-ingredient arti?cial tear formulation containing carboxymeth- ylcellulose (CMC) and hyaluronic acid (HA) in a murine dry eye model. Methods: Dry eye was induced in mice (C57BL/6) using an intelligently controlled environmental system (ICES). CMC+HA (Optive Fusion?), CMC-only (Refresh Tears? ), and HA-only (Hycosan? ) arti?cial tears and control phosphate-buffered saline (PBS) were administered

4 times daily and compared with no treatment (n =

64 eyes per group). During regimen

1 (prevention regimen), mice were administered arti?cial tears or PBS for

14 days (starting day 0) while they were exposed to ICES, and assessed on days

0 and 14. During regimen

2 (treatment regimen), mice exposed to ICES for

14 days with no intervention were administered arti?cial tears or PBS for

14 days (starting day 14) while continuing exposure to ICES, and assessed on days 0, 14, and 28. Corneal ?uorescein staining and conjunctival goblet cell density were measured. Results: Arti?cial tear-treated mice had signi?cantly better outcomes than control groups on corneal staining and goblet cell density (P<

0.01). Mice administered CMC+HA also showed signi?cantly lower corneal ?uorescein staining and higher goblet cell density, compared with CMC (P<

0.01) and HA (P

30 dots, grade 4). The total score from the

5 regions was recorded for analysis. Periodic acidCSchiff staining for goblet cells Mice from each treatment group were sacri?ced using an overdose mixture of ketamine and xylazine for histological evaluation on day

14 (during regimen 1) and day

28 (during regimen 2) after the start of ICES-induced dry eye. Mice in the treatment (CMC+HA, CMC, HA, PBS) and control (NT, nor- mal) groups were euthanized at the same time for comparison. Twelve enucleated eyes from sacri?ced mice (n=6) in each group were ?xed in 10% formalin, dehydrated, and embedded in paraf?n. To quantify goblet cells in the conjunctiva, 4-mm cross sections were stained with the periodic acidCSchiff reagent. The number of goblet cells was assessed under a microscope (Axio Imager Z1;

Carl Zeiss Meditec, Oberkochen, Germany). Statistical analyses The MannCWhitney U-test was used to compare the dif- ferences in ?uorescein staining scores and analysis of vari- ance was used to compare goblet cell density between the treatment and control groups;

P <

0.05 was considered sta- tistically signi?cant. Analyses were performed using SPSS 18.0 software (IBM, Armonk, NY). Results Corneal ?uorescein staining Figure

1 shows representative corneal ?uorescein staining in mice housed in the ICES chamber after receiving treatment with arti?cial tears. All arti?cial tear formulations (CMC+ HA, CMC, and HA) improved corneal staining compared with the control groups (PBS and NT) at day

14 during regimen

1 and day

28 (14 days after treatment initiation) during regimen 2. Quantitatively, there were no differences in corneal staining scores observed at the beginning of the ICES- induced dry eye process (day 0) during both regimens

1 and 2, and on day

14 when treatment with arti?cial tears was initi- ated during regimen

2 (Fig. 2a, b). After

14 days of treatment with arti?cial tears in both regimens

1 and 2, ?uorescein corneal staining was signi?cantly reduced in mice receiving CMC+HA, CMC only, or HA only compared with mice in the PBS-treated and NT control groups (P <