Polyoxyethylene sorbitan monooleate

Two kinds of ophthalmic solutions, a vehicle solution and a 0.05% fenofibrate hydrochloride solution, were compounded and used to compare the effect of the PPARα agonist. Ophthalmic vehicle solution was prepared using 0.1 mL polyoxyethylene sorbitan monooleate (Wako Pure Chemical Industries, Osaka, Japan) and 100 mL NaCl-based PBS (0.01 M; pH 7.4), which was prepared with disodium hydrogen phosphate dodecahydrate (232 g), sodium dihydrogen phosphate dihydrate (23.7 g), and distilled water (4000 mL). To prepare the 0.05% fenofibrate hydrochloride ophthalmic solution, we added 10 mg fenofibrate (Wako Pure Chemical Industries) to 20 mL vehicle solution. fenofibrate is poorly soluble in aqueous solutions, therefore, fenofibrate hydrochloride ophthalmic solution was stirred for 70 min. All ophthalmic solutions were recognized pH 3.7 to pH 3.75 and stored at 4 °C in a refrigerator, and used within a month of the initial compounding without sterile filtration.

Under general isoflurane anesthesia, a circular filter paper 3.2 mm in diameter was immersed in 1 N NaOH and placed on the central cornea of each rat for 1 min to create a corneal alkali burn. After alkali exposure, the ocular surface was washed with 40 mL physiological saline solution. The trimebutine ophthalmic solution or vehicle solution was administered twice daily to the alkali-burned corneas. The vehicle solution was filter-purified 100 mL NaCl-based phosphate buffered saline (0.01 M; pH 7.4), which was prepared with 232 g disodium hydrogen-phosphate 12-water, 23.7 g sodium dihydrogen phosphate dihydrate, 4000 mL distilled water, and 0.1 mL polyoxyethylene sorbitan monooleate (Wako Pure Chemical Industries, Osaka, Japan). The 0.2% trimebutine ophthalmic solution was prepared by adding 0.2 g of 2-(dimethylamino)-2-phenylbutyl 3,4,5-trimethoxybenzoate maleate (Tokyo Chemical Industry Co., Ltd.) to 100 mL of the vehicle solution. At each endpoint (1, 4, and 7 days after the alkali burn), the rats were sacrificed by exsanguination under isoflurane anesthesia. Pathological and molecular biological evaluations were performed on the enucleated eyes.

Under general isoflurane anesthesia, a circular filter paper 3.2 mm in diameter was immersed in 1 N NaOH and placed on the central cornea of each rat for 1 min to create a corneal alkali burn. After alkali exposure, the ocular surface was washed with 40 mL physiological saline. PPARβ/δ agonist ophthalmic solution or vehicle solution was administered twice a day to the alkali-burned cornea. The vehicle solution was filter purified 100mL NaCl-based PBS (0.01 M; pH 7.4), which was prepared with 232 g disodium hydrogen-phosphate 12-water, 23.7 g sodium dihydrogen phosphate dihydrate, 4000 mL distilled water, and 0.1 mL polyoxyethylene sorbitan monooleate (Wako Pure Chemical Industries, Osaka, Japan). The PPARβ/δ agonist ophthalmic solution was prepared by adding 10 mg GW501516 to 20 mL vehicle solution. At each endpoint (6 h, 1 day, 4 days, and 7 days after alkali burn), rats were sacrificed by exsanguination under isoflurane anesthesia. Pathological and molecular biological evaluations were performed on the enucleated eyes.