A study population of 30 female Wistar rats were anesthetized with an intraperitoneal injection of ketamine hydrochloride (25 mg/kg) and xylazine hydrochloride (5 mg/kg; both Sigma-Aldrich, St. Louis, MO, USA). All eyes were examined under a binocular microscope to exclude corneal scaring, opacity and NV prior to the study. Corneal injury was induced by placing a monolayer filter saturated with 1 mol/l NaOH onto the right eye of the rat for 2 min, as previously described (18 (link)–20 (link)). Following the establishment of the alkali burn corneal injury, the 30 alkali-injured rats were allocated at random into three groups: Alkali burn control group, which received, 3 drops of balanced salt solution (Sigma-Aldrich) 3 times a day for 7 days in the alkali-treated eyes; group 1, which received 1% cyclosporine (Sigma-Aldrich) from day 1 following alkali injury, 3 drops 3 times a day for 7 days in the alkali-treated eyes; and group 2, which received 90Sr-90Y β-irradiation from day 1 following alkali injury, 1 Gy once a day for 7 days in the alkali-treated eyes. In addition, 10 Wistar rats which did not receive any treatment were selected as the alkali burn control group, receiving 3 drops of the balanced salt solution, 3 times a day for 7 days).
Balanced salt solution
Manufactured by Merck Group
Balanced salt solution is a laboratory reagent that provides a balanced ionic and osmotic environment for cell cultures and other biological applications. It is a sterile, isotonic solution containing a mixture of salts, including sodium, potassium, calcium, and magnesium, with a pH and osmolarity designed to maintain the physiological conditions necessary for the survival and growth of cells.
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2 protocols using balanced salt solution
1
Alkali-Induced Corneal Injury in Rats
2
Production and Characterization of AAV2/9
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The D2Rs and GFP coding sequences were cloned into AAV genomes under the control of the chicken β-actin/cytomegalovirus (CBA/CMV) promoter for ubiquitous and robust expression. AAV 2/9 was produced via triple-transfection of HEK 293 T cells with the genome and helper plasmids. Virus was recovered from cells using freeze-thaw cycles, purified using an iodixanol gradient (Optiprep Density Gradient, Sigma-Aldrich, St. Louis, MO), followed by buffer exchange and concentration using concentrator columns (Orbital Biosciences, Topsfield, MA) as described previously [6 (link)]. The viral titer was determined using digital droplet PCR (ddPCR) and normalized to 1 × 1013 vector genomes (vg)/ml using Balanced Salt Solution (Sigma-Aldrich, St. Louis, MO).
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