Dimensions and Morphology of the Cornea in Three Strains of Mice

Purpose - To use a histologic approach to obtain dimensional and morphologic information on the cornea in three commonly used strains of mice. Methods - Adult mice (three each of 129/SVJ, C57BL/6, and BALB/c) were euthanatized, and the eyes were enucleated, immersed in 2% glutaraldehyde fixative, and prepared for light and transmission electron microscopy. The full corneal, epithelial, stromal, and posterior limiting lamina (PLL) with endothelium thicknesses were measured at the same location centrally and peripherally. Results - All three strains showed a statistically significant (P < 0.001) decrease in overall thickness in the peripheral compared with the central cornea. The decrease was due to a reduced thickness of both the epithelium and the stroma. The stroma and epithelium contributed to approximately two thirds and one third of the total corneal thickness, respectively. The epithelium had the classic stratified layout and consisted of 13.00 ± 1.41 layers centrally versus 10.33 ± 1.37 peripherally. Some adaptation of stromal tissue was found immediately adjacent to the epithelial basement membrane, but a clearly defined anterior limiting lamina did not exist. The stroma was organized into lamellae but lacked the anterior branching and interweaving reported in humans and had unmyelinated nerve fibers within micrometers of the endothelium. The PLL was 2.17 ± 0.3 μm thick and was divided into pre- and postnatal layers, with striated bodies in the postnatal portion. Conclusions - This study demonstrated that in the three strains of mice examined, the cornea becomes significantly thinner toward the periphery. Dimensionally, proportionally, and anatomically the three strains used appeared to be similar. However, morphologic differences were observed compared with other mammals, and awareness of these differences is important when using the mouse as an animal model applicable to the human

J.P.: Dimensions and morphology of the cornea in three strains of mice. Investigative ophthalmology &amp; visual science 50(8

PURPOSE. To use a histologic approach to obtain dimensional and morphologic information on the cornea in three commonly used strains of mice. METHODS. Adult mice (three each of 129/SVJ, C57BL/6, and BALB/c) were euthanatized, and the eyes were enucleated, immersed in 2% glutaraldehyde fixative, and prepared for light and transmission electron microscopy. The full corneal, epithelial, stromal, and posterior limiting lamina (PLL) with endothelium thicknesses were measured at the same location centrally and peripherally. RESULTS. All three strains showed a statistically significant (P Ͻ 0.001) decrease in overall thickness in the peripheral compared with the central cornea. The decrease was due to a reduced thickness of both the epithelium and the stroma. The stroma and epithelium contributed to approximately two thirds and one third of the total corneal thickness, respectively. The epithelium had the classic stratified layout and consisted of 13.00 Ϯ 1.41 layers centrally versus 10.33 Ϯ 1.37 peripherally. Some adaptation of stromal tissue was found immediately adjacent to the epithelial basement membrane, but a clearly defined anterior limiting lamina did not exist. The stroma was organized into lamellae but lacked the anterior branching and interweaving reported in humans and had unmyelinated nerve fibers within micrometers of the endothelium. The PLL was 2.17 Ϯ 0.3 m thick and was divided into pre-and postnatal layers, with striated bodies in the postnatal portion. CONCLUSIONS. This study demonstrated that in the three strains of mice examined, the cornea becomes significantly thinner toward the periphery. Dimensionally, proportionally, and anatomically the three strains used appeared to be similar. However, morphologic differences were observed compared with other mammals, and awareness of these differences is important when using the mouse as an animal model applicable to the human. (Invest Ophthalmol Vis Sci

Contact lens care solution killing efficacy against Acanthamoeba castellanii by in vitro testing and live-imaging

In the past decade there has been an increased incidence of Acanthamoeba keratitis, particularly in contact lens wearers. The aim of this study was to utilize in vitro killing assays and to establish a novel, time-lapse, live-cell imaging methodology to demonstrate the efficacy of contact lens care solutions in eradicating Acanthamoeba castellanii (A. castellanii) trophozoites and cysts. Standard qualitative and quantitative in vitro assays were performed along with novel time-lapse imaging coupled with fluorescent dye staining that signals cell death. Quantitative data obtained demonstrated that 3% non-ophthalmic hydrogen peroxide demonstrated the highest percent killing at 87.4% corresponding to a 4.4 log kill. The other contact lens care solutions which showed a 72.9 to 29.2% killing which was consistent with 4.3–2.8 log reduction in trophozoite viability. Both analytical approaches revealed that polyquaternium/PHMB-based was the least efficacious in terms of trophicidal activity. The cysticidal activity of the solutions was much less than activity against trophozoites and frequently was not detected. Live-imaging provided a novel visual endpoint for characterizing the trophocidal activity of the care solutions. All solutions caused rapid rounding or pseudocyst formation of the trophozoites, reduced motility and the appearance of different morphotypes. Polyquaternium/alexidine-based and peroxide-based lens care system induced the most visible damage indicated by significant accumulation of debris from ruptured cells. Polyquaternium/PHMB-based was the least effective showing rounding of the cells but minimal death. These observations are in keeping with care solution biocides having prominent activity at the plasma membrane of Acanthamoeba