Establishment of consistent section preparation method in Rhesus monkey eyeball for preclinical toxicology research
Objective To establish a consistent section preparation method that can fully cover anatomical and histological structures of normal eyes,and cornea and xeno-corneal endothelium grafts after implementing xeno-corneal grafts transplantation in Rhesus monkey for preclinical toxicology research.Methods There were ten adult rhesus monkeys in control group and ten adult rhesus monkeys in model group.Xenogenic corneal endothelial graft was performed on the right eye of the model group.Normal eyes and eyes that underwent xenograft transplantation of xeno-corneal endothelial grafts were sampled and fixed with Davidson's fluid.Normal eyes:Use a slicing blade to make a section along the optic nerve,the macula(fovea centralis),and the cornea at a distance of 1mm parallel to the temporal posterior long ciliary artery(LPCA)and above the optic nerve.The samples were routinely dehydrated,embedded,sectioned and stained before undergoing microscopic observation.Eyes that underwent xenograft transplantation of the xeno-corneal endothelial grafts:Cut the cornea and iris from behind the ciliary body,remove the lens,and then cut the corneal and iris with the corneal facing upwards at an interval of 2.5 mm.Sampling surface of the tissue were placed downward and sequentially into a grid embedding box.Three transverse plane sections of the remaining ocular tissues were cut and placed in an embedding box.After dehydration and embedding,the wax block was sectioned and stained with HE and Masson,respectively.Finally,the anatomical and histological characteristics of the normal eyes,the cornea,and the xeno-corneal endothelial grafts were confirmed.Results The prepared eyes sections can clearly display all tissue structures of the normal eyes,with bright staining and realistic morphology.After implementation of the xeno-corneal endothelial grafts transplantation,the structures of the cornea and the xeno-corneal endothelial grafts were clear,and the adhesion between the xeno-corneal endothelium and the cornea can be well displayed.The surface of the prepared tissue sections was flat and no artificial artifacts were observed.Conclusion By using a self-developed preparation method,we can obtain good eye section containing all structural features of the normal eye tissues,intact cornea and xeno-corneal endothelial grafts,and the other eye tissues.The method is easy,and has high consistency.It can be used for histopathological examination of ocular medication and pathological evaluation of the eyes after performing xeno-corneal endothelial grafts transplantation.We hoped that the section preparation method presented in this study will provide technical support for scientific assessment of the safety and efficacy of preclinical ophthalmic drugs/xeno-corneal endothelial grafts transplantation.
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