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T. Hisatomi, H. Enaida, Y. Hata, A. Ueno, H. Sakamoto, T. Sakamoto, T. Ishibashi; The Role of Cellular Migration in the Pathogenesis of a Macular Hole: A Critical New Method for Comprehensive Bird’s–Eye Analysis of the Internal Limiting Membrane . Invest. Ophthalmol. Vis. Sci. 2005;46(13):5428.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To investigate the role of cellular migration on the internal limiting membrane (ILM), we demonstrate a new method for a comprehensive bird’s–eye analysis of surgically excised ILM. We elucidate the pathogenesis of macular hole formation, focusing in particular on the role of cellular migration on the internal limiting membrane (ILM) around the macular hole with this new observation method.
Methods: In order to gain a comprehensive overview of ILM excised in macular hole surgery (n=36), the ILM was carefully unfolded and spread out onto a glass slide as a continuous flat sheet containing a macular hole. The specimens were observed by light and transmission electron microscopy (n=9), and the cellular distribution was analyzed by scanning electron microscopy in a quantitative manner (n=27). Immunohistochemistry for glial fibrillary acidic protein (GFAP) and cytokeratin 18 (CK18) was carried out for cellular characterization. Cellular proliferation was assessed by immunohistochemistry for proliferating cell nuclear antigen (PCNA) and Ki–67.
Results: Cellular migration was not apparent around the macular hole in the early stage of the macular hole’s development (stage 2, 0µm). Cellular migration developed around the macular hole (stage 3, 84µm) and the area of cellular migration gradually enlarged in the later stages (stage 4, 420µm). The immunophenotypic analysis showed that these cells were mainly GFAP+ glial cells and CK18+ retinal pigment epithelial cells. The PCNA and Ki–67 immunohistochemistry showed that part of these cells was proliferating on the ILM.
Conclusions: This new bird’s–eye–view observation enables us to carry out comprehensive analysis of cellular distribution from a temporal and spatial perspective. Cellular migration on ILM is not necessary for the initial formation of a macular break. Cellular migration developed from the macular hole and the migration and proliferation increased gradually with the development of the macular hole.
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