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Fundus autofluorescence and fate of glycoxidized particles injected into subretinal space in rabbit age-related macular degeneration model

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Abstract

Purpose

Abnormal fundus autofluorescence (FAF) is associated with the incidence or progression of dry and wet age-related macular degeneration (AMD). We previously developed a rabbit AMD model with drusen and type-1 choroidal neovascularization (CNV) that mimics the accumulation of lipofuscin using artificial glycoxidized particles. The objective of the current study was to investigate in vitro effects of glycoxidized particles on retinal pigment epithelial (RPE) cells, and the FAF and fate of injected particles in this model.

Methods

Glycoxidized particles were prepared by a 4-day incubation of water-in-oil emulsions of serum albumin and glycolaldehyde to allow glycoxidation and consequent cross-linking. After particles were added in the culture medium of confluent human RPE cells, cell viability, adhesion activity, and proliferation activity were assessed by cell counting. In anesthetized rabbits, 250 µg of glycoxidized particles were injected into the subretinal space to induce experimental AMD. FAF measurement and angiography with sodium fluorescein and indocyanine green were performed periodically using the Heidelberg Retina Angiograph 2 (HRA2). The eyes enucleated, and the lung and the spleen, excised at week 4 or 12, were histologically evaluated by light and fluorescence microscopy.

Results

Glycoxidized particles phagocytosed did not impair the cell viability, adhesion, and proliferation of RPE cells, as compared with RPE cells in controls. HRA2 showed different patterns of abnormal FAF in the area with the implanted glycoxidized particles, similar to pathological FAF patterns in aging human eyes with or without AMD. Histologic examination showed accumulated glycoxidized particles and large lipofuscin granules with green autofluorescence in and under the RPE and at the margins of or beneath drusen, possibly associated with abnormal FAF. In addition, some particles were detected in the lung and the spleen.

Conclusions

Glycoxidized particles phagocytosed might stay in RPE cells without any acute biological reaction. Our rabbit model of AMD simulated abnormal FAF patterns observed in aging human eyes with or without AMD. Glycoxidized particles phagocytosed by RPE cells could be deposited on Bruch’s membrane in rabbits, possibly excreted in part into choroidal circulation. This model may be useful for understanding various patterns of abnormal FAF histologically, and for elucidating the pathogenesis of AMD.

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Acknowledgements

Tsutomu Yasukawa, MD, PhD, conducted the current study as an Alexander von Humboldt Foundation scholar. We thank Grit Müller for help with animal surgery and Ute Weinbrecht, Karin Bartholomäus, and Silke Jantzen for technical assistance.

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Authors and Affiliations

  1. Department of Ophthalmology and Visual Science, Nagoya City University Graduate School of Medical Sciences, Mizuho-ku, Nagoya-shi, Aichi, 467–8601, Japan

    Megumi Hirata, Tsutomu Yasukawa, Noriyuki Kunou, Ayae Takase, Rina Sato & Yuichiro Ogura

  2. Department of Ophthalmology and Eye Clinic, University of Leipzig Medical Faculty, Leipzig, Germany

    Tsutomu Yasukawa, Peter Wiedemann & Wolfram Eichler

  3. Research and Development Center, Santen Pharmaceutical Co., Ltd, Ikoma, Japan

    Erika Kimura & Noriyuki Kunou

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  1. Megumi Hirata
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Correspondence to Tsutomu Yasukawa.

Additional information

Supported in part by German Research Community (DFG) grant WI 880/9–1 and by a 2006 Grant-in-Aid for Scientific Research No. 18591929 from Japan Society for the Promotion of Science.

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Hirata, M., Yasukawa, T., Wiedemann, P. et al. Fundus autofluorescence and fate of glycoxidized particles injected into subretinal space in rabbit age-related macular degeneration model. Graefes Arch Clin Exp Ophthalmol 247, 929–937 (2009). https://doi.org/10.1007/s00417-009-1070-1

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