Abstract
Purpose: To develop a model that can recapitulate key features of macular fibrosis in neovascular age-related macular degeneration (nAMD).
Methods: Adult C57BL/6J mice received 3 laser burns/eye to induce choroidal neovascularization (CNV). 7 days later, a second laser burn was directed to each of the neovascular lesions. Traditional laser-induced CNV was used as a control. Lesions were monitored at 10, 20, 30 and 40 days post-laser (p.l) treatment by fundus imaging, fundus fluorescein angiography (FFA), optical coherence tomography (OCT), and immunohistochemistry. The expression of collagen 1 (COL-1), fibronectin, alpha-smooth muscle actin (SMA), F4/80, complement factor B (CFB), C3, TGF- and FGF2 in retina and RPE/choroid was examined by immunofluorescence and RT-PCR.
Results: The two-stage laser protocol induced significantly larger lesions than the traditional laser-CNV by OCT and immunohistochemistry at all time points. Confocal microscopy detected collagen-1+ fibres and IBA1+/CD31+ blood vessels in lesions from the two-stage laser protocol 30 – 40 days p.l. Lesions from traditional laser-CNV contain only collagen-1+ fibres but not blood vessels at this time point. Higher levels of proinflammatory (iNOS, C3, CFB) and profibrotic (TGFβ, FGF2, and VEGF) genes were detected in the retinas from the two-stage laser-induced lesions compared to the traditional laser-CNV lesion. Higher number of infiltrating F4/80 macrophages was also observed in/around the two-stage laser induced fibrotic lesion.
Conclusion: The two-stage laser treatment induced subretinal fibrovascular membranes that persist over 40 days.
Methods: Adult C57BL/6J mice received 3 laser burns/eye to induce choroidal neovascularization (CNV). 7 days later, a second laser burn was directed to each of the neovascular lesions. Traditional laser-induced CNV was used as a control. Lesions were monitored at 10, 20, 30 and 40 days post-laser (p.l) treatment by fundus imaging, fundus fluorescein angiography (FFA), optical coherence tomography (OCT), and immunohistochemistry. The expression of collagen 1 (COL-1), fibronectin, alpha-smooth muscle actin (SMA), F4/80, complement factor B (CFB), C3, TGF- and FGF2 in retina and RPE/choroid was examined by immunofluorescence and RT-PCR.
Results: The two-stage laser protocol induced significantly larger lesions than the traditional laser-CNV by OCT and immunohistochemistry at all time points. Confocal microscopy detected collagen-1+ fibres and IBA1+/CD31+ blood vessels in lesions from the two-stage laser protocol 30 – 40 days p.l. Lesions from traditional laser-CNV contain only collagen-1+ fibres but not blood vessels at this time point. Higher levels of proinflammatory (iNOS, C3, CFB) and profibrotic (TGFβ, FGF2, and VEGF) genes were detected in the retinas from the two-stage laser-induced lesions compared to the traditional laser-CNV lesion. Higher number of infiltrating F4/80 macrophages was also observed in/around the two-stage laser induced fibrotic lesion.
Conclusion: The two-stage laser treatment induced subretinal fibrovascular membranes that persist over 40 days.
| Original language | English |
|---|---|
| Article number | 3 |
| Number of pages | 11 |
| Journal | Translational Vision Science & Technology |
| Volume | 9 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - 01 Mar 2020 |
Keywords
- retina
- fibrosis
- inflammation
- choroidal neovascularisation
- macrophage
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