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. 2013 Jun;182(6):2407–2417. doi: 10.1016/j.ajpath.2013.02.032

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© 2013 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

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Formation of a fibrovascular network after laser injury. A: At the site of laser injury (asterisk), F4/80⁺ macrophages (arrowheads), are seen within 24 to 40 hours. B: Adjacent to infiltrating F4/80⁺ macrophages (arrowhead), RPE cells exhibit increased SMA expression (arrow) at approximately 48 hours after laser injury (asterisk). C: SMA⁺ stellate-like cells (arrow) are seen originating from the center of laser spots at approximately 60 hours after injury (asterisk). At that time RPE cells (arrowhead) adjacent to the injury site exhibit strong SMA expression. D: At approximately 72 hours after laser injury, F4/80⁺ macrophages (arrowhead) are the predominant cell population within the area of injury (asterisk); SMA⁺ cells are also present (arrow). E: At approximately 72 hours, a SMA⁺ myofibroblastic scaffold is seen and adjacent RPE cells are SMA⁺, but endothelial cells have not yet infiltrated the site of laser injury. F: At approximately 96 hours, CD31⁺ neovessels (arrow) can be observed at the center of the site of laser injury. G and H: Between day 4 and 5 after laser injury, a fully formed fibrovascular lesion can be observed, with an extensive network of CD31⁺ neovessels (arrow). I and J: Confocal microscopy of a fully formed CNV lesion at day 10 after laser injury shows that neovessels are covered and demarcated by a SMA⁺ myofibroblastic network. Neovascularization does not extend beyond the SMA⁺ myofibroblastic network. K and L: Top (K) and midportion (L) of lesion at 10 days after laser injury. M: SMA expression is maintained in RPE cells adjacent to the site of laser injury, even 4 months after injury. N–Q: Staining of a CNV lesion at 72 hours after laser injury for the proliferation marker phospho-histone H3 (Ser10) exhibits extensive proliferation of multiple cell types within an early CNV lesion (N). Some F4/80⁺ cells stain strongly for this proliferation marker (O), as well as stellate SMA⁺ myofibroblastic cells (P) and adjacent SMA⁺ RPE cells (Q). Original magnification: ×10 (E–I, M, and N); ×20 (A–D); ×40 (K, L, O–Q).