Figure 1. Lodamin inhibits angiogenesis and inflammation in Matrigel plugs and inhibits corneal neovascularization after local or oral treatment.

(A) Matrigel containing VEGF and bFGF was injected subcutaneously (s.c., n = 5) to determine the effect of Lodamin on angiogenesis and macrophage infiltration. Upper panel shows representative plugs removed from Lodamin treated or untreated mice (bar = 1 cm). Bottom panel shows vessel staining with anti-CD31 antibody (brown) and nuclei staining with Hematoxilin Gill's (blue), bar = 100µm; arrows point to large vessel with open lumans (B) Quantification of infiltrating endothelial cell or macrophages in Matrigel plugs. Data are presented as percent of specific cell population out of the total cell population (n = 3–4, p<0.05). (C) Antiangiogenic activity of Lodamin following topical or systemic administration was evaluated in the mouse corneal micropocket assay using bFGF-induced neovascularization. Quantification of vessel area (mm²) in different corneal assays performed with Lodamin which were administered either orally (30 mg/kg q.d), by eye drops (30 mg/ml, q.d) or by subconjunctival injections (30 mg/ml, q.d). Graphs indicate the significant inhibition of vessel formation after 5 days of treatment. Vessel area was reduced by 38% after subconjunctival injection (P = 0.0002), by 30% after eye drops treatment (P = 0.003), and by 37% (P = 0.04) after oral administration. In each group n = 10. (D) Representative images taken from the different groups at day 5; bFGF pellet is detected as a white spot in the center of the cornea, blood vessels growing from limbal periphery are reduced in Lodamin treated group compared to controls. Vessel area in mm² is calculated by the following formula: π×clock hours×vessel length (mm)×0.2 mm.