Figure 4. FMOD induces endothelial cell migration in vivo.

(A) HVMECs combined with BM-MSC (10:1) with or without recombinant h-FMOD were mixed with Matrigel. Perfused vessels were labeled with human-specific rhodamine-labeled lectin and visualized with confocal microscopy; n = 6 plugs per group; experiment repeated 3 times. Original magnification, ×5. (B and C) Migration of host endothelial cells into Matrigel plugs containing nonpigmented melanocytes neutralized by anti FMOD or IgG as a control. Cells were stained for CD31 and CD45 and analyzed by FACS; n = 10 per group (C). (D) The corneal micropocket assay was performed with pellets containing 1.69 μM of FMOD, 1.69 μM GST, or 5.2 μM VEGF-A. FMOD versus control was significantly different. No significant difference in VA was found between FMOD and VEGF-A. The experiment was repeated 3 times and included 10 eyes per group in each experiment. (E) FMOD stimulated longer vessels than VEGF-A. (F) FMOD (2 μM) promotes angiogenesis in wound healing in ear skin of C57BL/6J mice; blood vessels visualized by FITC-conjugated dextran (green). Scale bar: 500 μm. (G) Quantification of blood vessel density; n = 5 mice per group; 2 punches per mouse. (H) Recombinant FMOD (1.19 μM) or control-GST injected into the vitreous of 4-day-old pups; n = 5 mice per group. On day 7, retinas were dissected and labeled (Alexa Fluor 594–conjugated isolectin). Scale bar: 1 nm. Original magnification, ×5. (I) Neovascular density was assessed by quantifying the capillary junctions within 4 equal areas (ImageJ). (J) Avascular retina was calculated as percentage of total retinal area quantified (ImageJ); n = 9 in each group, experiment was repeated twice. **P < 0.001; ***P < 0.0001.