Histological findings of ectopic bone formation after treatment with various concentrations of SDF-1 with/without BMP-2.

<p>(A) Representative histological sections 4 weeks after surgery (trichrome staining, ×40 in the box). (B) Bone formation ratio (%) and newly formed bone area (mm²) were analyzed histomorphometrically. Concomitant BMP-2 and SDF-1 treatment did not significantly activate ectopic bone formation compared to independent treatment with BMP-2 or SDF-1 alone (*, <i>p</i> < 0.05).</p

The effect of simultaneous SDF-1 and BMP-2 treatment on ectopic bone formation.

<p>Faxitron soft X-ray images after subcutaneous implantation of collagen sponge with/without BMP-2 (2.5μg) and SDF-1 (0, 0.1, 0.5, or 1 μg/collagen sponge) in mice. (A) Radiographic image of the harvested collagen sponges 4 weeks after implantation. (B) Comparison of radiological density of collagen sponges from different treatment groups (*, <i>p</i> < 0.05).</p

The effect of simultaneous treatment with SDF-1 and BMP-2 on orthotopic bone formation.

<p>Soft X-ray examination was carried out after implantation of collagen sponges to critical-size calvarial defects with/without BMP-2 (2.5μg) and SDF-1 (0, 0.1, 0.5, or 1 μg/collagen sponge) in mice. (A) Representative radiographic images, (B) Quantification of bone regeneration by radiographic density at 4 weeks post-implantation (*, <i>p</i> < 0.05).</p

Transwell migration assay.

<p>(A) Representative microscopic images of cells cultured in media containing SDF-1 alone, BMP-2 alone, SDF-1 + BMP-2, or neither factor. (B) Quantification of migration of C3H10T1/2 after 24h treatment with the indicated conditions. Treatment with SDF-1 (0.5μg/ml) and BMP-2 (0.5μg/ml) significantly promoted <i>in vitro</i> cell mobility (*, <i>p</i> < 0.05), while concomitant SDF-1 and BMP-2 application did not further enhance the cell migration.</p

Histological findings of orthotopic bone formation after treatment with various concentrations of SDF-1 with/without BMP2.

<p>(A) Representative histological sections 4 weeks post-surgery (trichrome staining, ×40 in the box). (B) Bone formation ratio (%) and newly formed bone area (mm²) were analyzed histomorphometrically. Concomitant BMP-2 and SDF-1 treatment did not significantly activate orthotopic bone formation compared to independent treatment with BMP-2 or SDF-1 alone (*, <i>p</i> < 0.05).</p

Tube formation assay for angiogenesis on Matrigel.

<p>Effect of SDF-1 and BMP-2 on <i>in vitro</i> angiogenesis was evaluated with HUVEC tube formation assay. The tube formation after treatment with SDF-1 alone, BMP-2 alone, combined SDF-1 & BMP-2 were microscopically compared (A). The number of branching point per field (B) and total length of tube per field (C) were quantified by counting 3 random fields/well under the microscope (x4). Representative photos of HUVEC tube formation assay reveal that BMP-2 or SDF-1 alone could significantly activate angiogenesis (*, <i>p</i> < 0.05). However, co-treatment of BMP-2 and SDF-1 did not show additive effect on antigenic tube formation compared to a single treatment of BMP-2.</p