Early angiogenesis is important to facilitate biomaterials’ osteogenic effects and avoid the bone regeneration failure for large-sized bone defects. Bioactive glasses (BG) have been widely utilized as a biomaterial for bone repair. However, the early angiogenesis of BG may be inadequate. In this study, we explored the effects of photobiomodulation (PBM) combined with BG on early angiogenesis to solve this bottleneck problem of insufficient early angiogenesis. In vitro, human umbilical vein endothelial cells (HUVECs) were cultured with BG extracts and treated with PBM using 1 J/cm2. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, real-time reverse transcription-polymerase chain reaction (real-time RT-PCR) and tubule formation assay were utilized to detect HUVECs’ proliferation, vascular growth factor genes expression and tubules formation. In vivo, bone defects at the femoral metaphysis in Sprague-Dawley rats were treated with BG particulates and PBM at 120 J/cm2. Immunohistochemical staining was applied to observe the vascular-like structure formation. In vitro results showed that PBM combined with BG significantly promoted HUVECs’ proliferation, genes expression and mature tubules formation. On days 2, 4 and 7, the VEGF gene expression in BG+PBM group was 2.70-, 2.59- and 3.05-fold higher than control (P<0.05), and higher than PBM and BG groups (P<0.05). On days 4 and 7, the bFGF gene expression in BG+PBM group was 2.42- and 1.82-fold higher than control (P<0.05), and also higher than PBM and BG groups (P<0.05). Tube formation assay showed that mature tubules formed in BG+PBM and PBM groups after 4 hours. The tubules number in BG+PBM group was significantly higher than other groups (P<0.05). In vivo results further confirmed that PBM induced early angiogenesis. More vascular-like structures were observed in BG+PBM and PBM groups 2-week post surgery. In conclusion, with the optimum PBM fluence and BG concentration, PBM combined with BG exerted additive effects on enhancing early angiogenesis.
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