Huang Lidong, Gong Weiyu, Huang Guibin, Li Jingyi, Wu Jilin, Wang Yuguang, Dong Yanmei
Department of Cariology and Endodontology, Peking University School and Hospital of Stomatology, Beijing, 100081, People's Republic of China.
National Engineering Laboratory for Digital and Material Technology of Stomatology, Peking University School and Hospital of Stomatology, Beijing 100081, People's Republic of China.
Biomed Mater. 2022 May 13;17(4). doi: 10.1088/1748-605X/ac6b07.
Bioactive glasses (BG) have been widely utilized as a biomaterial for bone repair. However, the early angiogenesis of BG may be inadequate, which weakens its osteogenic effects in large-sized bone defects and often leads to the failure of bone regeneration. 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., human umbilical vein endothelial cells (HUVECs) were cultured with BG extracts and treated with PBM using 1 J cm. 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., bone defects at the femoral metaphysis in Sprague-Dawley rats were treated with BG particulates and PBM at 120 J cm. Hematoxylin-eosin staining was used to observe the inflammatory response, tissue formation and biomaterial absorption of bone defects. Immunohistochemical staining was applied to observe the vascular-like structure formation. Theresults showed that PBM combined with BG significantly promoted HUVECs' proliferation, genes expression and mature tubules formation. On days 2, 4 and 7, the mRNA expression of VEGF in BG + PBM group was 2.70-, 2.59- and 3.05-fold higher than control (< 0.05), and significantly higher than PBM and BG groups (< 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 (< 0.05), and also higher than PBM and BG groups (< 0.05). Tube formation assay showed that mature tubules were formed in BG + PBM and PBM groups after 4 h, and the number in BG + PBM group was significantly higher than other groups (< 0.05).results further confirmed PBM induced early angiogenesis, with more vascular-like structures observed in BG + PBM and PBM groups 2 week post-surgery. With the optimum PBM fluence and BG concentration, PBM combined with BG exerted additive effects on enhancing early angiogenesis.
生物活性玻璃（BG）已被广泛用作骨修复的生物材料。然而，BG的早期血管生成可能不足，这削弱了其在大型骨缺损中的成骨作用，并常常导致骨再生失败。在本研究中，我们探索了光生物调节（PBM）联合BG对早期血管生成的影响，以解决早期血管生成不足这一瓶颈问题。用BG提取物培养人脐静脉内皮细胞（HUVECs），并使用1 J/cm²的能量进行PBM处理。采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法、实时逆转录-聚合酶链反应（实时RT-PCR）和小管形成试验检测HUVECs的增殖、血管生长因子基因表达和小管形成。对Sprague-Dawley大鼠股骨近端的骨缺损用BG颗粒和120 J/cm²的PBM进行处理。采用苏木精-伊红染色观察骨缺损处的炎症反应、组织形成和生物材料吸收情况。应用免疫组织化学染色观察血管样结构的形成。结果表明，PBM联合BG显著促进了HUVECs的增殖、基因表达和成熟小管的形成。在第2、4和7天，BG + PBM组中血管内皮生长因子（VEGF）的mRNA表达分别比对照组高2.70倍、2.59倍和3.05倍（P<0.05），且显著高于PBM组和BG组（P<0.05）。在第4和7天，BG + PBM组中碱性成纤维细胞生长因子（bFGF）基因表达分别比对照组高2.42倍和1.82倍（P<0.05），也高于PBM组和BG组（P<0.05）。小管形成试验显示，4小时后BG + PBM组和PBM组形成了成熟小管，且BG + PBM组的小管数量显著高于其他组（P<0.05）。结果进一步证实PBM可诱导早期血管生成，术后2周在BG + PBM组和PBM组观察到更多的血管样结构。通过最佳的PBM能量密度和BG浓度，PBM联合BG在增强早期血管生成方面发挥了协同作用。
