[生物活性玻璃对人脐静脉内皮细胞增殖、分化及血管生成的影响]
[Effects of bioactive glass on proliferation, differentiation and angiogenesis of human umbilical vein endothelial cells].
作者信息
Huang L D, Gong W Y, Dong Y M
机构信息
Department of Cariology and Endodontology, Peking University School and Hospital of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China.
出版信息
Beijing Da Xue Xue Bao Yi Xue Ban. 2021 Feb 22;53(2):371-377. doi: 10.19723/j.issn.1671-167X.2021.02.023.
OBJECTIVE
To investigate the effects of phytic acid derived bioactive PO-SiO-CaO gel-glasses (PSC) on the proliferation, differentiation and angiogenesis of human umbilical vein endothelial cells (HUVECs) .
METHODS
HUVECs were cultured in PSC extracts, which were prepared with endothelial cell medium (ECM) at a gradient concentration of 0.01, 0.1, 1 and 2 g/L. Cells cultured in ECM were used as the control. The effect of PSC on HUVECs proliferation was assessed on the 1st, 3rd, 5th, 7th and 10th days with (4, 5-dimethylthiazol-2-yl) 2, 5-diphenyltetrazolium bromide assay (MTT), and the optimum PSC concentration for HUVECs proliferation was used in the following experiments. The subsequent experiments were divided into two groups. The experimental group used PSC extracts to culture HUVECs (PSC group) and the control group used ECM to culture HUVECs (ECM group). Gene expression of angiogenic factors, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF), was detected on the 2nd, 4th and 7th days by real-time reverse transcription-polymerase chain reaction (real-time RT-PCR). The morphology and number of tubules formation were observed at the 4th and 10th hours. Image J software was used for counting and quantitative analysis.
RESULTS
The results of MTT assay showed that 0.1 g/L PSC group had the most significant effect on promoting HUVECs proliferation. The optical density values of 0.1 g/L PSC group on the 5th and 7th days were significantly higher than those of the other PSC groups and the control group ( < 0.05). The result of real-time RT-PCR showed that 0.1 g/L PSC extract up-regulated the mRNA expression of and significantly ( < 0.05). On the 4th day, the gene expressions of and in PSC group were 1.59 and 1.45 times higher than those in ECM group respectively, and on the 7th day, the gene levels of and in PSC group were 1.98 and 1.37 times higher than those in ECM group respectively. The tubule formation assay showed that the maturity and density of the tubules in 0.1 g/L PSC group was much better than that in the ECM group at the 10th hour. The quantitative analysis by Image J indicated that the tubules number in PSC group (29.63±2.29) was higher than in the ECM group (20.13±2.36), with statistical significance ( < 0.05).
CONCLUSION
PSC showed significant promoting effects on HUVECs' proliferation, differentiation and angiogenesis .
目的
研究植酸衍生的生物活性磷酸硅钙凝胶玻璃（PSC）对人脐静脉内皮细胞（HUVECs）增殖、分化和血管生成的影响。
方法
将HUVECs培养于PSC提取物中，该提取物用内皮细胞培养基（ECM）按0.01、0.1、1和2 g/L的梯度浓度制备。以在ECM中培养的细胞作为对照。在第1、3、5、7和10天用噻唑蓝比色法（MTT）评估PSC对HUVECs增殖的影响，并将促进HUVECs增殖的最佳PSC浓度用于后续实验。后续实验分为两组。实验组用PSC提取物培养HUVECs（PSC组），对照组用ECM培养HUVECs（ECM组）。在第2、4和7天通过实时逆转录聚合酶链反应（实时RT-PCR）检测血管生成因子血管内皮生长因子（VEGF）和碱性成纤维细胞生长因子（bFGF）的基因表达。在第4和10小时观察小管形成的形态和数量。使用Image J软件进行计数和定量分析。
结果
MTT分析结果显示，0.1 g/L PSC组对促进HUVECs增殖的作用最显著。0.1 g/L PSC组在第5和7天的光密度值显著高于其他PSC组和对照组（P<0.05）。实时RT-PCR结果显示，0.1 g/L PSC提取物显著上调了VEGF和bFGF的mRNA表达（P<0.05）。在第4天，PSC组中VEGF和bFGF的基因表达分别比ECM组高1.59倍和1.45倍，在第7天，PSC组中VEGF和bFGF的基因水平分别比ECM组高1.98倍和1.37倍。小管形成实验显示，在第10小时，0.1 g/L PSC组中小管的成熟度和密度远优于ECM组。Image J定量分析表明，PSC组中小管数量（29.63±2.29）高于ECM组（20.13±2.36），具有统计学意义（P<0.05）。
结论
PSC对HUVECs的增殖、分化和血管生成具有显著的促进作用。
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