Effect of bioactive glasses on angiogenic growth factors of human dental pulp cells

目的 体外观察、比较新型纳米生物活性玻璃nano-58S和传统生物活性玻璃45S5对人牙髓细胞增殖及血管内皮生长因子(vascular endothelial growth factor,VEGF)、碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)的作用,以期为进一步研究生物活性玻璃是否可在牙髓再生的组织工程学中发挥作用奠定基础.方法 实验组用45S5 (45S5组)、nano-58S(nano-58S组)生物活性玻璃浸提液培养人牙髓细胞,空白对照组用达尔伯克改良伊格尔培养基(Dulbecco's modified Eagle's medium,DMEM)培养,每组3孔.培养人牙髓细胞1、2、3d分别采用甲基噻唑基四唑法观察浸提液对细胞增殖的影响,用实时荧光定量反转录PCR和酶联免疫吸附测定法检测VEGF、bFGF基因表达水平和蛋白分泌量.结果 人牙髓细胞培养第2天,45S5组、nano-58S组细胞增殖率分别为空白对照组的(134.5±5.0)％和(146.3±19.8)％,与空白对照组相比差异均有统计学意义(P＜0.05)；45S5组和nano-58S组在第1天VEGF蛋白分泌量分别为(189.29±4.64)和(216.18±14.67) ng/L,均显著高于空白对照组[(159.03±11.69) ng/L] (P＜0.05),nano-58S组VEGF蛋白分泌量显著高于45S5组(P＜0.05)；两个实验组的bFGF蛋白分泌量亦显著高于空白对照组(P＜0.05)；45S5组和nano-58S组的VEGF基因表达量在第1天分别为(1.70±0.19)和(1.63±0.42),均显著高于空白对照组(1.00 +0.02) (P ＜0.05)；bFGF基因相对表达量在第3天分别为(1.49±0.02)和(2.30±0.04),亦均显著高于空白对照组(1.29 ±0.04) (P＜0.05).结论 生物活性玻璃可促进人牙髓细胞的增殖及VEGF、bFGF的基因表达和蛋白分泌,nano-58S比45S5的促进作用更显著.To investigate the effects of bioactive glasses (BG) including 45S5 and nano-58S on proliferation, angiogenic markers vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) secretion and gene expression of human dental pulp cells (HDPC).HDPC of 4th passage were cultured in Dulbecco's modified Eagle's medium (DMEM) which contained 0.1 g/L 45S5 or nano-58S ionic dissolution products. Meanwhile HDPC were cultured in DMEM without BG as control group. Proliferation of the cells was evaluated with methyl thiazolyl tetrazolium (MTT) assay on day 1, 2, 3. Quantitative real-time PCR and quantitative sandwich enzyme immunoassays were used to test VEGF and bFGF gene expression and protein secretion of HDPC on day 1, 2, 3.The relative growth rate (RGR) of 45S5 and nano-58S groups were (134.5 ± 5.0)% and (146.3 ± 19.8)%, which was significantly different from that of control group (P < 0.05). The quantity of VEGF secretion of two experimental groups were (189.29 ± 4.64) and (216.18 ± 14.67) ng/L, respectively, significantly higher than that of the control group [(159.03 ± 11.69) ng/L] (P < 0.05). Furthermore, the nano-58S group secreted much more VEGF than 45S5 group (P < 0.05).bFGF secretion of HDPC was also enhanced by both 45S5 and nano-58S bioactive glasses. The VEGF gene expression of 45S5 and nano-58S on day 1 were (1.70 ± 0.19) and (1.63 ± 0.42), while the bFGF gene expressin on day 3 were (1.49 ± 0.02) and (2.30 ± 0.04), all significantly higher than that of control group (P < 0.05).Bioactive glasses can enhance the proliferation, VEGF and bFGF secretion and gene expression of human dental pulp cells. Compared with 45S5, nano-58S showed a higher activation.国家自然科学基金PubMed中文核心期刊要目总览(PKU)中国科技核心期刊(ISTIC)中国科学引文数据库(CSCD)04234-2384