Aksel Hacer, Huang George T-J
Department of Bioscience Research, College of Dentistry University of Tennessee Health Science Center, Memphis, Tennessee.
Department of Bioscience Research, College of Dentistry University of Tennessee Health Science Center, Memphis, Tennessee.
J Endod. 2017 Jun;43(6):930-935. doi: 10.1016/j.joen.2017.01.036. Epub 2017 Apr 27.
The purpose of this study was to investigate whether combined and concerted delivery of vascular endothelial growth factor (VEGF) and bone morphogenetic protein 2 (BMP-2) enhances odonto/osteogenic differentiation of human dental pulp stem cells (DPSCs) in vitro.
Various concentrations of VEGF and/or BMP-2 with or without the presence of odonto/osteogenic medium (OM) were added into DPSC cultures for 21 days. The mineral formation in cultures was evaluated using alizarin red stain (ARS). Optimal concentrations of VEGF and BMP-2 were codelivered to DPSCs for total of 21 days with the following experimental groups: (1) group 1: OM only, (2) group 2: OM + VEGF, (3) group 3: OM + BMP-2, and (4) group 4: OM + VEGF + BMP-2 (subgroup 4a: VEGF present the first 7 days, 4b: BMP-2 present the last 14 days, and 4c, both present for 21 days). Cultures were then subjected to quantitative ARS analysis or harvested for quantitative polymerase chain reaction analysis for the expression of core-binding factor alpha 1 (CBFA1), alkaline phosphatase (ALP), and dentin matrix protein 1 (DMP-1).
No mineral formation was detected by ARS when VEGF and/or BMP-2 were used without OM. OM + VEGF, but not OM + BMP-2, formed more mineralization than OM (P < .05). In the codelivery groups, the highest mineralization was observed in OM + VEGF and subgroup 4a compared with OM or the other groups (P < .05). Quantitative polymerase chain reaction analysis showed that CBFA1, ALP, and DMP-1 levels were higher in groups 2, 3, and 4a compared with 4b and 4c (P < .05). CBFA1 expressed higher in groups 2, 3, and 4a compared with OM (P < .05). For ALP expression, only subgroup 4a expressed higher than OM (P < .05). No difference was detected between groups 2 and 3 (P > .05) in the expression of the 3 genes.
VEGF addition in the early phase rather than a continuous presence of both VEGF and BMP-2 enhances odonto/osteogenic differentiation of DPSCs.
本研究旨在探讨联合协同递送血管内皮生长因子（VEGF）和骨形态发生蛋白2（BMP-2）是否能在体外增强人牙髓干细胞（DPSC）的牙本质/成骨分化。
将不同浓度的VEGF和/或BMP-2在有或无牙本质/成骨培养基（OM）存在的情况下加入DPSC培养物中，培养21天。使用茜素红染色（ARS）评估培养物中的矿物质形成。将VEGF和BMP-2的最佳浓度共递送至DPSC，共21天，分为以下实验组：（1）组1：仅OM；（2）组2：OM + VEGF；（3）组3：OM + BMP-2；（4）组4：OM + VEGF + BMP-2（亚组4a：VEGF在前7天存在，4b：BMP-2在后14天存在，4c：两者均存在21天）。然后对培养物进行定量ARS分析，或收获进行定量聚合酶链反应分析，以检测核心结合因子α1（CBFA1）、碱性磷酸酶（ALP）和牙本质基质蛋白1（DMP-1）的表达。
当单独使用VEGF和/或BMP-2而无OM时，ARS未检测到矿物质形成。OM + VEGF组形成的矿化比OM组多，但OM + BMP-2组与OM组无差异（P <.05）。在共递送组中，与OM组或其他组相比，OM + VEGF组和亚组4a观察到最高的矿化（P <.05）。定量聚合酶链反应分析显示，与亚组4b和4c相比，组2、3和4a中的CBFA1、ALP和DMP-1水平更高（P <.05）。与OM组相比，组2、3和4a中的CBFA1表达更高（P <.05）。对于ALP表达，仅亚组4a高于OM组（P <.05）。组2和组3在这3种基因的表达上无差异（P >.05）。
早期添加VEGF而非VEGF和BMP-2持续存在可增强DPSC的牙本质/成骨分化。
