Huang S-C, Wu B-C, Kao C-T, Huang T-H, Hung C-J, Shie M-Y
School of Dentistry, Chung Shan Medical University, Taichung, Taiwan; Department of Dentistry, Chung Shan Medical University Hospital, Taichung, Taiwan.
Int Endod J. 2015 Mar;48(3):236-45. doi: 10.1111/iej.12305. Epub 2014 Jun 25.
To investigate the influence of mineral trioxide aggregate (MTA) on angiogenesis of primary human dental pulp cells (hDPCs) via the MAPK pathway, in particular p38.
Human dental pulp cells were cultured with MTA to angiogenesis, after which cell viability, ion concentration, osmolality, NO secretion, the von Willebrand factor (vWF) and angiopoietin-1 (Ang-1) protein expression were examined. PrestoBlue(®) was used for evaluating the proliferation of hDPCs. An enzyme-linked immunosorbent assay was employed to determine vWF and Ang-1 protein secretion in hDPCs cultured on MTA and the control. Cells cultured on the tissue culture plate without the cement were used as the control. The t-test was used to evaluate the significance of the differences between the mean values.
Mineral trioxide aggregate elicited a significant (P < 0.05) increased viability compared with the control (15%, 16% and 13% on days 1, 3 and 5 of cell seeding, respectively). MTA consumed calcium and phosphate ions, and released more Si ions in the medium. MTA significantly (P < 0.05) increased the osmolality of the medium to 313, 328 and 341 mOsm kg(-1) after 1, 3 and 5 days, respectively. P38 was activated through phosphorylation, and the phosphorylation kinase was investigated in the cell system after being cultured with MTA. Expression levels for Ang-1 and vWF in hDPCs on MTA were higher than those of the MTA + p38 inhibitor (SB203580) group (P < 0.05) at all of the time-points.
Mineral trioxide aggregate was able to activate the p38 pathway in hDPCs cultured in vitro. Moreover, Si increased the osmolality required to facilitate the angiogenic differentiation of hDPCs via the p38 signalling pathway. When the p38 pathway was blocked by SB203580, the angiogenic-dependent protein secretion decreased. These findings verify that the p38 pathway plays a key role in regulating the angiogenic behaviour of hDPCs cultured on MTA.
通过丝裂原活化蛋白激酶（MAPK）途径，特别是p38途径，研究三氧化矿物凝聚体（MTA）对原代人牙髓细胞（hDPCs）血管生成的影响。
将人牙髓细胞与MTA共同培养以促进血管生成，之后检测细胞活力、离子浓度、渗透压、一氧化氮（NO）分泌、血管性血友病因子（vWF）和血管生成素-1（Ang-1）蛋白表达。使用普洛麦格公司的碧色法细胞增殖检测试剂（PrestoBlue(®)）评估hDPCs的增殖情况。采用酶联免疫吸附测定法测定在MTA及对照组上培养的hDPCs中vWF和Ang-1蛋白的分泌。将在不含黏固剂的组织培养板上培养的细胞用作对照。采用t检验评估平均值之间差异的显著性。
与对照组相比，三氧化矿物凝聚体显著（P < 0.05）提高了细胞活力（在细胞接种第1、3和5天分别提高了15%、16%和13%）。MTA消耗了钙和磷酸根离子，并在培养基中释放了更多的硅离子。MTA分别在1、3和5天后显著（P < 0.05）提高了培养基的渗透压至313、328和341 mOsm kg⁻¹。p38通过磷酸化被激活，并在与MTA共同培养后的细胞体系中研究了磷酸化激酶。在所有时间点，MTA上hDPCs中Ang-1和vWF的表达水平均高于MTA + p38抑制剂（SB203580）组（P < 0.05）。
三氧化矿物凝聚体能够在体外培养的hDPCs中激活p38途径。此外，硅通过p38信号通路提高了促进hDPCs血管生成分化所需的渗透压。当p38途径被SB203580阻断时，血管生成相关蛋白的分泌减少。这些发现证实p38途径在调节在MTA上培养的hDPCs的血管生成行为中起关键作用。
