Khampatee V, Zhang C, Chou L
Department of Restorative Sciences and Biomaterials, Boston University Henry M. Goldman School of Dental Medicine, Boston, MA, USA.
Int J Dent. 2022 Aug 5;2022:3246811. doi: 10.1155/2022/3246811. eCollection 2022.
This study aimed to investigate the roles of aspirin (ASA) and its concentrations on the odontogenesis of human dental pulp cells (HDPCs) and to investigate the influence of ASA on TGF-1 liberation from dentin. . HDPCs were cultured in a culture medium with 25, 50, 75, 100, and 200 ·g/mL ASA and 0 ·g/mL ASA as a control. The mitochondrial activity of HDPCs was assessed using an MTT assay. Crystal violet staining and triton were used to evaluate cell proliferation rates. ALP activity was measured with a fluorometric assay. Expressions of DSP and RUNX2 were determined with the ELISA. DSP and RUNX2 mRNA levels were measured with RT-qPCR. Alizarin red staining was conducted to evaluate the mineralized nodule formation. Dentin slices were submerged in PBS (negative control), 17% EDTA (positive control), and ASA before collecting the solution for TGF-1 quantification by the ELISA. The data were analyzed by the tests and ANOVA, followed by the Tukey post hoc tests. values < 0.05 were considered statistically significant.
The results showed that 25-50 ·g/mL ASA promoted mitochondrial activity of HDPCs at 72 h ( < 0.05) and yielded significantly higher proliferation rates of HDPCs than the control at 14d and 21d ( < 0.001). All concentrations of ASA promoted odontogenic differentiation of HDPCs by enhancing the levels of DSP and RUNX2, their mRNA expression, and mineralization in a dose-dependent manner. Also, ASA yielded significantly higher TGF-1 liberation after conditioning dentin for 5 min (25, 200 ·g/mL; < 0.001) and 10 min (200 ·g/mL; < 0.05).
This study demonstrated that ASA, especially in high concentrations, promoted the odontogenesis of HDPCs and TGF-1 liberation from dentin, showing the potential of being incorporated into the novel pulp capping materials for dental tissue regeneration.
本研究旨在探究阿司匹林（ASA）及其浓度对人牙髓细胞（HDPCs）牙发生的作用，并研究ASA对牙本质中转化生长因子 -1（TGF-1）释放的影响。将HDPCs培养于含有25、50、75、100和200μg/mL ASA的培养基中，以0μg/mL ASA作为对照。使用MTT法评估HDPCs的线粒体活性。采用结晶紫染色和曲拉通来评估细胞增殖率。用荧光法测定碱性磷酸酶（ALP）活性。通过酶联免疫吸附测定（ELISA）法测定牙本质涎磷蛋白（DSP）和 Runt相关转录因子2（RUNX2）的表达。用逆转录定量聚合酶链反应（RT-qPCR）法检测DSP和RUNX2的mRNA水平。进行茜素红染色以评估矿化结节的形成。在收集用于ELISA法进行TGF-1定量的溶液之前，将牙本质切片浸入磷酸盐缓冲液（PBS，阴性对照）、17%乙二胺四乙酸（EDTA，阳性对照）和ASA中。数据通过检验和方差分析进行分析，随后进行Tukey事后检验。P值<0.05被认为具有统计学意义。
结果显示，25 - 50μg/mL ASA在72小时时促进了HDPCs的线粒体活性（P < 0.05），并且在第14天和第21天时，其HDPCs增殖率显著高于对照组（P < 0.001）。所有浓度的ASA均通过增强DSP和RUNX2的水平、它们的mRNA表达以及矿化，以剂量依赖的方式促进HDPCs的牙源性分化。此外，在牙本质处理5分钟（25、200μg/mL；P < 0.001）和10分钟（200μg/mL；P < 0.05）后，ASA产生的TGF-1释放量显著更高。
本研究表明，ASA，尤其是高浓度时，促进了HDPCs的牙发生以及牙本质中TGF-1的释放，显示出其被纳入新型牙髓盖髓材料用于牙组织再生的潜力。
