School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, UK.
School of Biomedical Sciences, University of Ulster, Derry~Londonderry, UK.
Int Endod J. 2020 Jun;53(6):834-845. doi: 10.1111/iej.13281. Epub 2020 Mar 20.
To use connectivity mapping, a bioinformatics approach, to identify compounds that could induce odontogenic differentiation of dental pulp cells (DPCs) and to experimentally validate this effect. A subsidiary aim was to investigate the anti-inflammatory effect of any identified compound.
The Gene Expression Omnibus (GEO) database was searched for microarray data sets assessing odontogenic differentiation of human DPCs. An odontogenic gene expression signature was generated by differential expression analysis. The statistical significant connectivity map (ssCMap) method was used to identify compounds with a highly correlating gene expression pattern. DPCs were treated with the compound identified, and osteo/odontogenic differentiation was assessed by Alizarin red staining, alkaline phosphatase activity and expression of osteo/odontogenic genes ALPL, RUNX2, COL1A1, DSPP, DMP1 and SPP1 by RT-PCR. The anti-inflammatory effect of the compound was assessed using an ex vivo pulpitis model, and cytokine levels were measured with multiplex assay. Means were compared using the t-test or ANOVA followed by a Bonferroni post hoc test with the level of significance set at P ≤ 0.05.
The GEO database search identified a specific gene expression signature for osteo/odontogenic differentiation. Analysis using ssCMap found that acetylsalicylic acid [(ASA)/aspirin] was the drug with the strongest correlation with that gene signature. The treatment of DPCs with 0.05 mmol L ASA showed increased alkaline phosphatase activity (P < 0.001), mineralization (P < 0.05), and increased the expression of the osteo/odontogenic genes, DMP1 and DSPP (P < 0.05). Low concentration (0.05 mmol L ) ASA reduced inflammatory cytokines IL-6 (P < 0.001), CCL21 (P < 0.05) and MMP-9 (P < 0.05) in an ex vivo pulpitis model.
Connectivity mapping, a web-based informatics method, was successfully used to identify aspirin as a candidate drug that could modulate the differentiation of DPCs. Aspirin was shown to induce odontogenic differentiation in DPCs in vitro and this, together with its anti-inflammatory effects, makes it a potential candidate for vital pulp therapies.
利用连接组学图谱分析，一种生物信息学方法，鉴定能诱导牙髓细胞（DPC）牙向分化的化合物，并进行实验验证。次要目的是研究鉴定化合物的抗炎作用。
从基因表达综合数据库（GEO）中搜索评估人 DPC 牙向分化的微阵列数据集。通过差异表达分析生成牙向分化基因表达谱。采用显著连接图谱（ssCMap）方法鉴定具有高度相关基因表达模式的化合物。用鉴定出的化合物处理 DPC，通过茜素红染色、碱性磷酸酶活性和 RT-PCR 检测成骨/牙向分化基因 ALPL、RUNX2、COL1A1、DSPP、DMP1 和 SPP1 的表达，评估 DPC 的成骨/牙向分化。采用牙髓炎模型评估化合物的抗炎作用，采用多重分析检测细胞因子水平。采用 t 检验或 ANOVA 分析，然后采用 Bonferroni 事后检验，以 P≤0.05 为差异有统计学意义。
GEO 数据库检索鉴定出了一个特定的成骨/牙向分化基因表达谱。ssCMap 分析发现，乙酰水杨酸（ASA）是与该基因特征相关性最强的药物。用 0.05mmol/L ASA 处理 DPC 可增加碱性磷酸酶活性（P<0.001）、矿化（P<0.05），并增加成骨/牙向分化基因 DMP1 和 DSPP 的表达（P<0.05）。低浓度（0.05mmol/L）ASA 可减少牙髓炎模型中促炎细胞因子 IL-6（P<0.001）、CCL21（P<0.05）和 MMP-9（P<0.05）的表达。
连接组学图谱分析，一种基于网络的信息学方法，成功地鉴定出阿司匹林是一种候选药物，可调节 DPC 的分化。阿司匹林能诱导 DPC 体外牙向分化，且具有抗炎作用，因此它是牙髓活力治疗的潜在候选药物。
