研究大麻二酚对人牙髓细胞体外成牙/成骨能力的影响。

Investigation of in vitro odonto/osteogenic capacity of cannabidiol on human dental pulp cell.

机构信息

Graduate Periodontics, School of Dentistry, University of Detroit Mercy, Detroit, MI, 48208, USA; Department of Periodontics, Hebei Key Laboratory of Stomatology, Hebei Clinical Research Center for Oral Diseases, School and Hospital of Stomatology, Hebei Medical University, Shijiazhuang, Hebei, China.

Graduate Periodontics, School of Dentistry, University of Detroit Mercy, Detroit, MI, 48208, USA; Department of Orthodontics, Hebei Key Laboratory of Stomatology, Hebei Clinical Research Center for Oral Diseases, School and Hospital of Stomatology, Hebei Medical University, Shijiazhuang, Hebei, China.

出版信息

J Dent. 2021 Jun;109:103673. doi: 10.1016/j.jdent.2021.103673. Epub 2021 Apr 16.

DOI:10.1016/j.jdent.2021.103673

PMID:33872753

Abstract

INTRODUCTION

Vital pulp treatment (VPT) maintains tooth vitality with certain dental materials by protecting pulp from noxious stimulation and promoting repair through enhancing cell proliferation/differentiation, migration, and inducing odontogenesis. As a non-psychotropic cannabis constituent, cannabidiol (CBD) possesses the properties of analgesic, anti-inflammation, and osteogenesis. Therefore, we hypothesize that CBD may induce the odonto/osteogenesis of human dental pulp cells (HDPCs), a critical feature using as effective pulp capping agent for VPT.

MATERIALS AND METHODS

In this in vitro study, the cytotoxicity of CBD on HDPCs was determined by MTT assay. Scratch assay was performed to analyze HDPC migration. The biomineralization was examined by collagen synthesis and calcium nodule formation and related odonto/osteogenic and angiogenic genes. Cannabinoid receptor (CB) specificity was evaluated by Western blotting and Von Kossa staining using specific antagonists AM251 for cannabinoid receptor 1 (CB1) and AM 630 targeted at cannabinoid receptor 2 (CB2). In addition, the underlying molecular mechanism of CBD-induced biomineralization were investigated by examining CB-dependent MAPK signaling pathways.

RESULTS

CBD demonstrated bi-phasic effects on HDPC viability in tested concentrations. We found CBD significantly promoted cell migration, enhanced collagen synthesis and mineralized deposits in HDPCs when treated by 1 μM CBD supplemented in the differentiation media. RT-PCR revealed CBD increased the expression of angiogenic and odontogenic genes, such as DSPP, DMP-1, OPN, ALP, Runx2, VEGFR1 and ICAM-1. These effects were via MAPK activation in a manner mainly mediated by CB2.

CONCLUSION

The results from this study suggested that CBD can induce odonto/osteogenesis from HDPCs and has the potential to develop new therapeutics in VPT in dentistry.

摘要

简介

活髓治疗（VPT）通过保护牙髓免受有害刺激并通过增强细胞增殖/分化、迁移以及诱导牙发生来促进修复，从而使用某些牙科材料维持牙齿活力。大麻的非精神活性成分大麻二酚（CBD）具有镇痛、抗炎和成骨作用。因此，我们假设 CBD 可能会诱导人牙髓细胞（HDPCs）的牙/骨发生，这是 VPT 中用作有效牙髓覆盖剂的关键特征。

材料和方法

在这项体外研究中，通过 MTT 测定法确定 CBD 对 HDPCs 的细胞毒性。划痕试验用于分析 HDPC 迁移。通过胶原蛋白合成和钙结节形成以及相关的牙/成骨和成血管基因来检查生物矿化。通过使用大麻素受体 1（CB1）的特异性拮抗剂 AM251 和针对大麻素受体 2（CB2）的 AM 630 进行 Western 印迹和 Von Kossa 染色来评估大麻素受体（CB）的特异性。此外，通过检查 CBD 诱导的生物矿化的 CB 依赖性 MAPK 信号通路来研究 CBD 诱导生物矿化的潜在分子机制。

结果

在测试浓度下，CBD 对 HDPC 活力表现出双相作用。我们发现，当在分化培养基中添加 1μM CBD 时，CBD 可显著促进细胞迁移，增强 HDPC 中的胶原蛋白合成和矿化沉积物。RT-PCR 显示 CBD 增加了血管生成和牙发生基因的表达，例如 DSPP、DMP-1、OPN、ALP、Runx2、VEGFR1 和 ICAM-1。这些作用是通过 MAPK 激活来实现的，主要是通过 CB2 介导的。