Department of Restorative Dentistry, School of Dentistry, University of São Paulo, São Paulo, Brazil.
Department of Endodontics, University of Texas Health Science Center at San Antonio, San Antonio, Texas.
J Endod. 2022 Dec;48(12):1511-1516. doi: 10.1016/j.joen.2022.09.005. Epub 2022 Sep 26.
Many mediators are produced during pulp inflammation and necrosis, including endocannabinoids (ECbs), which might affect the function of stem cells of the apical papilla (SCAP), cells of paramount importance for root formation, and regenerative endodontic treatment. The aim of this study was to evaluate the production of osteoclastogenesis-related mediators by SCAP modulated by ECbs and lipopolysaccharide (LPS) in vitro.
SCAP were cultured and treated with ECb anandamide (AEA), 2-arachidonoylglycerol, or N-arachidonoylaminophenol. All groups were incubated in the presence of a vehicle or LPS and the antagonist of transient receptor potential cation channel subfamily V member 1, capsazepine. After 24 hours, the culture medium supernatants were collected for further quantification of tumor necrosis factor alpha, CCL2, macrophage colony-stimulating factor, osteoprotegerin, and receptor activator of nuclear factor kappa B ligand.
Small amounts of tumor necrosis factor alpha and receptor activator of nuclear factor kappa B ligand were detected in SCAP supernatants, and none of the experimental conditions altered their production. A down-regulation in constitutive CCL2 production was observed in the AEA group compared with that in the LPS group. The production of macrophage colony-stimulating factor was significantly increased in all groups treated with AEA compared with the control and LPS-treated groups. Osteoprotegerin was significantly increased by AEA alone and by 2-arachidonoylglycerol and N-arachidonoylaminophenol in the presence of LPS and capsazepine.
AEA modulates some of the osteoclastogenic factors produced by SCAP in a bone resorption protective fashion.
牙髓炎症和坏死过程中会产生许多介质，包括内源性大麻素（ECbs），它们可能影响根尖乳头干细胞（SCAP）的功能，而 SCAP 对根形成和再生性牙髓治疗至关重要。本研究旨在评估 ECb 与脂多糖（LPS）共同作用下 SCAP 产生破骨细胞生成相关介质的情况。
SCAP 经培养并分别用 ECb 大麻素（AEA）、2-花生四烯酰甘油或 N-花生四烯酰氨基苯酚处理。所有组均在载体或 LPS 以及瞬时受体电位阳离子通道亚家族 V 成员 1 拮抗剂辣椒素存在的情况下孵育。24 小时后，收集培养物上清液，进一步定量检测肿瘤坏死因子-α、CCL2、巨噬细胞集落刺激因子、护骨素和核因子κB 受体激活剂配体。
SCAP 上清液中检测到少量肿瘤坏死因子-α和核因子κB 受体激活剂配体，但实验条件均未改变其产生。与 LPS 组相比，AEA 组的 CCL2 产生量呈下调。与对照组和 LPS 处理组相比，用 AEA 处理的所有组中巨噬细胞集落刺激因子的产生均显著增加。AEA 单独作用，以及在 LPS 和辣椒素存在的情况下，2-花生四烯酰甘油和 N-花生四烯酰氨基苯酚均可显著增加护骨素的产生。
AEA 以骨吸收保护方式调节 SCAP 产生的一些破骨细胞生成因子。
