Inserm, UMR 1229, Regenerative Medicine and Skeleton, University of Nantes, Nantes, France; Department of Endodontics, University of Nantes, Nantes, France; CHU de Nantes, Nantes University Hospital, PHU 4 OTONN, Nantes, France.
Department of Orthodontics, University of the Pacific School of Dentistry, San Francisco, California.
J Endod. 2020 Jun;46(6):818-826. doi: 10.1016/j.joen.2020.03.011. Epub 2020 Apr 21.
The aim of the present study was to assess the effects of different silicate-based sealers (ie, BioRoot RCS [Septodont, Saint Maur des Fosses, France], ProRoot ES [Dentsply Sirona, York, PA], and MTA Fillapex [Angelus, Londrina, PR, Brazil]) on cytokine production and viability of human periodontal ligament stem cells (PDLSCs). AH Plus (Dentsply DeTrey GmbH, Konstanz, Germany) was used as a reference material.
PDLSCs were cultured either in 2-dimensional or 3-dimensional conditions (in 0.15%-0.5% PuraMatrix [BD Biosciences, Bedford, MA]) for 24 hours with eluates from set endodontic sealers. Additionally, the toxicity of eluates from endodontic sealers was evaluated using an in vitro root model experimental procedure. PDLSC viability was determined using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay. PDLSC culture medium was used for cytokine quantification (interleukin [IL]-6, IL-8, growth-regulated oncogene, IL,-4 and IL-10) using the HCYTMAG-60K-PX41 Milliplex kit (EMD Millipore, Burlington, MA).
In 2-dimensional culture conditions, BioRoot RCS revealed a good PDLSC viability rate. ProRoot ES had no effect on PDLSC viability regardless of the dilution. MTA Fillapex was strongly cytotoxic even at the lowest extract dilutions (1:1, 1:2, and 1:4). Encapsulation of PDLSCs in PuraMatrix tended to decrease the cytotoxic effect of the sealers. In the 3-dimensional in vitro root model experimental procedure, BioRoot RCS, ProRoot ES, and MTA Fillapex revealed a cytocompatibility pattern. Different calcium silicate-based sealers exhibited different proinflammatory cytokine production. BioRoot RCS greatly stimulated the release of IL-10 and, to a lesser degree, IL-4 by PDLSCs (P < .05).
BioRoot RCS and ProRoot ES did not induce proinflammatory cytokines and promoted anti-inflammatory cytokine secretion by PDLSCs that may have a positive local impact by attenuating an initial inflammatory response.
本研究的目的是评估不同硅酸酯类密封剂（即 BioRoot RCS [Septodont，Saint Maur des Fosses，法国]、ProRoot ES [Dentsply Sirona，York，PA]和 MTA Fillapex [Angelus，Londrina，PR，巴西]）对人牙周膜干细胞（PDLSCs）细胞因子产生和活力的影响。AH Plus（Dentsply DeTrey GmbH，Konstanz，德国）用作参考材料。
将 PDLSCs 在二维或三维条件下（在 0.15%-0.5% PuraMatrix [BD Biosciences，Bedford，MA]中）培养 24 小时，并用凝固牙根管封闭剂的浸提液处理。此外，还通过体外根模型实验程序评估牙根管封闭剂浸提液的毒性。使用 3-（4,5-二甲基-2-噻唑基）-2,5-二苯基-2H-四唑溴盐测定法（3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay）来确定 PDLSC 的活力。使用 HCYTMAG-60K-PX41 Milliplex 试剂盒（EMD Millipore，Burlington，MA）测量 PDLSC 培养液中的细胞因子（白细胞介素[IL]-6、IL-8、生长调节致癌基因、IL-4 和 IL-10）的浓度。
在二维培养条件下，BioRoot RCS 显示出良好的 PDLSC 活力。ProRoot ES 对 PDLSC 活力没有影响，无论稀释度如何。即使在最低的浸提液稀释度（1:1、1:2 和 1:4）下，MTA Fillapex 也具有很强的细胞毒性。PDLSCs 的封装在 PuraMatrix 中倾向于降低密封剂的细胞毒性作用。在体外根模型实验程序的三维条件下，BioRoot RCS、ProRoot ES 和 MTA Fillapex 显示出细胞相容性模式。不同的硅酸钙基密封剂表现出不同的促炎细胞因子产生。BioRoot RCS 极大地刺激了 PDLSCs 释放 IL-10，并在较小程度上释放 IL-4（P <.05）。
BioRoot RCS 和 ProRoot ES 没有诱导促炎细胞因子的产生，并促进了 PDLSCs 抗炎细胞因子的分泌，这可能通过减轻初始炎症反应对局部产生积极影响。
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