Cellular Therapy and Hematopoietic Transplant Unit, Hematology Department, Virgen de la Arrixaca Clinical University Hospital, IMIB, University of Murcia, Murcia, Spain.
School of Dentistry, Faculty of Medicine, University of Murcia, Murcia, Spain.
Int Endod J. 2017 Dec;50 Suppl 2:e19-e30. doi: 10.1111/iej.12751. Epub 2017 Mar 17.
To investigate the cytotoxicity and bioactivity of several pulpotomy materials: Biodentine (Septodont, Saint-Maur-des-Fosses, France) MTA (Angelus, Londrina, PR, Brazil), Theracal LC (Bisco Inc., Schamburg, IL, USA) and IRM (Dentsply DeTrey GmbH, Konstanz, Germany), after contact with stem cells isolated from human exfoliated primary teeth (SHEDs).
SHEDs were cultured in the presence of the eluates of various pulpotomy materials for 24, 48 and 72 h. Cell viability was determined by mitochondrial dehydrogenase enzymatic (MTT) assay. Apoptosis and changes in cell phenotype were evaluated by flow cytometry. Also, an in vitro scratch wound-healing assay was used to determine their effects on cell migration. To assess cell morphology and attachment to the different pulpotomy materials, SHEDs were directly seeded onto the material surfaces and analysed by scanning electron microscopy (SEM). Finally, the deposition of a calcified matrix in presence of these materials was verified by Alizarin Red staining. Statistical analysis was performed with analysis of variance and Bonferroni or Tukey post-test (α = 0.05).
Cell viability in the presence of Biodentine eluates was significantly higher to that obtained using complete medium alone (control; P < 0.01) and was also significantly higher than using MTA Angelus from 48 h of incubation (P < 0.01). However, Theracal LC and IRM were associated with low rates of cell viability (P < 0.001). Similar results were obtained in an apoptosis assay. In addition, SHEDs maintained their mesenchymal phenotype in all conditions although their capacity to migrate was higher in the presence of Biodentine. SEM studies revealed a suitable proliferation rate, cell spreading and attachment, especially when using Biodentine and MTA Angelus discs. Finally, Biodentine eluates significantly induced calcified matrix deposition from 7 days of culture (P < 0.01).
Biodentine exhibited better cytocompatibility and bioactivity than MTA Angelus, Theracal LC and IRM.
研究几种盖髓材料的细胞毒性和生物活性：Biodentine（圣莫里斯德福斯，法国赛普敦）、MTA（朗德里纳，巴西安杰卢斯）、Theracal LC（比思科公司，芝加哥，美国）和 IRM（登士柏德特莱有限公司，康斯坦茨，德国）与从人脱落乳牙中分离出的干细胞（SHED）接触后的反应。
将 SHED 在各种盖髓材料的浸提液中培养 24、48 和 72 小时。通过线粒体脱氢酶酶（MTT）测定法来确定细胞活力。通过流式细胞术评估细胞凋亡和表型变化。此外，还使用体外划痕愈合试验来确定它们对细胞迁移的影响。为了评估 SHED 对不同盖髓材料的细胞形态和附着的影响，将 SHED 直接接种到材料表面，并通过扫描电子显微镜（SEM）进行分析。最后，通过茜素红染色来验证这些材料存在时钙化基质的沉积。使用方差分析和 Bonferroni 或 Tukey 后检验（α=0.05）进行统计分析。
Biodentine 浸提液中细胞活力明显高于单独使用完全培养基（对照；P<0.01），也明显高于从孵育 48 小时起使用 MTA 安杰卢斯（P<0.01）。然而，Theracal LC 和 IRM 与低细胞活力率相关（P<0.001）。在凋亡试验中也得到了类似的结果。此外，SHED 保持其间充质表型，但在 Biodentine 存在的情况下，其迁移能力更高。SEM 研究显示，Biodentine 和 MTA 安杰卢斯的细胞增殖率、细胞扩散和附着都合适。最后，Biodentine 浸提液从培养的第 7 天开始显著诱导钙化基质沉积（P<0.01）。
Biodentine 表现出比 MTA 安杰卢斯、Theracal LC 和 IRM 更好的细胞相容性和生物活性。
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