Abuarqoub Duaa, Aslam Nazneen, Jafar Hanan, Abu Harfil Zakariya, Awidi Abdalla
Cell Therapy Center, The University of Jordan, Amman 11180, Jordan.
Department of Pharmacology and Biomedical Sciences, Faculty of Pharmacy, University of Petra, Amman 11180, Jordan.
Dent J (Basel). 2020 Feb 8;8(1):17. doi: 10.3390/dj8010017.
Biodentine™ is a tricalcium silicate-based cement material that has a great impact on different biological processes of dental stem cells, compared to other biomaterials. Therefore, we aimed to investigate the optimum biocompatible concentration of Biodentine™ with stem cells derived from periodontal ligament (hPDLSCs) by determining cell proliferation, cytotoxicity, migration, adhesion and mineralization potential. hPDLSCs were treated with Biodentine™ extract at different concentrations; 20, 2, 0.2 and 0.02 mg/mL. Cells cultured without Biodentine™ were used as a blank control. The proliferation potential of hPDLSCs was evaluated by MTT viability analysis for 6 days. Cytotoxicity assay was performed after 3 days by using AnnexinV/7AAD. Migration potential was investigated by wound healing and transwell migration assays at both cellular and molecular levels. The expression levels of chemokines CXCR4, MCP-1 and adhesion molecules FGF-2, FN, VCAM and ICAM-1 were measured by qPCR. The communication potentials of these cells were determined by adhesion assay. In addition, mineralization potential was evaluated by measuring the expression levels of osteogenic markers; ALP, OCN, OPN and Collagen type1 by qPCR. Our results showed significant increase in the proliferation of hPDLSCs at low concentrations of Biodentine™ (2, 0.2 and 0.02 mg/mL) while higher concentration (20 mg/mL) exhibited cytotoxic effect on the cells. Moreover, 2 mg/mL Biodentine™ showed a significant increase in the migration, adhesion and mineralization potentials of the derived cells among all concentrations and when compared to the blank control. Our findings suggest that 2 mg/mL of Biodentine™ is the most biocompatible concentration with hPDLSCs, showing a high stimulatory effect on the biological processes.
BioDentine™是一种基于硅酸三钙的水门汀材料，与其他生物材料相比，它对牙干细胞的不同生物学过程有很大影响。因此，我们旨在通过测定细胞增殖、细胞毒性、迁移、黏附及矿化潜能，研究BioDentine™与牙周膜来源的干细胞（hPDLSCs）的最佳生物相容性浓度。用不同浓度（20、2、0.2和0.02mg/mL）的BioDentine™提取物处理hPDLSCs。未用BioDentine™培养的细胞用作空白对照。通过MTT活力分析评估hPDLSCs 6天的增殖潜能。3天后使用膜联蛋白V/7-氨基放线菌素D进行细胞毒性检测。通过伤口愈合实验和Transwell迁移实验在细胞和分子水平研究迁移潜能。通过qPCR检测趋化因子CXCR4、MCP-1以及黏附分子FGF-2、FN、VCAM和ICAM-1的表达水平。通过黏附实验确定这些细胞的通讯潜能。此外，通过qPCR测量成骨标志物（碱性磷酸酶、骨钙素、骨桥蛋白和Ⅰ型胶原）的表达水平来评估矿化潜能。我们的结果显示，低浓度（2、0.2和0.02mg/mL）的BioDentine™可显著促进hPDLSCs的增殖，而高浓度（20mg/mL）对细胞具有细胞毒性作用。此外，在所有浓度中，与空白对照相比，2mg/mL的BioDentine™可显著提高hPDLSCs的迁移、黏附及矿化潜能。我们的研究结果表明，2mg/mL的BioDentine™是与hPDLSCs最具生物相容性的浓度，对生物学过程具有高度刺激作用。
