Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea.
Dental Implantology, Graduate School of Clinical Dental Science, The Catholic University of Korea, Seoul 06591, Republic of Korea.
Medicina (Kaunas). 2023 Feb 15;59(2):377. doi: 10.3390/medicina59020377.
: A derivative of the enamel matrix was used to speed up periodontal regeneration, including the formation of new cementum, alveolar bone, and periodontal ligament. In this study, human gingiva-derived stem cell-derived cell spheroids were used to assess the effects of an enamel matrix derivative on cell viability, osteogenic differentiation, and mineralization. : Human gingiva-derived stem cells were used to create spheroids, which were then coupled with unloaded control groups and an enamel matrix derivative at a final concentration of 2.7, 27, 270, and 2700 μg/mL. The morphological examination of the created stem cell spheroids took place on days 1, 3, 5, and 7. The Live/Dead Kit assay was used to determine the qualitative viability of cells on days 3 and 7. Using the Cell Counting Kit-8, the quantitative vitality of the cell spheroids was assessed on days 1, 3, and 5. On days 7 and 14, alkaline phosphatase activity assays and Alizarin Red S staining were carried out to examine the osteogenic differentiation of the cell spheroids. RUNX2 and COL1A1 expression levels on days 7 and 14 were determined using real-time polymerase chain reaction. The added enamel matrix derivative at the tested concentrations did not significantly alter the morphology of the applied stem cells' well-formed spheroids on day 1. On days 3 and 7, the majority of the spheroids' cells fluoresced green while they were being cultivated. Alkaline phosphatase activity data revealed a substantial rise in the 2700 μg/mL group on day 7 when compared to the unloaded control ( < 0.05). On days 7 and 14, calcium deposits were distinctly seen in each group. In the 27 and 2700 μg/mL groups, the treatment with the enamel matrix derivative resulted in noticeably higher values for the Alizarin Red S staining ( < 0.05). qPCR results showed that adding an enamel matrix derivative to the culture of the 27 μg/mL group raised the level of RUNX2 mRNA expression. These results lead us to the conclusion that a derivative of the enamel matrix may be used to promote osteogenic differentiation in stem cell spheroids.
: 一种釉基质衍生物被用于加速牙周组织再生，包括新的牙骨质、牙槽骨和牙周韧带的形成。在这项研究中，用人牙龈来源的干细胞衍生的细胞球评估了釉基质衍生物对细胞活力、成骨分化和矿化的影响。: 用人牙龈来源的干细胞制备细胞球，然后将其与未加载的对照组和浓度分别为 2.7、27、270 和 2700μg/ml 的釉基质衍生物相耦合。在第 1、3、5 和 7 天对制备的干细胞球进行形态学检查。在第 3 和第 7 天使用 Live/Dead 试剂盒测定细胞的定性活力。使用细胞计数试剂盒-8 在第 1、3 和 5 天评估细胞球的定量活力。在第 7 和 14 天进行碱性磷酸酶活性测定和茜素红 S 染色，以检查细胞球的成骨分化。在第 7 和 14 天使用实时聚合酶链反应测定 RUNX2 和 COL1A1 的表达水平。在第 1 天，测试浓度的添加釉基质衍生物没有明显改变应用干细胞的良好成型球的形态。在第 3 和第 7 天，大多数球的细胞在培养过程中发绿光。碱性磷酸酶活性数据显示，与未加载的对照组相比，2700μg/ml 组在第 7 天有显著升高（<0.05）。在第 7 和 14 天，每个组都明显观察到钙沉积物。在 27 和 2700μg/ml 组中，用釉基质衍生物处理导致茜素红 S 染色的数值明显升高（<0.05）。qPCR 结果表明，在 27μg/ml 组的培养物中添加釉基质衍生物可提高 RUNX2 mRNA 表达水平。这些结果使我们得出结论，釉基质衍生物可用于促进干细胞球的成骨分化。
