Department of Restorative Dentistry, São Paulo State University (UNESP), School of Dentistry, Araraquara, São Paulo, Brazil.
Department of Dental Materials and Prosthodontics, São Paulo State University (UNESP), School of Dentistry, Araraquara, São Paulo, Brazil.
Arch Oral Biol. 2025 Jan;169:106095. doi: 10.1016/j.archoralbio.2024.106095. Epub 2024 Sep 24.
This study investigated the bioactive effects of retinoic acid and ascorbic acid on hSCAPs in vitro.
Cells were obtained from human third molars (n=4) and characterized for mesenchymal stem cell markers by flow cytometry. The experimental groups: control (α-MEM); vehicle control group (α-MEM + 0.17 % DMSO); retinoic acid 0.1, 1, and 10 µM; and ascorbic acid 3, 30, and 300 µM (n=8) were tested for cell viability (alamarBlue; 1, 3, and 7 days), total collagen synthesis (Sirius Red; 1 and 7 days), mineralized matrix formation (Alizarin red; 14 days), and the regulation of gene expression related to mineralization (ALPL and DSPP), cell migration (ITGAV and CXCL12) angiogenesis (VEGFA) and collagen synthesis (COL1A1 and COL3A1; RT-qPCR) on 1 and 7 days. ACTB and GAPDH were used as reference genes. Data were analyzed by ANOVA and complementary tests at a 5 % significance level.
Ascorbic acid 300 µM increased viability, and retinoic acid reduced it dose-dependently. Retinoic acid 0.1 µM and ascorbic acid 30 and 300 µM increased mineralized matrix formation and total collagen synthesis, and retinoic acid 10 µM decreased. On day 1, 0.1 µM retinoic acid upregulated the gene expression of COL1A1, COL3A1, VEGFA, CXCL12, ALPL, DSPP e ITGAV, and 300 µM ascorbic acid upregulated COL1A1, COL3A1 and DSPP. However, on day 7, retinoic acid downregulated ALPL, COL3A1, CXCL12, and VEGFA and downregulated ITGAV and VEGFA.
Retinoic acid 0.1 µM and ascorbic acid 300 µM biostimulated hSCAPs to differentiate into pro-regenerative phenotypes with potential application for REPs.
本研究旨在探讨维甲酸和抗坏血酸对体外 hSCAP 的生物活性作用。
从人第三磨牙中获得细胞（n=4），并通过流式细胞术对间充质干细胞标志物进行特征分析。实验组：对照组（α-MEM）；载体对照组（α-MEM+0.17% DMSO）；维甲酸 0.1、1 和 10µM；抗坏血酸 3、30 和 300µM（n=8）分别用于细胞活力检测（alamarBlue；1、3 和 7 天）、总胶原合成检测（Sirius Red；1 和 7 天）、矿化基质形成检测（茜素红；14 天）和与矿化相关的基因表达调控（ALPL 和 DSPP）、细胞迁移检测（ITGAV 和 CXCL12）、血管生成检测（VEGFA）和胶原合成检测（COL1A1 和 COL3A1；RT-qPCR），分别在 1 和 7 天进行。ACTB 和 GAPDH 用作参考基因。在 5%的显著性水平下，采用方差分析和补充检验对数据进行分析。
抗坏血酸 300µM 增加了细胞活力，而维甲酸则呈剂量依赖性地降低了细胞活力。0.1µM 维甲酸和 30 和 300µM 抗坏血酸增加了矿化基质形成和总胶原合成，而 10µM 维甲酸则减少了。在第 1 天，0.1µM 维甲酸上调 COL1A1、COL3A1、VEGFA、CXCL12、ALPL、DSPP 和 ITGAV 的基因表达，而 300µM 抗坏血酸上调 COL1A1、COL3A1 和 DSPP 的基因表达。然而，在第 7 天，维甲酸下调了 ALPL、COL3A1、CXCL12 和 VEGFA，并下调了 ITGAV 和 VEGFA。
0.1µM 维甲酸和 300µM 抗坏血酸刺激 hSCAP 分化为具有再生潜力的表型，可能应用于再生工程支架。
