Tuncer Budanur Damla, Zibandeh Noushin, Genç Deniz, Gökalp Muazzez, Kaşali Kamber, Akkoç Tunç, Sepet Elif
Pediatric Dentistry Department, Faculty of Dentistry, Istanbul University, Istanbul, Turkey.
Pediatric Allergy-Immunology Department, Faculty of Medicine, Marmara University, Istanbul, Turkey.
Dent Traumatol. 2018 Jun;34(3):188-200. doi: 10.1111/edt.12396. Epub 2018 Apr 18.
BACKGROUND/AIM: Ectoine is an amino acid that can preserve osmotic balance and has more preservative activity than other osmoregulators. There are no published reports on the osmoregulators' effects on viability or differentiation of dental stem cells. The aim of this study was to investigate the effect of Ectoine as a storage media on the viability and differentiation potential of human periodontal ligament mesenchymal stem cells (hPDLMSCs).
hPDLMSCs were obtained from impacted third molar teeth. The cells were isolated, submitted to trilineage differentiation, and characterized by flow cytometer (FC). hPDLMSCs were incubated with different media containing Ectoine, complete DMEM (cDMEM), Ectoine+cDMEM, milk, and tap water for 2, 6, 12, and 24 h. The cells were analyzed by FC for viability, early-apoptosis, late apoptosis, and necrosis rates. Osteogenic and fibroblastic differentiation in hPDLMSCs were investigated by Alizarin red stain and vimentin expression.
All treated groups showed significant decreases in cell viability after 2 h. Significant increases were detected in the number of dead cells between 2 and 12 h in all groups except the Ectoine+cDMEM group. The deposition of mineral matrix nodules was significantly higher in cells cultured with Ectoine+cDMEM compared with the other media. Higher vimentin expressions were detected in cells cultured with cDMEM and Ectoine+cDMEM media, respectively.
Ectoine added to cDMEM media, promoted cell survival plus osteogenic and fibroblastic differentiation of hPDLMSCs.
背景/目的：四氢嘧啶是一种能够维持渗透压平衡的氨基酸，其防腐活性高于其他渗透压调节剂。目前尚无关于渗透压调节剂对牙干细胞活力或分化影响的公开报道。本研究旨在探讨四氢嘧啶作为储存介质对人牙周膜间充质干细胞（hPDLMSCs）活力和分化潜能的影响。
hPDLMSCs取自阻生第三磨牙。分离细胞，进行三系分化，并通过流式细胞仪（FC）进行鉴定。将hPDLMSCs与含有四氢嘧啶、完全DMEM（cDMEM）、四氢嘧啶+cDMEM、牛奶和自来水的不同培养基孵育2、6、12和24小时。通过FC分析细胞的活力、早期凋亡、晚期凋亡和坏死率。通过茜素红染色和波形蛋白表达研究hPDLMSCs的成骨和成纤维细胞分化。
所有处理组在2小时后细胞活力均显著下降。除四氢嘧啶+cDMEM组外，所有组在2至12小时之间死亡细胞数量均显著增加。与其他培养基相比，用四氢嘧啶+cDMEM培养的细胞中矿化基质结节的沉积明显更高。在用cDMEM和四氢嘧啶+cDMEM培养基培养的细胞中分别检测到较高的波形蛋白表达。
向cDMEM培养基中添加四氢嘧啶可促进hPDLMSCs的细胞存活以及成骨和成纤维细胞分化。
