Department of Endodontology, Kyushu University Hospital, 3-1-1 Maidashi, Higashi-ku, Fukuoka-shi, Fukuoka, 812 8582, Japan.
Faculty of Dental Science, Division of Oral Rehabilitation, Department of Endodontology and Operative Dentistry, Kyushu University, Fukuoka, Japan.
Odontology. 2022 Jan;110(1):127-137. doi: 10.1007/s10266-021-00648-7. Epub 2021 Aug 11.
The aim of this study was to characterize a clonal human periodontal ligament (PDL) stem cell line (line 2-23 cells) cultured with root canal sealers based on methacrylate resin (SuperBond sealer; SB), bioactive glass (Nishika Canal Sealer BG; BG), or silicon (GuttaFlow 2; GF). The sealers were set in rubber molds to form sealer discs. Line 2-23 cells were cultured with or without the discs for 3 days. The cell viability was evaluated by direct cell counting and MTT assay. Inflammation-, PDL-, collagen-, and cell cycle-related gene expression was investigated by real-time RT-PCR. Collagen production was analyzed by Picro Sirius Red staining. Calcium ion concentration in the culture was measured by a QuantiChrom calcium assay kit. Line 2-23 cells survived when cultured with GF discs, but decreased cell viability was observed with SB and BG discs. The expression of inflammation-related genes was higher in cells cultured with SB discs, and expression of PDL-related genes was lower in cells exposed to SB and BG discs. These discs also down-regulated collagen production in line 2-23 cells. BG discs increased calcium ion concentration in the culture medium. Cells exposed to GF discs exhibited the same inflammation-, PDL-, collagen-, and cell cycle-related gene expression and collagen production as untreated cells. These results suggested that the characteristics of line 2-23 cells cultured with GF discs was highly resemble to untreated cells throughout the 3 days of the culture model.
本研究旨在基于甲基丙烯酸酯树脂(SuperBond 密封剂;SB)、生物活性玻璃(Nishika Canal Sealer BG;BG)或硅(GuttaFlow 2;GF)对一种克隆人牙周膜(PDL)干细胞系(2-23 细胞系)进行培养,并对其进行表征。将密封剂置于橡胶模具中形成密封剂圆盘。将 2-23 细胞系与或不与圆盘共培养 3 天。通过直接细胞计数和 MTT 测定评估细胞活力。通过实时 RT-PCR 研究炎症、PDL、胶原蛋白和细胞周期相关基因的表达。通过 Picrosirius Red 染色分析胶原蛋白的产生。通过 QuantiChrom 钙离子测定试剂盒测量培养物中的钙离子浓度。当用 GF 圆盘培养时,2-23 细胞系存活,但用 SB 和 BG 圆盘培养时细胞活力下降。用 SB 圆盘培养的细胞中炎症相关基因的表达较高,而用 SB 和 BG 圆盘培养的细胞中 PDL 相关基因的表达较低。这些圆盘还下调了 2-23 细胞中的胶原蛋白产生。BG 圆盘增加了培养基中的钙离子浓度。用 GF 圆盘暴露的细胞表现出与未经处理的细胞相同的炎症、PDL、胶原蛋白和细胞周期相关基因表达和胶原蛋白产生。这些结果表明,在用 GF 圆盘培养的 2-23 细胞系的特征在整个 3 天的培养模型中与未经处理的细胞高度相似。