Effects of D-galactose Induction on Aging Characteristics of the Human Dental Pulp Cell Culture Model: An In Vitro Study.

IF 1.6 Q3 DENTISTRY, ORAL SURGERY & MEDICINE
Suthasinee Saiyasilp, Savitri Vaseenon, Tanida Srisuwan, Patchanee Chuveera
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引用次数: 0

Abstract

Objective: This study aimed to investigate the effects of D-galactose (D-gal) on cellular senescence induction, cell proliferation, mineralization production, and odontogenic gene expression of isolated human dental pulp cells (HDPCs).

Methods: Isolated HDPCs were cultured and assigned to four groups: control, 1 g/L D-gal, 10 g/L D-gal, and 10 g/L D-gal with Biodentine (BD). Cell proliferation was evaluated at 24, 48, and 72 hours using Alamar Blue® assay. To evaluate cellular senescence at 48 hours, senescence-associated beta-galactosidase (SA-β-gal) activity and senescence-related genes (p16 and p21) were assessed with SA-β-gal staining assay and quantitative reverse-transcription polymerase chain reaction (qRT-PCR), respectively. To examine the mineralization potential under differentiating conditions, quantitative staining with Alizarin Red S and mineralization-related gene expression (dentine sialophosphoprotein, DSPP) were investigated at 14 days. One-way ANOVA was used for statistical analysis. The statistical significance level was set at 0.05.

Results: 1 g/L D-gal and 10 g/L of D-gal significantly decreased cell proliferation at 72 hours compared to the control group (p<0.05). SA-β-gal-positive cells were significantly more prevalent in both D-gal-treated groups than in the control group (p<0.05). The expressions of genes p16 and p21 were markedly increased in cells treated with 10 g/L D-gal compared to the control group (p<0.05). The addition of BD did not promote cell proliferation but significantly improved cellular senescence by reducing SA-β-gal activity, p16, and p21 expression (p<0.05) compared to the group without BD. For mineralization potential, the amount of mineralization was similar among groups under differentiating conditions. The reduction of DSPP gene expression was obvious only in the 10 g/L D-gal group (p<0.05). The addition of BD did not show a significant effect on mineralization.

Conclusion: Ten g/L of D-gal can effectively induce aging phenotypes and reduce DSPP gene expression in HDPCs. Co-incubation with BD extract reduced the expression of these aging phenotypes. Mineralization production was not altered in the presence of D-gal. The data support the development of in vitro model for aging dental pulp. (EEJ-2024-07-108).

d -半乳糖诱导人牙髓细胞培养模型衰老特性的体外研究
目的:研究d -半乳糖(D-gal)对离体人牙髓细胞(HDPCs)衰老诱导、细胞增殖、矿化产生和成牙基因表达的影响。方法:将分离的HDPCs细胞分为4组:对照组、1 g/L D-gal组、10 g/L D-gal组和10 g/L D-gal加Biodentine (BD)组。在24,48和72小时使用Alamar Blue®检测细胞增殖。采用SA-β-gal染色法和定量反转录聚合酶链反应(qRT-PCR)法分别检测衰老相关β-半乳糖苷酶(SA-β-gal)活性和衰老相关基因(p16和p21)。为了检测分化条件下的矿化潜力,在第14天用茜素红S定量染色和矿化相关基因(牙本质唾液磷蛋白,DSPP)的表达进行了研究。采用单因素方差分析进行统计分析。统计学显著性水平设为0.05。结果:与对照组相比,1 g/L D-gal和10 g/L D-gal在72 h时显著降低细胞增殖(p)。结论:10 g/L D-gal可有效诱导HDPCs衰老表型,降低DSPP基因表达。与BD提取物共孵育可降低这些衰老表型的表达。在D-gal的存在下,矿化产量没有改变。本研究结果支持牙髓老化体外模型的建立。(eej - 2024 - 07 - 108)。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
European Endodontic Journal
European Endodontic Journal DENTISTRY, ORAL SURGERY & MEDICINE-
CiteScore
3.40
自引率
5.60%
发文量
25
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