Anselmi Caroline, Mendes Soares Igor Paulino, Chang Sarah, Cardoso Lais M, de Carvalho Ana Beatriz Gomes, Dal-Fabbro Renan, de Souza Costa Carlos Alberto, Bottino Marco C, Hebling Josimeri
Department of Morphology and Pediatric Dentistry, School of Dentistry, São Paulo State University (UNESP), Araraquara, Brazil.
Department of Cariology, Restorative Sciences, and Endodontics, School of Dentistry, University of Michigan, Ann Arbor, Michigan, USA.
Int Endod J. 2025 Jul;58(7):1073-1090. doi: 10.1111/iej.14243. Epub 2025 Apr 26.
Tissue engineering can be applied to dentine regeneration, stimulating tissue repair by promoting mesenchymal cell migration and differentiation into odontoblast-like cells while modulating inflammation. This study aimed to investigate the effect of quercetin (QU) and calcium hydroxide (CH) incorporated into polycaprolactone (PCL)/polyethylene oxide (PEO) scaffolds on the differentiation of dental pulp stem cells (DPSCs) in a simulated inflammatory environment in vitro.
Dental pulp stem cells (DPSCs) were cultured and treated with different concentrations of quercetin (QU) to assess cell viability, mineralized matrix production and responses under inflammatory stimuli. Reactive oxygen and nitrogen species, as well as TNF-α synthesis, were quantified using fluorescence and ELISA methods. Scaffolds of PCL/PEO with calcium hydroxide and QU were fabricated via electrospinning, characterized and analysed for cell adhesion, viability, inflammatory and mineralisation-related genes in an artificial pulp chamber model. Statistical analysis was performed using anova, Kruskal-Wallis and confidence intervals with a significance level of 5%.
Polycaprolactone/polyethylene oxide scaffolds incorporated with CH and QU showed cytocompatibility and support for DPSC differentiation at concentrations of up to 5 M diluted in the culture medium. After 14 days of treatment, the scaffolds upregulated ALPL gene expression under the inflammatory stimulus, with no differences between the control group and the nonincorporated scaffold. The expression of osteocalcin (OCN) and dentine sialophosphoprotein (DSPP) genes was significantly upregulated for the scaffold-treated group when stimulated with LPS.
Incorporating QU and CH into PCL/PEO scaffolds modulated the inflammatory-related response and upregulated mineralisation-related genes of LPS-challenged dental pulp stem cells.
组织工程可应用于牙本质再生，通过促进间充质细胞迁移并分化为成牙本质样细胞，同时调节炎症反应来刺激组织修复。本研究旨在探讨聚己内酯（PCL）/聚环氧乙烷（PEO）支架中加入槲皮素（QU）和氢氧化钙（CH）对体外模拟炎症环境下牙髓干细胞（DPSC）分化的影响。
培养牙髓干细胞（DPSC），并用不同浓度的槲皮素（QU）进行处理，以评估细胞活力、矿化基质生成以及在炎症刺激下的反应。使用荧光和酶联免疫吸附测定（ELISA）方法对活性氧和氮物种以及肿瘤坏死因子-α（TNF-α）的合成进行定量。通过静电纺丝制备含有氢氧化钙和QU的PCL/PEO支架，在人工牙髓腔模型中对其进行表征，并分析细胞黏附、活力、炎症和矿化相关基因。使用方差分析（anova）、Kruskal-Wallis检验和置信区间进行统计分析，显著性水平为5%。
在培养基中稀释至浓度高达5 M时，含有CH和QU的聚己内酯/聚环氧乙烷支架表现出细胞相容性并支持DPSC分化。处理14天后，支架在炎症刺激下上调了碱性磷酸酶（ALPL）基因表达，对照组与未添加的支架之间无差异。用脂多糖（LPS）刺激时，支架处理组的骨钙素（OCN）和牙本质涎磷蛋白（DSPP）基因表达显著上调。
将QU和CH掺入PCL/PEO支架中可调节LPS刺激的牙髓干细胞的炎症相关反应并上调矿化相关基因。
