Mitigating lipopolysaccharide-induced impairment in human dental pulp stem cells with tideglusib-doped nanoparticles: Enhancing osteogenic differentiation and mineralization

IF 4.6 1区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE Dental Materials Pub Date : 2024-07-26 DOI:10.1016/j.dental.2024.07.012

Raquel Osorio , Francisco J. Rodríguez-Lozano , Manuel Toledano , Manuel Toledano-Osorio , David García-Bernal , Laura Murcia , Sergio López-García

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Abstract

Objective

Drug-loaded non-resorbable polymeric nanoparticles (NPs) are proposed as an adjunctive treatment for pulp regenerative strategies. The present in vitro investigation aimed to evaluate the effectiveness of tideglusib-doped nanoparticles (TDg-NPs) in mitigating the adverse effects of bacterial lipopolysaccharide endotoxin (LPS) on the viability, morphology, migration, differentiation and mineralization potential of human dental pulp stem cells (hDPSCs).

Methods

Cell viability, proliferation, and differentiation were assessed using a MTT assay, cell migration evaluation, cell cytoskeleton staining analysis, Alizarin Red S staining and expression of the odontogenic related genes by a real-time quantitative polymerase chain reaction (RT-qPCR) were also performed. Cells were tested both with and without stimulation with LPS at various time points. One-way ANOVA and Tukey's test were employed for statistical analysis (p < 0.05).

Results

Adequate cell viability was encountered in all groups and at every tested time point (24, 48, 72 and 168 h), without differences among the groups (p > 0.05). The analysis of cell cytoskeleton showed nuclear alteration in cultures with undoped NPs after LPS stimulation. These cells exhibited an in blue diffuse and multifocal appearance. Some nuclei looked fragmented and condensed. hDPSCs after LPS stimulation but in the presence of TDg-NPs exhibited less nuclei changes. LPS induced down-regulation of Alkaline phosphatase, Osteonectin and Collagen1 gene markers, after 21d. LPS half-reduced the cells production of calcium deposits in all groups (p < 0.05), except in the group with TDg-NPs (decrease about 10 %).

Significance

LPS induced lower mineral deposition and cytoskeletal disorganization in hDPSCs. These effects were counteracted by TDg-NPs, enhancing osteogenic differentiation and mineralization.

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用掺有tideglusib的纳米颗粒减轻脂多糖诱导的人牙髓干细胞损伤：增强成骨分化和矿化。

目的：有人提出将药物负载型不可吸收聚合物纳米粒子（NPs）作为牙髓再生策略的辅助治疗手段。本体外研究旨在评估掺杂潮霉素的纳米颗粒（TDg-NPs）在减轻细菌脂多糖内毒素（LPS）对人牙髓干细胞（hDPSCs）的活力、形态、迁移、分化和矿化潜能的不良影响方面的有效性：方法：使用 MTT 试验评估细胞活力、增殖和分化，还进行了细胞迁移评估、细胞骨架染色分析、茜素红 S 染色以及实时定量聚合酶链反应（RT-qPCR）检测牙源性相关基因的表达。在不同的时间点，对细胞进行了有 LPS 刺激和无 LPS 刺激的测试。采用单因素方差分析和 Tukey 检验进行统计分析（p 结果：所有组别在每个测试时间点（24、48、72 和 168 小时）都有足够的细胞活力，组间无差异（p > 0.05）。对细胞细胞骨架的分析表明，在 LPS 刺激后，未掺杂 NPs 的培养物中的细胞核发生了变化。这些细胞呈现蓝色弥漫和多灶外观。经过 LPS 刺激但有 TDg-NPs 存在的 hDPSCs 的细胞核变化较小。21 天后，LPS 诱导碱性磷酸酶、骨连接蛋白和胶原蛋白 1 基因标记下调。LPS 可使各组细胞产生的钙沉积减少一半（p）：LPS 可诱导 hDPSCs 减少矿物质沉积和细胞骨架紊乱。TDg-NPs 可抵消这些影响，促进成骨分化和矿化。

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来源期刊

Dental Materials 工程技术-材料科学：生物材料

CiteScore

9.80

自引率

10.00%

发文量

290

审稿时长

67 days

期刊介绍： Dental Materials publishes original research, review articles, and short communications. Academy of Dental Materials members click here to register for free access to Dental Materials online. The principal aim of Dental Materials is to promote rapid communication of scientific information between academia, industry, and the dental practitioner. Original Manuscripts on clinical and laboratory research of basic and applied character which focus on the properties or performance of dental materials or the reaction of host tissues to materials are given priority publication. Other acceptable topics include application technology in clinical dentistry and dental laboratory technology. Comprehensive reviews and editorial commentaries on pertinent subjects will be considered.