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从炎症牙髓和健康牙髓中分离并在炎症微环境中培养的牙髓干细胞的生物学特性

Biological Properties of Dental Pulp Stem Cells Isolated from Inflamed and Healthy Pulp and Cultured in an Inflammatory Microenvironment.

作者信息

Bucchi Cristina, Bucchi Ana, Martínez-Rodríguez Paulina

机构信息

Integral Adult Dentistry Department, Universidad de La Frontera, Temuco, Chile; Oral Biology Center, Faculty of Dentistry, Universidad de La Frontera, Temuco, Chile.

Centro de Investigación en Odontología Legal y Forense (CIO), Faculty of Dentistry, Universidad de La Frontera, Temuco, Chile.

出版信息

J Endod. 2023 Apr;49(4):395-401.e6. doi: 10.1016/j.joen.2023.02.002. Epub 2023 Feb 23.

Abstract

INTRODUCTION

The aim of this study was to assess whether the biological characteristics of dental pulp stem cells (DPSCs), such as viability, adhesion to dentin, mineralization, and release of immunomodulatory cytokines, are affected by the inflammatory status of the donor tissue and/or the sustained inflammatory environment.

METHODS

DPSCs were isolated from pulps from 3 caries-free teeth (healthy or hDPSCs), and from 3 teeth with irreversible pulpitis or deep caries (unhealthy DPSCs or uDPSCs). The cells were cultured in odontogenic and osteogenic media with or without lipopolysaccharides. Viability was analyzed by MTT assay at days 1, 3, 5, and 7; adhesion to dentin was evaluated through an environmental scanning electron microscope after 48 hours and through MTT assay; mineralization was analyzed with alizarin red staining after 21 days; and the release of proinflammatory (interleukin 6) and immunosuppressive cytokines (interleukin 10) was measured with the enzyme-linked immunosorbent assay after 24 hours and 7 days.

RESULTS

The inflammatory status of the pulp significantly reduced the viability and mineralization capacity of the DPSCs, although it did not affect the adhesion capacity to dentin or the secretion of the proinflammatory interleukin. The inflammatory microenvironment (lipopolysaccharide) only had a significant impact on the secretion of interleukin 6, which was augmented after 7 days.

CONCLUSIONS

The inflammatory status of the dental pulp should be taken into account when the use of DPSCs is intended either for research and/or for application in reparative or regenerative therapies.

摘要

引言

本研究的目的是评估牙髓干细胞(DPSCs)的生物学特性,如活力、对牙本质的黏附、矿化以及免疫调节细胞因子的释放,是否受供体组织的炎症状态和/或持续炎症环境的影响。

方法

从3颗无龋牙(健康牙髓干细胞或hDPSCs)以及3颗患有不可复性牙髓炎或深龋的牙齿(不健康牙髓干细胞或uDPSCs)的牙髓中分离出DPSCs。将细胞在有或无脂多糖的成牙本质细胞和成骨细胞培养基中培养。在第1、3、5和7天通过MTT法分析活力;48小时后通过环境扫描电子显微镜并通过MTT法评估对牙本质的黏附;21天后用茜素红染色分析矿化;24小时和7天后用酶联免疫吸附测定法测量促炎细胞因子(白细胞介素6)和免疫抑制细胞因子(白细胞介素10)的释放。

结果

牙髓的炎症状态显著降低了DPSCs的活力和矿化能力,尽管它不影响对牙本质的黏附能力或促炎白细胞介素的分泌。炎症微环境(脂多糖)仅对白细胞介素6的分泌有显著影响,7天后其分泌增加。

结论

当打算将DPSCs用于研究和/或用于修复或再生治疗时,应考虑牙髓的炎症状态。

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