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富血小板纤维蛋白通过细胞外信号调节蛋白激酶信号通路促进根尖乳头干细胞的成骨/成牙向分化。

Platelet-rich Fibrin Improves the Osteo-/Odontogenic Differentiation of Stem Cells from Apical Papilla via the Extracellular Signal-regulated Protein Kinase Signaling Pathway.

机构信息

Department of Pediatric Dentistry, School and Hospital of Stomatology, China Medical University, Shenyang, China; Deparrment of Oral Tissue Regeneration, Liaoning Provincial Key Laboratory of Oral Diseases, Shenyang, China.

Department of Pediatric Dentistry, School and Hospital of Stomatology, China Medical University, Shenyang, China; Deparrment of Oral Tissue Regeneration, Liaoning Provincial Key Laboratory of Oral Diseases, Shenyang, China.

出版信息

J Endod. 2020 May;46(5):648-654. doi: 10.1016/j.joen.2020.02.004. Epub 2020 Mar 20.

Abstract

INTRODUCTION

The aim of this study was to investigate the cytobiological effects of platelet-rich fibrin (PRF) on stem cells from the apical papilla (SCAPs) in vitro and to further explore the underlying molecular mechanisms.

METHODS

SCAPs were isolated from immature third molars. Different concentrations of PRF conditioned medium (one eighth, one quarter, and one half PRF) were prepared. After pretreatment with PRF, the proliferation rate and migration capacity of SCAPs were examined by the Cell Counting Kit-8 assay and wound healing assay, respectively. Alizarin red S staining was performed to examine mineralized nodule formation. Western blot analysis was used to detect the expression of osteo-/odontogenic markers and the extracellular signal-regulated kinase (ERK) pathway in SCAPs. Data were analyzed by one-way analysis of variance. P values <.05 were considered statistically significant.

RESULTS

The Cell Counting Kit-8 assay showed that one eighth PRF improved the proliferation rate and the migration capacity of SCAPs (P < .005), whereas one quarter PRF and one half PRF showed no significant difference compared with the control group. The expression of osteo-/odontogenic markers and the capacity to form mineralized nodules of SCAPs were promoted by one eighth PRF and one quarter PRF. In addition, PRF activated ERK signaling, and the ERK inhibitor attenuated PRF-induced osteo-/odontogenesis of SCAPs.

CONCLUSIONS

PRF improved the osteo-/odontogenic differentiation of SCAPs by activating the ERK pathway; meanwhile, PRF improved the proliferation and migration of SCAPs, and one eighth PRF achieved the most obvious promotion effect. The favorable cytobiological effects of PRF on SCAPs might serve as basis for PRF applications in regenerative endodontic treatment.

摘要

简介

本研究旨在探讨富血小板纤维蛋白(PRF)对体外根尖乳头干细胞(SCAP)的细胞生物学效应,并进一步探讨其潜在的分子机制。

方法

从未成熟的第三磨牙中分离出 SCAP。制备不同浓度的 PRF 条件培养基(PRF 的 1/8、1/4 和 1/2)。PRF 预处理后,通过细胞计数试剂盒-8 法和划痕愈合试验分别检测 SCAP 的增殖率和迁移能力。茜素红 S 染色检测矿化结节形成。Western blot 分析检测 SCAP 中骨/牙向标志物的表达和细胞外信号调节激酶(ERK)通路。采用单因素方差分析进行数据分析。P 值<0.05 被认为具有统计学意义。

结果

细胞计数试剂盒-8 法显示,PRF 的 1/8 提高了 SCAP 的增殖率和迁移能力(P<0.005),而 PRF 的 1/4 和 1/2 与对照组相比无显著差异。PRF 的 1/8 和 1/4 促进了 SCAP 中骨/牙向标志物的表达和矿化结节的形成。此外,PRF 激活了 ERK 信号通路,而 ERK 抑制剂减弱了 PRF 诱导的 SCAP 骨/牙向分化。

结论

PRF 通过激活 ERK 通路促进 SCAP 的骨/牙向分化;同时,PRF 促进了 SCAP 的增殖和迁移,PRF 的 1/8 达到了最明显的促进效果。PRF 对 SCAP 的良好细胞生物学效应可能为 PRF 在再生性牙髓治疗中的应用提供依据。

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