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姜黄素植物药质体处理对牙髓间充质干细胞免疫调节和牙髓再生基因调控的影响。

Effects of phytosomal curcumin treatment on modulation of immunomodulatory and pulp regeneration genes in dental pulp mesenchymal stem cells.

机构信息

Student Research Committee, Birjand University of Medical Sciences, Birjand, Iran.

Department of Immunology, Faculty of Medicine, Birjand University of Medical Sciences, Birjand, Iran.

出版信息

Odontology. 2022 Apr;110(2):287-295. doi: 10.1007/s10266-021-00659-4. Epub 2021 Sep 29.

Abstract

Dental pulp stem cells (DPSCs) are a new population of mesenchymal stem cells (MSCs) located in the oral cavity with potential capacities for tissue regeneration and immunomodulation. The purpose from this study was to determine effects of curcumin nanoparticle into phytosomal formulation (PC) on the relative expression of DSPP, VEGF-A, HLA-G5, VCAM1, RelA and STAT3 genes which are among the most important factors influencing processes of immunomodulatory and tissue regenerative by DPSCs. After isolation and culture of DPSCs, these cells were characterized according to predetermined criteria including flow cytometric analysis for detection of the most important cell surface markers and also evaluation of multilineage differentiation potential. Then, the MTT method was employed to check the cell viability in treatment with different concentrations of PC. Following DPSCs' treatment with an optimal-non-toxic dose of this nanoparticle, quantification of expression of target genes was performed using real-time PCR procedure. According to results of immunophenotyping analysis and cell differentiation experiments, the isolated cells were confirmed as MSCs as more than 99% of them expressed specific mesenchymal markers while only about 0.5% of them were positive for hematopoietic marker. The real-time PCR results indicated that PC significantly reduced the expression of RelA, STAT3, VCAM1 and HLA-G5 genes up to many times over while optimally enhanced the expression of DSPP and VEGF-A genes, although this enhance was statistically significant only for VEGF-A (all P < 0.001). The study suggests that PC affects the stemness capabilities of DPSCs and it may facilitate the development of MSCs-based therapeutics in regenerative dentistry.

摘要

牙髓干细胞(DPSCs)是一种位于口腔中的新型间充质干细胞(MSCs),具有组织再生和免疫调节的潜力。本研究的目的是确定姜黄素纳米粒子进入植物鞘氨醇配方(PC)对 DSPP、VEGF-A、HLA-G5、VCAM1、RelA 和 STAT3 基因相对表达的影响,这些基因是影响 DPSCs 免疫调节和组织再生过程的最重要因素之一。在分离和培养 DPSCs 后,根据预定标准对这些细胞进行特征描述,包括流式细胞术分析以检测最重要的细胞表面标志物,以及多谱系分化潜能的评估。然后,采用 MTT 法检查不同浓度 PC 处理对细胞活力的影响。在 DPSCs 用这种纳米颗粒的最佳非毒性剂量处理后,采用实时 PCR 程序检测目标基因的表达量。根据免疫表型分析和细胞分化实验的结果,分离的细胞被确认为 MSCs,因为它们中的 99%以上表达特定的间充质标志物,而只有约 0.5%的细胞对造血标志物呈阳性。实时 PCR 结果表明,PC 可显著降低 RelA、STAT3、VCAM1 和 HLA-G5 基因的表达,最高可达数倍,同时最佳地增强了 DSPP 和 VEGF-A 基因的表达,尽管这种增强在统计学上仅对 VEGF-A 具有显著意义(均 P<0.001)。研究表明,PC 影响 DPSCs 的干性能力,可能有助于再生牙科中基于 MSCs 的治疗方法的发展。

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