在牙周炎的体外模型中翻译牙周韧带干细胞来源的分泌组/条件培养基的蛋白质组和转录组动态。

Translating proteome and transcriptome dynamics of periodontal ligament stem cell-derived secretome/conditioned medium in an in vitro model of periodontitis.

机构信息

Korea Institute of Toxicology, 30 Baekhak1-Gil, Jeongeup, Jeollabuk-Do, 56212, South Korea.

Department of Maxillofacial Biomedical Engineering, College of Dentistry, Kyung Hee University, 26 Kyunghee-Daero, Dongdaemun-Gu, Seoul, 02447, South Korea.

出版信息

BMC Oral Health. 2024 Mar 27;24(1):390. doi: 10.1186/s12903-024-04167-z.

DOI:10.1186/s12903-024-04167-z

PMID:38539170

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10967149/

Abstract

BACKGROUND

Periodontal ligament stem cells (PDLSCs) have been proposed as therapeutic candidates in periodontal diseases and periodontium defects. Paracrine factors of PDLSCs, namely, secretome, can contribute to tissue regeneration comparable to direct stem cell application. This study explored restoration effects of PDLSC-derived secretome/conditioned medium (PDLSC-CM) on PDLSCs themselves in an inflammatory microenvironment and identified its action mechanisms using proteomics and transcriptomic profiling.

METHODS

PDLSC-CM was prepared from cells under healthy culture conditions. Mass spectrometry and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were then performed to analyze the PDLSC-CM proteome. Osteogenic differentiation of PDLSCs under inflammatory conditions or in the presence of PDLSC-CM was then characterized in assays of alkaline phosphatase activity, intracellular calcium levels, protein expression of osteogenic markers, and matrix mineralization. Furthermore, the transcriptomic profile was assessed to identify significantly enriched signaling pathways and associated molecular networks by RNA sequencing.

RESULTS

LC-MS/MS proteomics identified a total of 203 proteins and distinguished 187 significant protein changes in PDLSC-CM compared to control-CM. LPS-treated PDLSCs significantly attenuated osteogenic differentiation. When PDLSCs were treated with PDLSC-CM alone, their osteogenic activity was significantly upregulated compared to the control group. Moreover, the LPS-impaired osteogenesis of PDLSCs was reconstituted by PDLSC-CM treatment. RNA sequencing revealed 252, 1,326, and 776 differentially expressed genes in the control vs. LPS, control vs. PDLSC-CM, and LPS vs. LPS + PDLSC-CM groups, respectively.

CONCLUSION

This study suggest that PDLSC-CM restores the osteogenic potential of PDLSCs in an inflammatory environment through secretory functions representing potential repair and regenerative mechanisms.

摘要

背景

牙周膜干细胞（PDLSCs）已被提议作为牙周病和牙周缺损的治疗候选物。PDLSCs 的旁分泌因子，即分泌组，可促进与直接干细胞应用相当的组织再生。本研究使用蛋白质组学和转录组谱分析探索了 PDLSC 衍生的分泌组/条件培养基（PDLSC-CM）在炎症微环境中对 PDLSCs 本身的修复作用，并确定了其作用机制。

方法

从处于健康培养条件下的细胞中制备 PDLSC-CM。然后进行质谱和液相色谱-串联质谱（LC-MS/MS）分析以分析 PDLSC-CM 蛋白质组。在碱性磷酸酶活性、细胞内钙水平、成骨标志物的蛋白表达和基质矿化测定中，研究了炎症条件下或存在 PDLSC-CM 时 PDLSCs 的成骨分化。此外，通过 RNA 测序评估转录组谱，以鉴定显著富集的信号通路和相关分子网络。

结果

LC-MS/MS 蛋白质组学共鉴定出 203 种蛋白质，并区分出 PDLSC-CM 与对照-CM 相比有 187 种显著的蛋白质变化。LPS 处理的 PDLSCs 显著减弱成骨分化。当 PDLSCs 单独用 PDLSC-CM 处理时，与对照组相比，其成骨活性显著上调。此外，PDLSC-CM 处理恢复了 LPS 损害的 PDLSCs 的成骨作用。RNA 测序显示，在对照与 LPS、对照与 PDLSC-CM 以及 LPS 与 LPS+PDLSC-CM 组之间，分别有 252、1326 和 776 个差异表达基因。