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高度成骨/成牙骨质牙周膜细胞克隆的转录组谱。

Transcriptome profile of highly osteoblastic/cementoblastic periodontal ligament cell clones.

机构信息

Universidade Federal do Pará, Instituto de Ciências da Saude, Departmento de Saúde Pública, Belém, Pará, Brasil.

Universidade Estadual de Campinas, Instituto de Biologia (UNICAMP), Departamento de Genética e Evolução, Microbiologia e Imunologia, Laboratório de Genônica e Expressão, Campinas, SP, Brasil.

出版信息

J Appl Oral Sci. 2020 Oct 23;28:e20200242. doi: 10.1590/1678-7757-2020-0242. eCollection 2020.

Abstract

BACKGROUND

Heterogeneous cell populations of osteo/cementoblastic (O/C) or fibroblastic phenotypes constitute the periodontal dental ligament (PDL). A better understanding of these PDL cell subpopulations is essential to propose regenerative approaches based on a sound biological rationale.

OBJECTIVE

Our study aimed to clarify the differential transcriptome profile of PDL cells poised to differentiate into the O/C cell lineage.

METHODOLOGY

To characterize periodontal-derived cells with distinct differentiation capacities, single-cell-derived clones were isolated from adult human PDL progenitor cells and their potential to differentiate into osteo/cementoblastic (O/C) phenotype (C-O clones) or fibroblastic phenotype (C-F clones) was assessed in vitro. The transcriptome profile of the clonal cell lines in standard medium cultivation was evaluated using next-generation sequencing technology (RNA-seq). Over 230 differentially expressed genes (DEG) were identified, in which C-O clones showed a higher number of upregulated genes (193) and 42 downregulated genes.

RESULTS

The upregulated genes were associated with the Cadherin and Wnt signaling pathways as well as annotated biological processes, including "anatomical structure development" and "cell adhesion." Both transcriptome and RT-qPCR showed up-regulation of WNT2, WNT16, and WIF1 in C-O clones.

CONCLUSIONS

This comprehensive transcriptomic assessment of human PDL progenitor cells revealed that expression of transcripts related to the biological process "anatomical structure development," Cadherin signaling, and Wnt signaling can identify PDL cells with a higher potential to commit to the O/C phenotype. A better understanding of these pathways and their function in O/C differentiation will help to improve protocols for periodontal regenerative therapies.

摘要

背景

成骨/成牙骨质(O/C)或成纤维细胞表型的异质细胞群体构成牙周韧带(PDL)。更好地了解这些 PDL 细胞亚群对于基于合理生物学基础提出再生方法至关重要。

目的

我们的研究旨在阐明 PDLC 向 O/C 细胞系分化的差异转录组谱。

方法

为了表征具有不同分化能力的牙周来源细胞,从成人牙周膜祖细胞中分离出单细胞衍生的克隆,并在体外评估其分化为成骨/成牙骨质(O/C)表型(C-O 克隆)或成纤维表型(C-F 克隆)的潜能。使用下一代测序技术(RNA-seq)评估标准培养基培养的克隆细胞系的转录组谱。鉴定出超过 230 个差异表达基因(DEG),其中 C-O 克隆显示出更高数量的上调基因(193 个)和 42 个下调基因。

结果

上调基因与 Cadherin 和 Wnt 信号通路以及注释的生物学过程相关,包括“解剖结构发育”和“细胞黏附”。RNA-seq 和 RT-qPCR 均显示 C-O 克隆中 WNT2、WNT16 和 WIF1 的表达上调。

结论

这项对人牙周膜祖细胞的全面转录组评估表明,与“解剖结构发育”生物学过程、Cadherin 信号和 Wnt 信号相关的转录本的表达可以识别出具有更高向 O/C 表型分化潜能的 PDLC。更好地了解这些途径及其在 O/C 分化中的功能将有助于改善牙周再生治疗的方案。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f2e/9648949/6fbbaa4d5812/1678-7757-jaos-28-e20200242-gf01.jpg

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