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RNA sequencing of human periodontal ligament stem cells (PDLSCs) with Illumina HiSeq2000

ABSTRACT: To study the molecular mechanisms underlying PDLSC osteogenic differentiation, we developed RNA sequencing with Illumina HiSeq2000 to comprehensively identify RNA expression profiles in normal PDLSCs, osteogenic inductive PDLSCs and mechanical force-induced PDLSCs. Plenty of RNAs were found to be differentially expressed by RNA sequencing. Furthermore, the potential functions of these RNAs were investigated using bioinformatic analysis.

ORGANISM(S): Homo sapiens

PROVIDER: GSE106887 | GEO | 2018/03/09

REPOSITORIES: GEO

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The long noncoding RNA AC018926.2 plays a crucial role in palmitic acid exposure-compromised osteogenic potential of periodontal ligament stem cells

Project description:This study investigated the cellular response of human periodontal ligament stem cells (PDLSCs) to elevated levels of a specific fatty acid (FA), namely, palmitic acid (PA) and the role of long noncoding RNAs (lncRNAs) in regulating PA exposure-compromised osteogenic potential of PDLSCs. It was found that exposure of PDLSCs to abundant PA led to decreased osteogenic differentiation of cells. Through microarray analysis and gain/loss-of-function studies, AC018926.2 was identified and validated as the lncRNA that was most sensitive to PA and a regulator in the process of PDLSC osteogenic differentiation under a PA condition. Then, RNA sequencing and KEGG analysis demonstrated that AC018926.2 upregulated integrin α2 (ITGA2) expression at the transcriptional level and therefore activated ITGA2/FAK/AKT signaling. Further functional studies revealed that inactivation of ITGA2/FAK/AKT signaling by silencing ITGA2 counteracted the enhancement of the PDLSC osteogenic potential induced by AC018926.2 overexpression. Moreover, the results of bioinformatics analysis and the RNA immunoprecipitation (RIP) assay suggested that AC018926.2 might transcriptionally regulate ITGA2 expression by binding to poly (ADP-ribose) polymerase 1 (PARP1). Our data suggest that AC018926.2 plays a crucial role in PA exposure-compromised osteogenic potential of PDLSCs.

2022-11-26 | GSE218434 | GEO

Hyaluronic Acid Induction Promotes the Differentiation of Human Neural Crest-like Cells into Periodontal Ligament Stem-like Cells

Project description:Periodontal ligament (PDL) stem-like cells (PDLSCs) are considered to be promising for the regeneration of periodontium because they demonstrate multipotency, high proliferative capacity, and potential to regenerate bone, cementum, and PDL tissue. However, the transplantation of autologous PDLSCs is restricted by limited availability and the need for tooth extraction to isolate these cells. PDLSCs are derived from neural crest cells (NCs) in early vertebrate development and NCs persist in adult PDL tissue. Therefore, we devised to promote the regeneration of periodontium by activating NCs to differentiate into PDLSCs and increasing their total number. SK-N-SH cells cultured on the extracellular matrix derived of human PDL cells (SK-PDLSCs) strongly expressed PDL-related marker genes and highly differentiated into mesenchymal lineage cells compared to untreated SK-N-SH cells. Expression levels of various hyaluronic acid (HA)-related genes were upregulated in induced pluripotent stem cells (iPSCs)-derived PDLSCs and SK-PDLSCs compared to iPSCs-derived NCs and SK-N-SH cells, respectively. Magnetic-activated cell sorting-assisted CD44-positive enrichment population of SK-N-SH cells showed higher expression of PDL-related marker genes after SK-PDLSCs induction than the CD44-negative population. Knockdown of CD44 in SK-N-SH cells significantly inhibited their ability to differentiate into SK-PDLSCs, while lower molecular HA (LHA) induction enhanced SK-PDLSC differentiation. LHA-containing electrospun nanofibrous membranes also promoted their SK-PDLSC differentiation. Our findings suggest that SK-N-SH cells are applied as a new model to induce the differentiation of NCs into PDLSCs. The LHA-CD44 relation is important for the differentiation of NCs into PDLSCs. LHA-containing electrospun nanofibrous membranes would promote the regeneration of periodontium by activating NCs in PDL tissue and increasing the total number of PDLSCs.

2023-06-27 | GSE235864 | GEO

The long non-coding RNA landscape of periodontal ligament stem cells subjected to compressive force

Project description:To explore the lncRNA landscape of periodontal ligament stem cells (PDLSCs) subjected to compressive force.

2021-07-09 | GSE116787 | GEO

Comprehensive analysis of M2 macrophage-derived exosomes facilitating osteogenic differentiation of human periodontal ligament stem cells

Project description:As the primary seed cells in periodontal tissue engineering, the role of periodontal ligament stem cells (PDLSCs) in periodontal tissue regeneration and bone remodeling during orthodontic tooth movement (OTM) has been well documented. Nevertheless, the impact of different polarization states of macrophages on the osteogenic differentiation of PDLSCs is poorly understood. M0, M1 and M2 macrophage-derived exosomes (M0-exo, M1-exo and M2-exo) were treated with primary cultured human PDLSCs, respectively. Identification of differentially expressed microRNAs (DE-miRNA) in M0-exo and M2-exo by miRNA microarray. In summary, we have indicated for the first time that M2-exo can promote osteogenic differentiation of human PDLSCs, and have revealed the functions and pathways involved in the DE-miRNAs of M0-exo and M2-exo and their downstream targets.

2022-10-31 | GSE209957 | GEO

Single-Cell Transcriptomic Analysis of Dental Pulp and Periodontal Ligament Stem Cells [scRNA-seq]

Project description:In this study, we investigated the heterogeneity of DPSC and PDLSC cells using scRNA-seq and characterized the molecular features of each cell group. Combining the analyses of trajectory route and scoring, our results demonstrated that DPSCs displayed higher differentiation potentials, while PDLSCs exhibited more matured features. These findings provide a new insight into the complex landscape and functions of oral stem cells, which would enrich our understaning on their roles in embryonic development and regeneration of lost tissues due to dental and periodontal infections.

2023-12-11 | GSE227731 | GEO

microRNA expression of human periodontal ligament derived stem cell line exposed to 1 microMolar (uM) (-)-nicotine liquid or 1 uM nicotine electronic cigarette vapor extract against untreated control

Project description:Nicotine and cigarette smoking have been previously shown to inhibit the differentiation potential and miRNA expression of human periodontal ligament derived stem cells (PDLSCs). Electronic cigarette (EC) vapor also contains nicotine, as well as other non-nicotine compounds, and, like cigarette smoking, EC vaping results in similar routes of toxic exposure for PDLSCs. The effect of exposure on PDLSC miRNA expression is unknown. To determine these effects, cultured PDLSCs were exposed to media supplemented with 1 uM nicotine EC vapor extract for 72h, during which media was changed every 24h. To produce vapor extract, an EC filled with 36 mg/ml nicotine, 50%/50% (w/v) PG/VG, non-flavored e-liquid (American E-liquid) was vaped using an automated vaping robot programmed with experienced e-cigarette user patterns (i.e. puff volume: 70 ml, duration: 3s, frequency: 20s) and resulting vapor was extracted via a liquid impinger. 1 uM of pure (-)-nicotine liquid (Sigma Aldrich) was used as a positive control. Total miRNA was collected with the mirVana miRNA isolation kit. miRNA concentration and quality was measured by spectrophotometry and expression was analyzed with Affymetrix GeneChip miRNA 4.0 Arrays. Results were normalized against human-positive and negative chip controls using Robust Multi-Array Average Normalization.

2019-09-30 | E-MTAB-7893 | biostudies-arrayexpress

RNA sequencing of human periodontal ligament stem cells (PDLSCs) with Illumina HiSeq2000

Project description:RNA sequencing of human periodontal ligament stem cells (PDLSCs) with Illumina HiSeq2000

| PRJNA418326 | ENA

Gene expression profiling of KSHV-infected periodontal ligament cells

Project description:We reported the potential and mechnism of oral mesenchymal stem cell to become Kaposi Sarcoma progenitor cell. Human periodontal ligament cells were infected for 48 hours and 96 hours, then subjected to RNA extraction and sequencing. We found that KSHV-infected PDLSC shared a better similarity with KS biosies in cytokine production, including chemotaxis, angiogenesis, inflammation and differentiation. Furthermore, infection of PDLSC upregulated a spectum of genes involving mesenchymal-to-endothelial differentiation, a process which has been revealed in KS spindle cells. This study reveals the potential and mechnism in which oral mesenchymal stem cell is transformed by KSHV to KS progenitor cell.

2017-06-01 | GSE92341 | GEO

Proteomic and Phosphoproteomic Analyses of the Molecular Mechanisms of Periodontal Ligament Stem Cell Osteogenic Differentiation

Project description:Periodontal ligament stem cells (PDLSCs) are optimal seed cells for periodontal regeneration; thus, exploration of the mechanisms regulating PDLSCs osteogenic differentiation is critical. Here, tandem mass tag quantitative proteomics and phosphoproteomic analyses of human PDLSCs were carried out after osteogenic differentiation for 14 days. Proteomic analyses identified 314 differentially expressed proteins (DEPs) that were predicted to be mostly extracellular. Gene ontology and pathway enrichment analyses revealed that the mineral absorption signaling pathway was significantly involved. For phosphoproteomics, un-normalized data identified 1016 differentially expressed phosphorylation sites (DEPSs) corresponded to 380 DEPs, whereas in normalized data, 3030 DEPSs were identified, corresponding to 358 DEPs. Overlapping of these data identified 16 phosphorylated DEPs, among which tetratricopeptide repeat protein 17, stanniocalcin-1, and extracellular superoxide dismutase [Cu-Zn] were significantly related. Kyoto

2022-02-25 | PXD031905 | iProX

PAX9 regulates periodontal ligament stem cell-like differentiation of human induced pluripotent stem cells

Project description:Periodontal ligament stem cells (PDLSCs) play central roles in periodontal ligament (PDL) tissue homeostasis, repair, and regeneration. Previously, we established a protocol to differentiate human induced pluripotent stem cell-derived neural crest-like cells (iNCs) into PDLSC-like cells (iPDLSCs) using human PDL cell-derived extracellular matrix (ECM). However it remained unclear what factors principally regulate the differentiation of iNCs into iPDLSCs. In this study, we aimed to identify the transcription factor regulating production of human PDL cell-derived ECM, which is responsible for the generation of iPDLSCs. We cultured iNCs on two human PDL cell lines (HPDLC-3S and HPDLC-3U) and human dermal fibroblasts (HDF). iNCs cultured on HPDLC-3U showed higher iPDLSC-associated gene expression and mesenchymal differentiation capacity than cells cultured on HDF or HPDLC-3S. The transcription factor PAX9 was highly expressed in HPDLC-3U compared with HDF and HPDLC-3S. iNCs cultured on siPAX9-transfected HPDLC-3U displayed downregulation of iPDLSC-associated marker expression and adipocytic differentiation capacity relative to controls. Our findings suggest that PAX9 is one of the transcription factors regulating ECM production in human PDL cells, which is responsible for the differentiation of iNCs into iPDLSCs.

2022-07-18 | GSE208250 | GEO

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