Cald1 regulates osteogenic differentiation of PDLSCs.
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ABSTRACT:
PROVIDER: PRJNA988422 | ENA |
REPOSITORIES: ENA
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Project description:To explore underlying mechanisim of which Cald1 regulates osteogenic differentiation of PDLSCs, RNA -seq was performed on PDLSCs treated with NC shRNA and Cald1 shRNA respectively.
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Project description: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.
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Project description:Our previous study (Environ Pollut. 2023;323:121307) has indicated that Glucose-dependent insulinotropic polypeptide receptor (GIPR) and Glucagon-like peptide-1 receptor (GLP1R) are down-regulated in periodontal ligament stem cells (PDLSCs) by cigarette and E-cig. In the current study, we focus on investigating the functional roles of these two G protein-coupled receptors in PDLSCs, as well as their molecular mechanisms behind it. In brief, human PDLSCs treated with GIPR-siRNA, GIPR agonist (GIPRA) as well as GIPR, GLP1R dual agonist (2GRA) were used to study the effects of GIPR on cell growth of PDLSCs. Moreover, PDLSCs were induced to undergo osteogenic, adipogenic, chondrogenic, and neurogenic differentiation in vitro. GLP1R siRNA, GLP1R agonist (GLP1RA), and 2GRA were respectively administered to investigate the effects of GLP1R on the differentiation potential of PDLSCs.
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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.
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Project description:Tooth pulp contains various types of cells such as endothelial cells, neurons, fibroblasts, osteoblasts, osteoclasts, and odontoblasts. As well as cells that are called "postnatal dental pulp stem cells" (DPSCs). Also, four more types of dental MSC-like populations were identified and characterized: stem cells from human exfoliated deciduous teeth (SHED), periodontal ligament stem cells (PDLSCs), stem cells from the apical papilla (SCAP) and population of dental follicle-derived progenitor cells called "dental follicle progenitor cells" (DFPCs). Most of them might be used in wide range of biomedical applications. Nevertheless, they have systematic differences in their physiology, e.g. differences in proliferative and differentiation potential. These differences are not clearly defined on molecular level yet; therefore, we performed proteomics comparison of DPSCs and PDLSCs in control and osteogenic differentiation. Donor matched DPSCs and PDLSCs were isolated from two donors by standard protocol. Then, DPSCs and PDLSCs at passage 3 were seeded into 90 mm Petri dishes (Eppendorf) and cultured in standard conditions with DMEM (Gibco) supplemented with 15% fetal bovine serum (FBS), 37°C, 5% CO2. When cells reached 90-100% confluency, the medium was changed to osteogenic medium (DMEM supplemented with 10% FBS, 2 mM L-glutamine, 1% penicillin/streptomycin (HyClone), 50 mg/ml ascorbic acid (Sigma Aldrich), 0.1 mM dexamethasone (Sigma Aldrich) and 10 mM b–glycerophosphate (Sigma Aldrich).
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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
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Project description:Ability to perform osteogenic differentiation is one of the minimal criteria of mesenchymal stem cells (MSCs). Still, it is generally unknown whether osteogenic differentiation is universal cell fate or various phenotypically similar cell states. Besides this, MSCs and their secretomes are actively using for cell/cell-free therapy development, but systemic inter-source variation in MSCs secretomes, proteomes and differentiation mechanisms are still poorly understood. Therefore, here we compared proteomic and secretomic profiles of human mesenchymal cells from six sources: osteoblasts (bone), WJ-MSCs (Warton’s jelly), AD-MSCs (adipose), PDLSCs (tooth: Periodontal Ligament Stem Cells), DPSCs (tooth: Dental Pulp Stem Cells) and GFs (tooth: Gingival Fibroblasts). For experiments we used cells in early passages (3-5) isolated from 3-6 individuals. All cells were compared in standard cultivation and in the 10th day after induction of osteogenic differentiation.
2023-11-05 | PXD037320 | Pride
Project description:Although periodontal ligament stem cells (PDLSCs) are pivotal for periodontal tissue regeneration, their regenerative capacity diminishes with the PDL maturity. This study aims to elucidate the mechanisms underlying this phenomenon.Single-cell RNA sequencing (scRNA-seq) was conducted on immature and mature human PDL tissues to characterize the heterogeneity and predict critical regulators affecting osteogenic capacity of PDLSCs. Subsequently, the predicted role of RELB was experimentally validated both in vitro and in vivo using small interfering RNA.Our scRNA-seq analysis revealed significant heterogeneity within the PDLSC population. Notably, PDLSC-5 subtype, which exhibited osteogenic potential and positioned at the terminus of the pseudotime trajectory, was markedly reduced in mature PDL tissues. RELB was specifically overexpressed in PDLSC_5 and identified as a key transcription factor (TF) for this subcluster. Knockdown of RELB significantly inhibited the osteogenic differentiation capacity of PDLSCs in vitro and bone formation potential in vivo at both developmental stages. This inhibitory effect appears to be mediated, at least in part, through the downregulation of its target gene, superoxide dismutase 2 (SOD2).This study demonstrates that the proportion of PDLSCs with osteogenic ability significantly declines with maturity, accounting for the diminished regeneration capacity of mature periodontal tissue. Furthermore, RELB was identified as a crucial TF for maintaining the osteogenic potential of PDLSCs. These findings provide a theoretical basis for strategies aimed at optimizing the bone regeneration capacity of PDLSCs.
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Project description:Chromodomain helicase DNA-binding protein 7 (CHD7) is an ATP-dependent chromatin remodeling enzyme, functioning as chromatin reader to conduct epigenetic modification. Its effect on osteogenic differentiation of human dental follicle cells (hDFCs) remains unclear. Here we show CHD7 expression increases with osteogenic differentiation. knockdown of CHD7 impairs the osteogenic ability of hDFCs, characterized by reduced alkaline phosphatase activity and mineralization, and decreased expression of osteogenesis-related genes. Conversely, CHD7 overexpression enhances the osteogenic differentiation of hDFCs. Mechanically, RNA-seq analyses revealed the down-regulated enrichment of PTH (parathyroid hormone)/PTH1R (parathyroid hormone receptor-1) signaling pathway after CHD7 knockdown. We found the expression of PTH1R positively correlates with CHD7. Importantly, overexpression of PTH1R in CHD7-knockdown hDFCs partially rescued the impaired osteogenic differentiation. Our research demonstrates that CHD7 regulates the osteogenic differentiation of hDFCs by regulating the transcription of PTH1R.
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