Transcriptome characterization of human highly osteoblastic/cementoblastic periodontal ligament cell clones (OM medium)
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE94600 | GEO | 2024/12/15
REPOSITORIES: GEO
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Project description:we isolated single-cell-derived clones and evaluated their potential to differentiation into osteo/cementoblastic (O/C) phenotype (C-O clones) or fibroblastic phenotype (C-F clones) in vitro, and evaluated the transcriptomic profile of the groups of clones in standard medium cultivation (without O/C induction medium), using next-generation sequencing technology (RNA-seq).
2021-03-24 | GSE94599 | GEO
Project description:BackgroundHeterogeneous 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.ObjectiveOur study aimed to clarify the differential transcriptome profile of PDL cells poised to differentiate into the O/C cell lineage.MethodologyTo 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.ResultsThe 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.ConclusionsThis 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.
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Project description:Periodontal ligament is soft connective tissue between alveolar bone and cementum, the surface hard tissue of tooth. Periodontal ligament fibroblasts (PDLF) actively express osteo/cementogenic genes, which contribute in periodontal tissue homeostasis. However, the key factors by which PDLF retain such osteo/cementogenic abilities has not been clarified yet. Here, we find PPARg is expressed in vivo periodontal ligament tissue and its distribution pattern is highly correlated with the alkaline phosphate enzymatic activity. Knockdown of PPARg drastically deprives PDLF of osteo/cementogenic abilities in vitro. In contrast, PPARg agonists enhance it. PPARg is required for maintaining acetylation status of H3K9 and H3K27, active chromatin marks, and the supplementation of acetyl-coA, the donor of histone acetylation, is able to restore PPARg knockdown-induced decreased osteo/cementogenic abilities in PDLF. RNA-seq and ChIP-seq combined analyses identify 4 osteogenic transcripts, RUNX2, SULF2, RCAN2, and RGMA, in the PPARg-dependent active chromatin region marked by H3K27ac. Furthermore, the RUNX2 binding sites are selectively enriched in the PPARg-dependent active chromatin region. Together, these findings identify PPARg is the key transcriptional factor of PDLF to retain osteo/cementogenic abilities and reveal that global H3K27ac modification involves in the comprehensive osteo/cementogenic transcriptional alteration mediated by PPARg.
2021-08-31 | GSE178605 | GEO
Project description:The aim of this study is to clarify the biological functions of decorin (DCN) in the healing and regeneration of wounded periodontal tissue. We investigated the expression pattern of DCN during the healing of wounded periodontal tissue in rats by immunohistochemistry and the effects of DCN on the osteoblastic differentiation of human periodontal ligament (PDL) stem cells (HPDLSCs) and preosteoblasts by Alizarin red S staining, quantitative reverse transcription-polymerase chain reactions, and western blotting. The expression of DCN was increased around the wounded PDL tissue on day 5 after surgery compared with the nonwounded PDL tissue, whereas its expression was not changed in the osteoblastic layer around the wounded alveolar bone. Furthermore, DCN promoted the osteoblastic differentiation of HPDLSCs, but it did not affect the osteoblastic differentiation of preosteoblasts. ERK1/2 phosphorylation was upregulated during the DCN-induced osteoblastic differentiation of HPDLSCs. DCN did not affect proliferation, migration, or the PDL-related gene expression of HPDLSCs. In conclusion, this study demonstrates that DCN has a role in the healing of wounded periodontal tissue. Furthermore, DCN secreted from PDL cells may contribute to bone healing by upregulating osteoblastic differentiation through ERK1/2 signaling in HPDLSCs, indicating a therapeutic effect of DCN in periodontal tissue regeneration.
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Project description:Purpose : This study was performed to investigate the effects of knockdown of PPARg for osteo/cementogenic differentiation of PDLF through RNA sequencing. Method : PDLF were transfected wtih either control siRNA or siRNA for PPARg and then cultured in mineralization inducing medium for 6 days. Total RNAs were collected before and after the culture. Result : Gene ontology analyses of RNA-seq data revealed the genes functionally related with ECM such as collagen synthesis and trimerization, proteoglycans, and fibers were selectively increased by 6-day culture and suppressed biological processes by si-PPARg compared with si-control at day 6 exhibited 5 of 7 upper ranked biological processes were related with ECM organization and metabolism. Moreover, the upper rank of suppressed pathways by si-PPARg compared with si-control at day 6 showed `Endochondral Ossification` was ranked 1st. Conclusions:These results suggested that knockdown of PPARg was sufficient to eliminate osteo/cementogenic and ECM organizing potentials by comprehensive transcriptional dysregulation in PDLF.
2021-08-31 | GSE178606 | GEO
Project description:This study was designed to identify endoglin (ENG)-dependent BMP2-responsive genes in the mouse fibroblastic cell line PDL-L2.
2014-01-21 | GSE54220 | GEO
Project description:This study was designed to identify endoglin (ENG)-dependent BMP2-responsive genes in the mouse fibroblastic cell line PDL-L2. PDL-L2 cells were treated with an siRNA for endoglin (siENG) or a control siRNA (siCont). After 48hr, the cells were exposed to recombinant human BMP-2 (250 ng/ml) or vehicle and cultured for 12 hr.
2014-01-21 | E-GEOD-54220 | biostudies-arrayexpress
Project description:Hesperetin (3',5,7-trihydroxy-4-methoxyflavanone) is a metabolite of hesperidin (hesperetin-7-O-rutinoside), which belongs to the flavanone subgroup and is found mainly in citrus fruits. Hesperetin has been reported to be an effective osteoinductive compound in various in vivo and in vitro models. However, how hesperetin effects osteogenic differentiation is not fully understood. In this study, we investigated the capacity of hesperetin to stimulate the osteogenic differentiation of periodontal ligament stem cells (PDLSCs) and to relieve the anti-osteogenic effect of high glucose. Osteogenesis of PDLSCs was assessed by measurement of alkaline phosphatase (ALP) activity, and evaluation of the mRNA expression of ALP, runt-related gene 2 (Runx2), osterix (OSX), and FRA1 as osteogenic transcription factors, as well as assessment of protein expression of osteopontin (OPN) and collagen type IA (COLIA). When PDLSCs were exposed to a high concentration (30 mM) of glucose, osteogenic activity decreased compared to control cells. Hesperetin significantly increased ALP activity at doses of 1, 10, and 100 µM. Pretreatment of cells with hesperetin alleviated the high-glucose-induced suppression of the osteogenic activity of PDLSCs. Hesperetin scavenged intracellular reactive oxygen species (ROS) produced under high glucose condition. Furthermore, hesperetin increased the activity of the PI3K/Akt and ?-catenin pathways. Consistent with this, blockage of Akt or ?-catenin diminished the protective effect of hesperetin against high glucose-inhibited osteogenic differentiation. Collectively, our results suggest that hesperetin alleviates the high glucose-mediated suppression of osteogenic differentiation in PDLSCs by regulating ROS levels and the PI3K/Akt and ?-catenin signaling pathways.
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Project description:White mineral trioxide aggregate (WMTA) is a root canal treatment material, which is known to exhibit a dark brown color when in contact with sodium hypochlorite solution (NaOCl). This study aimed to investigate the effects of NaOCl on the surface properties of WMTA discs and WMTA-induced osteoblastic differentiation of periodontal ligament stem cells (PDLSCs). Mixed WMTA (ProRoot MTA) was filled into the molds to form WMTA discs. These discs were immersed in distilled water (D-WMTA) or 5% NaOCl (Na-WMTA). Their surface structures and Ca2+ release level was investigated. Moreover, they were cultured with a clonal human PDLSC line (line 1-17 cells). The main crystal structures of Na-WMTA were identical to the structures of D-WMTA. Globular aggregates with polygonal and needle-like crystals were found on D-WMTA and Na-WMTA, which included Ca, Si, Al, C and O. However, many amorphous structures were also identified on Na-WMTA. These structures consisted of Na and Cl, but did not include Ca. NaOCl immersion also reduced Ca2+ release level from whole WMTA discs. Line 1-17 cells cultured with D-WMTA formed many mineralized nodules and exhibited high expression levels of osteoblast-related genes. However, cells incubated with Na-WMTA generated a small number of nodules and showed low expression levels of osteoblast-related genes. These results indicated that NaOCl reduced Ca2+ release from WMTA by generating amorphous structures and changing its elemental distribution. NaOCl may also partially abolish the ability of WMTA to stimulate osteoblastic differentiation of PDLSCs.
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Project description:The structural and functional integrity of bone-periodontal ligament (PDL)-cementum complex stems from the load-bearing attachment sites (entheses) between soft (PDL) and hard (bone, cementum) tissues. These attachment sites are responsible for the maintenance of a bone-PDL-cementum complex biomechanical function. The objective was to investigate changes in spatiotemporal expression of key biomolecules in developing and functionally active entheses.Multilabeling technique was performed on hemimandibles of 3 wk and 3 mo-old scleraxis-GFP transgenic mice for CD146, CD31, NG2, osterix and bone sialoprotein. Regions of dominant stretch within the PDL were evaluated by identifying directionality of collagen fibrils, PDL fibroblasts and PDL cell cytoskeleton.CD146+ cells adjacent to CD31+ vasculature were identified at PDL-bone enthesis. NG2+ cells were located at coronal bone-PDL and apical cementum-PDL entheses in the 3-wk-old group, but at 3 mo, NG2 was positive at the entheses of the apical region and alveolar crest. NG2 and osterix were colocalized at the osteoid and cementoid regions of the PDL-bone and PDL-cementum entheses. Bone sialoprotein was prominent at the apical region of 3-wk-old mice. The directionality of collagen fibers, fibroblasts and their cytoskeleton overlapped, except in the apical region of 3 wk.Colocalization of biomolecules at zones of the PDL adjacent to attachment sites may be essential for the formation of precementum and osteoid interfaces at a load-bearing bone-PDL-tooth fibrous joint. Biophysical cues resulting from development and function can regulate recruitment and differentiation of stem cells potentially from a vascular origin toward osteo- and cemento-blastic lineages at the PDL-bone and PDL-cementum entheses. Investigating the coupled effect of biophysical and biochemical stimuli leading to cell differentiation at the functional attachment sites is critical for developing regeneration strategies to enable functional reconstruction of the periodontal complex.
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