Cannabidiol Modulates Odonto/Osteogenic Differentiation In Human Dental Pulp Stem Cells
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PROVIDER: PRJNA1132676 | ENA |
REPOSITORIES: ENA
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Project description:Canabidiol (CBD), derived from Cannabis sativa plant exhibits beneficial in alleviating a number of of oral and dental pathoses This study aimed to explore the impact of CBD on several traits of human dental pulp stem cell (hDPSC) such as their proliferation, apoptosis, migration, and odonto/osteogenic differentiation. hDPSCs were harvested from human dental pulp tissues. The cells were treated with 1.25, 2.5, 5, 10, 25, and 50 μg/mL CBD. Cell response in terms of cell proliferation, colony-forming unit, cell cycle progression, cell migration, apoptosis, and odonto/osteogenic differentiation of hDPSCs were assessed in normal and P. gingivalis lipopolysaccharide (LPS)-induced `inflammatory` milieus. RNA sequencing and proteomic analysis were performed to predict target pathways impacted by CBD. In a normal physiological growth milieu, exposure to 5 μg/mL CBD attenuated hDPSCs proliferation and reduced the number of colony-forming units. On the contrary, in LPS-exposed hDPSCs, a dose of 1.25 μg/mL CBD restored proliferation and odonto/osteogenic differentiation. Proteomic analysis demonstrated that several proteins involved in cell proliferation and differentiation were upregulated following CBD exposure, including CCL8, CDC42, and KFL5. RNA sequencing data indicated that CBD upregulated Notch signaling pathway. In an inhibitory experiment, DAPT, a Notch inhibitor, reduced the effect of CBD-rescued LPS-attenuated mineralization in hDPSCs, suggesting that CBD potentially mediates Notch activation to exert its effect on odonto/osteogenic differentiation of hDPSCs. CBD recovers the proliferation and survival of hDPSCs following exposure to LPS. Additionally,we report here, for the first time, that CBD-mediated Notch activation effectively restores the odonto/osteogenic differentiation ability of hDPSCs. These results while uncovers and underscore the potential role of CBD as a therapeutic option to enhance dentin regeneration.
2025-08-31 | GSE271641 | GEO
Project description:Msh homeobox 1 (MSX1) is a transcriptional factor regulating embryonic development of limbs and craniofacial tissues including bone and teeth. The purpose of this study was to investigate contribution of MSX1 to the osteogenic potential and calcification-related phenotypic expression of dental pulp stromal/mesenchymal cells isolated from human teeth. Immunohistochemisitry of a 3 week-old mouse molar showed that MSX1 protein was localized to odontoblasts and pulpal mesenchymal cells at different levels and in different manners depending upon the position of the cells in pulp tissue. When dental pulp stromal/mesenchymal cells were exposed to osteogenesis-induction medium, runt-related transcription factor-2 (RUNX2), bone morphogenetic protein-2 (BMP2), alkaline phosphatase (ALPL) and osteocalcin (OCN) mRNA levels, as well as alkaline phosphatase activity, increased on days 4-12, and, thereafter, the matrix was calcified on day 14. However, knockdown of MSX1 with small interfering RNA abolished this induction of the osteoblast-related gene expression, alkaline phosphatase activity and calcification. Interestingly, DNA microarray and quantative PCR analyses revealed that the MSX1 knockdown induced the sterol regulatory element-binding protein 2 (SREBP2) transcriptional factor and its downstream target genes in cholesterol-synthesis pathway. Inhibition of cholesterol synthesis enhances osteoblast differentiation of various mesenchymal cells. Thus, MSX1 may down-regulate the cholesterol synthesis-related genes to ensure osteoblast differentiation of dental pulp stromal/mesenchymal cells.
2017-01-25 | GSE69992 | GEO
Project description:Bone regeneration is currently a major clinical challenge. Adult human mesenchymal stem cells (MSCs) possess a unique advantage in organ tissue repair and physiological function recovery. Human dental pulp derived MSCs (hDPSCs) have the dual advantages of proliferation and osteogenic differentiation abilities. They can be induced to secrete bone matrix proteins to form osteogenic microtissues by constructing an appropriate osteogenic three-dimensional microenvironment. The biological characteristics of hDPSCs from microtissues and two-dimensional cultured cells are compared using RNA sequencing. According to RNA sequencing and subsequent functional verification, the osteogenic activations of hDPSCs might be related to cell adhesion related signaling pathways. In this process, cells could sense the tension from the environment to activate related signaling pathways, such as the FAK/RhoA/Rock signaling pathway. Further, the downstream osteogenic transcription factors could be activited into the nucleus, thereby promoting the expression of osteogenic proteins.
2024-06-30 | GSE246980 | GEO
Project description:This study investigated the efficacy of decellularized ECM derived from human dental pulp stem cells (dECM_DPSCs) and gingival-derived mesenchymal stem cells (dECM_GSCs) as an inductive scaffold for osteogenic differentiation of GSCs. ECM was constructed from DPSCs and GSCs that were cultured in a growth medium supplemented with L-ascorbic acid (N-ECM) or osteogenic induction medium (OM-ECM). The gene expression profile was examined using a high-throughput RNA sequencing technique.
2024-03-27 | GSE226347 | GEO
Project description:Although various sources of cMSCs show similar characteristics, they are different in osteogenic potential due to their original cellular sources. Thus, this study was designed to globally explore and analyze the in vitro differentiation potential and behavior of canine bone-marrow derived mesenchymal stem cells (cBM-MSCs) and canine dental pulp stem cells (cDPSCs) toward osteogenic lineage. Global study of an in vitro osteogenic differentiation potential of the isolated cells was performed using proteomic-based analysis through mass spectrometry with dimethyl labelling method at day 7 and 14 post-induction, comparing with undifferentiated cells. The obtained results could be used as a comprehensive data and principal knowledge of the osteogenic differentiation potential of cBM-MSCs and cDPSCs in vitro and the trend of MSC-based tissue engineering for osteogenic regenerative therapy, concentrating on cMSCs application.
2021-09-08 | PXD015512 | Pride
Project description:To evaluate the miRNA and mRNA expression profiles (miRNOME) we identified miRNAs during in vitro osteogenic differentiation of human dental pulp stem cells (DPSC). The DPSCs were cultured in the DMEM + beta-glycerol phosphate, ascorbic acid and dexamethasone for 2 dias to 21days. The microRNA or mRNA expression profiling during the differentiation process was analyzed through hybridizations with Agilent miRNA-microarray (8x15K format).
2014-11-28 | E-MTAB-3077 | biostudies-arrayexpress
Project description:This study aimed to investigate the effects of shear stress on osteogenic differentiation of human dental pulp stem cells (hDPSCs). The hDPSCs were subjected to shear stress for 24 hours before osteogenic induction for 21 days. The mRNA expression of osteogenic markers such as RUNX2, OSX, ALP, COL1A1, OCN, and OPN was evaluated by real-time RT-PCR. Alkaline Phosphatase (ALP) activity and Alizarin Red S (ARS) staining were investigated to confirm osteogenic differentiation and mineralization of hDPSCs, respectively. The protein expression of osterix was shown by immunofluorescence staining and western blotting. RNA sequencing was performed to investigate how shear stress affects the osteogenic differentiation of hDPSCs, which was validated through p38 inhibitor (SB203580) treatment. Real-time RT-PCR revealed that shear stress enhanced osteogenic marker gene expression. The increased Osterix protein expression was detected on Day 14 in the shear stress loading group compared to the static group. Shear stress enhanced ALP activity and mineralization, observed on Days 14 and 21. A volcano plot exhibited up- and downregulated genes, while the p38 inhibitor markedly inhibited osteogenic differentiation of hDPSCs triggered by shear stress. In conclusion, shear stress promotes the osteogenic differentiation of hDPSCs through the p38 mitogen-activated protein kinase signaling pathway.
2025-06-10 | GSE299041 | GEO
Project description:This study focuses on the impact of ECM (extra-cellular matrix) over cell processes such as proliferation, differentiation or mineralization which is more specifically related to our dental pulp cells. We cultured these dental pulp stem cells (DPSC) in mineralization growth or normal growth conditions. After 21 days, cells were incubated in decellularized solution until no intact cells are seen. After ECM was washed, we studied the mineralization associated to these two different ECM. Matrisome proteins were identified through proteomics. DPSC matrisome is composed of 225 individual different proteins. We classified them according to different categories, the 3 core matrisome categories: glycoproteins, collagens, proteoglycans and the 3 matrisome associated proteins categories: the regulators, affiliated and secreted. When comparing the proteins in the N-ECM and OM-ECM, most of the core matrisome proteins are downregulated in OM conditions, except 3 glycoproteins, as well as regulators and secreted factors. However, annexins were found to be upregulated in OM condition. Cell adhesion, tensile strength and growth factor binding are over-represented in NM ECM. The collagen group and glycoproteins were higher in N-ECM than OM-ECM Thereafter, gingival stem cells (GSC), the less inherit osteogenic potency cells, were seeded on these N-ECM and OM-ECM. When GSCs were seeded on DPSC-derived ECM, the OM-ECM dramatically promoted mineral deposition compared with N-ECM. We hypothesize that annexins could participate in the osteogenic inductive properties. ECM plays a pivotal role in many physiological processes, it regulates cells behavior and can orient cell differentiation. Dental pulp and oral mucosa share embryological origins but differ in their reactions to insults. Dental pulp can mineralize while oral mucosa heals ad integrum. We hypothesize that ECM participate in these characteristics.
2021-09-23 | PXD018951 | Pride
Project description:To evaluate the miRNA and mRNA expression profiles (miRNOME and transcriptome) we reconstructed networks identifying miRNAs and mRNA during in vitro osteogenic differentiation of human dental pulp stem cells (DPSC). The DPSCs were cultured in the DMEM + beta-glycerol phosphate, ascorbic acid and dexamethasone for 2 to 21 days. The microRNA or mRNA expression profiling during the differentiation process was analyzed through hybridizations with Agilent miRNA-microarray (8x15K format) or whole-human genome Agilent microarray (4x44K format).
2014-11-13 | E-MTAB-3010 | biostudies-arrayexpress