scholarly journals METTL3-mediated M6a Modification Regulates Cell Cycle Progression of Dental Pulp Stem Cells

2021 ◽  
Author(s):  
Haiyun Luo ◽  
Wenjing Liu ◽  
Yanli Zhang ◽  
Xiao Jiang ◽  
Shiqing Wu ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Tissue Engineering ◽  
Stem Cells ◽  
Flow Cytometry ◽  
Differentially Expressed Genes ◽  
Dental Pulp ◽  
Cell Cycle Control ◽  
Dental Pulp Stem Cells ◽  
Differentially Expressed ◽  
Expression Level

Abstract Background: Dental pulp stem cells (DPSCs) exhibited self-renewal, pluripotency capacity and served as promising cells source in endodontic regeneration and tissue engineering. Meanwhile, the regenerative capacity of DPSCs is limited and reduced in long lifespan. N6-methyladenosine (m6A) is the most prevalent, reversible internal modification in RNAs. The methyltransferases complex and demethylases mediated m6A methylation and cooperated to impact various biological processes associated with stem cell fate determination. However, the biological effect of m6A methylation in DPSCs remained unclear. Methods: Cell surface markers and differentiation potential of primary DPSCs were identified and m6A immunoprecipitation with deep sequencing (m6A RIP-seq) was used to uncover characteristics of m6A modifications in DPSCs transcriptome. Expression level of m6A-related genes were evaluated in immature/mature pulp tissues and cells. Lentiviral vectors were constructed to knockdown or overexpress methyltransferase like 3 (METTL3). Cell morphology, viability, senescence and apoptosis were further analyzed by β-galactosidase, TUNEL staining and flow cytometry. Bioinformatic analysis combing m6A RIP and shMETTL3 RNA-seq was used to functionally enrich overlapped genes and screen target of METTL3. Cell cycle distributions were assayed by flow cytometry and m6A RIP-qPCR was used to confirm METTL3 mediated m6A methylation in DPSCs. Results: Here, m6A peaks distribution, binding area and motif in DPSCs were first revealed by m6A RIP-seq. We also found a relative high expression level of METTL3 in immature DPSCs with superior regenerative potential and METTL3 knockdown induced cell apoptosis and senescence. Furthermore, Conjoint analysis of m6A RIP and RNA-sequencing showed differentially expressed genes affected by METTL3 depletion was mainly enriched in cell cycle, mitosis and alteration of METTL3 expression resulted in cell cycle arrest which indicated METTL3 make essential effect in cell cycle control. To further investigate underlying mechanisms, we explored proteins interaction network of differentially expressed genes and Polo-like Kinase 1 (PLK1), a critical cycle modulator was identified as target of METTL3-mediated m6A methylation in DPSCs. Conclusions: These results revealed m6A methylated hallmarks in DPSCs and a regulatory role of METTL3 in cell cycle control. Our study shed light on therapeutic approaches in vital pulp therapy and serve new insight in stem cells based tissue engineering.

scholarly journals METTL3-mediated m6A modification regulates cell cycle progression of dental pulp stem cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Haiyun Luo ◽  
Wenjing Liu ◽  
Yanli Zhang ◽  
Yeqing Yang ◽  
Xiao Jiang ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Tissue Engineering ◽  
Stem Cells ◽  
Flow Cytometry ◽  
Stem Cell ◽  
Cell Fate ◽  
Dental Pulp ◽  
Interaction Network ◽  
Dental Pulp Stem Cells ◽  
Tunel Staining

Abstract Background Dental pulp stem cells (DPSCs) are a promising cell source in endodontic regeneration and tissue engineering with limited self-renewal and pluripotency capacity. N6-methyladenosine (m6A) is the most prevalent, reversible internal modification in RNAs associated with stem cell fate determination. In this study, we aim to explore the biological effect of m6A methylation in DPSCs. Methods m6A immunoprecipitation with deep sequencing (m6A RIP-seq) demonstrated the features of m6A modifications in DPSC transcriptome. Lentiviral vectors were constructed to knockdown or overexpress methyltransferase like 3 (METTL3). Cell morphology, viability, senescence, and apoptosis were analyzed by β-galactosidase, TUNEL staining, and flow cytometry. Bioinformatic analysis combing m6A RIP and shMETTL3 RNA-seq functionally enriched overlapped genes and screened target of METTL3. Cell cycle distributions were assayed by flow cytometry, and m6A RIP-qPCR was used to confirm METTL3-mediated m6A methylation. Results Here, m6A peak distribution, binding area, and motif in DPSCs were first revealed by m6A RIP-seq. We also found a relatively high expression level of METTL3 in immature DPSCs with superior regenerative potential and METTL3 knockdown induced cell apoptosis and senescence. A conjoint analysis of m6A RIP and RNA sequencing showed METTL3 depletion associated with cell cycle, mitosis, and alteration of METTL3 resulted in cell cycle arrest. Furthermore, the protein interaction network of differentially expressed genes identified Polo-like kinase 1 (PLK1), a critical cycle modulator, as the target of METTL3-mediated m6A methylation in DPSCs. Conclusions These results revealed m6A methylated hallmarks in DPSCs and a regulatory role of METTL3 in cell cycle control. Our study shed light on therapeutic approaches in vital pulp therapy and served new insight into stem cell-based tissue engineering.

scholarly journals RNA Sequence Analyses throughout the Course of Mouse Cardiac Laminopathy Identify Differentially Expressed Genes for Cell Cycle Control and Mitochondrial Function

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Zhili Shao ◽  
Wonshill Koh ◽  
Ying Ni ◽  
Wei Li ◽  
Brendan Agatisa-Boyle ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Differentially Expressed Genes ◽  
Mitochondrial Function ◽  
Cell Cycle Control ◽  
Differentially Expressed ◽  
Sequence Analyses ◽  
Rna Sequence

scholarly journals Dental Pulp Stem Cells and Tissue Engineering Strategies for Clinical Application on Odontoiatric Field

10.5772/24871 ◽  
2011 ◽  
Author(s):  
Zavan Barbara ◽  
Bressan Eriberto ◽  
Sivolella Stefano ◽  
Brunello Giulia ◽  
Gardin Chiara ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Clinical Application ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells

scholarly journals Dental pulp stem cells express tendon markers under mechanical loading and are a potential cell source for tissue engineering of tendon-like tissue

2016 ◽  
Vol 8 (4) ◽  
pp. 213-222 ◽  
Author(s):  
Yu-Ying Chen ◽  
Sheng-Teng He ◽  
Fu-Hua Yan ◽  
Peng-Fei Zhou ◽  
Kai Luo ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mechanical Loading ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Cell Source

Evaluating adhesion and alignment of dental pulp stem cells to a spider silk substrate for tissue engineering applications

2017 ◽  
Vol 81 ◽  
pp. 104-112 ◽  
Author(s):  
Katherine Hafner ◽  
Dallas Montag ◽  
Hannah Maeser ◽  
Congyue Peng ◽  
William R. Marcotte ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Dental Pulp ◽  
Spider Silk ◽  
Dental Pulp Stem Cells ◽  
Engineering Applications

Proliferation rate of stem cells derived from human dental pulp and identification of differentially expressed genes

2014 ◽  
Vol 38 (5) ◽  
pp. 582-590 ◽  
Author(s):  
Muhammad Fawwaz Abdullah ◽  
Siti Fadilah Abdullah ◽  
Nor Shamsuria Omar ◽  
Zuliani Mahmood ◽  
Siti Noor Fazliah Mohd Noor ◽  
...  
Keyword(s):  
Stem Cells ◽  
Differentially Expressed Genes ◽  
Dental Pulp ◽  
Proliferation Rate ◽  
Differentially Expressed ◽  
Human Dental Pulp

scholarly journals Bladder Smooth Muscle Cells Differentiation from Dental Pulp Stem Cells: Future Potential for Bladder Tissue Engineering

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Bing Song ◽  
Wenkai Jiang ◽  
Amr Alraies ◽  
Qian Liu ◽  
Vijay Gudla ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Smooth Muscle ◽  
Dental Pulp ◽  
Transforming Growth Factor ◽  
Dental Pulp Stem Cells ◽  
Specific Marker ◽  
Bladder Tissue ◽  
Growth Environment ◽  
Gene And Protein Expression

Dental pulp stem cells (DPSCs) are multipotent cells capable of differentiating into multiple cell lines, thus providing an alternative source of cell for tissue engineering. Smooth muscle cell (SMC) regeneration is a crucial step in tissue engineering of the urinary bladder. It is known that DPSCs have the potential to differentiate into a smooth muscle phenotype in vitro with differentiation agents. However, most of these studies are focused on the vascular SMCs. The optimal approaches to induce human DPSCs to differentiate into bladder SMCs are still under investigation. We demonstrate in this study the ability of human DPSCs to differentiate into bladder SMCs in a growth environment containing bladder SMCs-conditioned medium with the addition of the transforming growth factor beta 1 (TGF-β1). After 14 days of exposure to this medium, the gene and protein expression of SMC-specific marker (α-SMA, desmin, and calponin) increased over time. In particular, myosin was present in differentiated cells after 11 days of induction, which indicated that the cells differentiated into the mature SMCs. These data suggested that human DPSCs could be used as an alternative and less invasive source of stem cells for smooth muscle regeneration, a technology that has applications for bladder tissue engineering.

Injectable thermosensitive hybrid hydrogel containing graphene oxide and chitosan as dental pulp stem cells scaffold for bone tissue engineering

2020 ◽  
Vol 162 ◽  
pp. 1338-1357 ◽  
Author(s):  
Nazanin Amiryaghoubi ◽  
Nader Noroozi Pesyan ◽  
Marziyeh Fathi ◽  
Yadollah Omidi
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Graphene Oxide ◽  
Bone Tissue Engineering ◽  
Bone Tissue ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Hybrid Hydrogel

scholarly journals Downregulation of miR-224-5p Promotes Migration and Proliferation in Human Dental Pulp Stem Cells

2019 ◽  
Vol 2019 ◽  
pp. 1-8 ◽  
Author(s):  
Zhihong Ke ◽  
Zailing Qiu ◽  
Tingting Xiao ◽  
Jianchai Zeng ◽  
Luning Zou ◽  
...  
Keyword(s):  
Stem Cells ◽  
Root Canal ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Differentially Expressed ◽  
Pulp Regeneration ◽  
Regenerative Endodontics ◽  
Differentially Expressed Mirna ◽  
And Migration ◽  
Proliferation And Migration

Introduction. Pulp regeneration, as a treatment for pulp necrosis, has significant advantages over root canal therapy for the preservation of living pulp. To date, research on pulp regeneration has mainly focused on the transplantation of pulp stem cells into the root canal, but there is still a lack of research on the migration of pulp cells into the root canal via cell homing. Stem cells from the apical tooth papilla (SCAP) are recognized as multidirectional stem cells, but these cells are difficult to obtain. MicroRNAs are small noncoding RNAs that play crucial roles in regulating normal and pathologic functions. We hypothesized that some types of microRNAs might improve the migration and proliferation function of dental pulp stem cells (DPSCs), which are easily obtained in clinical practice, and as a result, DPSCs might replace SCAP and provide valuable information for regenerative endodontics. Methods. Magnetic activated cell sorting of DPSCs and SCAP was performed. Next-generation sequencing was performed to examine DPSCs and SCAP miRNAs expression and to identify the most significant differentially expressed miRNA. CCK-8 and transwell assays were used to determine the impact of this miRNA on DPSCs proliferation and migration. Results. The most significant differentially expressed miRNA between DPSCs and SCAP was miR-224-5p. Downregulating miR-224-5p promoted DPSCs proliferation and migration; the opposite results were observed when miR-224-5p was upregulated. Conclusion. MiR-224-5p promotes proliferation and migration in DPSCs, a finding that is of great significance for further exploring the role of dental pulp stem cells in regenerative endodontics.

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