Hyaluronan based gel promotes human Dental Pulp Stem Cells bone differentiation by activating YAP/TAZ pathway

Abstract Background. Hyaluronic acid (HA) is the major component of the extracellular matrix of human tissue, where it regulates processes such as osmotic pressure, water retention, cell migration, and differentiation. For these reasons, hyaluronans are currently used in regenerative medicine in different areas. Nevertheless, hyaluronans exist in different forms accordingly with molecular weight and degree of crosslinking, which can have a different and context-depended effects on cellular processes. Thus, picking the most appropriate form of hyaluronan turn out to be fundamental as it can make a huge difference in tissue regeneration. MSCs have attracted attention in tissue regeneration for their proliferation potential and ability to differentiate in several cytotypes. Among MSCs, human Dental Pulp Stem Cells (hDPSCs) were shown to be remarkably suitable for bone differentiation.In this study, we tested the capability to induce osteogenic differentiation in hDPSCs of three hyaluronans forms: linear pharmaceutical-grade hyaluronans at high (HHA), low molecular weight (LHA), and the recently stabilized hybrid cooperative complexes (HCC), containing both sizes.Methods. hDPSCs were treated with HHA, LHA, HCC for 7, 14 and 21 days. The effects of hyaluronans on osteogenic differentiation were evaluated by qRT-PCR and WB of osteogenic markers and by Alizarin Red S staining. CD44, the main receptor of the HA on cell surface and an upstream regulator of YAP/TAZ signaling, was analyzed by immunofluorescence. YAP/TAZ expression was measured by qRT-PCR. To confirm the involvement of YAP/TAZ pathway, YAP/TAZ inhibitor-1 was used and the loss of function of YAP/TAZ was evaluated by qRT-PCR, WB and immunofluorescence.Results. HCC was found to be the most impacting in inducing osteogenesis, with significant effects already at 7-14 days of treatment. HCC induced strong overexpression of osteocalcin, osteopontin, and bone sialoprotein, calcification nodule formation, and CD44 up-regulation.In addition, we showed that this biological process is associated to the activation of YAP/TAZ pathway and its target genes CTGF, ANKDR-1, RUNX-1, and RUNX-2.Conclusions. In conclusion, in this study we show that HA’s molecular weight can have a tremendous impact on HA performance for bone regeneration, and we unveil a new molecular mechanism by which HA acts on stem cells.

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Evaluation of Resin-Based Material Containing Copaiba Oleoresin (Copaifera Reticulata Ducke): Biological Effects on the Human Dental Pulp Stem Cells

Biomolecules ◽

10.3390/biom10070972 ◽

2020 ◽

Vol 10 (7) ◽

pp. 972

Author(s):

Roberta Souza D’Almeida Couto ◽

Maria Fernanda Setubal Destro Rodrigues ◽

Leila Soares Ferreira ◽

Ivana Márcia Alves Diniz ◽

Fernando de Sá Silva ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Biological Effects ◽

Dental Pulp Stem Cells ◽

Nodule Formation ◽

Alizarin Red ◽

Pulp Capping ◽

Hsp 27 ◽

Dental Pulp Capping ◽

Human Dental Pulp

The purpose of this study was to analyze in vitro the biological effects on human dental pulp stem cells triggered in response to substances leached or dissolved from two experimental cements for dental pulp capping. The experimental materials, based on extracts from Copaifera reticulata Ducke (COP), were compared to calcium hydroxide [Ca(OH)2] and mineral trioxide aggregate (MTA), materials commonly used for direct dental pulp capping in restorative dentistry. For this, human dental pulp stem cells were exposed to COP associated or not with Ca(OH)2 or MTA. Cell cytocompatibility, migration, and differentiation (mineralized nodule formation (Alizarin red assay) and gene expression (RT-qPCR) of OCN, DSPP, and HSP-27 (genes regulated in biomineralization events)) were evaluated. The results showed that the association of COP reduced the cytotoxicity of Ca(OH)2. Upregulations of the OCN, DSPP, and HSP-27 genes were observed in response to the association of COP to MTA, and the DSPP and HSP-27 genes were upregulated in the Ca(OH)2 + COP group. In up to 24 h, cell migration was significantly enhanced in the MTA + COP and Ca(OH)2 + COP groups. In conclusion, the combination of COP with the currently used materials for dental pulp capping [Ca(OH)2 and MTA] improved the cell activities related to pulp repair (i.e., cytocompatibility, differentiation, mineralization, and migration) including a protective effect against the cytotoxicity of Ca(OH)2.

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miR-217 Suppresses Osteogenic Differentiation of Human Dental Pulp Stem Cells by Down-Regulating Sirtuin 1

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2020.2373 ◽

2020 ◽

Vol 10 (7) ◽

pp. 978-986

Author(s):

Haiquan Yue ◽

Yidan Guo ◽

Juan Song ◽

Ruimin Liu

Keyword(s):

Stem Cells ◽

Osteogenic Differentiation ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Sirtuin 1 ◽

Luciferase Reporter ◽

Induction Medium ◽

Alizarin Red ◽

Human Dental Pulp ◽

Osteogenic Induction

The paper is committed to uncovering the effect of miR-217 on osteogenic differentiation of human dental pulp stem cells (hDPSCs) and its mechanism. hDPSCs were separated from human dental pulp tissues for measurement of stemness. The osteogenic differentiation of hDPSCs was induced in an osteogenic induction medium. The hDPSCs were transfected with miR-217 mimic, miR-217 inhibitor and/or sh-SIRT1 accordingly. The expressions of miR-217 and SIRT1 were detected in hDPSCs after cell transfection and osteogenic differentiation. Calcium nodules were showed by alizarin red staining. Moreover, the expressions of osteogenic differentiation-related genes were also assessed. The binding of miR-217 to SIRT1 was predicted on starBase and further determined by dual-luciferase reporter assay. Down-regulated miR-217 and up-regulated SIRT1 were found during osteogenic differentiation of hDPSCs. The osteogenic differentiation of hDPSCs was suppressed after transfection of miR-217 mimic or sh-SIRT1 while promoted by miR-217 inhibition. Taken together, miR-217 can suppress osteogenic differentiation of hDPSCs by negatively regulating SIRT1.

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Biocompatibility and Bioactivity of Set Direct Pulp Capping Materials on Human Dental Pulp Stem Cells

Materials ◽

10.3390/ma13183925 ◽

2020 ◽

Vol 13 (18) ◽

pp. 3925

Author(s):

Yemi Kim ◽

Donghee Lee ◽

Dani Song ◽

Hye-Min Kim ◽

Sin-Young Kim

Keyword(s):

Stem Cells ◽

Cell Migration ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Nodule Formation ◽

Alizarin Red ◽

Pulp Capping ◽

Alp Activity ◽

Human Dental Pulp ◽

And Control

In this study, we assessed the biocompatibility and bioactivity of various pulp capping materials—ProRoot MTA (Dentsply Tulsa Dental Specialties), Biodentine (Septodont), TheraCal LC (Bisco), and Dycal (Dentsply Caulk)—on human dental pulp stem cells (hDPSCs). Experimental disks (diameter, 7 mm; height, 4 mm) were stored in a humified incubator at 37 °C for 48 h. Then, the pulp capping materials were tested for cytotoxic effects by methyl-thiazoldiphenyl-tetrazolium and scratch wound healing assays, and for mineralization potential by Alizarin red S (ARS) staining assay and alkaline phosphatase enzyme (ALP) activity. Cell viability and cell migration did not significantly differ between ProRoot MTA, Biodentine, and control (p > 0.05). TheraCal LC exhibited slower cell migration on days 2–4 compared to control (p < 0.05), and Dycal showed no cell migration. ALP activity was highest with Biodentine on days 10 and 14, and was lowered with TheraCal LC and Dycal (p < 0.05). In the ARS assay, hDPSCs grown in ProRoot MTA and TheraCal LC eluates showed significantly increased mineralized nodule formation on day 21 compared to Biodentine, Dycal, and control (p < 0.05). These findings indicate that ProRoot MTA, Biodentine, and TheraCal LC exhibit better biocompatibility and bioactivity than Dycal.

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Chrysin induces osteogenic differentiation of human dental pulp stem cells

Experimental Cell Research ◽

10.1016/j.yexcr.2020.112466 ◽

2021 ◽

Vol 400 (2) ◽

pp. 112466

Author(s):

J.F. Huo ◽

M.L. Zhang ◽

X.X. Wang ◽

D.H. Zou

Keyword(s):

Stem Cells ◽

Osteogenic Differentiation ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Human Dental Pulp

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Chlorogenic acid promotes osteogenic differentiation of human dental pulp stem cells via Wnt signaling

Stem Cells and Development ◽

10.1089/scd.2020.0193 ◽

2021 ◽

Author(s):

Xiaoping Hu ◽

Li Wang ◽

Yuanqiao He ◽

Minli Wei ◽

Huilin Yan ◽

...

Keyword(s):

Stem Cells ◽

Wnt Signaling ◽

Chlorogenic Acid ◽

Osteogenic Differentiation ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Human Dental Pulp

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Functionalization of Polycaprolactone Scaffolds with Hyaluronic Acid and β-TCP Facilitates Migration and Osteogenic Differentiation of Human Dental Pulp Stem Cells In Vitro

Tissue Engineering Part A ◽

10.1089/ten.tea.2014.0177 ◽

2015 ◽

Vol 21 (3-4) ◽

pp. 729-739 ◽

Cited By ~ 31

Author(s):

Jonas Jensen ◽

David Christian Evar Kraft ◽

Helle Lysdahl ◽

Casper Bindzus Foldager ◽

Muwan Chen ◽

...

Keyword(s):

Stem Cells ◽

Hyaluronic Acid ◽

Osteogenic Differentiation ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Human Dental Pulp

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Peroxiredoxin 6 Inhibits Osteogenic Differentiation and Bone Formation Through Human Dental Pulp Stem Cells and Induces Delayed Bone Development

Antioxidants and Redox Signaling ◽

10.1089/ars.2018.7530 ◽

2019 ◽

Vol 30 (17) ◽

pp. 1969-1982 ◽

Cited By ~ 2

Author(s):

Kyung-Ran Park ◽

Hyung-Mun Yun ◽

In Jun Yeo ◽

Sehyung Cho ◽

Jin Tae Hong ◽

...

Keyword(s):

Stem Cells ◽

Bone Formation ◽

Osteogenic Differentiation ◽

Dental Pulp ◽

Bone Development ◽

Dental Pulp Stem Cells ◽

Peroxiredoxin 6 ◽

Human Dental Pulp

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A Novel Hypoxic lnc-RNA, HRL-SC, Promotes Proliferation and Migration of Human Dental Pulp Stem Cells Through PI3K/AKT Signalling Pathway

10.21203/rs.3.rs-1143046/v1 ◽

2021 ◽

Author(s):

Junkai Zeng ◽

Ming Chen ◽

Yeqing Yang ◽

Buling Wu

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Differential Expression Analysis ◽

Dental Pulp Stem Cells ◽

Signalling Pathway ◽

Gene Set Enrichment Analysis ◽

Qrt Pcr ◽

Human Dental Pulp ◽

And Migration ◽

Proliferation And Migration

Abstract Background: Human dental pulp stem cells (hDPSCs) are critical for pulp generation. hDPSCs proliferate faster under hypoxia, but the regulatory mechanism of long noncoding RNAs (lncRNAs) in this process is not fully understood.Methods: Novel lncRNAs were obtained by reanalysis of transcriptome datasets coming from RNA-Seq under hypoxia compared with normoxia, and differential expression analysis of target genes were performed. Bioinformatics analyses including Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and Gene Set Enrichment Analysis (GSEA) analysis were used to understand the function of key novel lncRNA. hDPSCs were isolated from dental pulp tissue. EdU test and scratch healing test were used to detect the proliferation and migration of hDPSCs. qRT-PCR was used to detect the RNA level expression changes of selected genes. RNA fluorescence in situ hybridization (FISH), small interfering RNA (siRNA), qRT-PCR and western blot analysis were used to explore the function of key novel lncRNA. Results: We identified 496 novel lncRNAs in hDPSCs under hypoxia, including 45 expressed differentially novel lncRNAs. Of them, we focused on a key novel lncRNA, which we named HRL-SC (hypoxia related lncRNA in stem cells). Functional annotation revealed that HRL-SC was associated with hypoxic conditions and PI3K/AKT signalling pathway. HRL-SC was mainly located in the cytoplasm of hDPSCs and had stably high expression under hypoxia. Knockdown of HRL-SC inhibited proliferation and migration of hDPSCs and expression levels of PI3K/AKT related marker proteins. Furthermore, AKT activator SC79 partially offset the inhibitory effect caused by the knockdown, indicating that HRL-SC promoted hDPSCs through PI3K/AKT signalling pathway.Conclusion: Hypoxia related lncRNA HRL-SC promotes proliferation and migration of hDPSCs through PI3K/AKT signalling pathway and it may provide a better understanding for regenerative application of hDPSCs.

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miR-6807-5p Inhibited the Odontogenic Differentiation of Human Dental Pulp Stem Cells Through Directly Targeting METTL7A

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.759192 ◽

2021 ◽

Vol 9 ◽

Author(s):

Ning Wang ◽

Xiao Han ◽

Haoqing Yang ◽

Dengsheng Xia ◽

Zhipeng Fan

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Binding Protein ◽

Dental Pulp Stem Cells ◽

Luciferase Reporter ◽

Control Group ◽

Alizarin Red ◽

Odontogenic Differentiation ◽

Human Dental Pulp ◽

Tooth Tissue

Background: Tooth tissue regeneration mediated by mesenchymal stem cells (MSCs) has become the most ideal treatment. Although the known regulatory mechanism and some achievements have been discovered, directional differentiation cannot effectively induce regeneration of tooth tissue. In this study, we intended to explore the function and mechanism of miR-6807-5p and its target gene METTL7A in odontogenic differentiation.Methods: In this study, human dental pulp stem cells (DPSCs) were used. Alkaline phosphatase (ALP), Alizarin red staining (ARS), and calcium ion quantification were used to detect the odontogenic differentiation of miR-6807-5p and METTL7A. Real-time RT-PCR, western blot, dual-luciferase reporter assay, and pull-down assay with biotinylated miRNA were used to confirm that METTL7A was the downstream gene of miR-6807-5p. Protein mass spectrometry and co-immunoprecipitation (Co-IP) were used to detect that SNRNP200 was the co-binding protein of METTL7A.Results: After mineralized induction, the odontogenic differentiation was enhanced in the miR-6807-5p-knockdown group and weakened in the miR-6807-5p-overexpressed group compared with the control group. METTL7A was the downstream target of miR-6807-5p. After mineralized induction, the odontogenic differentiation was weakened in the METTL7A-knockdown group and enhanced in the METTL7A-overexpressed group compared with the control group. SNRNP200 was the co-binding protein of METTL7A. The knockdown of SNRNP200 inhibited the odontogenic differentiation of DPSCs.Conclusion: This study verified that miR-6807-5p inhibited the odontogenic differentiation of DPSCs. The binding site of miR-6807-5p was the 3′UTR region of METTL7A, which was silenced by miR-6807-5p. METTL7A promoted the odontogenic differentiation of DPSCs. SNRNP200, a co-binding protein of METTL7A, promoted the odontogenic differentiation of DPSCs.

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