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      Analysis of the characteristics and expression profiles of coding and noncoding RNAs of human dental pulp stem cells in hypoxic conditions

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          Abstract

          Background

          Human dental pulp stem cell (DPSC)-mediated regenerative endodontics is a promising therapy for damaged teeth; however, the hypoxic environment in root canals can affect tissue regeneration. In this study, we investigate the characteristics and possible regulatory mechanisms of DPSC function under hypoxic conditions.

          Methods

          Human DPSCs were cultured under normoxia (20% O 2) and hypoxia (3% O 2). DPSC proliferation and osteo/odontogenic differentiation potential were assessed by Cell Counting Kit-8 (CCK8) assay, carboxyfluorescein succinimidyl ester (CFSE) assay, alkaline phosphatase (ALP) activity, Alizarin red staining, real-time RT-PCR assays, and western blot analysis. Microarray and bioinformatic analyses were performed to investigate the differences in the mRNA, lncRNA, and miRNA expression profiles of DPSCs.

          Results

          DPSCs exhibited a more powerful proliferation ability and lower osteo/odontogenic differentiation potential in hypoxic conditions. A total of 60 mRNAs (25 upregulated and 35 downregulated), 47 lncRNAs (20 upregulated and 27 downregulated), and 14 miRNAs (7 upregulated and 7 downregulated) in DPSCs were differentially expressed in the hypoxia group compared with the normoxia group. Bioinformatic analysis identified that 7 mRNAs (GRPR, ERO1L, ANPEP, EPHX1, PGD, ANGPT1, and NQO1) and 5 lncRNAs (AF085958, AX750575, uc002czn.2, RP3-413H6.2, and six-twelve leukemia (STL)) may be associated with DPSCs during hypoxia according to CNC network analysis, while 28 mRNAs (including GYS1, PRKACB, and NQO1) and 13 miRNAs (including hsa-miR-3916 and hsa-miR-192-5p) may be involved according to miRNA target gene network analysis. The depletion of one candidate lncRNA, STL, inhibited the osteo/odontogenic differentiation potentials of DPSCs.

          Conclusions

          Our results revealed that hypoxia could enhance the proliferation ability and impair the osteo/odontogenic differentiation potential of DPSCs in vitro. Furthermore, our results identified candidate coding and noncoding RNAs that could be potential targets for improving DPSC function in regenerative endodontics and lead to a better understanding of the mechanisms of hypoxia’s effects on DPSCs.

          Electronic supplementary material

          The online version of this article (10.1186/s13287-019-1192-2) contains supplementary material, which is available to authorized users.

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          Most cited references61

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          Pulp revascularization of immature teeth with apical periodontitis: a clinical study.

          Rui Ding, Gary Cheung, Jie Chen … (2009)
          The purpose of this study was to examine the effect of a pulpal revascularization procedure for immature necrotic teeth with apical periodontitis. Twelve patients, each with an immature permanent tooth with chronic or acute apical periodontitis, were recruited. A triantibiotic mix (ciprofloxacin, metronidazole, and minocycline) was used to disinfect the pulp for 1 week. Then a blood clot was created in the canal, over which grey mineral trioxide aggregate was placed. Patients were recalled periodically. Six patients dropped from the study (as a result of pain or failure to induce bleeding after canal disinfection) and instead received a standard apexification procedure. Another 3 patients did not attend any recall appointments. The remaining teeth (n = 3) were found to exhibit complete root development, with a positive response to pulp testing. Revascularization could be effective for managing immature permanent teeth with apical periodontitis with appropriate case selection.
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            Curcumin inhibits cell proliferation and induces apoptosis of human non-small cell lung cancer cells through the upregulation of miR-192-5p and suppression of PI3K/Akt signaling pathway.

            Hai Jin, Fan Qiao, Yan-Yan Wang … (2015)
            Curcumin is the main active ingredient extracted from the traditional Chinese medicine, turmeric, which acts against non-small cell lung cancer cell (NSCLC), lowers blood pressure, is anti-inflammatory, choleretic, and exerts anti‑oxidant effects, without any obvious toxicity in the long term. The aim of the present study was to investigate whether the anticancer effect of curcumin inhibited cell proliferation and induced apoptosis of human NSCLC through the upregulation of microRNA‑192-5p (miR-192-5p) and suppression of the PI3K/Akt signaling pathway. In the present study, treatment with curcumin inhibited cell proliferation, induced cell apoptosis and increased the caspase-3 activity of A549 cells. The results also showed that, miR-192-5p relative expression of NCL-H460 cells was relatively lower than that of A549 cells, which was higher, with that of BEAS-2E cells being the highest. miR-192-5p mimics suppressed cell proliferation and increased cell apoptosis of A549 cells. However, anti-miR-192-5p mimics increased cell proliferation and inhibited cell apoptosis of A549 cells. Curcumin treatment effectively increased the relative miR‑192-5p expression and suppressed the PI3K/Akt signaling pathway. miR-192-5p mimics enhanced the effect of curcumin on cell viability and apoptosis and suppressed the PI3K/Akt signaling pathway in A549 cells. Anti-miR-192-5p mimics reversed the effect of curcumin on A549 cells and PI3K/Akt expression. Collectively, our findings suggested that curcumin inhibited cell proliferation and induced apoptosis of human non-small cell lung cancer cells through the upregulation of miR-192-5p and suppression of the PI3K/Akt signaling pathway.
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              Histologic characterization of regenerated tissues in canal space after the revitalization/revascularization procedure of immature dog teeth with apical periodontitis.

              Martin Trope, Xiaojing Wang, Blayne Thibodeau … (2009)
              Recently, it has been shown that it is possible to treat an immature tooth with an infected pulp space and apical periodontitis in such a way as to heal and promote the ingrown of new vital tissue into the pulp space. However, the type of new-grown tissue is unclear. Based on the samples of a previously reported study, we further investigated histologically the types of tissues that had grown into the canal space. The canal dentinal walls were thickened by the apposition of newly generated cementum-like tissue termed herein "intracanal cementum (IC)." One case showed partial survival of pulp tissue juxtaposed with fibrous connective tissue that formed IC on canal dentin walls. The IC may also form a bridge at the apex, in the apical third or midthird of the canal. The root length in many cases was increased by the growth of cementum. The generation of apical cementum or IC may occur despite the presence of inflammatory infiltration at the apex or in the canal. These cementum or cementum-like tissues were similar to cellular cementum. Bone or bone-like tissue was observed in the canal space in many cases and is termed intracanal bone (IB). Connective tissue similar to periodontal ligament was also present in the canal space surrounding the IC and/or IB. Our findings explained in part why many clinical cases of immature teeth with apical periodontitis or abscess may gain root thickness and apical length after conservative treatment with the revitalization procedure.
              Article 0 comments Cited 81 times – based on 0 reviews     Review now
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                Author and article information

                Contributors
                Ruitang Shi: shiruitang@126.com
                Haoqing Yang: 444167838@qq.com
                Xiao Lin: dailyl@163.com
                Yangyang Cao: 2385465168@qq.com
                Chen Zhang: 550856635@qq.com
                Zhipeng Fan: +86 10 5709 9313 , zpfan@ccmu.edu.cn
                Benxiang Hou: +86 10 5709 9230 , houbenxiang@gmail.com
                Journal
                Journal ID (nlm-ta): Stem Cell Res Ther
                Journal ID (iso-abbrev): Stem Cell Res Ther
                Title: Stem Cell Research & Therapy
                Publisher: BioMed Central (London )
                ISSN (Electronic): 1757-6512
                Publication date (Electronic): 12 March 2019
                Publication date PMC-release: 12 March 2019
                Publication date Collection: 2019
                Volume: 10
                Electronic Location Identifier: 89
                Affiliations
                [1 ]ISNI 0000 0004 0369 153X, GRID grid.24696.3f, Department of Endodontics, Beijing Stomatological Hospital, School of Stomatology, , Capital Medical University, ; Beijing, China
                [2 ]ISNI 0000 0004 0369 153X, GRID grid.24696.3f, Laboratory of Molecular Signaling and Stem Cells Therapy, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Beijing Stomatological Hospital, School of Stomatology, , Capital Medical University, ; Beijing, China
                [3 ]ISNI 0000 0004 0369 153X, GRID grid.24696.3f, Department of Implant Dentistry, Beijing Stomatological Hospital, School of Stomatology, , Capital Medical University, ; Beijing, China
                Article
                Publisher ID: 1192
                DOI: 10.1186/s13287-019-1192-2
                PMC ID: 6417198
                PubMed ID: 30867055
                SO-VID: 1154e731-aaa8-4671-bbf7-a01be6645bc8
                Copyright © © The Author(s). 2019
                License:

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                Date received : 31 October 2018
                Date revision received : 8 January 2019
                Date accepted : 25 February 2019
                Categories
                Subject: Research
                Custom metadata
                issue-copyright-statement © The Author(s) 2019

                ScienceOpen disciplines: Molecular medicine
                Keywords: hypoxia,dental pulp stem cells (dpscs),coding rna,long noncoding rna (lncrna),microrna (mirna)
                Data availability:
                ScienceOpen disciplines: Molecular medicine
                Keywords: hypoxia, dental pulp stem cells (dpscs), coding rna, long noncoding rna (lncrna), microrna (mirna)

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