In Vitro Cellular & Developmental Biology - Animal

, Volume 51, Issue 8, pp 797–807 | Cite as

Integrated miRNA and mRNA expression profiling of tension force-induced bone formation in periodontal ligament cells

  • Maolin Chang
  • Heng Lin
  • Meng Luo
  • Jie Wang
  • Guangli Han
Article

Abstract

Tension force-induced bone formation is a complex biological process altered by various factors, for example miRNAs and gene regulatory network. However, we know little about critical gene regulators and their functional consequences on this complex process. The aim of this study was to determine the integrated relation between microRNA and mRNA expression in tension force-induced bone formation in periodontal ligament cells by a system biological approach. We identified 818 mRNAs and 32 miRNAs differentially expressed between cyclic tension force-stimulated human periodontal ligament cells and control cells by microarrays. By using miRNA/mRNA network analysis, protein-protein interactions network analysis, and hub analysis, we found that miR-195-5p, miR-424-5p, miR-1297, miR-3607-5p, miR-145-5p, miR-4328, and miR-224-5p were core microRNAs of tension force-induced bone formation. WDR33, HSPH1, ERBB3, RIF1, IKBKB, CREB1, FGF2, and PAG1 were identified as hubs of the PPI network, suggesting the biological significance in this process. The miRNA expression was further examined in human PDLC and animal samples by using quantitative real-time PCR. Thus, we proposed a model of tension force-induced bone formation which is co-regulated through integration of the miRNA and mRNA. This study illustrated the benefits of system biological approaches in the analysis of tension force-induced bone formation as a complex biological process. We used public information and our experimental data to do comprehensive analysis and revealed the coordination transcriptional control of miRNAs of tension force-induced bone formation.

Keywords

MiRNA Gene expression profiling Bone formation Network Orthodontic tooth movement 

Supplementary material

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ESM 1(XLS 3517 kb)
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Supplementary Fig. 1

Description of in vivo orthodontic force applying system. (a) Mice positioned in the supine position on a surgical table. (b) Surgical table placed under a stereomicroscope with light system. (c) Use of a mouth-opener in a mouse. (d) Occlusal view of the maxillary molar region. The distal end of a nickel–titanium open-coil spring (Tomy, Japan) was bonded to the occlusal surface of the right first maxillary molar using a light-cured resin. (e) A tension gauge was attached to the surgical table to show the force magnitude. (f) The coil spring bonded to both upper incisors. Stereomicroscope (S), mouth-opener (O), coil spring (C), upper first molar (M), tension gauge (T). (JPEG 1223 kb)

11626_2015_9892_Fig7_ESM.jpg (354 kb)
Supplementary Fig. 2

Validation of differentially expressed mRNAs in vivo. (a-h) The mRNAs expression of WDR33, RIF1, IKBKB, CREB1, FGF2, PAG1 and HSPH1 at tension sites of orthodontic tooth movement mice molar at day 3 was determined by real-time PCR. β-actin was used as a normalization control. All data were based on three independent experiments. Asterisks indicate statistical significance between experimental and control groups at P < 0.05. (JPEG 354 kb)

11626_2015_9892_Fig8_ESM.jpg (915 kb)
Supplementary Fig. 3

Strongly expression of FGF2 and CREB1 at tension sites of orthodontic tooth movement mice molar. (a) FGF2 expression was detected by immunofluorescence staining. FGF2 was strongly localized at tension site of first molar but were weakly localized at the contralateral sites at day 3. TS: tension site; CS: contralateral site. (Original magnification, 200×) (b) CREB1 expression was detected by immunohistochemistry staining. CREB1 was strongly localized at tension sites of first molar but were weakly localized at the compression sites at day 3. Scale bar for A, 200 μm; scale bar for B and C, 100 μm. TS: tension site; CS: compression site. (JPEG 914 kb)

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Copyright information

© The Society for In Vitro Biology 2015

Authors and Affiliations

  • Maolin Chang
    • 1
  • Heng Lin
    • 1
  • Meng Luo
    • 1
  • Jie Wang
    • 1
  • Guangli Han
    • 1
  1. 1.State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory for Oral Biomedicine of Ministry of Education (KLOBM), School and Hospital of StomatologyWuhan UniversityWuhanChina

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