Intermittent compressive force induces cell cycling and reduces apoptosis in embryoid bodies of mouse induced pluripotent stem cells / 国际口腔科学杂志·英文版

In vitro manipulation of induced pluripotent stem cells (iPSCs) by environmental factors is of great interest for three-dimensional (3D) tissue/organ induction. The effects of mechanical force depend on many factors, including force and cell type. However, information on such effects in iPSCs is lacking. The aim of this study was to identify a molecular mechanism in iPSCs responding to intermittent compressive force (ICF) by analyzing the global gene expression profile. Embryoid bodies of mouse iPSCs, attached on a tissue culture plate in 3D form, were subjected to ICF in serum-free culture medium for 24 h. Gene ontology analyses for RNA sequencing data demonstrated that genes differentially regulated by ICF were mainly associated with metabolic processes, membrane and protein binding. Topology-based analysis demonstrated that ICF induced genes in cell cycle categories and downregulated genes associated with metabolic processes. The Kyoto Encyclopedia of Genes and Genomes database revealed differentially regulated genes related to the p53 signaling pathway and cell cycle. qPCR analysis demonstrated significant upregulation of Ccnd1, Cdk6 and Ccng1. Flow cytometry showed that ICF induced cell cycle and proliferation, while reducing the number of apoptotic cells. ICF also upregulated transforming growth factor β1 (Tgfb1) at both mRNA and protein levels, and pretreatment with a TGF-β inhibitor (SB431542) prior to ICF abolished ICF-induced Ccnd1 and Cdk6 expression. Taken together, these findings show that TGF-β signaling in iPSCs enhances proliferation and decreases apoptosis in response to ICF, that could give rise to an efficient protocol to manipulate iPSCs for organoid fabrication.

Dysregulation of Notch signaling related genes in oral lichen planus

Objective To investigate a dysregulation of Notch signaling in oral lichen planus (OLP) using public available microarray dataset. Methods A mRNA expression profiling dataset from Gene Expression Omnibus was downloaded. Differential gene expression between OLP and normal oral epithelium was examined using NetworkAnalyst. The dysregulated genes related to Notch signaling were identified. Results Thirteen genes in Notch signaling pathway were significantly differential expressed between OLP and normal epithelium. OLP samples significantly increased the mRNA levels of HEYL, APH1B, CNTN1 and PSEN2. Whilst, ITCH, HES1, TLE2, DLK2, DTX2, NOTCH3, JAG2, RFNG, and SPEN were downregulated in OLP groups. Conclusions Notch signaling was dysregulated and may participate in pathophysiologic process in OLP.

Hypoxia Enhances Osteogenic Differentiation in Retinoic Acid-Treated Murine-Induced Pluripotent Stem Cells

Hypoxic condition influences biological responses in various cell types. However, a hypoxic regulating osteogenic differentiation remains controversy. Here, an influence of short-term culture in hypoxic condition on osteogenic marker gene expression by retinoic acid-treated murine gingival fibroblast-derived induced pluripotent stem cells (RA-miPS) was investigated. Results demonstrated that hypoxic condition significantly upregulated Vegf, Runx2, Osx, and Ocn mRNA expression by RA-miPS in normal culture medium at day 3. Further, desferrioxamine significantly downregulated pluripotent marker (Nanog and Oct4) and enhanced osteogenic marker (Runx2, Osx, Dlx5, and Ocn) gene expression as well as promoted in vitro mineral deposition. However, the effect of cobalt chloride on osteogenic differentiation of RA-miPS was not robust. In summary, the results imply that hypoxic condition may be useful in the enhancement of osteogenic differentiation in RA-miPS.

Regulation of osteoprotegerin expression by Notch signaling in human oral squamous cell carcinoma cell line

Objective To investigate the influence of Notch signaling on osteoprotegerin (OPG) expression in a human oral squamous cell carcinoma cell line. Methods Activation of Notch signaling was performed by seeding cells on Jagged1 immobilized surfaces. In other experiments, a γ-secretase inhibitor was added to the culture medium to inhibit intracellular Notch signaling. OPG mRNA and protein were determined by real-time PCR and ELISA, respectively. Finally, publicly available microarray database analysis was performed using connection up- or down-regulation expression analysis of microarrays software. Results Jagged1-treatment of HSC-4 cells enhanced HES1 and HEY1 mRNA expression, confirming the intracellular activation of Notch signaling. OPG mRNA and protein levels were significantly suppressed upon Jagged1 treatment. Correspondingly, HSC-4 cells treated with a γ-secretase inhibitor resulted in a significant reduction of HES1 and HEY1 mRNA levels, and a marked increase in OPG protein expression was observed. These results implied that Notch signaling regulated OPG expression in HSC-4 cells. However, Jagged1 did not alter OPG expression in another human oral squamous cell carcinoma cell line (HSC-5) or a human head and neck squamous cell carcinoma cell line (HN22). Conclusions Notch signaling regulated OPG expression in an HSC-4 cell line and this mechanism could be cell line specific.

Transplantation of cryopreserved teeth: a systematic review / 国际口腔科学杂志·英文版

The aim of this article was to examine the research articles regarding biological and mechanical properties of cryopreserved teeth for potential use in tooth transplantation. A systematic review of literatures was performed by Pubmed searching with assigned key words from January 1, 1990 to June 8, 2009. All articles were examined for inclusion criteria. Secondary search was conducted by hand-search through references of included articles from primary search. A total of 24 articles were obtained from both primary and secondary search and used as fundamental articles in this review. Periodontal ligament tissues of cryopreserved teeth were able to maintain their biological properties resulted in a satisfactory healing of periodontium. Dental pulp tissues, however, may be compromised by limitation of permeability of cryopreservative agent into pulp cavity. Therefore, an endodontic treatment of transplanted cryopreserved teeth was recommended. Cryopreserved teeth had comparable mechanical properties to those of normal teeth. Importantly, the success of cryopreserved tooth transplantation treatment in orthodontic patients was reported. The cryopreserved teeth for tooth banking have a potential clinical application for treatment of missing teeth. Case selection, however, is critical for treatment success. More studies and data regarding masticatory function and periodontal healing of transplanted cryopreserved teeth are needed.