Project description:Human periodontal ligament (HPDL) is continuously exposed to mechanical stress in vivo. In this study, in utilizing DNA chips, we analyzed the influences of mechanical stress on the gene expression profile of HPDL cells in vitro. HPDL cells were obtained from extracted first premolars of individuals undergoing tooth extraction for orthodontic treatment. Then, HPDL cells were applied to a stretch apparatus. They were constantly stretched and relaxed at 0.5 Hz for 48hr with 110ï¼ force elongation. After the application of the cyclic tension force, total RNA was extracted. Then, in utilizing the DNA chips (Human Oligo 30K DNA Chip containing almost all human genes in the genome). We analyzed the differences of gene expression between the stretched and the non-stretched control HPDL cells The DNA chip analysis identified 17 up-regulated genes that showed at least 2-fold difference in their relative intensities between the stretched and the control. This result included the genes for the glutamate receptor binding protein: HOMER1, for the growth factor receptor: CNTFR, for the ECM remodeling protein: MMP15, for the protein interacting with calcineurin A: DSCR1, for the cytoskeletal protein: LRRFIP1, for the glutamate receptor: GRIN3A and some novel genes. On the basis of these data, we suggest that mechanical stress, in other words in vivo occlusal force, may affect the functions of HPDL cells Gene expression profiles were constructed using Human Oligo DNA Chip 30K (Hitachi software engineering) in human periodontal ligament cells (The 48-h stretched and the 48-h non-stretched, control).
Project description:This study aimed to investigate the microRNA expression profile of mechnically strained human periodontal ligament-derived stem cells, and SurePrint G3 Human v16 miRNA Array (Agilent) was employed as a screening platform. We discovered 39 differentially expressed microRNAs between the stretched and the static control group.
Project description:This study aimed to investigate the microRNA expression profile of mechnically strained human periodontal ligament-derived stem cells, and SurePrint G3 Human v16 miRNA Array (Agilent) was employed as a screening platform. We discovered 39 differentially expressed microRNAs between the stretched and the static control group. Human periodontal ligament-derived stem cells were cultured on elastic silicone membranes and either subjected to a dynamic mechanical strain protocol (2hr, 5% elongation, 0.5Hz) or left undisturbed. Total RNA was collected and extracted by TRIzol. RNA samples with 28S/18S ratios in the range of 1.4 to 1.8 were used for microRNA microarray analysis using the Agilent SurePrint G3 Human v16 miRNA Array Kit. The samples in each group were triplicated.
Project description:In this study, the effect of a storage medium (hK-HTCM) in which hair keratin was dissolved in a 1:1 mixed solution of Histidine-Tryptophan-Ketoglutarate and Culture media solution (HTCM) was evaluated on the viability and proliferation of human periodontal ligament cells. There was no difference in cytotoxicity between 0.1% and 0.25% hK-HTCM against 0% hK-HTCM and human periodontal ligament cells. Human periodontal ligament cells were cultured in 0.1% and 0.25% hK-HTCM for 48 hours, and after removing hair keratin from the culture medium, the cells resumed proliferation. When exposed to 0.25% hK-HTCM, human periodontal ligament cells showed differential expression of genes related to cell cycle and cell division regulation. On the other hand, differential expression of genes related to phosphorylation and ubiquitination related to cell cycle resumption was observed in human periodontal ligament cells after removal of 0.25% hK-HTCM. 0.25% hK-HTCM showed the ability to regulate the cell cycle of human periodontal ligament cells without showing cytotoxicity, and its potential to be used as a long-term storage medium for avulsed teeth was confirmed.
Project description:Differentially expressed long noncoding RNA expression between periodontal ligament stem cells from healthy periodontal tissue and periodontal ligament stem cells from inflammatory periodontal tissue.
Project description:Periodontitis can impair the osteogenic differentiation of human periodontal mesenchymal stem cells, but the underlying molecular mechanisms are still poorly understood. Long noncoding RNAs (lncRNAs) have been demonstrated to play significant roles under both physiologic and pathological conditions. We performed comprehensive lncRNAs profiling by lncRNA microarray to identify differentially expressed long noncoding RNA expression between Periodontal ligament stem cells from healthy Periodontal tissue and periodontal ligament stem cells from inflammatory periodontal tissue. Our analysis identified 233 lncRNAs and 423 mRNAs that were differently expressed (fold change >2.0, p-value < 0.05) between the two groups of cells. The GO analysis revealed that the significantly down-regulated biological processes included multicellular organismal process, developmental process and multicellular organismal development and the significantly up-regulated biological processes included cellular process, biological regulation and response to stimulus in periodontal ligament stem cells from inflammatory periodontal tissue. The Pathway analysis revealed that the differentially expressed mRNAs may involved in Focal adhesion, ECM-receptor interaction, Bacterial invasion of epithelial cells, Long-term depression, Circadian entrainment and HIF-1 signaling pathway. Two-condition experiment, periodontal ligament stem cells from healthy periodontal tissue (hPDLSCs) vs. periodontal ligament stem cells from inflammatory periodontal tissue (pPDLSCs), Biological replicates: 3 control replicates (hPDLSCs), 3 testing replicates (pPDLSCs).
Project description:We used microarrays to detect the differences in gene-expression of the periontal ligament between patients with healthy periodontal ligament and patients with periodontitis RNA was extracted directly from the middle third of the human periodontal ligament
Project description:Irisin is recognized as a myokine produced by muscles, regulating metabolism and energy homeostasis, however, it may play a role in many other biological functions. Little is known about its effect on periodontal ligament cells. We employed Affymetrix to profile mRNA expression patterns between 3D human periodontal ligament cell spheroids treated with and without irisin. The mRNA expression profiling identified approximately 1000 mRNAs to be differentially expressed between the two groups, which suggests that irisin is involved in gene regulation in human periodontal ligament cells.