Fibulin-5 contributes to microfibril assembly in human periodontal ligament cells.
Ontology highlight
ABSTRACT:
SUBMITTER: Hisanaga Y
PROVIDER: S-EPMC2775106 | biostudies-literature | 2009 Oct
REPOSITORIES: biostudies-literature
Publications
Fibulin-5 contributes to microfibril assembly in human periodontal ligament cells.
Hisanaga Yutaka Y Nakashima Kazuki K Tsuruga Eichi E Nakatomi Yuka Y Hatakeyama Yuji Y Ishikawa Hiroyuki H Sawa Yoshihiko Y
Acta histochemica et cytochemica 20091001 5
The elastic system fibers comprise oxytalan, elaunin and elastic fibers, which differ in their relative microfibril and elastin content. Human periodontal ligaments (PDL) contain only oxytalan fibers (pure microfibrils) among them. Since fibulin-5 regulates the organization of elastic fibers to link the fibers to cells, we hypothesized that fibulin-5 may contribute to the formation of oxytalan fibers. We used siRNA for fibulin-5 in PDL cell culture to examine the extracellular deposition of fibr ...[more]
Similar Datasets
Project description:Understanding the sequence of events from undifferentiated stem cells to neuron is not only important for the basic knowledge of stem cell biology, but also for therapeutic applications. In this study we examined the sequence of biological events during neural differentiation of human periodontal ligament stem cells (hPDLSCs). Here, we show that hPDLSCs-derived neural-like cells display a sequence of morphologic development highly similar to those reported before in primary neuronal cultures derived from rodent brains. We observed that cell proliferation is not present through neurogenesis from hPDLSCs. Futhermore, we may have discovered micronuclei movement and transient cell nuclei lobulation coincident to in vitro neurogenesis. Morphological analysis also reveals that neurogenic niches in the adult mouse brain contain cells with nuclear shapes highly similar to those observed during in vitro neurogenesis from hPDLSCs. Our results provide additional evidence that it is possible to differentiate hPDLSCs to neuron-like cells and suggest the possibility that the sequence of events from stem cell to neuron does not necessarily requires cell division from stem cell.
| S-EPMC6888846 | biostudies-literature
Project description:BackgroundThe periodontal ligament is essential for homeostasis of periodontal tissue. A hypoxic milieu of the periodontal tissue is generated under periodontitis or during orthodontic treatment, which affects the periodontal and bone remodelling process. Here, we provide a comprehensive proteomic characterization of periodontal ligament cells under hypoxic conditions, aiming to reveal previously unappreciated biological changes and to help advance hypoxia-based therapeutic strategies for periodontal diseases.MethodsHuman periodontal ligament cells (hPDLCs) were characterized using immunohistochemistry (IHC) and flow cytometry (FACS). Successful hypoxia treatment of hPDLCs with 1% O2 was confirmed by qRT-PCR. Proliferation was evaluated using an MTT assay. The proteomic expression profile under hypoxia was studied with the isobaric tags for relative and absolute quantification (iTRAQ) approach followed by protein identification and bioinformatic analysis, and western blot verification was performed.ResultsThe hPDLCs were positive for vimentin, CD73 and CD105 and negative for keratin, CD34 and CD45. After hypoxia treatment, the mRNA expression of hypoxia-inducible factor 1a (HIF1a) was upregulated. The proliferation rate was elevated during the first 6 h but decreased from 6 h to 72 h. A total of 220 differentially expressed proteins were quantified in hPDLCs under hypoxia (1% O2, 24 h), including 153 upregulated and 67 downregulated proteins, five of which were verified by western blot analysis. The Gene Ontology enriched terms included the energy metabolic process, membrane-bound organelle and vesicle, and protein binding terms. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated several involved pathways, including glycolysis/gluconeogenesis, biosynthesis of amino acids, the HIF-1 signalling pathway, and focal adhesion. A protein-protein interaction (PPI) network demonstrated the dominant role of autophagy over apoptosis under hypoxia.ConclusionThe proteomic profile of hPDLCs under hypoxia was mainly related to energy metabolism, autophagy, and responses to stimuli such as adhesion and inflammation. Previously unrecognized proteins including solute carrier family proteins, heat shock proteins, ubiquitination-related enzymes, collagen and S100 family proteins are involved in adaptive response to hypoxia in hPDLCs and are thus of great research interest in future work.
| S-EPMC6717648 | biostudies-literature
Project description:The periodontal ligament (PDL) is highly ordered connective tissue located between the alveolar bone and cementum. An aligned and organized architecture is required for its physiological function. We applied micropatterning technology to arrange PDL cells in 10- or 20-μm-wide extracellular protein patterns. Cell and nuclear morphology, cytoskeleton, proliferation, differentiation, and matrix metalloproteinase system expression were investigated. Micropatterning clearly elongated PDL cells with a low cell-shape index and low spreading area. The nucleus was also elongated as nuclear height increased, but the nuclear volume remained intact. The cytoskeleton was rearranged to form prominent bundles at cells' peripheral regions. Moreover, proliferation was promoted by 10- and 20-μm micropatterning. Osteogenesis and adipogenesis were each inhibited, but micropatterning increased PDL cells' stem cell markers. β-catenin was expelled to cytoplasm. YAP/TAZ nuclear localization and activity both decreased, which might indicate their role in micropatterning-regulated differentiation. Collagen Ι expression increased in micropatterned groups. It might be due to the decreased expression of matrix metalloproteinase-1, 2 and the tissue inhibitor of metalloproteinase-1 gene expression elevation in micropatterned groups. The findings of this study provide insight into the effects of a micropatterned surface on PDL cell behavior and may be applicable in periodontal tissue regeneration.
| S-EPMC5937357 | biostudies-literature
Project description:Human periodontal ligament stem cells are easily accessible and can differentiate into Schwann cells. We hypothesized that human periodontal ligament stem cells can be used as an alternative source for the autologous Schwann cells in promoting the regeneration of injured peripheral nerve. To validate this hypothesis, human periodontal ligament stem cells (1 × 10(6)) were injected into the crush-injured left mental nerve in rats. Simultaneously, autologous Schwann cells (1 × 10(6)) and PBS were also injected as controls. Real-time reverse transcriptase polymerase chain reaction showed that at 5 days after injection, mRNA expression of low affinity nerve growth factor receptor was significantaly increased in the left trigeminal ganglion of rats with mental nerve injury. Sensory tests, histomorphometric evaluation and retrograde labeling demonstrated that at 2 and 4 weeks after injection, sensory function was significantly improved, the numbers of retrograde labeled sensory neurons and myelinated axons were significantly increased, and human periodontal ligament stem cells and autologous Schwann cells exhibited similar therapeutic effects. These findings suggest that transplantation of human periodontal ligament stem cells show a potential value in repair of mental nerve injury.
| S-EPMC4146018 | biostudies-literature
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 110％ 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
2009-11-21 | GSE19118 | GEO
Project description:Abstract Background Periodontitis is a common oral inflammatory disease, and lipopolysaccharide (LPS) is a key risk factor in periodontitis pathology. Here, we used LPS‐induced periodontal ligament cells (PDLCs) to explore the molecular mechanism of periodontitis. Methods Cell viability, proliferation, and apoptosis were analyzed by Cell Counting Kit‐8, 5‐ethynyl‐20‐deoxyuridine (EDU), and flow cytometry assays, respectively. Apart from that, their targeting relationship was validated using dual‐luciferase reporter and RNA‐pull down. Results Circular RNA_0138960 (circ_0138960) was notably upregulated in periodontitis sufferers (p < .001) and LPS‐disposed PDLCs (p < .05). LPS exposure dampened PDLC proliferation, and promoted apoptosis and inflammation (p < .05). Circ_0138960 acted as a microRNA sponge for miR‐518a‐5p to affect histone deacetylase 6 (HDAC6) expression. Circ_0138960 absence‐mediated protective effects in LPS‐induced PDLCs were largely abrogated via silencing miR‐518a‐5p or HDAC6 overexpression (p < .05). Conclusion Circ_0138960 promoted LPS‐induced dysfunction in PDLCs by targeting miR‐518a‐5p/HDAC6 axis, which provided novel potential therapeutic targets for periodontitis. Please designate which figure should be used for the graphical table of contents and considered for the publication cover.
| S-EPMC9639461 | biostudies-literature
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).
2010-06-19 | E-GEOD-19118 | biostudies-arrayexpress
Project description:Periodontal ligament (PDL) stem cells (PDLSCs) have been reported as a useful cell source for periodontal tissue regeneration. However, one of the issues is the difficulty of obtaining a sufficient number of PDLSCs for clinical application because very few PDLSCs can be isolated from PDL tissue of donors. Therefore, we aimed to identify a specific factor that converts human PDL cells into stem-like cells. In this study, microarray analysis comparing the gene profiles of human PDLSC lines (2-14 and 2-23) with those of a cell line with a low differentiation potential (2-52) identified the imprinted gene mesoderm-specific transcript (MEST). MEST was expressed in the cytoplasm of 2-23 cells. Knockdown of MEST by siRNA in 2-23 cells inhibited the expression of stem cell markers, such as CD105, CD146, p75NTR, N-cadherin, and NANOG; the proliferative potential; and multidifferentiation capacity for osteoblasts, adipocytes, and chondrocytes. On the other hand, overexpression of MEST in 2-52 cells enhanced the expression of stem cell markers and PDL-related markers and the multidifferentiation capacity. In addition, MEST-overexpressing 2-52 cells exhibited a change in morphology from a spindle shape to a stem cell-like round shape that was similar to 2-14 and 2-23 cell morphologies. These results suggest that MEST plays a critical role in the maintenance of stemness in PDLSCs and converts PDL cells into PDLSC-like cells. Therefore, this study indicates that MEST may be a therapeutic factor for periodontal tissue regeneration by inducing PDLSCs.
| S-EPMC7366229 | biostudies-literature
Project description:Periodontal ligament cells (PDLCs), which have potential for multilineage differentiation, are candidates for use in regeneration of periodontal tissue defects; however, our understanding of the mechanisms underlying the lineage commitment of PDLCs remains limited. C4orf7, which is specifically expressed in the periodontal ligament (PDL) tissue, may be crucial in deciding the fate of PDLCs and regulating the periodontal bone balance. In this study, we examined the expression of C4orf7 in PDL tissue, using immunohistochemical staining. We transfected PDLCs with lentiviral vectors expressing C4orf7 and examined the effect of C4orf7 on the balance of PDLC osteogenic and osteoclastogenic differentiation. Osteogenic induction resulted in the downregulation of mRNA and protein expression levels of the osteogenic/cementoblastic markers: ALP, RUNX2, COL1, OPN, OPG, OSX, IBSP, CAP, and CEMP1. Transfected cells also exhibited an increased RANKL/OPG ratio, which is an indicator of osteoclastogenic differentiation. ALP activity assays and Alizarin red staining confirmed the negative effect of C4orf7 on PDLC osteogenic differentiation. Finally, we investigated the effect of C4orf7 on the lineage commitment of PDLCs to adipocytes. We observed increased expression levels of PPARγ2, GLUT4, ZFP423, FABP4, and LPL mRNAs, as well as a gradual accumulation of lipid droplets in the C4orf7-overexpressing group compared with controls. In summary, our data confirm that C4orf7 has an important role in the regulation of periodontal bone remodeling through promotion of the adipogenic/osteoclastogenic, and inhibition of the osteogenic/cementoblastic, differentiation of PDLCs. Therefore, C4orf7 is a potential therapeutic target for the treatment of periodontal disease and other bone metabolic disorders.
| S-EPMC5752921 | biostudies-literature
Project description:ObjectivesIt has recently been shown that enamel matrix derivative (EMD) components (Fraction C, containing <6 kDa peptides (mainly a 5.3 kDa tyrosine-rich amelogenin peptide (TRAP)), and Fraction A, containing a mixture of >6 kDa peptides (including a leucine-rich amelogenin peptide (LRAP))) differentially regulate osteogenic differentiation of periodontal ligament (PDL) cells. The present study examined whether EMD and the EMD Fractions (i) bind and internalize into PDL cells and (ii) precipitate and form insoluble complexes on PDL cells.Materials and methodsBiotin-labelled EMD/EMD Fractions were incubated with PDL cells under various different culture conditions and confocal and electron microscopies were carried out to examine the binding and intracellular trafficking of these proteins.ResultsThe results reported here show, for the first time, that at least some components in Fraction A and the TRAP peptide in Fraction C can bind and be internalized by human PDL cells via receptor-mediated endocytosis. In addition, Fraction A was found to form insoluble aggregate-like structures on PDL cells, whereas Fraction C was soluble in culture media.ConclusionSoluble amelogenin isoform TRAP appears to be internalizing into a subset of PDL cells. Moreover, TRAP uptake is most likely controlled by receptor-mediated endocytosis.Clinical relevanceInformation on interaction between PDL cells and EMD/TRAP might prove useful in designing targeted interventions (i.e. use of chemically prepared soluble amelogenin peptides) to repair/regenerate periodontal tissues. Such interventions can also (i) avoid the use of rather crude animal-derived enamel matrix protein (EMP)/EMD preparation and (ii) preparation of cost-effective and more controlled chemically synthesized amelogenin peptides for the clinical use.
| S-EPMC4762925 | biostudies-literature