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TGF-β2 downregulates osteogenesis under inflammatory conditions in dental follicle stem cells

International Journal of Oral Science 2018年第4期10卷 232-243页

作者： Soyoun Um Joo-Hee Lee Byoung-Moo Seo Biotooth Engineering Lab Dental Research InstituteDental Regenerative BiotechnologyDepartment of Dental ScienceSchool of DentistrySeoul National University Biotooth Engineering Lab Department of Oral and Maxillofacial Surgery and Craniomaxillofacial Life ScienceDental Research InstituteSchool of DentistrySeoul ational University

Bone formation is important for the reconstruction of bone-related structures in areas that have been damaged by *** conditions such as those that occur in patients with rheumatoid arthritis, cystic fibrosis, and periodontitis have been shown to inhibit osteoblastic differentiation. This study focussed on dental follicle stem cells(DFSCs), which are found in developing tooth germ and participate in the reconstruction of alveolar bone and periodontal tissue in periodontal disease. After bacterial infection of inflamed dental tissue, the destruction of bone was observed. Currently, little is known about the relationship between the inflammatory environment and bone formation. Osteogenic differentiation of inflamed DFSCs resulted in decreased alkaline phosphatase(ALP) activity and alizarin red S staining compared to normal DFSCs. Additionally, in vivo transplantation of inflamed and normal DFSCs demonstrated severe impairment of osteogenesis by inflamed DFSCs. Protein profile analysis via liquid chromatography coupled with tandem mass spectrometry was performed to analyse the differences in protein expression in inflamed and normal tissue. Comparison of inflamed and normal DFSCs showed significant changes in the level of expression of transforming growth factor(TGF)-β2. Porphyromonas gingivalis(P.g.)-derived lipopolysaccharide(LPS) was used to create in vitro inflammatory conditions similar to periodontitis. The osteogenic differentiation of LPS-treated DFSCs was suppressed, and the cells displayed low levels of TGF-β1 and high levels of TGF-β2. DFSCs treated with TGF-β2 inhibitors showed significant increases in alizarin red S staining and ALP activity. TGF-β1 expression was also increased after inhibition of TGF-β2. By examining inflamed DFSCs and LPS-triggered DFSCs, these studies showed both clinically and experimentally that the increase in TGF-β2 levels that occurs under inflammatory conditions inhibits bone formation.

关键词： TGF downregulates osteogenesis under inflammatory conditions in dental follicle stem cells

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Micromolar sodium fluoride mediates anti-osteoclastogenesis in Porphyromonas gingivalis-induced alveolar bone loss

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International Journal of Oral Science 2015年第4期7卷 242-249页

作者： Ujjal K Bhawal Hye-Jin Lee Kazumune Arikawa Michiharu Shimosaka Masatoshi Suzuki Toshizo Toyama Takenori Sato Ryota Kawamata Chieko Taguchi Nobushiro Hamada Ikuo Nasu Hirohisa Arakawa Koh Shibutani department of Biochemistry and Molecular Biology Nihon University School of Dentistry at Matsudo research institute of Oral Health Nihon University School of Dentistry at Matsudo department of Oral Health Graduate School of DentistryKanagawa Dental University department of dental Hygiene Howon University department of Preventive and Public Oral Health Nihon University School of Dentistry at Matsudo department of Anesthesiology Nihon University School of Dentistry at Matsudo department of Infection Control Division of MicrobiologyKanagawa Dental University department of Radiopraxis Science Kanagawa Dental University

Osteoclasts are bone-specific multinucleated cells generated by the differentiation of monocyte/macrophage lineage precursors. Regulation of osteoclast differentiation is considered an effective therapeutic approach to the treatment of bone-lytic diseases. Periodontitis is an inflammatory disease characterized by extensive bone resorption. In this study, we investigated the effects of sodium fluoride （NaF） on osteoclastogenesis induced by Porphyromonas gingivalis, an important colonizer of the oral cavity that has been implicated in periodontitis. NaF strongly inhibited the P. gingivalis-induced alveolar bone loss. That effect was accompanied by decreased levels of cathepsin K, interleukin （IL）-1β, matrix metalloproteinase 9 （MMP9）, and tartrate-resistant acid phosphatase, which were up-regulated during P. gingivalis-induced osteoclastogenesis. Consistent with the in vivo anti-osteoclastogenic effect, NaF inhibited osteoclast formation caused by the differentiation factor RANKL （receptor activator of nuclear factor KB ligand） and macrophage colony-stimulating factor （M-CSF）. The RANKL-stimulated induction of the transcription factor nuclear factor of activated T cells （NFAT） cl was also abrogated by NaF. Taken together, our data demonstrate that NaF inhibits RANKL-induced osteoclastogenesis by reducing the induction of NFATcl, ultimately leading to the suppressed expression of cathepsin K and MMP9. The in vivo effect of NaF on the inhibition of P. gingivalis-induced osteoclastogenesis strengthens the potential usefulness of NaF for treating periodontal diseases.

关键词： alveolar bone loss osteoclasts Porphyromonas gingivalis sodium fluoride

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Chondrogenesis of periodontal ligament stem cells by transforming growth factor-β3 and bone morphogenetic protein-6 in a normal healthy impacted third molar

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International Journal of Oral Science 2013年第1期5卷 7-13页

作者： Sunyoung Choi Tae-Jun Cho Soon-Keun Kwon Gene Lee Jaejin Cho Laboratory of Developmental Biology and Stem Cell Differentiation/Transplantation Department of Dental Regenerative BiotechnologySchool of DentistrySeoul National University dental research institute School of DentistrySeoul National University Laboratory of Molecular Genetics School of DentistrySeoul National University

The periodontal ligament-derived mesenchymal stem cell is regarded as a source of adult stem cells due to its ***, the proof of chondrogenic potential of the cells is ***,we investigated the chondrogenic differentiation capacity of periodontal ligament derived mesenchymal stem cells induced by transforming growth factor(TGF)-p3 and bone morphogenetic protein(BMP)-*** isolation of periodontal ligament stem cells(PDLSCs) from human periodontal ligament,the cells were cultured in Dulbecco’s modified Eagle’s medium(DMEM) with 20%fetal bovine serum(FBS).A mechanical force initiated chondrogenic differentiation of the *** chondrogenic differentiation,10μg·LTGF-β3 or 100μg·LBMP-6 and the combination treating group for synergistic effect of the growth *** analyzed the PDLSCs by fluorescence-activated cell sorting and chondrogenesis were evaluated by glycosaminoglycans assay,histology,immunohistochemistry and genetic *** showed mesenchymal stem cell properties proved by FACS *** contents were increased 217%by TGF-β3 and 220%by BMP-6. The synergetic effect of TGF-β3 and BMP-6 were shown up to 281%compared to *** combination treatment increased Sox9, aggrecan and collagen II expression compared with not only controls,but also TGF-P3 or BMP-6 single treatment *** histological analysis also indicated the chondrogenic differentiation of PDLSCs in our *** results of the present study demonstrate the potential of the dental stem cell as a valuable cell source for chondrogenesis,which may be applicable for regeneration of cartilage and bone fracture in the field of cell therapy.

关键词： bone morphogenetic protein-6 chondrogenesis growth factor periodental ligament cell stem cell transforming growth factor-β3

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The photodynamic therapy on Streptococcus mutans biofilms using erythrosine and dental halogen curing unit

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International Journal of Oral Science 2012年第4期4卷 196-201页

作者： Young-Ho Lee Ho-Won Park Ju-Hyun Lee Hyun-Woo Seo Si-Young Lee department of pediatric dentistry Graduate SchoolCollege of DentistryGangneung-Wonju National University department of pediatric dentistry College of DentistryGangneung-Wonju National University department of Microbiology and Immunology College of DentistryGangneung-Wonju National University

The purpose of our study was to evaluate the effect of photodynamic therapy （PDT）, using erythrosine as a photosensitizing agent and a dental halogen curing unit as a light source, on Streptococcus mutans in a biofilm phase. The S. mutans biofilms were formed in a 24-well cell culture cluster. Test groups consisted of biofilms divided into four groups： group 1： no photosensitizer or light irradiation treatment （control group）; group 2： photosensitizer treatment alone; group 3： light irradiation alone; group 4： photosensitizer treatment and light irradiation. After treatments, the numbers of colony-forming unit （CFU） were counted and samples were examined by confocal laser scanning fluorescence microscopy （CLSM）. Only group 4 （combined treatment） resulted in significant increases in cell death, with rates of 75% and 55% after 8 h of incubation, and 74% and 42% at 12 h, for biofilms formed in brain-heart infusion （BHI） broth supplemented with 0% or O. 1% sucrose, respectively. Therefore, PDT of S. mutans biofilms using a combination of erythrosine and a dental halogen curing unit, both widely used in dental clinics, resulted in a significant increase in cell death. The PDT effects are decreased in biofilms that form in the presence of sucrose.

关键词： biofilms dental halogen curing unit erythrosine photodynamic therapy Streptococcus mutans

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Influence of glass-based dental ceramic type and thickness with identical shade on the light transmittance and the degree of conversion of resin cement

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International Journal of Oral Science 2018年第1期10卷 44-49页

作者： Soram Oh Su-Mi Shin Hyun-Jung Kim Janghyun Paek Sung-Joon Kim Tai Hyun Yoon Sun-Young Kim department of Conservative dentistry Graduate School Kyung Hee University Seoul Korea department of Prosthodontics School of Dentistry Kyung Hee University Seoul Korea department of dentistry School of Medicine Jeju National University Jeju Korea department of Physics Korea University Seoul Korea and SDepartment of Conservative Dentistry dental research institute School of Dentistry Seoul National University Seoul Korea

The purpose of this study was to assess the influence of the types and thicknesses of glass ceramic plates on light transmittance and compare the degrees of conversion(DC) of resin cement under the ceramic materials. Three ceramic plates with thicknesses of 0.5, 1.0, 2.0, and 4.0 mm were fabricated from each of five commercial ceramic blocks in shade A2: high-translucency and low-translucency IPS Empress CAD(EmpHT and EmpLT); high-translucency and low-translucency IPS *** CAD(EmxHT and EmxLT); and Vita Mark Ⅱ(Vita). The translucency parameter was obtained using a colorimeter. The light transmittance rate was measured using a photodetector attached to an optical power meter. The DC of a resin cement(Variolink N) underneath the ceramic plates was examined by Fourier transform infrared spectroscopy. The translucency parameter, light transmittance rate, and DC showed significant differences by ceramic type and thickness(P 0.05). Both type and thickness of the glass ceramics significantly influenced the light transmittance and DC of the light-cured resin cement beneath the ceramic of the same shade.

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Dopaminergic effects on in vitro osteogenesis

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Bone research 2015年第3期3卷 130-139页

作者： Dong Joon Lee Henry C Tseng Sing Wai Wong Zhengyan Wang Meng Deng Ching-Chang Ko NC Oral Health institute School of Dentistry University of North Carolina Duke Eye Center and department of Ophthalmology Duke University Medical Center Oral Biology Program School of Dentistry University of North Carolina department of pediatric dentistry School of Dentistry University of North Carolina department of Orthodontics School of Dentistry University of North Carolina

Multiple growth factors（e.g., BMP2, TGF-b1, FGF2） and isolated genes have been shown to improve osteoblastic proliferation and mineralization, advancing bone tissue engineering. Among these factors, both polydopamine（PDA） and dopamine（DA） monomer have recently been reported to increase osteoblast proliferation and mineralization in vitro. Although a well-characterized neurotransmitter, DA＇s role in the bone is unknown. We hypothesize that DA can directly act on osteoblasts, and examined whether osteoblasts express DA receptors that respond to exogenous DA. m RNAs and protein cell lysates were obtained from MC3T3-E1 cells during osteogenic differentiation phase. Reverse transcription polymerase chain reaction and western blot analysis were used to examine the expression of DA receptors, D1–D5. Dose-response effect and time course of DA treatment on cell proliferation, mineralization, and osteogenic differentiation were investigated at pre-determined days. Real-time PCR was performed to investigate whether DA affects osteogenic gene expression（ALP, BSP, OC, OSX, RUNX2, and Collagen1a2） with or without receptor antagonists（SCH233390 and GR103691）. Two-way ANOVA was used for statistical analysis. All five DA receptors（D1, D2, D3, D4, and D5） m RNAs and proteins were expressed in MC3T3-E1 cells. DA treatment increased cell proliferation for up to 7 days（P, 0.05）. Osteogenic mineralization was significantly greater in the DA-treated group than control group（P, 0.05）. Finally, expression of all the osteogenic genes was inhibited by DA receptor antagonists for D1, D3, and D5. Our findings suggest that MC3T3-E1 osteoblasts express functional DA receptors that enhance proliferation and mineralization. PDA is not biologically inert and has important implications in orthopedic applications. Furthermore, osteoblast differentiation might be regulated by the nervous system, presumably during bone development, remodeling, or repair.

关键词： osteogenic differentiation cell proliferation polymerase chain reaction bone tissue engineering western blot analysis statistical analysis bone development gene expression nervous system growth factors in vitro

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A simple,sensitive and non-destructive technique for characterizing bovine dental enamel erosion:attenuated total reflection Fourier transform infrared spectroscopy

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International Journal of Oral Science 2016年第1期8卷 54-60页

作者： In-Hye Kim Jun Sik Son Bong Ki Min Young Kyoung Kim Kyo-Han Kim Tae-Yub Kwon department of dental Science Graduate SchoolKyungpook National UniversityDaeguKorea Korea Textile Development institute DaeguKorea Center for research Facilities Yeungnam UniversityGyeongsanKorea department of Conservative dentistry School of DentistryKyungpook National UniversityDaeguKorea department of dental Biomaterials School of DentistryKyungpook National UniversityDaeguKorea

Although many techniques are available to assess enamel erosion in vitro, a simple, non-destructive method with sufficient sensitivity for quantifying dental erosion is required. This study characterized the bovine dental enamel erosion induced by various acidic beverages in vitro using attenuated total reflection Fourier transform infrared （ATR-FTIR） spectroscopy. Deionized water （control） and 10 acidic beverages were selected to study erosion, and the pH and neutralizable acidity were measured. Bovine anterior teeth （110） were polished with up to 1 200-grit silicon carbide paper to produce flat enamel surfaces, which were then immersed in 20 mL of the beverages for 30 min at 37 ℃. The degree of erosion was evaluated using ATR-FTIR spectroscopy and Vickers＇ microhardness measurements. The spectra obtained were interpreted in two ways that focused on the ~1, ~3 phosphate contour： the ratio of the height amplitude of ~3 P04 to that of/11 P04 （Method 1） and the shift of the v3 P04 peak to a higher wavenumber （Method 2）. The percentage changes in microhardness after the erosion treatments were primarily affected by the pH of the immersion media. Regression analyses revealed highly significant correlations between the surface hardness change and the degree of erosion, as detected by ATR-FTIR spectroscopy （P〈0.001）. Method 1 was the most sensitive to these changes, followed by surface hardness change measurements and Method 2. This study suggests that ATR- FTIR spectroscopy is potentially advantageous over the microhardness test as a simple, non-destructive, sensitive technique for the quantification of enamel erosion.

关键词： acidic beverage enamel erosion Fourier transform infrared spectroscopy microhardness sensitivity

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Distribution and relative activity of matrix metalloproteinase-2 in human coronal dentin

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International Journal of Oral Science 2011年第4期3卷 192-199页

作者： Lee W. Boushell Masaru Kaku Yoshiyuki Mochida Mitsuo Yamauchi department of Operative dentistry School of Dentistry University of North Carolina department of Bio-Prosthodontics Medical and Dental Science Niigata University Graduate School Chuo-ku department of Periodontology and Oral Biology Division of Oral Biology Boston University Henry M. Goldman School of Dental Medicine Center for Advanced Biomedical Research dental research Center School of Dentistry University of North Carolina

The presence of matrix metalloproteinase-2 （MMP-2） in dentin has been reported, but its distribution and activity level in mature human coronal dentin are not well understood. The purpose of this study was to determine the MMP-2 distribution and relative activity in demineralized dentin. Crowns of twenty eight human molars were sectioned into inner （ID）, middle （MD）, and outer dentin （OD） regions and demineralized. MMP-2 was extracted with 0.33 mol·L-1 EDTA/2 mol·L-1 guanidine-HCl, pH 7.4, and MMP-2 concentration was estimated with enzyme-linked immunoabsorbant assay （ELISA）. Further characterization was accomplished by Western blotting analysis and gelatin zymography. The mean concentrations of MMP-2 per mg dentin protein in the dentin regions were significantly different （P=0.043）： 0.9 ng （ID）, 0.4 ng （MD）, and 2.2 ng （OD）, respectively. The pattern of MMP-2 concentration was OD〉ID〉MD. Western blotting analysis detected -66 and -72 kDa immunopositive proteins corresponding to pro- and mature MMP-2, respectively, in the ID and MD, and a -66 kDa protein in the OD. Gelatinolytic activity consistent with MMP-2 was detected in all regions. Interestingly, the pattern of levels of Western blot immunodetection and gelatinolytic activity was MD〉ID〉OD. The eoneentration of MMP-2 in human coronal dentin was highest in the region of dentin that contains the dentinoenamel junction and least in the middle region of dentin. However, levels of Western blot immunodetection and gelatinolytic activity did not correlate with the estimated regional concentrations of MMP-2, potentially indicating region specific protein interactions.

关键词： matrix metalloproteinase-2 human coronal dentin distribution gelatinolytic activity

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Magnesium/Nano-hydroxyapatite Composite for Bone Reconstruction: The Effect of Processing Method

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Journal of Bionic Engineering 2020年第1期17卷 92-99页

作者： Mohammad Khodaei Farahnaz Nejatidanesh Mohammad Javad Shirani Srinivasan Iyengar Hossein Sina Omid Savabi department of Materials Science and Engineering Golpayegan University of TechnologyGolpayegan ***Iran dental Materials research Center Dental Research InstituteSchool of DentistryIsfahan University of Medical SciencesIsfahan ***Iran Post graduate student of Prosthodontics Student Research CommitteeSchool of DentistryIsfahan University of Medical SciencesIsfahan ***Iran Materials Engineering Division Faculty of EngineeringLund UniversityLund 11822100Sweden European Spallation Source ERIC Lund 17622100Sweden dental research Center Dental Research InstituteSchool of DentistryIsfahan University of Medical SciencesIsfahan ***Iran

Nano-ceramic particles can serve as reinforcing agents for metallic materials to improve their mechanical ***,it is important to ensure chemical compatibility between the matrix and *** the present study,magnesium composites with and without nano-hydroxyapatite(nHA)particles were fabricated for bone reconstruction *** different techniques were used,Conventional Sintering(CS)of powder compacts and Spark Plasma Sintering(SPS)of pre-compacted *** showed that a 10 wt%addition of nHA particles to magnesium,followed by SPS improved the compression strength by 27%.CS did not lead to any significant improvement compared to SPS processing.X-ray diffraction data after CS revealed the formation of unfavorable phases due to chemical reactions between nHA particles and the magnesium matrix,while these phases were absent after SPS *** mechanical properties of the specimens fabricated by CS were much inferior to those processed using *** shorter processing time associated with SPS leaded to reduced interaction between nHA particles and the Mg-matrix,compared to CS.

关键词： magnesium nano-hydroxyapatite composite materials in situ reaction biomedical-materials

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Human periodontal ligament stem cells repair mental nerve injury

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Neural Regeneration research 2013年第30期8卷 2827-2837页

作者： Bohan Li Hun-Jong Jung Soung-Min Kim Myung-Jin Kim Jeong Won Jahng Jong-Ho Lee department of Oral and Maxillofacial Surgery School of Dentistry Binzhou Medical College department of Occupation and Environment Konkuk Postgraduate Medical School department of Oral and Maxillofacial Surgery School of Dentistry Seoul National University dental research institute Seoul National University

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 × 106） were injected into the crush-injured left mental nerve in rats. Simultaneously, autologous Schwann cells （1 × 106） 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 sig-nificantaly 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 in-jection, 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.

关键词： neural regeneration peripheral nerve injury stem cells periodontal ligament stem cells mentalnerve Schwann cells cell transplantation sensory nerve neurotrophic factor neuroregeneration

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