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Bone-conditioned medium contributes to initiation and progression of osteogenesis by exhibiting synergistic TGF-β1/BMP-2 activity

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International Journal of Oral Science 2018年第4期10卷 203-211页

作者： Maria B.Asparuhova Jordi Caballé-Serrano Daniel Buser Vivianne Chappuis Laboratory of Oral Cell Biology School of Dental MedicineUniversity of Bern department of Oral surgery and Stomatology School of Dental MedicineUniversity of Bern department of Oral and maxillofacial surgery School of Dental MedicineUniversitat Internacional de Catalunya

Guided bone regeneration (GBR) often utilizes a combination of autologous bone grafts, deproteinized bovine bone mineral(DBBM), and collagen membranes. DBBM and collagen membranes pre-coated with bone-conditioned medium (BCM) extracted from locally harvested autologous bone chips have shown great regenerative potential in GBR. However, the underlying molecular mechanism remains largely unknown. Here, we investigated the composition of BCM and its activity on the osteogenic potential of mesenchymal stromal cells. We detected a fast and significant (P <0.001) release of transforming growth factor-β1 (TGF-β1) from autologous bone within 10 min versus a delayed bone morphogenetic protein-2 (BMP-2) release from 40 min onwards. BCMs harvested within short time periods (10, 20, or 40 min), corresponding to the time of a typical surgical procedure, significantly increased the proliferative activity and collagen matrix production of BCM-treated cells. Long-term (1, 3, or 6 days)-extracted BCMs promoted the later stages of osteoblast differentiation and maturation. Short-term-extracted BCMs, in which TGF-β1 but no BMP-2was detected, reduced the expression of the late differentiation marker osteocalcin. However, when both growth factors were present simultaneously in the BCM, no inhibitory effects on osteoblast differentiation were observed, suggesting a synergistic TGF-β1/BMP-2 activity. Consequently, in cells that were co-stimulated with recombinant TGF-β1 and BMP-2, we showed a significant stimulatory and dose-dependent effect of TGF-β1 on BMP-2-induced osteoblast differentiation due to prolonged BMP signaling and reduced expression of the BMP-2 antagonist noggin. Altogether, our data provide new insights into the molecular mechanisms underlying the favorable outcome from GBR procedures using BCM, derived from autologous bone grafts.

关键词： Bone-conditioned medium contributes to initiation and progression of osteogenesis by exhibiting synergistic TGF BMP-2 activity

Low-frequency pulsed electromagnetic field pretreated bone marrow-derived mesenchymal stem cells promote the regeneration of crush-injured rat mental nerve

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Neural Regeneration Research 2018年第1期13卷 145-153页

作者： NaRi Seo Sung-Ho Lee Kyung Won Ju JaeMan Woo BongJu Kim SoungMin Kim Jeong Won Jahng Jong-Ho Lee department of Oral and maxillofacial surgery Graduate School of Dentistry Seoul National University Seoul South Korea department of Oral and maxillofacial surgery Seoul National University Dental Hospital Seoul South Korea dental Research Institute Seoul National University Seoul South Korea Clinical Translational Research Center for dental Science (CTRC) Seoul National University Dental Hospital Seoul South Korea

Bone marrow-derived mesenchymal stem cells （BMSCs） have been shown to promote the regeneration of injured peripheral nerves. Pulsed electromagnetic field （PEMF） reportedly promotes the proliferation and neuronal differentiation of BMSCs. Low-frequency PEMF can induce the neuronal differentiation of BMSCs in the absence of nerve growth factors. This study was designed to investigate the effects of low-frequency PEMF pretreatment on the proliferation and function of BMSCs and the effects of low-frequency PEMF pre-treated BMSCs on the regeneration of injured peripheral nerve using in vitro and in vivo *** in vitro experiments, quantitative DNA analysis was performed to determine the proliferation of BMSCs, and reverse transcription-polymerase chain reaction was performed to detect S100 （Schwann cell marker）, glial fibrillary acidic protein （astrocyte marker）, and brain-derived neurotrophic factor and nerve growth factor （neurotrophic factors） mRNA expression. In the in vivo experiments, rat models of crush-injured mental nerve established using clamp method were randomly injected with low-frequency PEMF pretreated BMSCs, unpretreated BMSCs or PBS at the injury site （1 × 106 cells）. DiI-labeled BMSCs injected at the injury site were counted under the fluorescence microscope to determine cell survival. One or two weeks after cell injection, functional recovery of the injured nerve was assessed using the sensory test with von Frey filaments. Two weeks after cell injection, axonal regeneration was evaluated using histomorphometric analysis and retrograde labeling of trigeminal ganglion neurons. In vitro experiment results revealed that low-frequency PEMF pretreated BMSCs proliferated faster and had greater mRNA expression of growth factors than unpretreated BMSCs. In vivo experiment results revealed that compared with injection of unpretreated BMSCs, injection of low-frequency PEMF pretreated BMSCs led to higher myelinated axon count and axon density and more Di

关键词： nerve regeneration mesenchymal stem cells low-frequency pulsed electromagnetic field peripheral nerve injury crush-injured mental nerve

Detection accuracy for epithelial dysplasia using an objective autofluorescence visualization method based on the luminance ratio

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International Journal of Oral Science 2017年第4期9卷 200-209页

作者： nanami yamamoto koji kawaguchi hisako fujihara mitsuhiko hasebe yuta kishi masaaki yasukawa kenichi kumagai yoshiki hamada department of oral and maxillofacial surgery school of dental medicinetsurumi universityYokohamaJapan department of oral hygiene tsurumi junior collegeYokohamaJapan

The autofluorescence visualization method （AVM） uses blue excitation light to assist in the diagnosis of epithelial dysplasia. It detects epithelial dysplasia as a black area, which is known as fluorescence visualization loss （FVL）. In this study, we evaluated the detection accuracy for epithelial dysplasia of the tongue using the objective AVM and assessed its possible clinical utility. Seventy-nine tongue specimens clinically suspected to have leukoplakia or squamous cell carcinoma （SCC） were analyzed. First, the AVM was subjectively performed using the Visually Enhanced Lesion scope （VELscope）, and the iodine-staining method was then performed. After biopsy, the histopathological results and the luminance ratio between the lesion and healthy tissue were compared, and a receiver operating characteristic curve was created. The cutoff value for the objective AVM was determined; the lesion was considered FVL-positive or -negative when the luminance ratio was higher or lower than the cutoff value, respectively. The histopathological diagnoses among the 79 specimens were SCC （n = 30）, leukoplakia with dysplasia （n = 34）, and leukoplakia without dysplasia （n= 15）. The cutoff value of the luminance ratio was 1.62, resulting in 66 FVL-positive and 13 FVL-negative specimens. The luminance ratio was significantly higher in the epithelial dysplasia-positive than -negative group （P〈O.O00 1）. The objective AVM showed much higher consistency between histopathological results than did the two methods （kappa statistic = 0.656）. In conclusion, objective autofluorescence visualization has a potential as an auxiliary method for diagnosis of epithelial dysplasia.

关键词： autofluorescence epithelial dysplasia oral cancer Visually Enhanced Lesion scope

ERK phosphorylation functions in invadopodia formation in tongue cancer cells in a novel silicate fibre-based 3D cell culture system

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International Journal of Oral Science 2018年第4期10卷 253-262页

作者： Masaharu Noi Ken-Ichi Mukaisho Saori Yoshida Shoko Murakami Shinya Koshinuma Takeshi Adachi Yoshisato Machida Masashi Yamori Takahisa Nakayama Gaku Yamamoto Hiroyuki Sugihara department of Oral and maxillofacial surgery Shiga University of Medical Science department of Pathology Division of Molecular Diagnostic PathologyShiga University of Medical Science dental Oral surgery Nagahama Red Cross Hospital

To screen for additional treatment targets against tongue cancer, we evaluated the contributions of extracellular signal-related kinase(ERK), AKT and ezrin in cancer development. Immunohistochemical staining showed that ERK and ezrin expressions were significantly higher in invasive squamous cell carcinoma than in carcinoma in situ. To investigate the roles of ERK and ezrin in cancer development, we used the non-woven silica fibre sheet Cellbedwith a structure resembling the loose connective tissue morphology in a novel 3 D culture system. We confirmed that the 3 D system using CellbedTMaccurately mimicked cancer cell morphology in vivo. Furthermore, cell projections were much more apparent in 3 D-cultured tongue cancer cell lines than in 2 D cultures. Typically, under conventional 2 D culture conditions, F-actin and cortactin are colocalized in the form of puncta within ***, in the 3 D-cultured cells, colocalization was mainly observed at the cell margins, including the projections. Projections containing F-actin and cortactin colocalization were predicted to be invadopodia. Although suppressing ezrin expression with small interfering RNA transfection caused no marked changes in morphology, cell projection formation was decreased, and the tumour thickness in vertical sections after 3 D culture was markedly decreased after suppressing ERK activity because both the invasion ability and proliferation were inhibited. An association between cortactin activation as well as ERK activity and invadopodia formation was detected. Our novel 3 D culture systems using Cellbed? are simple and useful for in vitro studies before conducting animal experiments. ERK contributes to tongue cancer development by increasing both cancer cell proliferation and migration via cortactin activation.

关键词： ERK phosphorylation functions in invadopodia formation in tongue cancer cells in a novel silicate fibre-based 3D cell culture sy HSC

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

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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

Clinical significance of MET in gastric cancer

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World Journal of Gastrointestinal Oncology 2015年第11期7卷 317-327页

作者： Mikito Inokuchi Sho Otsuki Yoshitaka Fujimori Yuya Sato Masatoshi Nakagawa Kazuyuki Kojima department of Gastrointestinal surgery Tokyo Medical and Dental University department of Minimally Invasive surgery Tokyo Medical and Dental University

Chemotherapy has become the global standard treatment for patients with metastatic or unresectable gastric cancer(GC),although outcomes remain *** molecular-targeted therapies inhibiting signaling pathways of various tyrosine kinase receptors have been developed,and monoclonal antibodies targeting human epidermal growth factor receptor 2 or vascular endothelial growth factor receptor 2 have become standard therapy for *** growth factor and its receptor,c-MET(MET),play key roles in tumor growth through activated signaling pathways from receptor in GC *** amplification of MET leads to the aberrant activation found in GC tumors and is related to survival in patients with *** review discusses the clinical significance of MET in GC and examines MET as a potential therapeutic target in patients with *** studies in animal models have shown that MET antibodies or smallmolecule MET inhibitors suppress tumor-cell proliferation and tumor progression in MET-amplified GC *** drugs are now being evaluated in clinical trials as treatments for metastatic or unresectable GC.

关键词： MET Gastric cancer Genomic amplification Immunohis

Feasibility of laparoscopic total gastrectomy in overweight patients: Implications of less impact of overweight on laparoscopic versus open approach

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World Journal of Clinical Cases 2018年第16期6卷 1094-1100页

作者： Masatoshi Nakagawa Kazuyuki Kojima Mikito Inokuchi Kenta Kobayashi Toshiro Tanioka Keisuke Okuno Kentaro Gokita department of Gastric surgery Tokyo Medical and Dental University Center for Minimally Invasive surgery Tokyo Medical and Dental University

AIM To investigate safety and oncological feasibility of laparoscopic total gastrectomy(LTG) in overweight(OW) *** Patients who underwent total gastrectomy(110 laparoscopic, 211 open) for gastric cancer between January 1999 and July 2016 were included. Propensity score matching selected 152 patients(76 laparoscopic, 76 open), which were subsequently divided into the OW(≥ 25) or non-OW(< 25) group by body mass index. Postoperative outcomes of laparoscopic versus open approaches were compared between OW and non-OW groups. RESULTS In the propensity-matched population, baseline characteristics were comparable between the OW and nonOW groups for the laparoscopy and open groups. In the laparoscopy group, operative time was longer(P = 0.01) in the OW group, however, other perioperative results including complication rates were comparable between the non-OW and OW groups. In the open group, number of retrieved lymph nodes were less(P = 0.03) and local complication rate was more frequent(P = 0.03) in the OW group. CONCLUSION LTG in OW patients remains technically challenging but can be performed safely. Our findings imply that OW has a lesser effect on the laparoscopic versus open approach to total gastrectomy.

关键词： Gastric cancer Laparoscopy Overweight Total gastrectomy

Basal cell carcinoma stem cells exhibit osteogenic and chondrogenic differentiation potential

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BIOCELL 2021年第6期45卷 1543-1550页

作者： MAJA MILOSEVIC MILOS LAZAREVIC BOSKO TOLJIC MILAN PETROVIC MIROSLAV VUKADINOVIC ZORAN JEZDIC BOBAN ANICIC DRAGO JELOVAC SVETLANA JOVANOVIC JELENA MILASIN department of Human Genetics School of Dental MedicineUniversity of BelgradeBelgrade11000Serbia Clinic for maxillofacial surgery School of Dental MedicineUniversity of BelgradeBelgrade11000Serbia department of Public Health School of Dental MedicineUniversity of BelgradeBelgrade11000Serbia

Specific cell subpopulations identified as cancer stem cells(CSCs)can be found in basal cell carcinoma(BCC).Generally,CSCs have a marked trans-differentiation potential that could potentially be used in differentiation ***,there are no studies regarding BCC CSCs *** aim of the study was to analyze the characteristic of CSCs of BCC with emphasis on their differentiation potential upon specific *** staining and cell morphology were used for differentiation confirmation,along with the expression analysis of osteogenic(ALP,BSP,Runx2,OCN,BMP2),chondrogenic(COL1 and COL2A1),adipogenic(PPAR-γ)and neurogenic(Nestin and MAP2)*** CSCs differentiated into osteogenic and chondrogenic lineages,as judged by staining and high expression of specific markers(from 2-to 92-fold higher upon induction).Concomitantly with differentiation,the levels of cancer stem cell markers decreased in the ***-differentiation and neuro-differentiation were *** conclusion,BCC CSCs exhibit the capacity to trans-differentiate,a characteristic that may potentially be useful in the development of new strategies for the treatment of aggressive BCCs.

关键词： Basal cell carcinoma Cancer stem cells Differentiation potential Osteo-differentiation Chondro-differentiation

Tooth fragment embedded in the lower lip for 10 months following dentoalveolar trauma:A case report with literature review

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Burns & Trauma 2014年第3期2卷 141-145页

作者： NB Nagaveni KV Umashankara department of Pedodontics and Preventive Dentistry College of Dental SciencesDavangere department of Oral and maxillofacial surgery Bapuji Dental College and HospitalDavangereKarnatakaIndia

Traumatic injuries to maxillary anterior teeth are a common finding in children because of falls while playing. Sequelae of trauma to dental hard tissue include broken, lost, aspirated and swallowed teeth. One additional hazard is the embedding of fractured tooth fragments in the soft tissues, particularly in the lip. A 10-year-old male patient complained of pain in the lower lip. There was a history of trauma to the upper anterior tooth 10 months previously. Clinical examination showed scarring and discoloration over the lower lip, and the presence of a hard mass was felt on palpation. Intraoral examination revealed an Ellis and Davey class II fracture of number 11. A radiograph of the lip was taken, which showed a radiopaque structure similar to the shape of the missing tooth fragment. Under local anesthesia, the tooth fragment was removed successfully, and the class II fracture was restored with composite. Therefore, proper clinical and complete radiographic examination of both hard and soft tissues following dental trauma is essential to rule out such occurrences.

关键词： Fractured incisor lip laceration tooth embedding