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Background: An observer-rated questionnaire for alexithymia based on the original 17-item Beth Israel Hospital Psychosomatic Questionnaire for Alexithymia (BIQ) was developed by Sifneos in 1973 and modified into a 12-item version of BIQ by Taylor et al. in 1997. However, it has rarely been used in a clinical or research context and studies have not given satisfactory inter-rater reliability for the 12-item version. Objective: To develop a structured interview in Japanese for the12-item modified version of BIQ (m-SIBIQ) to determine the reliability and validity of the m- scores and its factor structure model. Methods: Ninety-two Japanese young adults were interviewed. The inter-rater reliability of the m-SIBIQ was assessed by exploratory factor analysis. For the concurrent and convergent validities, correlation analysis was done between the scores of m-SIBIQ and the self-reported questionnaires: 20-Item Toronto Alexithymia Scale (TAS-20), NEO Five-Factor Inventory (NEO-FFI), Emotional Empathy Scale (EES), Interpersonal Reactivity Index (IRI), Beck Depression Inventory-II (BDI-II), and State-Trait Anxiety Inventory (STAI). Goodness of fit of the structure model of the m-SIBIQ was evaluated using confirmatory factor analysis, and the results were examined through stepwise multiple regression analysis. Results: Good reliability was obtained for the total score of m-SIBIQ: Cronbach's α.950 (p<.001) and ICC.75(p<.05). The validity of the factor structure was obtained by confirmatory factor analysis using covariance. The model of the alexithymia constructs was configured by the operative thinking (la pensée opératoire) and affect awareness components. The stepwise multiple regression analysis extracted the total score of m-SIBIQ as significantly, negatively correlated with the Openness to experience score of NEO-FFI and significantly, positively correlated with the emotionally chilly score of EES and the score of difficulty describing feelings (DDF) of TAS-20. There were no correlations between the m-SIBIQ and BDI-II scores. Conclusion: For Japanese young adults, the m-SIBIQ is a reliable and valid instrument for overcoming weaknesses of the self-reported procedures by bringing to light the alexithymia construct and principal dimensions.
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In this study, the effect of defects on the acceptor properties of nitrogen-doped ZnO nanoparticles (NPs) was investigated through the fabrication of light-emitting diodes (LEDs). Nitrogen-doped ZnO NPs were synthesized by an arc discharge in-gas evaporation method and post-annealed at 800 °C in an oxygen and nitrogen atmosphere. The annealed ZnO NPs were characterized by X-ray diffraction, scanning electron microscopy, Raman spectroscopy, and photoluminescence spectroscopy. It was found that the annealing of nitrogen-doped ZnO NPs in a nitrogen environment increased the number of zinc vacancies, while annealing in an oxygen environment increased the number of oxygen vacancies due to nitrogen desorption. The output characteristics of LEDs fabricated with oxygen-annealed NPs were degraded, while those with nitrogen-annealed NPs were significantly improved. From these results, the contribution of zinc vacancies to acceptor formation in ZnO NPs was confirmed for the first time in actual pn junction devices.
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The importance of the parent-progeny relationship tracking technique in single-cell analysis has grown with the passage of time. In this study, fundamental image-processing techniques were combined to develop software capable of inferring cell cycle alterations in fission yeast cells, which exhibit equipartition during division. These methods, exclusively relying on bright-field images as input, could track parent-progeny relationships after cell division by assessing the temporal morphological transformation of these cells. In the application of this technique, the software was employed for calculating intracellular fluorescent dots during every stage of the cell cycle, using a yeast strain expressing EGFP-fused Swi6, which binds to chromatin. The results obtained with this software were consistent with those of previous studies. This software facilitated single-cell-level tracking of parent-progeny relationships in cells exhibiting equipartition during division and enabled the monitoring of spatial fluctuations in a cell cycle-dependent protein. This method, expediting the analysis of extensive datasets, may also empower large-scale screening experiments that cannot be conducted manually.
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Schizosaccharomyces , Schizosaccharomyces/genética , Schizosaccharomyces/metabolismo , Imagem com Lapso de Tempo , Ciclo Celular , Divisão Celular , Proteínas de Ciclo Celular/metabolismoRESUMO
Heterojunction light-emitting diodes (LEDs), based on p-type ZnO and n-type ZnMgO nanoparticles, have been demonstrated. ZnMgO nanoparticles were prepared by the thermal diffusion of Mg onto ZnO nanoparticles. p-ZnO/GZO homostructure LEDs and p-ZnO/n-ZnMgO/GZO heterostructure LEDs have been fabricated using ZnO and ZnMgO nanoparticles. By comparing the characteristic results of these diodes, it can be seen that LEDs with the p-ZnO/n-ZnMgO/GZO structure showed better I-V characteristics with a lower current density leakage than those with the p-ZnO/GZO LED structure. Moreover, the emission intensity was improved by adding the ZnMgO NP layer to the LEDs. These results show that the ZnMgO NP layer acts as a hetero-barrier layer that suppresses the diffusion of holes into the n-type layer and confines holes to the p-type layer.
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Periodontal disease is a chronic inflammatory condition caused by periodontal pathogens in the gingival sulcus. Short-chain fatty acids (SCFAs) produced by causal bacteria are closely related to the onset and progression of periodontal disease and have been reported to proliferate in the periodontal sulcus of patients experiencing this pathology. In such patients, propionic acid (C3), butyric acid (C4), isobutyric acid (IC4), valeric acid (C5), isovaleric acid (IC5), and caproic acid (C6), henceforth referred to as [C3-C6], has been reported to have a detrimental effect, while acetic acid (C2) exhibits no detrimental effect. In this study, we established an inexpensive and simple enzymatic assay that can fractionate and measure these acids. The possibility of applying this technique to determine the severity of periodontal disease by adapting it to specimens collected from humans has been explored. We established an enzyme system using acetate kinase and butyrate kinase capable of measuring SCFAs in two fractions, C2 and [C3-C6]. The gingival crevicular fluid (GCF) and saliva of 10 healthy participants and 10 participants with mild and severe periodontal disease were measured using the established enzymatic method and conventional gas chromatography-mass spectrometry (GC-MS). The quantification of C2 and [C3-C6] in human GCF and saliva was well correlated when using the GC-MS method. Furthermore, both C2 and [C3-C6] in the GCF increased with disease severity. However, while no significant difference was observed between healthy participants and periodontal patients when using saliva, [C3-C6] significantly differed between mild and severe periodontal disease. The enzymatic method was able to measure C2 and [C3-C6] separately as well as using the GC-MS method. Furthermore, the C2 and [C3-C6] fractions of GCF correlated with disease severity, suggesting that this method can be applied clinically. In contrast, the quantification of C2 and [C3-C6] in saliva did not differ significantly between healthy participants and patients with periodontal disease. Future studies should focus on inflammation rather than on tissue destruction.
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Ácidos Graxos Voláteis , Doenças Periodontais , Ácido Acético , Ácido Butírico , Ácidos Graxos Voláteis/análise , Líquido do Sulco Gengival/química , Humanos , Doenças Periodontais/diagnósticoRESUMO
In this work, nitrogen-doped ZnO nanoparticles were synthesized in various conditions by the gas evaporation method with DC arc plasma. Nitrogen concentrations of 6.38 × 1018 cm-3 to 2.6 × 1019 cm-3 were obtained at a chamber pressure of 150 torr, using arc currents of 20 A to 70 A. The intensities of local vibrational modes at 275 cm-1 and 581 cm-1 in the Raman spectra of ZnO nanoparticles showed a dependency on the nitrogen concentration in the ZnO nanoparticles. The ratios of donor-acceptor pair and exciton emissions in the photoluminescence spectra of nitrogen-doped ZnO nanoparticles, and the electroluminescence of light-emitting diodes based on these nanoparticles, were nearly proportional to the Raman peak's intensity at 275 cm-1. The results indicated that the nitrogen dopants in the ZnO nanoparticles were acting as an acceptor.
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OBJECTIVE: The Eating Disorder Examination-Questionnaire (EDE-Q) is one of the most widely used tools to assess the core psychopathology of eating disorders (ED). However, recent empirical findings did not support the original four-factor structure. The aims of the present study were to investigate the factor structure of the EDE-Q in Japanese ED patients, to test the reliability and convergent validity of the EDE-Q, to examine group differences between various ED groups and healthy participants, and to explore the main behavioral features of Japanese ED patients using the newly developed Japanese version of EDE-Q. METHOD: A total of 148 ED patients and 469 healthy participants completed the EDE-Q, Eating Attitudes Test-26 (EAT-26), and Eating Disorder Inventory-2 (EDI-2). The factor structure, reliability, and validity of the EDE-Q were assessed in ED patients. Group differences were assessed using the new Japanese version of the EDE-Q (EDE-Q-J). RESULTS: The EDE-Q-J had three factors. Cronbach's alphas ranged from 0.83 to 0.93. Total score and subscale scores of "Dieting" and "Bulimia and Food Preoccupation" of EAT-26 and of "Drive for Thinness," "Body Dissatisfaction," and "Bulimia" of EDI-2 correlated with the global score and three subscale scores of the EDE-Q-J. DISCUSSION: For Japanese female ED patients, the EDE-Q-J had three subscales that were not consistent with the original subscales, but were interpretable. It demonstrated sufficient reliability and validity. Japanese female patients with restricting-type anorexia nervosa (AN-R) displayed less dissatisfaction with shape and weight than healthy participants. AN-R patients in Japan might present with a non-fat-phobic symptom profile.
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Transtornos da Alimentação e da Ingestão de Alimentos , Inquéritos e Questionários , Estudos de Casos e Controles , Transtornos da Alimentação e da Ingestão de Alimentos/diagnóstico , Transtornos da Alimentação e da Ingestão de Alimentos/psicologia , Transtornos da Alimentação e da Ingestão de Alimentos/terapia , Feminino , Humanos , Japão , Psicometria , Psicopatologia , Reprodutibilidade dos TestesRESUMO
BACKGROUND: The Eating Disorder Quality of Life (ED-QOL) scale is a 25-item self-report measure that assesses health-related quality of life (HRQoL) of eating-disorder patients. Although the ED-QOL is one of the most widely used questionnaires in many countries, no prior research has addressed the psychometric properties of the Japanese translation of the ED-QOL. Therefore, the aim of the present study was to assess its reliability and validity. METHODS: A total of 99 Japanese female eating disorder patients and 469 female healthy university undergraduate students completed the Japanese translation of the ED-QOL in addition to the Eating Attitudes Test-26 (EAT-26) and Eating Disorder Inventory-2 (EDI-2). The patient group consisted of 37 patients with anorexia nervosa restricting type (AN-R), 35 patients with binge-eating/purge type (AN-BP), and 27 patients with bulimia nervosa (BN). We performed confirmatory factor analyses on the ED-QOL subscales both for Japanese eating disorder patients and for healthy university undergraduate students. Reliability was assessed using internal consistency indicated by Cronbach alpha coefficients and convergent validity was assessed using Pearson's correlation coefficients. To assess group differences between the eating disorder patients and healthy university undergraduate students, Student's t-tests were conducted. RESULTS: The CFA showed that the CFI was .90 and RMSEA was .084 (90% confidence interval = .079-.088). The internal consistency of the ED-QOL varied from good to excellent. The EAT-26 total score and three subscales and the EDI-2 subscales had significant correlations with the ED-QOL global QOL score and four subscales. There were no significant correlations between the EDI-2 subscale "Body Dissatisfaction" and the ED-QOL subscales "Physical/Cognitive" and "Work/School". Eating disorder patients scored significantly higher than healthy university undergraduate students on all ED-QOL subscales and the global QOL score. CONCLUSIONS: Based on this study, the Japanese translation of the ED-QOL can be regarded as reliable, valid, and functional for female eating-disorder patients and female healthy university undergraduate students.
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Recent studies have revealed that tracking single cells using time-lapse fluorescence microscopy is an optimal tool for spatiotemporal evaluation of proteins of interest. Using this approach with Saccharomyces cerevisiae as a model organism, we previously found that heterochromatin regions involved in epigenetic regulation differ between individual cells. Determining the regularity of this phenomenon requires measurement of spatiotemporal epigenetic-dependent changes in protein levels across more than one generation. In past studies, we conducted these analyses manually to obtain a dendrogram, but this required more than 15 h, even for a single set of microscopic cell images. Thus, in this study, we developed a software-based analysis system to analyze time-lapse cellular images of S. cerevisiae, which allowed automatic generation of a dendrogram from a given set of time-lapse cell images. This approach is divided into two phases: a cell extraction and tracking phase, and an analysis phase. The cell extraction and tracking phase generates a set of necessary information for each cell, such as geometrical properties and the daughter-mother relationships, using image processing-based analysis techniques. Then, based on this information, the analysis phase allows generation of the final dendrogram by analyzing the fluorescent characteristics of each cell. The system is equipped with manual error correction to correct for the inevitable errors that occur in these analyses. The time required to obtain the final dendrograms was drastically reduced from 15 h in manual analysis to 0.8 h using this novel system.
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Processamento de Imagem Assistida por Computador/métodos , Saccharomyces cerevisiae/citologia , Imagem com Lapso de Tempo/métodos , Divisão Celular , Microscopia de Fluorescência/métodos , Saccharomyces cerevisiae/fisiologia , SoftwareRESUMO
We aimed to assess the efficacy of a reablement program in improving the independence from long-term care services of older adults with mild disability. This parallel, two-arm, randomized controlled, superiority trial was conducted in Neyagawa, a local government area in Osaka, Japan. Eligible participants were community-dwelling individuals aged ≥65 years certified as support-required level. They were assigned in a 1:1 ratio to receive either a community-based, multicomponent, multidisciplinary, individualized goal-directed, and time-limited intervention (the CoMMIT program) plus standard care or standard care alone. The primary outcome was independence, that is, the nonuse of long-term care services during the three-month follow-up period. The study was terminated early due to slow enrollment. A total of 375 participants were enrolled and randomized to either the intervention (n = 190) or control (n = 185) group. The proportions of independence were 11.1% and 3.8% in the intervention and control groups, respectively (absolute difference: 7.3; 95% confidence interval: 2.0-12.5). There was no difference in the risk of serious adverse events between the groups. The CoMMIT program plus standard care was found superior to standard care alone in enhancing the independence from long-term care services of older adults with mild disability.
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Pessoas com Deficiência , Vida Independente , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Japão , Masculino , Índice de Gravidade de Doença , Fatores de TempoRESUMO
INTRODUCTION: We presented a rare case of caval lobular capillary hemangioma. CASE PRESENTATION: A 66-year-old female visited our department complaint with shadow defect in vena cava of right renal hilum appeared on computed tomography for periodically checking 3 years after radical hysterectomy with bilateral ovariectomy. Abdominal computed tomography identified a shadow defect of 35 mm in diameter in the inferior vena cava continuing posteriorly to a 35 mm mass of retroperitoneum. During the total removal of this lesion, we identified the lesion was connected to right ovarian vein. The specimen consisted of microcapillaries which formed reticular structure. Immunostaining of specimens identified positive CD31, CD34, and Factor 8 in all cells. Ki67 antibody was positive at 2-3% of all cells. These findings suggested the tumor was intravenous lobular papillary hemangioma. CONCLUSION: This is the first report of intravenous lobular papillary hemangioma originated from right ovarian vein and extended to inferior vena cava.
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BACKGROUND: Periodontitis results in the destruction of tooth-supporting periodontal tissues and does not have the ability to heal spontaneously. Various approaches have been introduced to regenerate periodontal tissues; however, these approaches have limited efficacy for treating severe defects. Cytotherapies combine stem cell biology and tissue engineering to form a promising approach for overcoming these limitations. In this study, we isolated periodontal ligament (PDL)-derived cells from patients and created cell sheets with "Cell Sheet Engineering Technology", using temperature responsive culture dishes, in which all the cultured cells can be harvested as an intact transplantable cell sheet by reducing the temperature of the culture dish. Subsequently, the safety and efficacy of autologous PDL-derived cell sheets were evaluated in a clinical setting. METHODS: A single-arm and single-institute clinical study was performed to verify the safety and efficacy of autologous PDL-derived cell sheets in patients with periodontitis. Wisdom teeth were extracted from patients diagnosed with chronic periodontitis, ranging in age from 33 to 63 years (mean [±SD], 46 ± 12), and periodontal tissues were scraped for cell sources. Three-layered PDL-derived cell sheets were constructed using temperature-responsive culture dishes and transplanted in an autologous fashion following standard flap surgeries. Bony defects were filled with beta-tricalcium phosphate granules. Clinical variables were evaluated at baseline, 3 months, and 6 months. Cone-beam computed tomography was performed at baseline and 6 months. Additionally, mid-long-term follow-up has been performed with patients' agreements. RESULTS: Our method was found to be safe and no severe adverse events were identified. All the findings, including reduction of periodontal probing depth (mean ± SD, 3.2 ± 1.9 mm), clinical attachment gain (2.5 ± 2.6 mm), and increase of radiographic bone height (2.3 ± 1.8 mm), were improved in all 10 cases at 6 months after the transplantation. These therapeutic effects were sustained during a mean follow-up period of 55 ± 19 months, and there were no serious adverse events. CONCLUSIONS: The results of this study validate the safety and efficacy of autologous PDL-derived cell sheets in severe periodontal defects, and the stability of this efficacy during mid-long-term follow up. This cytotherapeutic approach, based on cell sheet engineering, offers an innovative strategy to treat the recognized unmet need of treating severe periodontal defects.
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Carcinosarcomas of the urinary bladder are rare biphasic neoplasms, consisting of both malignant epithelial and malignant mesenchymal components, and the prognosis of this tumor is unfavorable in most patients with even possibility of resection of disease. A 77-year-old male with a history of transurethral resection (TUR) of urothelial carcinoma (UC) of the bladder and adjuvant intravesical chemotherapy with pirarubicin 10 years ago revisited our department with a gross hematuria. Cystoscopy demonstrated an approximately 2.5 cm nonpapillary tumor on the right wall of the bladder. Pelvic MRI showed the tumor without extending the base of the bladder wall. The tumor could be completely removed with TUR. The malignant epithelial elements consisted of high-grade UC and the majority of mesenchymal components were fibrosarcomatous differentiation based on immunohistochemical studies. The tumor could be pathologically also suspected to be an early stage on TUR specimens. Although he has received no additional intervention due to the occurrence of myocardial infarction at three weeks after the TUR, he has been alive with no evidence of recurrence of the disease 27 months after the TUR. Some early stages of bladder carcinosarcoma might have a favorable prognosis without aggressive treatments.
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Artificial skin has achieved considerable therapeutic results in clinical practice. However, artificial skin treatments for wounds in diabetic patients with impeded blood flow or with large wounds might be prolonged. Cell-based therapies have appeared as a new technique for the treatment of diabetic ulcers, and cell-sheet engineering has improved the efficacy of cell transplantation. A number of reports have suggested that adipose-derived stem cells (ASCs), a type of mesenchymal stromal cell (MSC), exhibit therapeutic potential due to their relative abundance in adipose tissue and their accessibility for collection when compared to MSCs from other tissues. Therefore, ASCs appear to be a good source of stem cells for therapeutic use. In this study, ASC sheets from the epididymal adipose fat of normal Lewis rats were successfully created using temperature-responsive culture dishes and normal culture medium containing ascorbic acid. The ASC sheets were transplanted into Zucker diabetic fatty (ZDF) rats, a rat model of type 2 diabetes and obesity, that exhibit diminished wound healing. A wound was created on the posterior cranial surface, ASC sheets were transplanted into the wound, and a bilayer artificial skin was used to cover the sheets. ZDF rats that received ASC sheets had better wound healing than ZDF rats without the transplantation of ASC sheets. This approach was limited because ASC sheets are sensitive to dry conditions, requiring the maintenance of a moist wound environment. Therefore, artificial skin was used to cover the ASC sheet to prevent drying. The allogenic transplantation of ASC sheets in combination with artificial skin might also be applicable to other intractable ulcers or burns, such as those observed with peripheral arterial disease and collagen disease, and might be administered to patients who are undernourished or are using steroids. Thus, this treatment might be the first step towards improving the therapeutic options for diabetic wound healing.
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Tecido Adiposo/citologia , Técnicas de Cultura de Células/métodos , Diabetes Mellitus Tipo 2/complicações , Transplante de Células-Tronco/métodos , Cicatrização/fisiologia , Animais , Células Cultivadas , Diabetes Mellitus Experimental/terapia , Diabetes Mellitus Tipo 2/terapia , Masculino , Células-Tronco Mesenquimais/fisiologia , Obesidade/complicações , Ratos Endogâmicos Lew , Ratos Zucker , Pele ArtificialRESUMO
BACKGROUND: Although the Eating Disorder Examination Questionnaire version 6.0 (EDE-Q) is one of the most widely used questionnaires for eating disorders in Western countries, no research has addressed the psychometric properties of the EDE-Q in a Japanese sample. METHODS: We explored the factor structure of the EDE-Q and examined the internal consistency of the derived scales for Japanese participants (Study I), the convergent validity with other eating disorder-related psychological measures (Study II) and the distinction between the derived two body image-related factors with psychological measures (StudyIII). The EDE-Q was administered to 1,430 undergraduate students in Study I and in Study II was subsequently assessed by two self-report measures of eating pathology, the Eating Attitudes Test (EAT-26) for 558 undergraduate students and the Eating Disorders Inventory-II (EDI-II) 111. In StudyIII, another 225 undergraduate students participated in an examination of the relationships of the derived body image-related subscales of the EDE-Q with the psychological measures of the Rosenberg Self-Esteem Scale, Beck Depression Inventory, Public Self-Consciousness Scale, and Multidimensional Perfectionism Scale. RESULTS: Exploratory factor analysis of the EDE-Q identified four meaningful factors. Of the original four EDE-Q factors, "Restriction" and "Eating Concern" were retained. However, the other two factors, "Shape" and "Weight" Concerns, were combined into two different factors: "Fear of Obesity" and "Self-Esteem Based on Shape and Weight". Internal consistency of the derived four factors was adequate, and the relationships with EDI-II and EAT-26 measures demonstrated convergent validity. Analysis of the distinction between "Fear of Obesity" and "Self-Esteem Based on Shape and Weight" revealed that only "Self-Esteem Based on Shape and Weight" was significantly associated with the measures assessing psychopathology related to eating disorders. CONCLUSIONS: This study describes restructured factors of the EDE-Q that were tested with undergraduate students. The distinction between two factors, "Fear of Obesity" and "Self-Esteem Based on Shape and Weight", may further the understanding of the psychopathology of the eating disorders of adolescent Japanese subjects to facilitate future developments in research and treatment.
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The reported incidence rate of iatrogenic ureteral injury is 0.5 to 3% among abdominal surgery. We report a case of ureterouterine fistula after caesarean section. A 38-year-old woman visited our department with a complaint of urinary incontinence without dry time after caesarean section. Several examinations revealed right ureterouterine fistula.Ureteroneocystostomy using psoas hitch and hysterectomy was performed. We found a firm adhesion and stitches around right lower ureter over the uterus, which lead to an additional hysterectomy. After surgery, urinary incontinence had improved. Following two years after surgery, we observed no urinary incontinence or renal dysfunction.
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Periodontitis is a chronic inflammatory disease that induces the destruction of tooth-supporting tissues, followed by tooth loss. Although several approaches have been applied to periodontal regeneration, complete periodontal regeneration has not been accomplished. Tissue engineering using a combination of cells and scaffolds is considered to be a viable alternative strategy. We have shown that autologous transplantation of periodontal ligament-derived multipotent mesenchymal stromal cell (PDL-MSC) sheets regenerates periodontal tissue in canine models. However, the indications for autologous cell transplantation in clinical situations are limited. Therefore, this study evaluated the safety and efficacy of allogeneic transplantation of PDL-MSC sheets using a canine horizontal periodontal defect model. Canine PDL-MSCs were labeled with enhanced green fluorescent protein (EGFP) and were cultured on temperature-responsive dishes. Three-layered cell sheets were transplanted around denuded root surfaces either autologously or allogeneically. A mixture of ß-tricalcium phosphate and collagen gel was placed on the bone defects. Eight weeks after transplantation, dogs were euthanized and subjected to microcomputed tomography and histological analyses. RNA and DNA were extracted from the paraffin sections to verify the presence of EGFP at the transplantation site. Inflammatory markers from peripheral blood sera were quantified using an enzyme-linked immunosorbent assay. Periodontal regeneration was observed in both the autologous and the allogeneic transplantation groups. The allogeneic transplantation group showed particularly significant regeneration of newly formed cementum, which is critical for the periodontal regeneration. Serum levels of inflammatory markers from peripheral blood sera showed little difference between the autologous and allogeneic groups. EGFP amplicons were detectable in the paraffin sections of the allogeneic group. These results suggest that allogeneic PDL-MSC sheets promoted periodontal tissue regeneration without side effects. Therefore, allogeneic transplantation of PDL-MSC sheets has a potential to become an alternative strategy for periodontal regeneration.
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Periodontitis is a inflammation induced by a bacterial infection that causes the destruction of the attachment apparatus of dental roots. Several materials, such as bone graft materials, barrier membranes and protein products have been developed and used to treat periodontal defects clinically; however, it is difficult to regenerate the complete periodontal tissue structure. Recently, cytotherapeutic approaches have been introduced to overcome the limitation of conventional procedures. The in vitro-expanded autologous cells derived from several kinds of tissues have already been used in several clinical trials. These cytotherapeutic treatments have been shown to be safe and effective for the treatment of periodontitis. Our strategy has been to integrate stem cell biology and cell sheet engineering, in which a temperature-responsive intelligent polymer is grafted onto the surface of cell culture dish to create a 'cell sheet', to achieve a novel treatment method for periodontitis. By simple reduction of the temperature to below 32°C, a contiguous cell sheet, which is capable of keeping extracellular matrix proteins and cell-cell interactions intact, can be harvested for transplantation without the use of scaffolds. This technology has already been employed in clinical trials, confirming the safety and efficacy of the treatment. In this review, we introduce recent progress in the engineering of cell sheets and review the potential of cell sheet technology for periodontal regenerative medicine.
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Periodontite/terapia , Regeneração , Medicina Regenerativa/métodos , Engenharia Tecidual/métodos , Alicerces Teciduais , Humanos , Periodontite/patologiaRESUMO
Afadin is a filamentous actin-binding protein and a mediator of nectin signaling. Nectins are Ig-like cell adhesion molecules, and the nectin family is composed of four members, nectin-1 to nectin-4. Nectins show homophilic and heterophilic interactions with other nectins or proteins on adjacent cells. Nectin signaling induces formation of cell-cell junctions and is required for the development of epithelial tissues, including skin. This study investigated the role of afadin in epithelial tissue development and established epithelium-specific afadin-deficient (CKO) mice. Although showing no obvious abnormality in the skin development and homeostasis, the mice showed the reduced neutrophil infiltration into the epidermis during chemical-induced inflammation with 12-O-tetradecanoylphorbol 13-acetate (TPA). Immunohistochemical and quantitative real-time PCR analyses showed that the expression levels of cytokines including Cxcl2, Il-1ß and Tnf-α were reduced in CKO keratinocytes compared with control keratinocytes during TPA-induced inflammation. Primary-cultured skin keratinocytes from CKO mice also showed reduced expression of these cytokines and weak activation of Rap1 compared with those from control mice after the TPA treatment. These results suggested a remarkable function of afadin, which was able to enhance cytokine expression through Rap1 activation in keratinocytes during inflammation.
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Quimiocina CXCL2/metabolismo , Interleucina-1beta/metabolismo , Queratinócitos/metabolismo , Proteínas dos Microfilamentos/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Animais , Epiderme/metabolismo , Inflamação/induzido quimicamente , Inflamação/metabolismo , Camundongos , Camundongos Knockout , Proteínas dos Microfilamentos/genética , Neutrófilos/metabolismo , Cultura Primária de Células , Acetato de Tetradecanoilforbol , Proteínas rap1 de Ligação ao GTP/metabolismoRESUMO
Addressing the problem of vascularization is of vital importance when engineering three-dimensional (3D) tissues. Endothelial cells are increasingly used in tissue-engineered constructs to obtain prevascularization and to enhance in vivo neovascularization. Rat bone marrow stromal cells were cultured in thermoresponsive dishes under osteogenic conditions with human umbilical vein endothelial cells (HUVECs) to obtain homotypic or heterotypic cell sheets (CSs). Cells were retrieved as sheets from the dishes after incubation at 20 °C. Monoculture osteogenic CSs were stacked on top of homotypic or heterotypic CSs, and subcutaneously implanted in the dorsal flap of nude mice for 7 days. The implants showed mineralized tissue formation under both conditions. Transplanted osteogenic cells were found at the new tissue site, demonstrating CS bone-inductive effect. Perfused vessels, positive for human CD31, confirmed the contribution of HUVECs for the neovascularization of coculture CS constructs. Furthermore, calcium quantification and expression of osteocalcin and osterix genes were higher for the CS constructs, with HUVECs demonstrating the more robust osteogenic potential of these constructs. This work demonstrates the potential of using endothelial cells, combined with osteogenic CSs, to increase the in vivo vascularization of CS-based 3D constructs for bone tissue engineering purposes.