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1.
J Appl Oral Sci ; 27: e20180564, 2019 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-31365710

RESUMO

OBJECTIVE: Studies on the oral health of individuals with intellectual disability (ID) have identified problems that include a high prevalence of periodontal disease. The use of probiotics to treat periodontal disease has been the focus of considerable research, and bovine milk fermented with Lactobacillus rhamnosus L8020 (L8020 yogurt) has been shown to reduce the oral prevalence of four periodontal pathogens. The aim of this randomized, double-blind, placebo-controlled trial was to compare the effects of L8020 yogurt (test group) with those of placebo yogurt (placebo group) on the papillary-marginal-attached (PMA) index, gingival index (GI), and probing depth (PD) in 23 individuals with ID. METHODOLOGY: All patients were required to consume the allocated yogurt after breakfast for 90 days. PMA index and GI scores as well as PDs were assessed before the start of yogurt consumption (baseline), after 45 and 90 days of consumption, and 30 days after the cessation of consumption. Student's t-test, Mann-Whitney U test or Fisher's exact test was used for inter-group comparisons, and the mixed effect model of repeated measurements was used for data analysis. RESULTS: The decrease in PMA index score was significantly greater in the test group than in the placebo group (p<0.001). The GI score also decreased during the study, with a tendency for greater decrease in the test group. Furthermore, decreases in PD between baseline, 45 and 90 days tended to be greater in the test group than in the placebo group. CONCLUSION: These results suggest that regular consumption of bovine milk fermented with L. rhamnosus L8020 can lower the risk of periodontal disease in individuals with ID.


Assuntos
Deficiência Intelectual/fisiopatologia , Lactobacillus rhamnosus , Leite , Doenças Periodontais/prevenção & controle , Iogurte , Adulto , Animais , Método Duplo-Cego , Feminino , Humanos , Deficiência Intelectual/complicações , Masculino , Pessoa de Meia-Idade , Doenças Periodontais/patologia , Índice Periodontal , Valores de Referência , Reprodutibilidade dos Testes , Fatores de Risco , Estatísticas não Paramétricas , Fatores de Tempo , Resultado do Tratamento , Adulto Jovem
2.
J. appl. oral sci ; 27: e20180564, 2019. tab, graf
Artigo em Inglês | LILACS-Express | ID: biblio-1012518

RESUMO

Abstract Studies on the oral health of individuals with intellectual disability (ID) have identified problems that include a high prevalence of periodontal disease. The use of probiotics to treat periodontal disease has been the focus of considerable research, and bovine milk fermented with Lactobacillus rhamnosus L8020 (L8020 yogurt) has been shown to reduce the oral prevalence of four periodontal pathogens. Objective The aim of this randomized, double-blind, placebo-controlled trial was to compare the effects of L8020 yogurt (test group) with those of placebo yogurt (placebo group) on the papillary-marginal-attached (PMA) index, gingival index (GI), and probing depth (PD) in 23 individuals with ID. Methodology All patients were required to consume the allocated yogurt after breakfast for 90 days. PMA index and GI scores as well as PDs were assessed before the start of yogurt consumption (baseline), after 45 and 90 days of consumption, and 30 days after the cessation of consumption. Student's t-test, Mann-Whitney U test or Fisher's exact test was used for inter-group comparisons, and the mixed effect model of repeated measurements was used for data analysis. Results The decrease in PMA index score was significantly greater in the test group than in the placebo group (p<0.001). The GI score also decreased during the study, with a tendency for greater decrease in the test group. Furthermore, decreases in PD between baseline, 45 and 90 days tended to be greater in the test group than in the placebo group. Conclusion These results suggest that regular consumption of bovine milk fermented with L. rhamnosus L8020 can lower the risk of periodontal disease in individuals with ID.

3.
J Prosthodont Res ; 61(2): 185-192, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27363289

RESUMO

PURPOSE: Studies of effective methods for the easy removal of denture adhesives from a denture base are not well represented in the literature. We previously assessed the removability of denture adhesives by immersing within denture cleaners, showing that some cleaners have a weak effect, insufficiently effective in daily use. In this study, we prepared a cellulase, as a potential component for denture adhesive removers, and we examined whether the addition of cellulase to denture cleaners is effective in the removal of cream denture adhesives. METHODS: We prepared the cellulase Meicelase as one component for the liquefaction of denture adhesives. We used two denture cleaners and two cream adhesives. After the immersion of plates in sample solutions, we evaluated the area of the sample plate still covered with adhesives. Biofilm removal assay was also performed using denture cleaners containing cellulase. RESULTS: The addition of cellulase accelerated the removal of cream adhesives in immersion experiments to a rate faster than that of water and denture cleaners. However, it did not influence the removability of Candida albicans biofilms from acrylic resin specimens. CONCLUSION: Cellulase hastened the liquefaction of cream adhesives.


Assuntos
Celulase , Cimentos Dentários , Bases de Dentadura , Higienizadores de Dentadura , Resinas Acrílicas , Biofilmes , Candida albicans/fisiologia , Bases de Dentadura/microbiologia , Higienizadores de Dentadura/química , Resinas Sintéticas
4.
Int J Dev Biol ; 60(1-3): 21-8, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26934293

RESUMO

Neural crest (NC) cells are a group of cells located in the neural folds at the boundary between the neural and epidermal ectoderm. Cranial NC cells migrate to the branchial arches and give rise to the majority of the craniofacial region, whereas trunk and tail NC cells contribute to the heart, enteric ganglia of the gut, melanocytes, sympathetic ganglia, and adrenal chromaffin cells. Positional information is indispensable for the regulation of cranial or trunk and tail NC cells. However, the mechanisms underlying the regulation of positional information during human NC induction have yet to be fully elucidated. In the present study, supplementation of bone morphogenetic protein (BMP) 4 in defined serum-free culture conditions including fibroblast growth factor-2 and Wnt3a from day 8 after NC specification induced the expression of cranial NC markers, AP2alpha, MSX1, and DLX1, during NC cell differentiation from human pluripotent stem cells. On the other hand, the proportion of cells expressing p75(NTR) or HNK1 decreased compared with that of cells cultured without BMP4, whereas gene expression analysis demonstrated that the expression levels of cranial NC-associated genes increased in BMP4-treated NC cells. These BMP4-treated NC cells were capable of differentiation into osteocytes and chondrocytes. The results of the present study indicate that BMP4 regulates cranial positioning during NC development.


Assuntos
Proteína Morfogenética Óssea 4/farmacologia , Diferenciação Celular/efeitos dos fármacos , Crista Neural/citologia , Células-Tronco Pluripotentes/citologia , Ativação Transcricional/efeitos dos fármacos , Diferenciação Celular/genética , Linhagem Celular , Condrócitos/citologia , Condrócitos/metabolismo , Ossos Faciais/citologia , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Imuno-Histoquímica , Fator de Transcrição MSX1/genética , Fator de Transcrição MSX1/metabolismo , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Crista Neural/metabolismo , Osteócitos/citologia , Osteócitos/metabolismo , Células-Tronco Pluripotentes/metabolismo , Receptores de Fator de Crescimento Neural/genética , Receptores de Fator de Crescimento Neural/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Crânio/citologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Ativação Transcricional/genética , Transcriptoma/efeitos dos fármacos , Transcriptoma/genética
5.
In Vitro Cell Dev Biol Anim ; 51(8): 769-75, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25898826

RESUMO

Neural differentiation is an important target of human embryonic stem cells, which provide a source for cell-based therapy, developmental biology, and pharmaceutical research. Previous studies revealed that inhibition of the bone morphogenetic protein is required for neural induction from human embryonic stem cells. On the contrary, the functions of fibroblast growth factors and Activin/Nodal signaling are controversial. Fibroblast growth factor-2 and Activin/Nodal pathways exert divergent influences on human embryonic stem cell concerning the maintenance of both pluripotency and cellular differentiation. We hypothesized that the combination of fibroblast growth factor-2 and Activin A at various concentrations synergistically exerts diverse effects on cell differentiation. To determine the effects of fibroblast growth factor-2 and Activin A on cellular differentiation into neural lineages, we examined the expression of neural differentiation markers in human embryonic stem cells treated with fibroblast growth factor-2 and/or Activin A at various concentrations in a growth factor-defined serum-free medium in short-term culture. In this study, we provide evidence that fibroblast growth factor-2 and Activin A synergistically regulated the initiation of human embryonic stem cell differentiation into neural cell lineages even though human embryonic stem cells autonomously differentiate into neural cell lineages.


Assuntos
Ativinas/farmacologia , Diferenciação Celular/efeitos dos fármacos , Fator 2 de Crescimento de Fibroblastos/farmacologia , Células-Tronco Pluripotentes/efeitos dos fármacos , Diferenciação Celular/fisiologia , Linhagem da Célula/efeitos dos fármacos , Linhagem da Célula/fisiologia , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Humanos , Neurônios/citologia , Neurônios/efeitos dos fármacos , Neurônios/fisiologia , Células-Tronco Pluripotentes/fisiologia
6.
Stem Cells Dev ; 23(18): 2180-9, 2014 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-24410631

RESUMO

Epithelial-mesenchymal transition (EMT) has been thought to occur during early embryogenesis, and also the differentiation process of human embryonic stem (hES) cells. Spontaneous differentiation is sometimes observed at the peripheral of the hES cell colonies in conventional culture conditions, indicating that EMT occurs in hES cell culture. However, the triggering mechanism of EMT is not yet fully understood. The balance between self-renewal and differentiation of human pluripotent stem (hPS) cells is controlled by various signal pathways, including the fibroblast growth factor (FGF)-2. However, FGF-2 has a complex role for self-renewal of hES cells. FGF-2 activates phosphatidylinositol-3 kinase/AKT, mitogen-activated protein kinase/extracellular signal-regulated kinase-1/2 kinase, and also protein kinase C (PKC). Here, we showed that a PKC rapidly induced an early growth response protein-1 (EGR-1) in hES cells, which was followed by upregulation of EMT-related genes. Before the induction of EMT-related genes, EGR-1 was translocated into the nucleus, and then bound directly to the promoter region of SNAIL, which is a master regulator of EMT. SNAIL expression was attenuated by knockdown of EGR-1, but upregulated by ectopic expression of EGR-1. EGR-1 as the downstream signal of PKC might play a key role in EMT initiation during early differentiation of hES cells. This study would lead to a more robust understanding of the mechanisms underlying the balance between self-renewal and initiation of differentiation in hPS cells.


Assuntos
Proteína 1 de Resposta de Crescimento Precoce/metabolismo , Células-Tronco Embrionárias/metabolismo , Transição Epitelial-Mesenquimal , Regiões Promotoras Genéticas , Proteína Quinase C/metabolismo , Fatores de Transcrição/genética , Linhagem Celular , Células-Tronco Embrionárias/efeitos dos fármacos , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Transição Epitelial-Mesenquimal/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Ligação Proteica/efeitos dos fármacos , Fatores de Transcrição da Família Snail , Acetato de Tetradecanoilforbol/farmacologia , Fatores de Transcrição/metabolismo
7.
PLoS One ; 8(1): e54122, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23349801

RESUMO

BACKGROUND: The self-renewal of human pluripotent stem (hPS) cells including embryonic stem and induced pluripotent stem cells have been reported to be supported by various signal pathways. Among them, fibroblast growth factor-2 (FGF-2) appears indispensable to maintain self-renewal of hPS cells. However, downstream signaling of FGF-2 has not yet been clearly understood in hPS cells. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we screened a kinase inhibitor library using a high-throughput alkaline phosphatase (ALP) activity-based assay in a minimal growth factor-defined medium to understand FGF-2-related molecular mechanisms regulating self-renewal of hPS cells. We found that in the presence of FGF-2, an inhibitor of protein kinase C (PKC), GF109203X (GFX), increased ALP activity. GFX inhibited FGF-2-induced phosphorylation of glycogen synthase kinase-3ß (GSK-3ß), suggesting that FGF-2 induced PKC and then PKC inhibited the activity of GSK-3ß. Addition of activin A increased phosphorylation of GSK-3ß and extracellular signal-regulated kinase-1/2 (ERK-1/2) synergistically with FGF-2 whereas activin A alone did not. GFX negated differentiation of hPS cells induced by the PKC activator, phorbol 12-myristate 13-acetate whereas Gö6976, a selective inhibitor of PKCα, ß, and γ isoforms could not counteract the effect of PMA. Intriguingly, functional gene analysis by RNA interference revealed that the phosphorylation of GSK-3ß was reduced by siRNA of PKCδ, PKCε, and ζ, the phosphorylation of ERK-1/2 was reduced by siRNA of PKCε and ζ, and the phosphorylation of AKT was reduced by PKCε in hPS cells. CONCLUSIONS/SIGNIFICANCE: Our study suggested complicated cross-talk in hPS cells that FGF-2 induced the phosphorylation of phosphatidylinositol-3 kinase (PI3K)/AKT, mitogen-activated protein kinase/ERK-1/2 kinase (MEK), PKC/ERK-1/2 kinase, and PKC/GSK-3ß. Addition of GFX with a MEK inhibitor, U0126, in the presence of FGF-2 and activin A provided a long-term stable undifferentiated state of hPS cells even though hPS cells were dissociated into single cells for passage. This study untangles the cross-talk between molecular mechanisms regulating self-renewal and differentiation of hPS cells.


Assuntos
Proliferação de Células , Células-Tronco Pluripotentes/metabolismo , Proteína Quinase C-delta/metabolismo , Proteína Quinase C-épsilon/metabolismo , Proteína Quinase C/metabolismo , Ativinas/farmacologia , Fosfatase Alcalina/metabolismo , Western Blotting , Carbazóis/farmacologia , Linhagem Celular , Cromonas/farmacologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Fator 2 de Crescimento de Fibroblastos/farmacologia , Quinase 3 da Glicogênio Sintase/metabolismo , Glicogênio Sintase Quinase 3 beta , Humanos , Imuno-Histoquímica , Indóis/farmacologia , Maleimidas/farmacologia , Morfolinas/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação/efeitos dos fármacos , Células-Tronco Pluripotentes/citologia , Proteína Quinase C/genética , Proteína Quinase C-delta/genética , Proteína Quinase C-épsilon/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Interferência de RNA , Transdução de Sinais/efeitos dos fármacos , Acetato de Tetradecanoilforbol/farmacologia
8.
Int J Dev Biol ; 55(2): 181-7, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21305471

RESUMO

Human bone marrow-derived mesenchymal stem cells (hMSCs) are potential cellular sources of therapeutic stem cells as they have the ability to proliferate and differentiate into a wide array of mesenchymal cell types such as osteoblasts, chondroblasts and adipocytes. hMSCs have been used clinically to treat patients with graft vs. host disease, osteogenesis imperfect, or alveolar cleft, suggesting that transplantation of hMSCs is comparatively safe as a stem cell-based therapy. However, conventional culture medium for hMSCs contains fetal bovine serum (FBS). In the present study, we developed a growth factor-defined, serum-free medium for culturing hMSCs. Under these conditions, TGF-beta1 promoted proliferation of hMSCs. The expanded hMSC population expressed the human pluripotency markers SSEA-3, -4, NANOG, OCT3/4 and SOX2. Furthermore, double positive cells for SSEA-3 and a mesenchymal cell marker, CD105, were detected in the population. The potential to differentiate into osteoblasts and adipocytes was confirmed. This work provides a useful tool to understand the basic biological properties of hMSCs in culture.


Assuntos
Proliferação de Células , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Pluripotentes/metabolismo , Técnicas de Cultura de Tecidos , Antígenos CD/biossíntese , Antígenos Glicosídicos Associados a Tumores/biossíntese , Diferenciação Celular , Células Cultivadas , Meios de Cultura Livres de Soro , Endoglina , Proteínas de Homeodomínio/biossíntese , Humanos , Proteína Homeobox Nanog , Fator 3 de Transcrição de Octâmero/biossíntese , Receptores de Superfície Celular/biossíntese , Fatores de Transcrição SOXB1/biossíntese , Antígenos Embrionários Estágio-Específicos/biossíntese , Fator de Crescimento Transformador beta/farmacologia
9.
J Investig Clin Dent ; 2(3): 187-96, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25426790

RESUMO

AIM: The aim of this study was to find the oral isolate of lactobacilli, which has the potential to inhibit either periodontal, cariogenic, or fungal pathogens in vitro, and to examine the effects of bovine milk fermented with the isolate on the oral carriage of cariogenic and periodontal pathogens. METHODS: The inhibitory effects of the supernatant of Man-Rogosa-Sharpe broth, in which each of 42 oral isolates of lactobacilli grown, was examined. One isolate, Lactobacillus rhamnosus L8020, that showed the potential to inhibit either periodontal, cariogenic, or fungal pathogens in vitro, was used to examine the effects of fermented milk on the oral carriage of cariogenic and periodontal pathogens, which was examined by a placebo-controlled and cohort trial using 50 participants. RESULTS: Edible yogurt containing Lactobacillus rhamnosus L8020 significantly reduced the oral carriage of mutans streptococci (P < 0.01) and four periodontal pathogens examined: Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, and Fusobacterium spp. (P < 0.01), but the phenomenon were not observed with the placebo yogurt (P > 0.05). CONCLUSION: These results suggest that yogurt with Lactobacillus rhamnosus L8020 could reduce the risk of dental caries and periodontal disease.


Assuntos
Antibiose/fisiologia , Bactérias Gram-Negativas/fisiologia , Lactobacillus rhamnosus/metabolismo , Boca/microbiologia , Streptococcus mutans/fisiologia , Iogurte/microbiologia , Animais , Carga Bacteriana , Técnicas Bacteriológicas , Bacteroides/fisiologia , Candida albicans/fisiologia , Bovinos , Estudos de Coortes , Método Duplo-Cego , Feminino , Fusobacterium/fisiologia , Humanos , Lactobacillus rhamnosus/fisiologia , Masculino , Placebos , Porphyromonas gingivalis/fisiologia , Prevotella intermedia/fisiologia , Saliva/microbiologia , Streptococcus sobrinus/fisiologia , Adulto Jovem
10.
J Prosthodont Res ; 54(1): 1-6, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19733525

RESUMO

PURPOSE: To investigate the effects of titanium (Ti) ions on the cell viability, the cell differentiation and the gene expressions related to bone resorption including Receptor Activator of NF-kappaB Ligand (RANKL) and Osteoprotegerin (OPG) in the tissues around dental implants, the osteoblast-, osteoclast-, and gingival epithelial-like cells were exposed to Ti ions. METHODS: An MTS assay was carried out to evaluate the viabilities of osteoblast-like MC3T3-E1, osteoclast-like RAW264.7 and epithelial cell-like GE-1 cells. The gene expressions in these cells were analyzed by the use of RT-PCR and real-time quantitative RT-PCR. RESULTS: Ti ions in the concentration range 1-9 ppm had little effect on the viabilities of MC3T3-E1, RAW264.7 and GE-1, whereas 20 ppm Ti ions significantly decreased the viabilities of all cells. Analyses of RT-PCR and real-time quantitative RT-PCR data revealed that Ti ions at 9 ppm remarkably inhibited the expressions of Runx2, Osterix and type I collagen in MC3T3-E1. In RAW264.7, Ti ions showed no effects on the levels of mRNAs for TRAP and cathepsin K enhanced by RANKL. Ti ions at the range of 1-9 ppm showed no effects on the levels of mRNAs for RANKL and OPG in GE-1, while Ti ions at 9 ppm enhanced the expression of these genes in MC3T3-E1. CONCLUSIONS: These results, taken together, suggested that Ti ions show the biological effects, both on the viabilities of osteoblast and osteoclast and on the differentiation of either the osteoblastic or osteoclastic cells, which may influence the prognosis of dental implants.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Implantes Dentários , Células Epiteliais/citologia , Gengiva/citologia , Osteoblastos/citologia , Osteoclastos/citologia , Titânio/efeitos adversos , Animais , Remodelação Óssea/efeitos dos fármacos , Reabsorção Óssea/genética , Diferenciação Celular/genética , Células Cultivadas , Depressão Química , Relação Dose-Resposta a Droga , Expressão Gênica/efeitos dos fármacos , Íons , Camundongos , Osteoprotegerina , Ligante RANK
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