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1.
J Prosthodont Res ; 66(1): 184-192, 2022 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-34053972

RESUMO

PATIENT: A 54-year-old woman presenting with anterior alveolar ridge resorption was submitted to a connective tissue graft (CTG) for esthetic improvement before rehabilitation with a fixed partial denture. Palate-harvested connective tissue was used as a graft after extra-oral removal of the epithelium. Unexpectedly, complete wound healing was not observed. Moreover, 6 months post-surgery, a white discharge was detected at the grafted site. The adjacent tooth showing a root fracture was initially associated with the symptoms and was then extracted. Concomitantly, the unhealed tissue at the grafted site was also excised, leading to temporary symptom resolution. However, the white discharge reappeared after 2 months. The excision area was expanded to remove the grafted tissue entirely, and the wound was completely healed. Since the alveolar ridge resorption had become larger compared to the preoperative condition, the patient was subjected to a second CTG, now using a connective tissue harvested from the palate by a single incision technique. The wound healed uneventfully, and the final prosthesis was delivered 6 months after soft tissue stabilization. The patient has been followed-up for more than 28 months without any recurrence of white discharge. DISCUSSION: Histopathological and cytological examination detected keratinized epithelial tissues and cells, respectively, in excised tissues and white discharge specimens. Consequently, a possible relationship between white discharge and residual epithelium in the harvested graft was strongly suspected. CONCLUSION: Success of the CTG procedure requires careful method selection for tissue transplantation and treatment execution.


Assuntos
Perda do Osso Alveolar , Aumento do Rebordo Alveolar , Tecido Conjuntivo , Prótese Parcial Fixa , Feminino , Gengiva , Humanos , Pessoa de Meia-Idade
2.
Sci Rep ; 7: 44522, 2017 03 16.
Artigo em Inglês | MEDLINE | ID: mdl-28300208

RESUMO

Whole-organ regeneration has great potential for the replacement of dysfunctional organs through the reconstruction of a fully functional bioengineered organ using three-dimensional cell manipulation in vitro. Recently, many basic studies of whole-tooth replacement using three-dimensional cell manipulation have been conducted in a mouse model. Further evidence of the practical application to human medicine is required to demonstrate tooth restoration by reconstructing bioengineered tooth germ using a postnatal large-animal model. Herein, we demonstrate functional tooth restoration through the autologous transplantation of bioengineered tooth germ in a postnatal canine model. The bioengineered tooth, which was reconstructed using permanent tooth germ cells, erupted into the jawbone after autologous transplantation and achieved physiological function equivalent to that of a natural tooth. This study represents a substantial advancement in whole-organ replacement therapy through the transplantation of bioengineered organ germ as a practical model for future clinical regenerative medicine.


Assuntos
Medicina Regenerativa , Engenharia Tecidual , Germe de Dente/transplante , Dente/transplante , Animais , Engenharia Biomédica/tendências , Cães , Humanos , Odontogênese/fisiologia , Regeneração/fisiologia , Células-Tronco , Dente/crescimento & desenvolvimento , Erupção Dentária , Germe de Dente/fisiologia , Reimplante Dentário , Transplante Autólogo/métodos
3.
J Prosthodont Res ; 59(3): 178-84, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26077377

RESUMO

PURPOSE: Many studies have identified risk factors for dental implant failure, although few have investigated the correlation among implant fixtures within single patients. A better analytical method may include repeated measures analysis including generalized estimating equations (GEE). This retrospective cohort study aimed to (1) identify the risk factors for failure of dental implantation and (2) evaluate an analytical method using GEE analysis. METHODS: We analyzed data on early and late implant failures in 296 patients providing 721 rough surface dental implants (2.44 implants per patient). Potential predictors of implant failure included age, gender, smoking, location of implant, use of bone augmentation, number of remaining teeth, opposing tooth condition, fixture length, fixture diameter and type of suprastructure (fixed or removable partial denture). The likelihood of early and late implant failure was estimated by GEE. RESULTS: The early failure rate was 1.5% (11/721 implants, 7/296 patients) and the 10-year cumulative survival rate was 94.0% (7/710 implants, 5/293 patients). The GEE analysis revealed that a significant risk factor for early implant failure was smoking (p<0.01), whereas significant risk factors for late failure were maxillary implant (p=0.02), posterior implant (p<0.01), number of remaining teeth (≥20) (p<0.01), opposing unit being a removable partial denture or nothing (p=0.04) and having a removable type suprastructure (p<0.01). CONCLUSIONS: GEE analysis showed that smoking was a risk factor for early implant failure, and several risk factors were identified for late implant failure.


Assuntos
Implantes Dentários , Falha de Restauração Dentária/estatística & dados numéricos , Idoso , Aumento do Rebordo Alveolar/efeitos adversos , Estudos de Coortes , Prótese Parcial Removível/efeitos adversos , Feminino , Humanos , Estudos Longitudinais , Masculino , Maxila/cirurgia , Estudos Retrospectivos , Fatores de Risco , Fumar/efeitos adversos , Fatores de Tempo
4.
J Bone Miner Res ; 30(9): 1585-96, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25753754

RESUMO

Articular cartilage repair remains a challenging problem. Based on a high-throughput screening and functional analysis, we found that fluocinolone acetonide (FA) in combination with transforming growth factor beta 3 (TGF-ß3) strongly potentiated chondrogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs). In an in vivo cartilage defect model in knee joints of immunocompromised mice, transplantation of FA/TGF-ß3-treated hBMSCs could completely repair the articular surface. Analysis of the intracellular pathways revealed that FA enhanced TGF-ß3-induced phosphorylation of Smad2 and Smad3. Additionally, we performed a pathway array and found that FA activates the mTORC1/AKT pathway. Chemical inhibition of mTORC1 with rapamycin substantially suppressed FA effect, and inhibition of AKT completely repressed chondrogenesis of hBMSCs. Inhibition of glucocorticoid receptor with mifepristone also suppressed FA effect, suggesting that FA involves binding to the glucocorticoid receptor. Comparative analysis with other glucocorticoids (triamcinolone acetonide [TA] and dexamethasone [DEX]) revealed the unique ability of FA to repair articular cartilage surgical defects. Analysis of intracellular pathways showed that the mTORC1/AKT pathway and the glucocorticoid receptor was highly activated with FA and TA, but to a lesser extent with DEX. Collectively, these results show a unique ability of FA to enhance TGF-ß3-associated chondrogenesis, and suggest that the FA/TGF-ß3 combination may be used as major inducer of chondrogenesis in vitro. Additionally, FA/TGF-ß3 could be potentially applied in a clinical setting to increase the efficiency of regenerative approaches based on chondrogenic differentiation of stem cells.


Assuntos
Células da Medula Óssea/citologia , Cartilagem Articular/efeitos dos fármacos , Condrogênese/fisiologia , Fluocinolona Acetonida/química , Células-Tronco Mesenquimais/citologia , Fator de Crescimento Transformador beta3/metabolismo , Animais , Anti-Inflamatórios/química , Cartilagem Articular/metabolismo , Diferenciação Celular , Células Cultivadas , Condrogênese/efeitos dos fármacos , Dexametasona/química , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Camundongos SCID , Mifepristona/metabolismo , Receptores de Glucocorticoides/metabolismo , Regeneração , Transdução de Sinais , Triancinolona Acetonida/química
5.
J Prosthodont Res ; 59(2): 152-8, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25623362

RESUMO

PURPOSE: The aim of this article was to investigate the accuracy in the reproducibility of full-arch implant provisional restorations to final restorations between a 3D Scan/CAD/CAM technique and the conventional method. METHODS: We fabricated two final restorations for rehabilitation of maxillary and mandibular complete edentulous area and performed a computer-based comparative analysis of the accuracy in the reproducibility of the provisional restoration to final restoration between a 3D scanning and CAD/CAM (Scan/CAD/CAM) technique and the conventional silicone-mold transfer technique. Final restorations fabricated either by the conventional or Scan/CAD/CAM method were successfully installed in the patient. The total concave/convex volume discrepancy observed with the Scan/CAD/CAM technique was 503.50mm(3) and 338.15 mm(3) for maxillary and mandibular implant-supported prostheses (ISPs), respectively. On the other hand, total concave/convex volume discrepancy observed with the conventional method was markedly high (1106.84 mm(3) and 771.23 mm(3) for maxillary and mandibular ISPs, respectively). CONCLUSIONS: The results of the present report suggest that Scan/CAD/CAM method enables a more precise and accurate transfer of provisional restorations to final restorations compared to the conventional method.


Assuntos
Desenho Assistido por Computador , Implantação Dentária/métodos , Implantes Dentários , Planejamento de Prótese Dentária/métodos , Imageamento Tridimensional , Boca Edêntula/reabilitação , Idoso , Prótese Dentária Fixada por Implante , Humanos , Masculino , Reprodutibilidade dos Testes
7.
Bone ; 69: 165-73, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25240457

RESUMO

Bone morphogenetic protein 2 (BMP2) is a potent osteoinductive cytokine that plays crucial roles in bone repair. However, large amounts of BMP2 are required to induce sufficient bone formation in humans possibly due to a feedback response of BMP antagonists. The engineered BMP2 variant L51P is deficient in BMP receptor type I activation but maintains affinity for BMP antagonists and can allow for the inactivation of BMP antagonists, and eventually enhance BMP2 action. As hypothesized, simultaneous addition of L51P enhanced the BMP2-induced osteogenesis. To test the ability of L51P to competitively inactivate BMP antagonists, cell binding affinity of BMP2 ligands was investigated in the presence or absence of L51P. Because the BMP antagonists were highly expressed 3 days after exogenous BMP2 stimulation, we collected supernatants from 3-day stimulated cell cultures and used as condition culture media (CM). The results showed a significant decrease in the cell binding of BMP2 ligands when cells were incubated with exogenous BMP2 and CM, whereas L51P addition competitively rescued the suppression of BMP2-to-cell binding induced by CM incubation. In a delayed experimental model, L51P was applied 3 days after exogenous BMP2 stimulation and we could observe a striking enhancement of the BMP2-induced SMAD-1/5/8 phosphorylation and luciferase activity of the Id1 promoter compared to the simultaneous addition of the two factors. These findings provide a deeper insight into the cellular and molecular mechanisms involved in the effect of L51P in suppressing the BMP antagonists and enhancing BMP activity. Additionally, these results demonstrate that L51P is a promising down regulator of BMP-induced negative feedback, which could have a significant impact in future applications of BMP2 in research and clinical settings.


Assuntos
Proteína Morfogenética Óssea 2/antagonistas & inibidores , Osteogênese/efeitos dos fármacos , Animais , Western Blotting , Proteína Morfogenética Óssea 2/metabolismo , Linhagem Celular , Retroalimentação Fisiológica/efeitos dos fármacos , Feminino , Camundongos , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Proteínas Recombinantes/farmacologia , Microtomografia por Raio-X
8.
Stem Cell Res Ther ; 5(1): 31, 2014 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-24580841

RESUMO

INTRODUCTION: During normal pulp tissue healing, inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-α) or interleukins, act in the initial 48 hours (inflammatory phase) and play important roles not only as chemo-attractants of inflammatory cells and stem/progenitor cells but also in inducing a cascade of reactions toward tissue regeneration or reparative dentin formation or both. Previous reports have shown that inflammatory cytokines regulate the differentiation capacity of dental pulp stem/progenitor cells (DPCs), but none has interrogated the impact of these cytokines on the stem cell phenotype of stem/progenitor cells. This study investigated the effects of a short-term treatment with TNF-α on the stem cell phenotype and differentiation ability of human DPCs. METHODS: An in vivo mouse model of pulp exposure was performed for analysis of expression of the mesenchymal stem cell marker CD146 in DPCs during the initial stage of inflammatory response. For in vitro studies, human DPCs were isolated and incubated with TNF-α for 2 days and passaged to eliminate TNF-α completely. Analysis of stem cell phenotype was performed by quantification of cells positive for mesenchymal stem cell markers SSEA-4 (stage-specific embryonic antigen 4) and CD146 by flow cytometry as well as by quantitative analysis of telomerase activity and mRNA levels of OCT-4 and NANOG. Cell migration, colony-forming ability, and differentiation toward odontogenesis and adipogenesis were also investigated. RESULTS: The pulp exposure model revealed a strong staining for CD146 during the initial inflammatory response, at 2 days after pulp exposure. In vitro experiments demonstrated that a short-term (2-day) treatment of TNF-α increased by twofold the percentage of SSEA-4+ cells. Accordingly, STRO-1, CD146, and SSEA-4 protein levels as well as OCT-4 and NANOG mRNA levels were also significantly upregulated upon TNF-α treatment. A short-term TNF-α treatment also enhanced DPC function, including the ability to form cell colonies, to migrate, and to differentiate into odontogenic and adipogenic lineages. CONCLUSIONS: A short-term treatment with TNF-α enhanced the stem cell phenotype, migration, and differentiation ability of DPCs.


Assuntos
Adipogenia , Células-Tronco Adultas/citologia , Polpa Dentária/citologia , Odontogênese , Fator de Necrose Tumoral alfa/farmacologia , Adulto , Células-Tronco Adultas/efeitos dos fármacos , Células-Tronco Adultas/metabolismo , Animais , Antígenos de Superfície/genética , Antígenos de Superfície/metabolismo , Antígeno CD146/genética , Antígeno CD146/metabolismo , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Camundongos , Proteína Homeobox Nanog , Fator 3 de Transcrição de Octâmero/genética , Fator 3 de Transcrição de Octâmero/metabolismo , Fenótipo , Antígenos Embrionários Estágio-Específicos/genética , Antígenos Embrionários Estágio-Específicos/metabolismo
9.
Cells Tissues Organs ; 199(4): 249-55, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25614023

RESUMO

Several preclinical studies have shown that Escherichia coli-derived bone morphogenetic protein-2 (E-BMP-2) is as effective as mammalian cell-derived bone morphogenetic protein-2 (C-BMP-2) in the treatment of bone defects. However, further investigation of the effectiveness and determination of the optimal dosage of E-BMP-2 in large animals are still necessary before its full application in humans. This study investigated the efficiency of different concentrations of E-BMP-2 adsorbed in ß-TCP for bone augmentation and osseointegration of immediate dental implants in a swine socket lift model. Following exposure of the maxillary sinus lateral wall, a 3.4-mm (diameter) cavity was drilled and filled with 0.1 g of ß-TCP containing different doses of E-BMP-2 (0, 10, 30, or 100 µg/site) to lift the Schneiderian membrane. A dental implant was then immediately inserted. Bone-to-implant contact (BIC) and bone density (BD) examined via histological analysis were used as parameters to assess E-BMP-2 efficiency in bone formation. The implant stability quotient (ISQ) was measured using Osstell to determine the effect of E-BMP-2/ß-TCP on implant stability. After 8 weeks, the groups that received 30 and 100 µg of E-BMP-2 showed substantial new bone formation in the elevated space, while no bone formation was observed with ß-TCP alone. Accordingly, BIC and BD presented a dose-dependent response to increasing doses of E-BMP-2. However, there was no increase in implant stability with E-BMP-2 treatment. In conclusion, the E-BMP-2/ß-TCP combination was efficient in bone formation and osseointegration of dental implants in a socket lift model in mini-pigs.


Assuntos
Proteína Morfogenética Óssea 2/metabolismo , Fosfatos de Cálcio/metabolismo , Escherichia coli/patogenicidade , Osteogênese/efeitos dos fármacos , Fator de Crescimento Transformador beta/metabolismo , Animais , Regeneração Óssea , Humanos , Masculino , Proteínas Recombinantes/metabolismo , Suínos
10.
J Prosthodont Res ; 57(4): 262-7, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24126107

RESUMO

PURPOSE: This study aimed to compare the various complication-free rates and survival rates of remaining teeth among implant-supported fixed dentures (IFDs), removable partial dentures (RPDs) and no-restoration (NR) patients with unilateral free-end edentulism. MATERIAL AND METHODS: The study subjects were selected among those who received prosthodontic treatment at Okayama University Dental Hospital for their unilateral free-end edentulism (2 or 3 missing teeth). Thirty-three patients were included in the IFD group, 41 matched patients in the RPD group, and 10 patients who received RPDs but refused their use were regarded as NR group. The remaining dentition was classified into five subcategories in relation to the missing portion: adjacent teeth to the missing portion (AD), contralateral posterior dentition in the same jaw (CS) and in the opposite jaw (CO), ipsilateral opposing posterior dentition (IO), and anterior dentition (AN). Complications were defined as tooth extraction, periodontal lesions, periapical lesions or loss of retention of the prosthesis and were assessed by one examiner based on the hospital chart records. RESULTS: The cumulative complication-free rates in the remaining teeth were significantly different among each of the three groups (p<0.01), with a significantly lower incidence rate in the IFD group. Regarding the cumulative survival rate of the remaining teeth, there was a significant difference only between IFD and NR group (p=0.01), especially in the CO region (p=0.04). CONCLUSIONS: Stable posterior occlusal support obtained with IFD treatment for unilateral free-end edentulism may reduce the incidence of complications in the remaining teeth, by decreasing the adverse mechanical stress.


Assuntos
Prótese Dentária Fixada por Implante , Dentição , Prótese Parcial Fixa , Arcada Edêntula/reabilitação , Dente/fisiologia , Adulto , Idoso , Força de Mordida , Dente Suporte/efeitos adversos , Prótese Dentária Fixada por Implante/efeitos adversos , Prótese Parcial Fixa/efeitos adversos , Prótese Parcial Removível , Feminino , Humanos , Arcada Edêntula/fisiopatologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Estresse Mecânico , Perda de Dente/etiologia , Perda de Dente/prevenção & controle
11.
J Prosthodont Res ; 57(3): 156-61, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23838063

RESUMO

PURPOSE: This study aimed to compare the survival rates of remaining teeth between implant-supported fixed dentures (IFDs) and removable partial dentures (RPDs) in patients with large edentulous cases. The second goal was to assess the risk factors for remaining tooth loss. MATERIALS AND METHODS: The study subjects were selected among those who received prosthodontic treatment at Okayama University Dental Hospital for their edentulous space exceeding at least four continuous missing teeth. Twenty-one patients were included in the IFD group and 82 patients were included in the RPD group. Survival rates of remaining teeth were calculated in three subcategories: (1) whole remaining teeth, (2) adjacent teeth to intended edentulous space, and (3) opposing teeth to intended edentulous space. RESULTS: The ten-year cumulative survival rate of the whole remaining teeth was significantly higher in the IFD group (40.0%) than in the RPD group (24.4%). On the other hand, there was no significant difference between two groups in the survival rate of teeth adjacent or opposing to intended edentulous space. A Cox proportional hazard analysis revealed that RPD restoration and gender (male) were the significant risk factors for remaining tooth loss (whole remaining teeth). CONCLUSIONS: These results suggest that IFD treatment can reduce the incidence of remaining tooth loss in large edentulous cases.


Assuntos
Implantes Dentários/efeitos adversos , Prótese Dentária Fixada por Implante/efeitos adversos , Prótese Parcial Fixa/efeitos adversos , Prótese Parcial Removível/efeitos adversos , Arcada Parcialmente Edêntula/terapia , Perda de Dente/epidemiologia , Perda de Dente/etiologia , Adulto , Idoso , Força de Mordida , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Fatores de Tempo
12.
Int J Prosthodont ; 26(3): 260-4, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23626980

RESUMO

PURPOSE: To compare the complication rate of natural teeth adjacent to implant supported dentures (IFDs) with that of teeth serving as abutments for fixed partial dentures (FPDs). The second goal was to assess the risk factors for complications in teeth adjacent to bounded edentulous spaces. MATERIALS AND METHODS: The study subjects were selected from patients who received prosthodontic treatment for their bounded edentulous space not exceeding two missing teeth between February 1990 and March 2007. Sixty-one patients were included in the IFD group and 66 patients were included in the FPD group. Tooth complications were defined as tooth extraction, periodontal lesion, periapical lesion, and loss of prosthesis and were assessed by one examiner based on dental records. RESULTS: The 8-year cumulative complication rate for the IFD group (7.9%) was significantly lower than for the FPD group (40.7%). Additionally, the 8-year cumulative complication rate of vital teeth (6%) was significantly lower than that of nonvital teeth (45.9%). A cox proportional hazard analysis revealed that nonvitality of dental pulp was a significant risk factor for tooth complications, whereas treatment modality was not. CONCLUSIONS: Teeth adjacent to IFD-treated edentulous spaces presented fewer complications than natural teeth serving as abutments for FPDs. Conservation of teeth adjacent to edentulous spaces as vital teeth was the key finding to limit further tooth loss.


Assuntos
Prótese Dentária Fixada por Implante/efeitos adversos , Falha de Restauração Dentária , Prótese Parcial Fixa/efeitos adversos , Doenças Periapicais/etiologia , Periodontite/etiologia , Perda de Dente/etiologia , Dente não Vital/etiologia , Adulto , Idoso , Distribuição de Qui-Quadrado , Feminino , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Modelos de Riscos Proporcionais , Estudos Retrospectivos , Fatores de Risco , Estatísticas não Paramétricas
13.
Biochimie ; 95(2): 374-81, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23116713

RESUMO

A significant number of natural compounds have been shown to regulate the behavior of the cells, in collaboration with cellular proteins. CCN2/connective tissue growth factor (CTGF) has been reported to have essential roles in cartilage development, chondrocyte proliferation and differentiation as well as regulation of the extracellular matrix metabolism. Previous studies demonstrated the capability of CCN2 to regenerate surgical defects in articular cartilage of rat knee. Also, transgenic mice over-expressing cartilage-specific CCN2 were shown to be more resistant to aging-related cartilage degradation. We hypothesized that small molecules that induce CCN2 in chondrocytes could be novel candidates to increase the resistance to aging-related cartilage degradation, or even to correct cartilage degenerative changes incurred in OA. Therefore, this study screened a compound library and identified the ß-carboline alkaloid harmine as a novel inducer of CCN2 in human chondrocytic HCS-2/8 cells and osteoarthritic articular chondrocytes. Harmine increased the expression of the cartilage markers aggrecan and COL2α1, as well as that of the master regulator of chondrogenesis, SOX-9. Moreover, harmine notably induced chondrogenesis of prechondrocytic ATDC5 cells in micromass cultures. The chondroprotective effect of harmine was investigated under inflammatory condition by stimulation with TNFα, and harmine was shown to ameliorate TNFα-induced decrease in expression of CCN2 and cartilage markers. These findings uncover novel chondrogenic effects of harmine and indicate harmine as a potential drug for prevention and/or repair of cartilage degradation.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Cartilagem Articular/efeitos dos fármacos , Condrócitos/efeitos dos fármacos , Matriz Extracelular/efeitos dos fármacos , Harmina/farmacologia , Substâncias Protetoras/farmacologia , Bibliotecas de Moléculas Pequenas/farmacologia , Agrecanas/genética , Agrecanas/metabolismo , Envelhecimento/efeitos dos fármacos , Envelhecimento/metabolismo , Envelhecimento/patologia , Biomarcadores/metabolismo , Cartilagem Articular/citologia , Cartilagem Articular/metabolismo , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Condrócitos/citologia , Condrócitos/metabolismo , Condrogênese/efeitos dos fármacos , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Fator de Crescimento do Tecido Conjuntivo/genética , Fator de Crescimento do Tecido Conjuntivo/metabolismo , Matriz Extracelular/metabolismo , Expressão Gênica/efeitos dos fármacos , Humanos , Osteoartrite/metabolismo , Osteoartrite/patologia , Osteoartrite/prevenção & controle , Fatores de Transcrição SOX9/genética , Fatores de Transcrição SOX9/metabolismo , Fator de Necrose Tumoral alfa/farmacologia
14.
PLoS One ; 8(12): e83545, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24386225

RESUMO

Dental pulp cells (DPCs) are known to be enriched in stem/progenitor cells but not well characterized yet. Small non-coding microRNAs (miRNAs) have been identified to control protein translation, mRNA stability and transcription, and have been reported to play important roles in stem cell biology, related to cell reprogramming, maintenance of stemness and regulation of cell differentiation. In order to characterize dental pulp stem/progenitor cells and its mechanism of differentiation, we herein sorted stem-cell-enriched side population (SP) cells from human DPCs and periodontal ligament cells (PDLCs), and performed a locked nucleic acid (LNA)-based miRNA array. As a result, miR-720 was highly expressed in the differentiated main population (MP) cells compared to that in SP cells. In silico analysis and a reporter assay showed that miR-720 targets the stem cell marker NANOG, indicating that miR-720 could promote differentiation of dental pulp stem/progenitor cells by repressing NANOG. Indeed, gain-and loss-of-function analyses showed that miR-720 controls NANOG transcript and protein levels. Moreover, transfection of miR-720 significantly decreased the number of cells positive for the early stem cell marker SSEA-4. Concomitantly, mRNA levels of DNA methyltransferases (DNMTs), which are known to play crucial factors during stem cell differentiation, were also increased by miR-720 through unknown mechanism. Finally, miR-720 decreased DPC proliferation as determined by immunocytochemical analysis against ki-67, and promoted odontogenic differentiation as demonstrated by alizarin red staining, as well as alkaline phosphatase and osteopontin mRNA levels. Our findings identify miR-720 as a novel miRNA regulating the differentiation of DPCs.


Assuntos
Diferenciação Celular/genética , Polpa Dentária/citologia , Polpa Dentária/fisiologia , MicroRNAs/genética , Fenótipo , Células-Tronco/citologia , Células-Tronco/metabolismo , Regiões 3' não Traduzidas , Adulto , Pareamento de Bases , Sequência de Bases , Biomarcadores , Linhagem Celular , Proliferação de Células , Análise por Conglomerados , DNA (Citosina-5-)-Metiltransferases/metabolismo , DNA Metiltransferase 3A , Citometria de Fluxo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Proteínas de Homeodomínio/genética , Humanos , MicroRNAs/química , MicroRNAs/metabolismo , Proteína Homeobox Nanog , Células da Side Population/metabolismo , Transcrição Gênica , DNA Metiltransferase 3B
15.
Cells Tissues Organs ; 198(5): 367-76, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24434422

RESUMO

OBJECTIVE: Escherichia coli-derived recombinant human bone morphogenetic protein-2 (E-BMP-2) has been shown to be as effective as mammalian cell-derived BMP-2. However, several in vitro and in vivo experiments are still necessary to validate the effectiveness of E-BMP-2 due to the difference in synthesis process, mainly related to protein nonglycosylation. The objective of this study was to investigate whether biodegradable polylactide-co-glycolide (PLGA) membrane is a suitable carrier for E-BMP-2 delivery for bone regeneration of critical-sized defects in rat calvaria. MATERIALS AND METHODS: First, the osteoinductive effect of E-BMP-2 was confirmed in vitro in mouse bone marrow stromal cells by analysis of osteocalcin mRNA levels, and calcium deposition was detected by alizarin red staining. Before in vivo experiments, the release profile of E-BMP-2 from PLGA membranes was determined by ELISA. E-BMP-2 (0, 1, 5 and 10 µg/µl) was applied for ectopic and orthotopic bone formation and was analyzed by X-ray, micro-CT and histology. RESULTS: Release-profile testing showed that PLGA membrane could retain 94% of the initially applied E-BMP-2. Ectopic bone formation assay revealed that combination of E-BMP-2/PLGA membrane strongly induced bone formation. Stronger osteoinductivity with complete repair of critical-sized defects was observed only with PLGA membranes adsorbed with 5 and 10 µg/µl of E-BMP-2, whereas no bone formation was observed in the groups that received no membrane or 0-µg/µl dose of E-BMP-2. CONCLUSION: PLGA membrane was shown to be a suitable carrier for sustained release of E-BMP-2, and the E-BMP-2/PLGA membrane combination was demonstrated to be efficient in bone regeneration in a model of critical-sized defects.


Assuntos
Proteína Morfogenética Óssea 2/administração & dosagem , Proteína Morfogenética Óssea 2/química , Regeneração Óssea/efeitos dos fármacos , Ácido Láctico/química , Ácido Poliglicólico/química , Crânio/fisiologia , Fator de Crescimento Transformador beta/administração & dosagem , Fator de Crescimento Transformador beta/química , Animais , Escherichia coli/genética , Escherichia coli/metabolismo , Feminino , Humanos , Membranas Artificiais , Camundongos , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Ratos , Ratos Wistar , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/química , Crânio/efeitos dos fármacos
16.
J Dent Educ ; 76(12): 1580-8, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23225677

RESUMO

This educational trial was an eight-day problem-based learning (PBL) course for fourth-year predoctoral students at Okayama University's dental school who interviewed elderly residents living in a nursing home. The purpose of this PBL course was to introduce geriatric dentistry to the students by allowing them, independently, to discover the clinical problems of elderly patients as well as the solutions. The sixty-five students were divided into nine small groups and received patient information (age, gender, degree of care needed, medical history, food type, medications, and oral condition) in datasheets before visiting the nursing home. Each group of students directly interviewed one patient and the caregivers and identified the patient's medical, psychological, and social problems. After the interview, the students participated in a PBL tutorial to delineate a management approach for the patient's problems. To measure the efficacy of this program, the students completed a questionnaire before and after the course regarding their level of understanding of and attitudes toward geriatric dentistry, clinical research, and self-study. The results showed that student's perceptions of their knowledge about and attitudes toward oral health care for the elderly significantly increased after the PBL course, which suggests that such tutorials should be an option for dental curricula.


Assuntos
Avaliação Geriátrica/métodos , Odontologia Geriátrica/educação , Avaliação das Necessidades , Planejamento de Assistência ao Paciente , Aprendizagem Baseada em Problemas , Idoso , Idoso de 80 Anos ou mais , Competência Clínica , Odontologia Comunitária/educação , Assistência Odontológica para Idosos , Educação em Odontologia/métodos , Feminino , Humanos , Pacientes Internados , Japão , Masculino , Casas de Saúde , Padrões de Prática Odontológica , Avaliação de Programas e Projetos de Saúde
17.
J Prosthodont Res ; 56(4): 229-48, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23137671

RESUMO

New technologies that facilitate solid alveolar ridge augmentation are receiving considerable attention in the field of prosthodontics because of the growing requirement for esthetic and functional reconstruction by dental implant treatments. Recently, several studies have demonstrated potential advantages for stem-cell-based therapies in regenerative treatments. Mesenchymal stem/stromal cells (MSCs) are now an excellent candidate for tissue replacement therapies, and tissue engineering approaches and chair-side cellular grafting approaches using autologous MSCs represent the clinical state of the art for stem-cell-based alveolar bone regeneration. Basic studies have revealed that crosstalk between implanted donor cells and recipient immune cells plays a key role in determining clinical success that may involve the recently observed immunomodulatory properties of MSCs. Part II of this review first overviews progress in regenerative dentistry to consider the implications of the stem cell technology in dentistry and then highlights cutting-edge stem-cell-based alveolar bone regenerative therapies. Factors that affect stem-cell-based bone regeneration as related to the local immune response are then discussed. Additionally, pre-clinical stem cell studies for the regeneration of teeth and other oral organs as well as possible applications of MSC-based immunotherapy in dentistry are outlined. Finally, the marketing of stem cell technology in dental stem cell banks with a view toward future regenerative therapies is introduced.


Assuntos
Transplante de Células-Tronco , Engenharia Tecidual , Perda do Osso Alveolar/cirurgia , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/administração & dosagem , Medicina Regenerativa , Dente/fisiologia
18.
J Prosthodont Res ; 56(4): 249-55, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23083963

RESUMO

PURPOSE: Current clinical procedures to control or regenerate bone loss due to peri-implantitis are not predictable neither accomplish complete resolution. Therefore, early detection of the onset and the active periods of bone loss are crucial for prevention of extensive peri-implant bone resorption. This study aimed to determine a possible association between the presence of collagenases (MMP-1, MMP-8 and MMP-13) in peri-implant sulcular fluid (PISF) and active periods of bone loss by annually adjusted vertical bone loss (AVBL) measurements. METHODS: Intended sample consisted of 76 consecutive patients who received oral implant treatment at the Fixed Prosthodontic Clinic of Okayama University Hospital from 1990 to 2000. Twelve subjects were lost to follow-up or refused to participate. Consequently, the actual sample consisted of 64 patients who were followed-up for at least one year. Those patients with AVBL>0.6mm were included in the severe peri-implantitis group, and randomly selected, age-, gender- and implantation site-matched healthy patients (AVBL<0.3mm) comprised the control group. PISF samples were collected from both groups and further analyzed by western blot for detection of collagenases. RESULTS: Four patients presented severe peri-implantitis. MMP-8 was the only collagenase detected in peri-implant sites with ongoing bone loss. PISF samples from control group showed no positive reactions to any collagenase. CONCLUSION: This study showed MMP-8 as a possible marker for progressive bone loss in peri-implantitis.


Assuntos
Perda do Osso Alveolar/enzimologia , Implantes Dentários , Metaloproteinase 8 da Matriz/análise , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
19.
J Prosthodont Res ; 56(3): 151-65, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22796367

RESUMO

Stem cells can self-renew and produce different cell types, thus providing new strategies to regenerate missing tissues and treat diseases. In the field of dentistry, adult mesenchymal stem/stromal cells (MSCs) have been identified in several oral and maxillofacial tissues, which suggests that the oral tissues are a rich source of stem cells, and oral stem and mucosal cells are expected to provide an ideal source for genetically reprogrammed cells such as induced pluripotent stem (iPS) cells. Furthermore, oral tissues are expected to be not only a source but also a therapeutic target for stem cells, as stem cell and tissue engineering therapies in dentistry continue to attract increasing clinical interest. Part I of this review outlines various types of intra- and extra-oral tissue-derived stem cells with regard to clinical availability and applications in dentistry. Additionally, appropriate sources of stem cells for regenerative dentistry are discussed with regard to differentiation capacity, accessibility and possible immunomodulatory properties.


Assuntos
Odontologia/tendências , Células-Tronco , Humanos , Células-Tronco Pluripotentes Induzidas , Células-Tronco Mesenquimais , Transplante de Células-Tronco , Engenharia Tecidual
20.
J Prosthodont Res ; 54(1): 36-41, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19819208

RESUMO

PURPOSE: To examine gene expression profile changes in the mouse masseter muscle tissue after repetitive electrical stimulation by using a DNA microarray technique. METHODS: Nine male ICR mice aged 10 weeks were used. Each anesthetized mouse was secured on a platform in a supine position and the masseter muscle tissues on both sides were exposed. Bipolar electrodes were set on the right masseteric fascia to electrically stimulate the masseter muscle (8 V, 10 Hz, 20 ms) for 30 min. After cessation of stimulation bilateral masseter muscle tissues were sampled at 0 h (n=3), 1h (n=3), 2h (n=3). Total RNA was isolated from the homogenized muscle tissues and purified mRNA samples (50 microg) were processed and hybridized with microarray slides. Probe arrays were then scanned and analyzed to calculate the signal density. Gene expression profiles were compared at each time point between the right (stimulation side) and left (control side) masseter. When the gene expression levels were different more than 2-fold, the difference was regarded as positive. RESULTS: Of the 6400 genes assessed, 1733 genes were up-regulated and 515 genes were down-regulated in the stimulation side at least once during the experimental time course. These up- or down-regulated genes were associated with autoimmune/inflammatory disease (28/114), cardiovascular disease (17/61), neuroscience (12/50), apoptosis (27/93), diabetes/obesity (9/28), signal transduction (66/250) and others. 28 genes were up-regulated and 25 genes were down-regulated at all time points. CONCLUSIONS: Dramatic gene expression changes were induced by the repetitive electrical muscle stimulation in mouse masseter.


Assuntos
Estimulação Elétrica , Perfilação da Expressão Gênica , Músculo Masseter , Animais , Apoptose/genética , Doenças Autoimunes/genética , Doenças Cardiovasculares/genética , Diabetes Mellitus/genética , Regulação para Baixo , Inflamação/genética , Masculino , Camundongos , Camundongos Endogâmicos ICR , Obesidade/genética , Análise de Sequência com Séries de Oligonucleotídeos , RNA/isolamento & purificação , RNA Mensageiro , Transdução de Sinais/genética , Regulação para Cima
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