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1.
Clin Implant Dent Relat Res ; 23(5): 735-744, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34436812

RESUMO

AIM: To compare the 1-year outcomes of immediate loading (IL) and delayed loading (DL) protocols for implants placed by fully guided surgery in partially edentulous jaws. MATERIALS AND METHODS: This study included 72 patients who received implant surgery with either IL (93 implants, 36 patients) or DL (94 implants, 26 patients). A prefabricated provisional prosthesis was delivered immediately for the IL group (86 implants, 32 patients) with the exception of 4 subjects in whom an initial torque of >20 Ncm and an implant stability quotient of >65 were not achieved, while all DL-group implants were loaded after 3 months. The 1-year implant survival rate estimated by intention-to-treat (ITT) and per-protocol (PP) analyses, and the marginal bone loss (MBL) estimated by cone-beam computed tomography were statistically evaluated (p < 0.05). RESULTS: The survival rate in the DL group was 100% at both patient and implant levels. With only 26 subjects with 78 implants surviving in the IL group, the survival rates were 69.4% and 83.4% at the patient and implant levels, respectively, in the ITT analysis, and 78.1% and 90.2% in the PP analysis. All intergroup differences in survival rates were statistically significant (p < 0.01). MBL was less than 0.1 mm in both groups (p > 0.05). CONCLUSIONS: IL for implants placed by fully guided surgery in the partially edentulous jaws increased the probability of failure compared to 3-month DL. Regardless of when loading occurred, marginal bone levels remained stable.


Assuntos
Perda do Osso Alveolar , Implantes Dentários , Carga Imediata em Implante Dentário , Arcada Edêntula , Perda do Osso Alveolar/etiologia , Implantação Dentária Endóssea , Prótese Dentária Fixada por Implante , Falha de Restauração Dentária , Humanos , Arcada Edêntula/cirurgia , Resultado do Tratamento
2.
Clin Oral Implants Res ; 31(1): 93-102, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31724239

RESUMO

OBJECTIVES: This study histologically analyzed biopsy samples obtained from sites of damaged extraction socket grafting using deproteinized bovine bone mineral (DBBM) or deproteinized porcine bone mineral (DPBM) with coverage by a collagen membrane. MATERIAL AND METHODS: One hundred patients participated in this randomized controlled clinical trial of extraction socket grafts performed in cases of periodontally compromised teeth. All participants were blinded to their group allocations, and each material was grafted with coverage by collagen membranes after extraction of the tooth and removal of granulation tissue. At implant placement at 4 months, a biopsy was harvested at the implant site using a trephine was analyzed histologically. RESULTS: Eighty-five biopsy samples were acquired, of which 81 were finally included in the histologic analysis (42 in DBBM and 39 in DPBM group). Both DBBM and DPBM groups showed comparable proportions of residual biomaterial (12.37 ± 5.67% and 12.21 ± 5.75%, respectively), newly formed bone (15.07 ± 10.52% and 18.47 ± 11.47%, respectively), and nonmineralized tissue (72.56 ± 10.07% and 71.55 ± 15.47%, respectively). There were no significant differences in these histologic parameters between the two groups with different biomaterials. CONCLUSION: Comparable histologic bone formation was found in both socket grafted groups with DBBM or DPBM covered by collagen membranes in periodontally damaged extraction sockets. However, a wide variation in new bone formation was found after 4 months of postsurgical healing and a tendency of higher new bone formation was shown at damaged sockets that had an intact unilateral residual wall regardless of buccal or lingual side.


Assuntos
Substitutos Ósseos , Alvéolo Dental , Animais , Bovinos , Colágeno , Humanos , Minerais , Suínos , Extração Dentária
3.
Implant Dent ; 28(5): 421-429, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31306296

RESUMO

PURPOSE: To investigate clinical factors and cellular responses of in situ human alveolar bone-derived mesenchymal stromal cells involved in early periimplant marginal bone loss. MATERIALS AND METHODS: Thirty-seven completely or partially edentulous patients were enrolled in this study. Periapical radiographs were taken at the time of implant surgery, at 3-month follow-up, and at 1-year follow-up. Univariate analysis and multiple logistic regression were performed to investigate the associations between marginal bone loss and study variables. The mRNA expression levels of 21 bone-remodeling- and tissue-healing-associated genes were analyzed by subgroup. RESULTS: Thirty-one patients with 98 implants were followed. The incidence and mean amount of bone loss were higher for overdentures than for other prosthesis and higher for the maxilla than for the mandible. The bone loss group showed lower mRNA expression levels of runt-related transcription factor-2, bone morphogenetic protein-2, and peroxisome proliferator-activated receptor gamma-2 and higher receptor activator of NKκB ligand/osteoprotegerin (RANKL/OPG) ratio. CONCLUSION: Within the limitations of the study, certain genes involved in bone remodeling (runt-related transcription factor-2 [Runx-2], bone morphogenetic protein-2 [BMP-2], and peroxisome proliferator-activated receptor gamma-2 [PPARγ-2]) and RANKL/OPG are correlated with early periimplant bone loss, with the type of suprastructure and the involved jaw being significant clinical factors.


Assuntos
Perda do Osso Alveolar , Implantes Dentários , Células-Tronco Mesenquimais , Implantação Dentária Endóssea , Prótese Dentária Fixada por Implante , Falha de Restauração Dentária , Seguimentos , Humanos , Mandíbula , Projetos Piloto , Estudos Prospectivos
4.
Clin Implant Dent Relat Res ; 20(5): 729-737, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30051954

RESUMO

BACKGROUNDS: Clinical benefits in bone grafting of intact extraction socket have been widely known, but limited evidence is available for the procedure in damaged extraction sockets due to periodontal disease. PURPOSE: This study aimed to determine the dimensional alteration of alveolar ridge following bone grafting of damaged extraction sockets, and compare the outcomes of using deproteinized bovine (DBBM) versus porcine bone mineral (DPBM) in the damaged sockets. MATERIALS AND METHODS: One hundred patients (n = 50 for each group) with periodontitis-induced damaged extraction socket were included in this randomized, single-blind trial. After removal of tooth and granulation tissue, sites were grafted with either DBBM (DBBM group) or DPBM (DPBM group), and covered with collagen membrane. Linear/volumetric analyses of hard and soft-tissue dimensions were performed on reconstructed/superimposed computed tomography and scanned cast images, taken immediately and 4 months after surgery. RESULTS: The two groups showed comparable hard tissue augmentation with minimal reductions in the grafted volume, as well as in vertical (1.22 ± 2.16 and 1.45 ± 1.92 mm for DPBM and DBBM group, respectively) and horizontal (1.43 ± 3.40 and 1.83 ± 2.85 mm on the central section, respectively) dimensions at 4 months after surgery. However, several cases showed large variations in maintenance of the grafted volume. None of the measured parameters in hard and soft tissue dimensions differed significantly between DBBM and DPBM sites. CONCLUSIONS: DBBM and DPBM can comparably augment damaged extraction sockets with minimal postoperative reduction of the grafted volume. However, the large variations in the results should be further evaluated for application in routine dental clinics.


Assuntos
Aumento do Rebordo Alveolar/métodos , Substitutos Ósseos/uso terapêutico , Periodontite/cirurgia , Alvéolo Dental/cirurgia , Processo Alveolar/diagnóstico por imagem , Animais , Bovinos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Periodontite/etiologia , Radiografia Dentária , Método Simples-Cego
5.
Stem Cell Res ; 28: 87-95, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29448134

RESUMO

This study aimed to determine the cellular characteristics and behaviors of human bone marrow stromal cells (hBMSCs) expanded in media in a hypoxic or normoxic condition and with or without fibroblast growth factor-2 (FGF-2) treatment. hBMSCs isolated from the vertebral body and expanded in these four groups were evaluated for cellular proliferation/migration, colony-forming units, cell-surface characterization, in vitro differentiation, in vivo transplantation, and gene expression. Culturing hBMSCs using a particular environmental factor (hypoxia) and with the addition of FGF-2 increased the cellular proliferation rate while enhancing the regenerative potential, modulated the multipotency-related processes (enhanced chondrogenesis-related processes/osteogenesis, but reduced adipogenesis), and increased cellular migration and collagen formation. The gene expression levels in the experimental samples showed activation of the hypoxia-inducible factor-1 pathway and glycolysis in the hypoxic condition, with this not being affected by the addition of FGF-2. The concurrent application of hypoxia and FGF-2 could provide a favorable condition for culturing hBMSCs to be used in clinical applications associated with bone tissue engineering, due to the enhancement of cellular proliferation and regenerative potential.


Assuntos
Técnicas de Cultura de Células/métodos , Fator 2 de Crescimento de Fibroblastos/farmacologia , Células-Tronco Mesenquimais/citologia , Adulto , Animais , Biomarcadores/metabolismo , Diferenciação Celular , Hipóxia Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Senescência Celular/efeitos dos fármacos , Colágeno/metabolismo , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Camundongos , Pessoa de Meia-Idade , Células-Tronco
6.
J Oral Maxillofac Surg ; 75(2): 297.e1-297.e13, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27288839

RESUMO

PURPOSE: This study evaluated a commercially available, 3-dimensional gel-type polyethylene glycol (PEG) membrane as a carrier for recombinant human bone morphogenetic protein-2 (rhBMP-2) using a rat calvarial defect model. Another gel-type carrier, fibrin-fibronectin system (FFS), was used as a positive control. MATERIALS AND METHODS: Critical-size defects were made in the rat calvarium, which were allocated to 1 of 10 groups comprising 2 healing periods and biomaterial conditions: 1) sham control, 2) FFS only, 3) FFS plus BMP-2, 4) PEG only, and 5) PEG plus BMP-2. Radiographic and histologic analyses were performed at 2 and 8 weeks after surgery. RESULTS: After 2 weeks, some parts of the FFS were biodegraded and extensive cellular infiltration was observed at sites that received FFS or FFS plus BMP-2. The PEG membrane retained its augmented volume without cellular infiltration at sites that received PEG or PEG plus BMP-2. After 8 weeks, the FFS was completely degraded and replaced by new bone and connective tissues. In contrast, the volume of residual PEG was similar to that at 2 weeks, with slight cellular infiltration. In particular, there was progressive bone regeneration around micro-cracks and resorbed outer surface in the PEG + BMP-2 group. Although the PEG + BMP-2 group showed increased area and percentage of new bone, there was no statistical relevance after 2 and 8 weeks in histomorphometric analyses. However, the appearance of the healing differed (with new bone formation along micro-cracks in the PEG + BMP-2 group), and further studies with longer healing periods are needed to draw conclusions about clinical applications. CONCLUSION: Evidence of mechanical stability and new bone formation along micro-cracks when using PEG plus BMP-2 might support the PEG membrane as a candidate carrier material for rhBMP-2.


Assuntos
Proteína Morfogenética Óssea 2/administração & dosagem , Portadores de Fármacos/administração & dosagem , Polietilenoglicóis/administração & dosagem , Crânio/lesões , Fator de Crescimento Transformador beta/administração & dosagem , Animais , Desenvolvimento Ósseo/efeitos dos fármacos , Géis/administração & dosagem , Masculino , Membranas Artificiais , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes/administração & dosagem , Crânio/diagnóstico por imagem , Crânio/efeitos dos fármacos , Crânio/crescimento & desenvolvimento , Microtomografia por Raio-X
7.
J Adv Prosthodont ; 8(6): 494-503, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28018568

RESUMO

PURPOSE: The aim of this study was to compare the efficacies of two-implant splinting (2-IS) and single-implant restoration (1-IR) in the first and second molar regions over a mean functional loading period (FLP) of 40 months, and to propose the appropriate clinical considerations for the splinting technique. MATERIALS AND METHODS: The following clinical factors were examined in the 1-IR and 2-IS groups based on the total hospital records of the patients: sex, mean age, implant location, FLP, bone grafting, clinical crown-implant ratio, crown height space, and horizontal distance. The mechanical complications [i.e., screw loosening (SL), screw fracture, crown fracture, and repeated SL] and biological complications [i.e., peri-implant mucositis (PM) and peri-implantitis (PI)] were also evaluated for each patient. In analysis of two groups, the chi-square test and Student's t-test were used to identify the relationship between clinical factors and complication rates. The optimal cutoff value for the FLP based on complications was evaluated using receiver operating characteristics analysis. RESULTS: In total, 234 patients with 408 implants that had been placed during 2005 - 2014 were investigated. The incident rates of SL (P<.001), PM (P=.002), and PI (P=.046) differed significantly between the 1-IR and 2-IS groups. The FLP was the only meaningful clinical factor for mechanical and biological complication rates in 2-IS. CONCLUSION: The mechanical complication rates were lower for 2-IS than for 1-IR, while the biological complication rates were higher for 2-IS. FLP of 39.80 and 46.57 months were the reference follow-up periods for preventing biological and mechanical complications, respectively.

8.
Tissue Eng Part C Methods ; 22(10): 941-951, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-27609497

RESUMO

Centrifugation based on density gradients is a general methodology for isolating human bone marrow (hBM)-derived mesenchymal stem cells (hBMSCs). The mononuclear cell (MNC) layer can be obtained using a density gradient solution in the conventional protocol, but it is not suitable for direct transplantation due to the possible toxicity of this solution. The results obtained are also influenced by the skill level when applying the technique, which involves time-consuming processes. We have developed a novel protocol for isolating hBMSCs using hyaluronic acid (HA), which is the most widely used injectable biomaterial in clinical settings and a major component of the extracellular matrix. Laying hBM over the HA and then applying centrifugation yielded three separate layers, with the HA layer, including MNCs being the most superficial one. Increasing the volume of HA and/or its crosslinking rate enhanced the yield of MNCs from hBM, and the cell yield was also significantly higher for a lower centrifugal acceleration (530 g) than for a higher one (1500 g). Isolated hBMSCs by HA exhibited similar biological characteristics such as in terms of their proliferation rate, fibroblast-like morphology, cell-cycle status, immunophenotype, and multipotency. The use of either type of hBMSC confirmed the regenerative potential of bone and bone marrow-like tissue in ectopic transplantation models. This is the first report of a novel protocol for isolating hBMSCs that utilize HA. We suggest that this novel isolation technique can be used for the direct application of autogenous MSCs with advantages of being less time-consuming and involving steps that are easier to perform.


Assuntos
Células da Medula Óssea/citologia , Diferenciação Celular , Separação Celular/métodos , Ácido Hialurônico/química , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Células-Tronco Mesenquimais/citologia , Engenharia Tecidual/métodos , Adulto , Células Cultivadas , Matriz Extracelular/química , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Regeneração
9.
Cell Transplant ; 25(10): 1819-1832, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-27075568

RESUMO

Several in vivo studies have found that transplanting mesenchymal stem cells (MSCs) into degenerative intervertebral discs (IVDs) leads to regeneration of disc cells. Since the exact underlying mechanisms are not understood, we investigated the mechanisms of action of MSCs in regeneration of degenerative IVDs via paracrine actions. Human MSCs and degenerative disc cells from the same donor vertebrae were directly or indirectly cocultured. The multidifferentiation potential, cell proliferation, collagen synthesis, and mRNA expression levels were assessed. The proliferation rates of MSCs and degenerative disc cells were higher in the coculture system than in the monolayer cultures or in the conditioned medium of each cell type. During coculturing with nucleus pulposus (NP) cells, mRNA expression of the extracellular matrix (ECM) components aggrecan, versican (VCAN), SOX9, and type II and type VI collagen was significantly increased in MSCs, whereas mRNA expression for type V collagen was increased in MSCs cocultured with annulus fibrosus (AF) cells. In addition, the accumulation of total ECM collagen was greater in cocultured degenerative disc cells than in monocultured cells. During coculturing, MSCs downregulated the expression levels of various proinflammatory cytokine genes in degenerative NP [interleukin-1α ( IL-1α), IL-1ß, IL-6, and tumor necrosis factor-α ( TNF-α)] and AF cells ( IL-1α and IL-6), which are involved in the degradation of ECM molecules. In association with the trophic effect of MSCs on degenerative disc cells, upregulation of growth factor mRNA expression was shown in MSCs cocultured with degenerative NP cells [epidermal growth factor ( EGF), insulin-like growth factor-1 ( IGF-1), osteogenic protein-1 ( OP-1), growth and differentiation factor-7 ( GDF-7), and transforming growth factor-ß ( TGF-ß)] or degenerative AF cells ( IGF-1, OP-1, and GDF-7). In terms of MSC-based clinical approaches to IVD regeneration, implanting MSCs into a degenerative IVD may both stimulate MSC differentiation into an NP- or AF-like phenotype and stimulate the biological activation of degenerative disc cells for self-repair.


Assuntos
Anel Fibroso/citologia , Células-Tronco Mesenquimais/metabolismo , Núcleo Pulposo/citologia , Comunicação Parácrina/fisiologia , Adulto , Anel Fibroso/metabolismo , Proteína Morfogenética Óssea 7/genética , Proteína Morfogenética Óssea 7/metabolismo , Diferenciação Celular , Proliferação de Células , Técnicas de Cocultura , Matriz Extracelular/metabolismo , Feminino , Humanos , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/metabolismo , Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/terapia , Masculino , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Pessoa de Meia-Idade , Núcleo Pulposo/metabolismo , Projetos Piloto , Regeneração/fisiologia , Fatores de Transcrição SOX9/genética , Fatores de Transcrição SOX9/metabolismo
10.
J Biomed Mater Res B Appl Biomater ; 104(3): 515-24, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25939881

RESUMO

OBJECTIVES: The aim of this study was to evaluate HA coated with different ratios of TCP as a carrier for hABMSCs obtained during implant osteotomy in comparison to slowly-resorbing biomaterial, Bio-Oss, as a negative control, using in vitro and in vivo experiments. MATERIALS AND METHODS: Human ABMSCs (hABMSCs) harvested during implant osteotomy were transplanted using HA/TCP or Bio-Oss as carriers in a murine ectopic transplantation model (n = 12). Pore size and cell affinity were evaluated in vitro. The area of newly formed bone was analyzed histometrically, the number of osteocytes was counted, and immunohistochemical staining was conducted against several markers of osteogenesis, including alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX-2), osteocalcin (OCN), and osteopontin (OPN). Osteoclast formation was evaluated by tartrate-resistant acid phosphatase staining. RESULTS: The carrier materials had comparable pore sizes. The cell affinity assay resulted in a high proportion of cell adhesion (>90%) in all experimental groups. Substantial new bone and osteocyte formation was observed on both HA/TCP carriers, whereas it was minimal with Bio-Oss. Positive immunostaining for ALP, RUNX-2, OCN, and OPN was observed with HA/TCP, but only limited expression of osteogenic markers with Bio-Oss. Conversely, there was a minimal osteoclast presence with Bio-Oss, but a significant presence of osteoclasts with both HA/TCP carriers. CONCLUSIONS: Both types of scaffolds, BCP and Bio-Oss, showed high stem cell-carrying potential, but the in vivo healing patterns of their complexes with hABMSC could be affected by the microenvironment on the surfaces of the scaffolds.


Assuntos
Matriz Óssea/metabolismo , Cerâmica , Hidroxiapatitas , Células-Tronco Mesenquimais/metabolismo , Minerais , Osteogênese/efeitos dos fármacos , Matriz Óssea/citologia , Células Cultivadas , Cerâmica/química , Cerâmica/farmacologia , Humanos , Hidroxiapatitas/química , Hidroxiapatitas/farmacologia , Células-Tronco Mesenquimais/citologia , Minerais/química , Minerais/farmacologia
11.
Bone ; 78: 34-45, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25952967

RESUMO

Human bone marrow mesenchymal stem cells (hBMSCs) were isolated from bone marrow of the vertebral body. The hBMSCs were cultured under either hypoxic (1% O2) or normoxic (21% O2; control) conditions and the characteristics as mesenchymal stem cells were compared. Results revealed that hypoxia reduced proliferative potential and colony-forming efficiency of hBMSCs, and significantly enhanced osteogenic and chondrogenic differentiation. The hBMSCs enhanced the regenerative potential of bone in vivo. In vitro synthesis of soluble and insoluble collagen was significantly increased in the hypoxic condition. In vivo collagen tissue regeneration was also enhanced under the hypoxic condition, with concomitant increased expressions of various subtypes of collagen and lysyl-oxidase family mRNA. MicroRNA assays revealed that miR-155-5p, which negatively regulates HIF-1α, was significantly highly expressed. These observations demonstrate that hBMSCs obtained from human vertebrae exhibit altered characteristics under hypoxic conditions, and each factor contributing to hBMSC-mediated tissue healing should be evaluated with the goal of allowing their clinical application.


Assuntos
Células da Medula Óssea/citologia , Células-Tronco Mesenquimais/citologia , Regeneração/fisiologia , Adipócitos/citologia , Adulto , Diferenciação Celular , Hipóxia Celular , Proliferação de Células , Células Cultivadas , Condrócitos/citologia , Condrogênese , Colágeno/química , Feminino , Citometria de Fluxo , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Masculino , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Osteogênese , Oxigênio/química , Coluna Vertebral/citologia , Células-Tronco , Cicatrização
12.
J Clin Periodontol ; 42(7): 678-87, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25946650

RESUMO

OBJECTIVES: This study evaluated the dimensional ridge alteration in a buccal-bone-deficient extraction socket, and ridge regeneration following socket grafting accompanied by recombinant human bone morphogenetic protein 2 (rhBMP-2) or a collagen membrane covering. MATERIAL AND METHODS: In five beagle dogs, entire buccal bone of the extracted sockets of premolars was surgically removed and immediately grafted using one of the following graft protocols: (1) sham surgery without any grafting, and grafting with (2) deproteinized bovine bone mineral (DBBM), (3) DBBM/rhBMP-2 and (4) DBBM covered with a collagen membrane (DBBM/Membrane). Quantitative/qualitative analyses were performed radiographically/histologically after 8 weeks. RESULTS: Buccal-deficient extraction sockets healed with significant reduction in buccolingual dimension along the entire length of the socket, but all grafting techniques reduced the dimensional changes compared to the non-grafted control sites. Histologically, sites received DBBM only exhibited minimal regeneration, whereas sites grafted with DBBM/rhBMP-2 or DBBM/Membrane exhibited greater new bone formation extending the entire augmented area. CONCLUSIONS: Buccal-bone-deficiency may lead to significant volume reduction after tooth extraction along the entire length of the socket, and socket grafting accompanied by rhBMP-2 or covered with a membrane can be candidate therapies for preservation of the buccolingual dimension and successful ridge regeneration.


Assuntos
Perda do Osso Alveolar/tratamento farmacológico , Proteína Morfogenética Óssea 2/uso terapêutico , Regeneração Óssea/efeitos dos fármacos , Alvéolo Dental/efeitos dos fármacos , Fator de Crescimento Transformador beta/uso terapêutico , Perda do Osso Alveolar/cirurgia , Aumento do Rebordo Alveolar/métodos , Animais , Regeneração Óssea/fisiologia , Substitutos Ósseos/uso terapêutico , Bovinos , Colágeno , Cães , Humanos , Masculino , Mandíbula/efeitos dos fármacos , Mandíbula/patologia , Mandíbula/cirurgia , Membranas Artificiais , Minerais/uso terapêutico , Osteogênese/efeitos dos fármacos , Osteogênese/fisiologia , Distribuição Aleatória , Proteínas Recombinantes/uso terapêutico , Extração Dentária/efeitos adversos , Alvéolo Dental/patologia , Alvéolo Dental/cirurgia , Microtomografia por Raio-X/métodos
13.
Cell Transplant ; 24(9): 1767-79, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25290076

RESUMO

This study aimed to enhance the attachment of periodontal ligament stem cells (PDLSCs) onto the decellularized dental root surface using surface coating with fibronectin and/or calcium phosphate (CaP) and to evaluate the activity of PDLSCs attached to a coated dental root surface following tooth replantation. PDLSCs were isolated from five dogs, and the other dental roots were used as a scaffold for carrying PDLSCs and then assigned to one of four groups according to whether their surface was coated with CaP, fibronectin, CaP/fibronectin, or left uncoated (control). Fibronectin increased the adhesion of PDLSCs onto dental root surfaces compared to both the control and CaP-coated groups, and simultaneous surface coating with CaP and fibronectin significantly accelerated and increased PDLSC adhesion compared to the fibronectin-only group. On in vivo tooth replantation, functionally oriented periodontal new attachment was observed on the CaP/fibronectin-coated dental roots to which autologous PDLSCs had adhered, while in the control condition, dental root replantation was associated only with root resorption and ankylosis along the entire root length. CaP and fibronectin synergistically enhanced the attachment of PDLSCs onto dental root surfaces, and autologous PDLSCs could produce de novo periodontal new attachment in an experimental in vivo model.


Assuntos
Fosfatos de Cálcio/química , Fibronectinas/química , Ligamento Periodontal/citologia , Células-Tronco/citologia , Animais , Antígenos CD/metabolismo , Adesão Celular , Células Cultivadas , Cães , Masculino , Transplante de Células-Tronco , Células-Tronco/metabolismo , Dente/patologia , Raiz Dentária/citologia , Raiz Dentária/diagnóstico por imagem , Raiz Dentária/metabolismo , Transplante Autólogo , Microtomografia por Raio-X
14.
Tissue Eng Part A ; 21(3-4): 451-62, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25138561

RESUMO

Human bone marrow stem cells (hBMSCs) represent a promising regenerative material because of their mutipotency, including their ability to regenerate collagenous soft tissues. We previously found that water-soluble chitin (WSC) enhances the ability of human periodontal ligament stem cells (hPDLSCs) to synthesize collagen tissue. The aim of this study was to determine the effects of WSC on hBMSCs and hPDLSCs for the collagen synthesis both in vitro and in vivo. hBMSCs and hPDLSCs were isolated and expanded with or without 0.3 mg/mL WSC. A series of in vitro and in vivo analyses were performed to evaluate their characteristics as stem cell populations. Then, collagen and hydroxyproline assays were conducted using both in vitro and in vivo assay models, and the real-time polymerase chain reaction was performed to analyze the expression of collagen-related markers. WSC-treated and nontreated hBMSCs and hPDLSCs were transplanted into immunocompromised mice, and histology and immunohistochemistry analyses were conducted after 8 weeks. The in vitro results showed that those cells possessed the characteristics of mesenchymal stem cells. The amount of soluble collagen synthesized was significantly greater in WSC-treated hBMSCs than in the nontreated group; conversely, treatment of hPDLSCs with WSC decreased the formation of soluble collagen. The amount of insoluble collagen synthesized was greater in the WSC-treated groups than in the nontreated groups for both hBMSCs and hPDLSCs. The hydroxyproline contents of the regenerated soluble and insoluble collagens were similar. The expressions of mRNA for collagen types I-V, hyaluronic acid synthase 1 (HAS1), HAS2, and HAS3, and the LOX family were higher in WSC-treated hPDLSCs than in the nontreated group, whereas WSC increased the expression of collagen type III and decreased that of collagen type I in hBMSCs. The histology and immunohistochemistry results revealed that WSC significantly increased the amount of collagen formed in vivo by both types of stem cells. Collectively, treatment with WSC significantly enhanced the collagen-forming potentials of hBMSCs and hPDLSCs, but the collagen they produced exhibited distinctively different characteristics. These findings suggest that the appropriate stem-cell source should be chosen based on the purpose of the required regenerated tissue.


Assuntos
Quitosana/farmacologia , Colágeno/biossíntese , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Ligamento Periodontal/citologia , Ligamento Periodontal/metabolismo , Adulto , Animais , Materiais Biocompatíveis/farmacologia , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Células Cultivadas , Quitina , Feminino , Humanos , Masculino , Teste de Materiais , Células-Tronco Mesenquimais/efeitos dos fármacos , Camundongos , Camundongos Nus , Pessoa de Meia-Idade , Ligamento Periodontal/efeitos dos fármacos
15.
Stem Cells Dev ; 24(2): 228-43, 2015 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-25122057

RESUMO

The effects of fibroblast growth factor-2 (FGF-2) on collagen tissue regeneration by human bone marrow stem cells (hBMSCs) were investigated. hBMSCs were isolated from human vertebral body bone marrow during vertebral surgery and a population of hBMSCs with the characteristics of mesenchymal stem cells was observed. The FGF-2 treatment (5 ng/mL) affected on the colony-forming efficiency, proliferation, and in vitro differentiation of hBMSCs. Insoluble/soluble collagen and hydroxyproline synthesis was significantly enhanced in hBMSCs expanded with FGF-2 and the treatment of FGF-2 caused a reduction in the mRNA expression of collagen type I, but an increase of collagen types II and III along with lysyl oxidase family genes. Collagen formation was also examined using an in vivo assay model by transplanting hBMSCs into immunocompromised mice (n=4) and the histologic and immunohistochemical results revealed that significantly more collagen with a well-organized structure was formed by FGF-2-treated hBMSCs at 8 weeks posttransplantation (P<0.05). The DNA microarray assay demonstrated that genes related to extracellular matrix formation were significantly upregulated. To elucidate the underlying mechanism, chemical inhibitors against extracellular-signal-regulated kinase (ERK) and phosphoinositide 3-kinase (PI3K) were treated and following downstream expression was observed. Collectively, FGF-2 facilitated the collagen-producing potency of hBMSCs both in vitro and in vivo, rendering them more suitable for use in collagen regeneration in the clinical field.


Assuntos
Células da Medula Óssea/metabolismo , Diferenciação Celular/efeitos dos fármacos , Colágeno/biossíntese , Fator 2 de Crescimento de Fibroblastos/farmacologia , Células-Tronco Mesenquimais/metabolismo , Regeneração/efeitos dos fármacos , Coluna Vertebral/metabolismo , Adulto , Animais , Células da Medula Óssea/citologia , Células Cultivadas , Feminino , Xenoenxertos , Humanos , Masculino , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Camundongos , Camundongos SCID , Pessoa de Meia-Idade , Proteína-Lisina 6-Oxidase/biossíntese , Coluna Vertebral/citologia
16.
J Biomed Mater Res A ; 103(2): 545-54, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24764177

RESUMO

There is growing concern about unwanted effects associated with the clinical use of recombinant human bone morphogenetic protein-2 (rhBMP-2) at high concentrations, including cyst-like bone formation and excessive fatty marrow formation. We, therefore, evaluated the induction of mineralized/adipose tissue formation and the bone-healing pattern associated with the controlled release of E. coli-derived rhBMP-2 (ErhBMP-2) by a heparin-conjugated fibrin (HCF) system using ectopic and orthotopic in vivo models, respectively. In the ectopic transplantation model, mineralized tissue formed at the most superficial layer of the transplanted area and on the surfaces of grafted materials, and most of the interstitial space within the transplanted area was filled with excessive adipose tissue specifically at sites that received ErhBMP-2. However, sites that received ErhBMP-2 and HCF showed significantly increased mineralized tissue formation and decreased adipose tissue formation compared to the normal fibrin system with ErhBMP-2. In the orthotopic (calvarial defect) model, controlled release of ErhBMP-2 induced by HCF significantly reduced adipose tissue formation within the defect area compared to the clinically approved absorbable collagen sponge. From these results, it can be concluded that the use of a HCF system loaded with ErhBMP-2 may reduce adipose tissue formation and enhance mineralized tissue formation.


Assuntos
Adipogenia/efeitos dos fármacos , Tecido Adiposo/metabolismo , Proteína Morfogenética Óssea 2 , Calcificação Fisiológica/efeitos dos fármacos , Portadores de Fármacos , Heparina , Animais , Proteína Morfogenética Óssea 2/química , Proteína Morfogenética Óssea 2/farmacologia , Preparações de Ação Retardada/química , Preparações de Ação Retardada/farmacologia , Portadores de Fármacos/química , Portadores de Fármacos/farmacologia , Heparina/química , Heparina/farmacologia , Humanos , Masculino , Camundongos , Ratos , Ratos Sprague-Dawley
17.
J Periodontal Implant Sci ; 44(3): 109-17, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24921054

RESUMO

PURPOSE: The aim of this study was to determine the effects of nutritional supplements on periodontal health and tooth mobility after surgery. METHODS: Patients were randomly assigned to an intervention group who consumed nutritional supplement drinks for 8 weeks, while the placebo group did not receive any such supplements. The gingival index (GI) and tooth mobility were measured at baseline and at 1, 4, and 8 weeks. In addition, the oral health impact profile-14 and anthropometric measurements along with loss of appetite and dietary intake were assessed at baseline and 8 weeks. RESULTS: At 1 week, GI values were reduced in the intervention group (P<0.05), and tooth mobility had increased, but to a lesser extent in the intervention group (P<0.05). At 8 weeks, the intakes of protein, vitamins A and B1, and niacin were increased in the intervention group. CONCLUSIONS: These results demonstrate that nutritional supplementation improves early periodontal healing after surgery.

18.
J Periodontol ; 85(9): e339-47, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24678850

RESUMO

BACKGROUND: The aims of this study are to determine whether the antigen-inexperienced (naive, CD45RB high-density) T-cell (CD4(+)CD45RB(High) T-cell) transfer model is associated with alveolar bone resorption, to elucidate the local osteogenic/adipogenic potential of alveolar bone marrow stromal cells (ABCs) from T-cell-transferred animals, and to investigate the systemic osteogenic potential by transplanting human periodontal ligament stem cells (hPDLSCs) into these animals. METHODS: CD4(+)CD45RB(High) and CD4(+)CD45RB(Low) (antigen-experienced [memory, CD45RB low-density]) T cells were sorted and transferred into severe combined immunodeficiency (SCID) mice to induce inflammatory bowel disease-like syndrome (n = 8). hPDLSCs were transplanted into T-cell-transferred SCID mice to examine ectopic cementum formation 8 weeks after T-cell transfer. The mandibles and tibias of these mice were retrieved for microcomputed tomography (micro-CT), histomorphometric analysis, and isolation of ABCs 16 weeks after T-cell transfer. The in vitro osteogenic and adipogenic potentials of the ABCs were evaluated. RESULTS: Histologic and micro-CT analysis revealed that the transfer of CD4(+)CD45RB(High) T-cell subset was sufficient for alveolar bone resorption and affected the osteogenic/adipogenic potential of ABCs. Furthermore, it was found that CD4(+)CD45RB(High) T-cell-transferred animals have decreased systemic osteogenic potential, as evidenced using the in vivo ectopic hPDLSC transplantation model. CONCLUSION: CD4(+)CD45RB(High) T-cell transfer induced both alveolar bone resorption and reduced systemic osteogenic potential, with a concomitant downregulation of the osteogenic potential of ABCs.


Assuntos
Transferência Adotiva/métodos , Perda do Osso Alveolar/imunologia , Linfócitos T CD4-Positivos/imunologia , Hospedeiro Imunocomprometido , Antígenos Comuns de Leucócito/imunologia , Adipogenia/imunologia , Adulto , Perda do Osso Alveolar/patologia , Processo Alveolar/patologia , Animais , Densidade Óssea/imunologia , Linfócitos T CD4-Positivos/transplante , Técnicas de Cultura de Células , Diferenciação Celular/imunologia , Células Cultivadas , Cementogênese/imunologia , Feminino , Humanos , Doenças Inflamatórias Intestinais/imunologia , Masculino , Mandíbula/imunologia , Mandíbula/patologia , Células-Tronco Mesenquimais/imunologia , Camundongos , Camundongos Endogâmicos ICR , Camundongos SCID , Osteogênese/imunologia , Ligamento Periodontal/citologia , Ligamento Periodontal/imunologia , Transplante de Células-Tronco/métodos , Tíbia/imunologia , Tíbia/patologia , Microtomografia por Raio-X/métodos , Adulto Jovem
19.
J Biomed Mater Res B Appl Biomater ; 102(1): 80-8, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23852942

RESUMO

Various synthetic bone substitutes have been developed to reconstruct the bony defects that clinicians often encounter during surgical procedures. Among various synthetic bone substitutes, calcium phosphate (Ca-P) ceramics have been investigated because their composition and structure are similar to those of human bone. We evaluated the bone healing and biodegradation patterns of three types of Ca-P ceramic particle with various hydroxyapatite (HA)/ß-tricalcium phosphate (ß-TCP) weight ratio: pure ß-TCP, biphasic Ca-P (BCP) with a HA/ß-TCP weight ratio of 60/40 (BCP 60/40), and BCP with an HA/ß-TCP weight ratio of 20/80 (BCP 20/80). Four 8-mm-diameter defects were created in ten rabbits. Three of the defects in each rabbit were separately and randomly filled with one of the three experimental Ca-P ceramic particles, and the fourth was filled with blood clots (control). The specimens were harvested at 2 and 8 weeks post-surgery. The histologic and histometric findings revealed that the augmented space and new bone formation were significantly better for all three Ca-P ceramics than for the control group at both 2 and 8 weeks (p < 0.05). Compared to the pure ß-TCP, the two BCP groups were found to provide a larger amount of newly formed bone and bone density at the 2- and 8-week post-operative periods (p < 0.05). Throughout the observation period, BCP 60/40 and BCP 20/80 exhibited a similar bone healing and biodegradation patterns with regard to both individual particles and the total augmented area in vivo.


Assuntos
Substitutos Ósseos/química , Fosfatos de Cálcio/química , Implantes Absorvíveis , Animais , Regeneração Óssea , Substitutos Ósseos/metabolismo , Fosfatos de Cálcio/metabolismo , Humanos , Masculino , Teste de Materiais , Microscopia Eletrônica de Varredura , Osseointegração , Coelhos , Crânio/anatomia & histologia , Crânio/fisiologia , Crânio/cirurgia
20.
J Periodontal Implant Sci ; 44(6): 293-9, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25568810

RESUMO

PURPOSE: Systemic disease can manifest oral signs at an early phase, which may be crucial for the diagnosis and timing of treatment. This report describes two patients who presented with gingival enlargement as an early sign of acute leukemia. METHODS: Two patients presented with oral symptoms including severe gingival enlargement. The progress of their symptoms was associated with underlying systemic disease. RESULTS: The patients were transferred to the Department of Hematology and diagnosed with acute myelomonocytic leukemia. They received appropriate treatment and survived. CONCLUSIONS: Gingival enlargement can be caused by underlying systemic diseases. Accurate diagnosis and timely referral are important for preventing a fatal situation. It must be emphasized that some oral signs and symptoms may be closely correlated with systemic diseases.

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