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The aim of the present study was to investigate the physiological role of nicotinamide phosphoribosyltransferase (NAMPT) associated with odontogenic differentiation during tooth development in mice. Mouse dental papilla cell-23 (MDPC-23) cells cultured in differentiation media were stimulated with the specific NAMPT inhibitor, FK866, and Visfatin (NAMPT) for up to 10 days. The cells were evaluated after 0, 4, 7, and 10 days. Cell viability was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The mineralization assay was performed by staining MDPC-23 cells with Alizarin Red S solution. After cultivation, MDPC-23 cells were harvested for quantitative PCR or Western blotting. Analysis of variance was performed using StatView 5.0 software (SAS Institute Inc., Cary, NC, USA). Statistical significance was set at p < 0.05. The expression of NAMPT increased during the differentiation of murine odontoblast-like MDPC-23 cells. Furthermore, the up-regulation of NAMPT promoted odontogenic differentiation and accelerated mineralization through an increase in representative odontoblastic biomarkers, such as dentin sialophosphoprotein, dentin matrix protein-1, and alkaline phosphatase in MDPC-23 cells. However, treatment of the cells with the NAMPT inhibitor, FK866, attenuated odontogenic differentiation, as evidenced by the suppression of odontoblastic biomarkers. These data indicate that NAMPT regulated odontoblastic differentiation through the regulation of odontoblastic biomarkers. The increase in NAMPT expression in odontoblasts was closely related to the formation of the extracellular matrix and dentin via the Runx signaling pathway. Therefore, these data suggest that NAMPT is a critical regulator of odontoblast differentiation during tooth development.
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Alpha-lipoic acid (ALA) is a naturally occurring antioxidant and has been previously used to treat diabetes and cardiovascular disease. However, the autophagy effects of ALA against oxidative stress-induced dopaminergic neuronal cell injury remain unclear. The aim of this study was to investigate the role of ALA in autophagy and apoptosis against oxidative stress in the SH-SY5Y human dopaminergic neuronal cell line. We examined SH-SY5Y phenotypes using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay (cell viability/proliferation), 4′,6-diamidino-2-phenylindole dihydrochloride nuclear staining, Live/Dead cell assay, cellular reactive oxygen species (ROS) assay, immunoblotting, and immunocytochemistry. Our data showed ALA attenuated hydrogen peroxide (H2O2)-induced ROS generation and cell death. ALA effectively suppressed Bax up-regulation and Bcl-2 and BclxL down-regulation. Furthermore, ALA increased the expression of the antioxidant enzyme, heme oxygenase-1. Moreover, the expression of Beclin-1 and LC-3 autophagy biomarkers was decreased by ALA in our cell model. Combined, these data suggest ALA protects human dopaminergic neuronal cells against H2O2-induced cell injury by inhibiting autophagy and apoptosis.
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Alpha-lipoic acid (ALA) is a naturally occurring antioxidant and has been previously used to treat diabetes and cardiovascular disease. However, the autophagy effects of ALA against oxidative stress-induced dopaminergic neuronal cell injury remain unclear. The aim of this study was to investigate the role of ALA in autophagy and apoptosis against oxidative stress in the SH-SY5Y human dopaminergic neuronal cell line. We examined SH-SY5Y phenotypes using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay (cell viability/proliferation), 4′,6-diamidino-2-phenylindole dihydrochloride nuclear staining, Live/Dead cell assay, cellular reactive oxygen species (ROS) assay, immunoblotting, and immunocytochemistry. Our data showed ALA attenuated hydrogen peroxide (H2O2)-induced ROS generation and cell death. ALA effectively suppressed Bax up-regulation and Bcl-2 and BclxL down-regulation. Furthermore, ALA increased the expression of the antioxidant enzyme, heme oxygenase-1. Moreover, the expression of Beclin-1 and LC-3 autophagy biomarkers was decreased by ALA in our cell model. Combined, these data suggest ALA protects human dopaminergic neuronal cells against H2O2-induced cell injury by inhibiting autophagy and apoptosis.
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The bone and dentin have distinct healing processes. The healing process of bones is regenerative, as newly formed tissues are morphologically and functionally similar to the original bone structures. In contrast, the healing process of dentin is reparative due to its failure to replicate some of its key morphological features. In this review, we compare and contrast the healing processes of bone and dentin. We describe how distinct morphological and physiological structures of the 2 tissues translate into different signaling molecules, growth factors, and matrix protein secretion.
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Purpose@#The aims of this study were to examine the salivary microbiota in conditions of periodontal health and disease and to explore microbial changes following nonsurgical periodontal treatment. @*Methods@#Non-stimulated saliva samples were collected from 4 periodontally healthy participants at baseline and from 8 patients with chronic periodontitis at baseline and 3 months following nonsurgical periodontal therapy. The V3 and V4 regions of the 16S rRNA gene from the DNA of saliva samples were amplified and sequenced. The salivary microbial compositions of the healthy participants and patients with periodontitis prior to and following nonsurgical treatment of periodontitis were compared based on the relative abundance of various taxa. @*Results@#On average, 299 operational taxonomic units were identified in each sample. The phylogenetic diversity in patients with periodontitis was higher than that in healthy participants and decreased following treatment. The abundance of the phylum Spirochaetes and the genus Treponema in patients with periodontitis was 143- and 134-fold higher than in the healthy control group, respectively, but decreased significantly following treatment. The species that were overabundant in the saliva of patients with periodontitis included the Peptostreptococcus stomatis group, Porphyromonas gingivalis, the Fusobacterium nucleatum group, Parvimonas micra, Porphyromonas endodontalis, Filifactor alocis, and Tannerella forsythia. The phylum Actinobacteria, the genus Streptococcaceae_uc, and the species Streptococcus salivarius group were more abundant in healthy participants than in those with periodontitis. There was a trend toward a decrease in disease-associated taxa and an increase in health-associated taxa following treatment. @*Conclusions@#Our results revealed differences in the taxa of salivary microbiota between conditions of periodontal health and disease. The taxa found to be associated with health or disease have potential for use as salivary biomarkers for periodontal health or disease.
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PURPOSE: The purpose of this study was to evaluate the effects of platelet-rich plasma (PRP) on periodontal healing of replanted root surfaces in dogs histologically and histomorphometrically. METHODS: A total of 36 roots of mandibular incisors and premolars from 6 mongrel dogs were used. The roots were randomly divided into 3 groups: 1) a positive control group (n=12), in which the periodontal ligament (PDL) and cementum were retained and the roots were soaked in saline; 2) a negative control group (n=12), in which the PDL and cementum were removed and the roots were soaked in saline; and 3) an experimental group (n=12), in which the PDL and cementum were removed and the roots were soaked in PRP. After soaking the root surfaces, the extracted roots were replanted into the extraction sockets. The roots were covered using a coronally repositioned flap RESULTS: Histologically, irregular-thickness PDL-like and cementum-like tissues were observed in the 4-week experimental group and the positive control group. PDL-like tissue and cementum-like tissue with a more uniform thickness were observed at 8 weeks. In the negative control group, PDL-like tissue and cementum-like tissue were rarely found, and root resorption and ankylosis were observed. In the cross-sectional histomorphometric analysis, the experimental group demonstrated a higher rate of formation of cementum-like tissue and a lower tooth ankylosis rate than the positive and negative control groups at 4 and 8 weeks. Although there was a significant difference in the tooth ankylosis rate and the formation of cementum-like tissue across the 3 groups (P 0.017). CONCLUSIONS: Applying PRP to root surfaces during tooth replantation in dogs can reduce tooth ankylosis and increase PDL-like and cementum-like tissue formation.
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Animales , Perros , Anquilosis , Diente Premolar , Vestuario , Cemento Dental , Incisivo , Ligamento Periodontal , Periodontitis , Plasma Rico en Plaquetas , Resorción Radicular , Anquilosis del Diente , Reimplante Dental , DienteRESUMEN
Resveratrol (3,4′,5,-trihydroxystilbene), a phytoalexin present in grapes, exerts a variety of actions to reduce superoxides, prevents diabetes mellitus, and inhibits inflammation. Resveratrol acts as a chemo-preventive agent and induces apoptotic cell death in various cancer cells. However, the role of resveratrol in odontoblastic cell differentiation is unclear. In this study, the effect of resveratrol on regulating odontoblast differentiation was examined in MDPC-23 mouse odontoblastic cells derived from mouse dental papilla cells. Resveratrol significantly accelerated mineralization as compared with the control culture in differentiation of MDPC-23 cells. Resveratrol significantly increased expression of ALP mRNA as compared with the control in differentiation of MDPC-23 cells. Resveratrol significantly accelerated expression of ColImRNA as compared with the control in differentiation of MDPC-23 cells. Resveratrol significantly increased expressions of DSPP and DMP-1 mRNAs as compared with the control in differentiation of MDPC-23 cells. Treatment of resveratrol did not significantly affect cell proliferation in MDPC-23 cells. Results suggest resveratrol facilitates odontoblast differentiation and mineralization in differentiation of MDPC-23 cells, and may have potential properties for development and clinical application of dentin regeneration materials.
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Animales , Ratones , Muerte Celular , Diferenciación Celular , Proliferación Celular , Papila Dental , Dentina , Diabetes Mellitus , Inflamación , Mineros , Odontoblastos , Regeneración , ARN Mensajero , Superóxidos , VitisRESUMEN
Metformin (1,1-dimethylbiguanide hydrochloride), derived from French lilac (Galega officinalis), is a first-line anti-diabetic drug prescribed for patients with type 2 diabetes. However, the role of metformin in odontoblastic cell differentiation is still unclear. This study therefore undertook to examine the effect of metformin on regulating odontoblast differentiation in MDPC-23 mouse odontoblastic cells derived from mouse dental papilla cells. As compared to controls, metformin significantly accelerated the mineralization, significantly increased and accelerated the expressions of ALP and Col I mRNAs, and significantly increased the accelerated expressions of DSPP and DMP-1 mRNAs, during differentiation of MDPC-23 cells. There was no alteration in cell proliferation of MDPC-23 cells, on exposure to metformin. These results suggest that the effect of metformin on MDPC-23 mouse odontoblastic cells derived from mouse dental papilla cells, facilitates the odontoblast differentiation and mineralization, without altering the cell proliferation.
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Animales , Humanos , Ratones , Diferenciación Celular , Proliferación Celular , Papila Dental , Metformina , Mineros , Odontoblastos , ARN MensajeroRESUMEN
Nuclear factor I-C (NFI-C) plays a pivotal role in various cellular processes such as odontoblast and osteoblast differentiation. Nfic-deficient mice showed abnormal tooth and bone formation. The transplantation of Nfic-expressing mouse bone marrow stromal cells rescued the impaired bone formation in Nfic(-/-) mice. Studies suggest that NFI-C regulate osteogenesis and dentinogenesis in concert with several factors including transforming growth factor-β1, Krüppel-like factor 4, and β-catenin. This review will focus on the function of NFI-C during tooth and bone formation and on the relevant pathways that involve NFI-C.
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Animales , Ratones , Desarrollo Óseo , Dentinogénesis , Células Madre Mesenquimatosas , Factores de Transcripción NFI , Odontoblastos , Osteoblastos , Osteogénesis , Osteoporosis , DienteRESUMEN
PURPOSE: In osteoclast differentiation, actin-rich membrane protrusions play a crucial role in cell adhesion. Odontogenic ameloblast-associated protein (Odam) contributes to cell adhesion by inducing actin rearrangement. Odam-mediated RhoA activity may play a significant role in multinucleation of osteoclasts. However, the precise function of Odam in osteoclast cell adhesion and differentiation remains largely unknown. Here, we identify a critical role for Odam in inducing osteoclast adhesion and differentiation. MATERIALS AND METHODS: The expression of Odam in osteoclasts was evaluated by immunohistochemistry. Primary mouse bone marrow and RAW264.7 cells were used to test the cell adhesion and actin ring formation induced by Odam. RESULT: Odam was expressed in osteoclasts around alveolar bone. Odam transfection induced actin filament rearrangement and cell adhesion compared with the control or collagen groups. Overexpression of Odam promoted actin stress fiber remodeling and cell adhesion, resulting in increased osteoclast fusion. CONCLUSION: These results suggest that Odam expression in primary mouse osteoclasts and RAW264.7 cells promotes their adhesion, resulting in the induction of osteoclast differentiation.
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Animales , Ratones , Citoesqueleto de Actina , Actinas , Médula Ósea , Adhesión Celular , Colágeno , Inmunohistoquímica , Membranas , Osteoclastos , Fibras de Estrés , TransfecciónRESUMEN
Molecular interactions between epithelium and mesenchyme are important for root formation. Nuclear factor I-C (Nfic) has been identified as a key regulator of root formation. However, the mechanisms of root formation and their interactions between Hertwig's epithelial root sheath (HERS) and mesenchyme remain unclear. In this study, we investigated the role of Nfic in root patterning and growth during molar root development. The molars of Nfic knockout mice exhibited an enlarged pulp chamber and apical displacement of the pulpal floor, characteristic features of taurodontism, due to delayed furcation formation. In developing molar roots of mutant mice at P14, BrdU positive cells decreased in the apical mesenchyme of the elongation region whereas those cells increased in the dental papilla of the furcation region. Whereas cytokeratin 14 and laminin were localized in HERS cells of mutant molars, Smoothened (Smo) and Gli1 were downregulated in preodontoblasts. In contrast, cytokeratin 14 and Smo were localized in the cells of the furcation region of mutant molars. These results indicate that Nfic regulates cell proliferation in the dental mesenchyme and affects the fate of HERS cells in a site-specific manner. From the results, it is suggested that Nfic is required for root patterning and growth during root morphogenesis.
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Animales , Ratones , Bromodesoxiuridina , Proliferación Celular , Papila Dental , Cavidad Pulpar , Epitelio , Queratina-14 , Laminina , Mesodermo , Ratones Noqueados , Diente Molar , Morfogénesis , Factores de Transcripción NFI , Raíz del Diente , DienteRESUMEN
MicroRNA (miRNA, miR) is essential in regulating cell differentiation either by inhibiting mRNA translation or by inducing its degradation. However, the role of miRNA in odontoblastic cell differentiation is still unclear. In this study, we examined the molecular mechanism of miR-27-mediated regulation of odontoblast differentiation in MDPC-23 mouse odontoblastic cells derived from mouse dental papilla cells. The results of the present study demonstrated that the miR-27 expression increases significantly during MDPC-23 odontoblastic cell differentiation. Furthermore, miR-27 up-regulation promotes the differentiation of MDPC-23 cells and accelerates mineralization without cell proliferation. The over-expression of miR-27 significantly increased the expression levels of Wnt1 mRNA and protein. In addition, the results of target gene prediction revealed that Wnt1 mRNA has an miR-27 binding site in its 3'UTR, and is increased by miR-27. These results suggested that miR-27 promotes MDPC-23 odontoblastic cell differentiation by targeting Wnt1 signaling. Therefore, miR-27 is a critical odontoblastic differentiation molecular target for the development of miRNA based therapeutic agents in dental medicine.
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Animales , Ratones , Regiones no Traducidas 3' , Sitios de Unión , Diferenciación Celular , Proliferación Celular , Papila Dental , MicroARNs , Odontoblastos , Biosíntesis de Proteínas , ARN Mensajero , Regulación hacia ArribaRESUMEN
PURPOSE: The purpose of this animal study was to perform a histological and histomorphometric analysis in order to elucidate the effect of fibroblast growth factor-2 (FGF-2) on injured periodontal ligament (PDL) and cementum after tooth replantation in dogs. METHODS: The roots of 36 mandibular premolars from six mongrel dogs were used in this study. The roots were randomly divided into three groups: (1) a positive control group (n=12), in which the PDL was retained; (2) a negative control group (n=12), in which the PDL and the cementum between the notches were removed; and (3) an experimental group (n=12), in which the PDL and the cementum between the notches were removed and the roots were soaked in an FGF-2 solution (30 microg/0.1 mL). After treating the root surfaces, the extracted roots were replanted into extraction sockets. The animals were sacrificed four and eight weeks after surgery for histologic and histomorphometric evaluation. RESULTS: At four and eight weeks, normal PDLs covered the roots in the positive control group. In the negative control group, most replanted roots showed signs of replacement resorption. In the experimental group, new PDL-like tissue and cementum-like tissue were observed to partially occupy the region between the root surfaces and the newly formed bone. Histomorphometric analysis showed that the mean length of the newly formed cementum-like tissue on the roots treated with FGF-2 was significantly greater than that of the tissue on the roots in the negative control group (four weeks, P=0.008; eight weeks, P=0.042). However, no significant differences were observed between the roots treated with FGF-2 and the negative control roots with respect to newly formed PDL-like tissue. CONCLUSIONS: The results of this study suggest that use of FGF-2 on injured root surfaces promotes cementogenesis after tooth replacement in dogs.
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Animales , Perros , Diente Premolar , Cementogénesis , Cemento Dental , Factor 2 de Crecimiento de Fibroblastos , Factores de Crecimiento de Fibroblastos , Ligamento Periodontal , Periodoncio , Regeneración , Diente , Reimplante DentalRESUMEN
NFI-C null mice demonstrate aberrant odontoblast differentiation and abnormal dentin formation, and thus develop molars lacking roots. However, other tissues and organs in the body including ameloblasts appear to be unaffected. Abnormal dentin in NFI-C null mice shares morphological similarities to the osteodentin that is formed in dental caries. However, little is known about the relationship between NFI-C and osteodentin formation. In this study, to elucidate the molecular characteristics of abnormal odontoblast in NFI-C null mice, we examined the levels of Ask-1, Cdc-2, Smad2/3, and TGF-betaR1 in cell culture and tissue sections from NFI-C null mice using immunofluorescence and immunohistochemistry. NFI-C protein was localized in the nucleus and cytoplasm of normal odontoblasts in vitro. Ask-1 and Cdc-2 proteins were shown in the perinuclear cytoplasm of both normal and NFI-C null mice. There were no differences in the pattern of production of Ask-1 and Cdc-2 proteins between normal and NFI-C null mice. Smad2/3 was not found in the odontoblast and subodontoblastic cells of the normal mice, whereas NFI-C null mice showed Smad2/3 immunoreactivity in the odontoblasts and subodontoblastic cells of the tooth pulp. TGF-betaR1 was weakly immunopositive in the odontoblast and subodontoblastic cells of normal mice, whereas it was detected strongly in the subodontoblastic cells of the NFI-C null mice. These results suggest that disruption of NFI-C increased the expression of Smad2/3 and TGF-betaR1 in developing odontoblasts and consequently caused abnormal dentin formation, similar to osteodentin.
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Animales , Ratones , Ameloblastos , Técnicas de Cultivo de Célula , Citoplasma , Caries Dental , Dentina , Técnica del Anticuerpo Fluorescente , Inmunohistoquímica , Diente Molar , Odontoblastos , DienteRESUMEN
The aim of this study was to examine the possibility of periodontal ligament regeneration when autotransplantation was used by the periodontal ligament fibroblasts cultured on the acellular dermal matrix in teeth without a periodontal ligament. One minipig was used in this study. The mandibular and maxillary permanent incisors were extracted for the culture of the periodontal ligament cells. The roots of the unextracted teeth were classified into a positive control group, in which the normal periodontal ligament was preserved. The roots of the extracted teeth were divided into the following two groups: The negative control group, in which the periodontal ligament had been removed and the acellular dermal matrix was not applied; and an experimental group, in which the periodontal ligament had been removed and periodontal ligament fibroblast cultured on an acellular dermal matrix was applied. The prepared teeth were transplanted, and completely submerged using physical barrier membranes. The animal was sacrificed 4 weeks after the autotransplant. The transplanted teeth were examined histologically. In this study, the periodontal ligament was normal in the positive control group, and ankylosis was discovered on the denuded root surface in the negative control group. Periodontal ligament-like connective tissue was found adjacent to the denuded root and the new cementum-like layer of hard tissue was formed in the experimental group. These results suggest that the periodontal ligament fibroblasts cultured on the acellular dermal matrix may play a role in regenerating the periodontal ligament-like tissue with new cememtum-like tissue formation.
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Animales , Dermis Acelular , Anquilosis , Autoinjertos , Tejido Conectivo , Fibroblastos , Incisivo , Membranas , Ligamento Periodontal , Regeneración , Porcinos Enanos , DienteRESUMEN
This study evaluated the possibility of the 3-dimensional attachment of human periodontal ligament fibroblasts to a periodntally involved root surface after an EDTA treatment in vitro. The human PDL fibroblasts were isolated from the middle third of the root of periodontally healthy teeth extracted for orthodontic reasons. The cells were cultured in a medium containing Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at 37degrees C in humidified air containing 5% CO2. Eight single-rooted teeth were obtained from patients diagnosed with periodotitis. After scaling and root planing, four teeth were etched with 24% ethylenediaminetetracetic acid (EDTA) for two minutes (Experimental group). The other four teeth were not treated with EDTA and were used as the control group. The human PDL fibroblasts were placed in the total root surface and cultured for 4 weeks. The teeth were fixed in 2.5% glutaraldehyde in PBS before preparation for the scanning electron microscopy (SEM) examination. The human PDL fibroblasts showed a healthy morphology on the root surfaces treated with EDTA (Experimental group) and a relatively unhealthy appearance on the treated root surfaces (Control group). This suggests that EDTA favorably affects the 3-dimensional attachment of human PDL fibroblasts cultured on the root surfaces, which may play an important role in periodontal healing and regeneration.
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Humanos , Águilas , Ácido Edético , Fibroblastos , Glutaral , Microscopía Electrónica de Rastreo , Ligamento Periodontal , Regeneración , Aplanamiento de la Raíz , DienteRESUMEN
Periodontal ligament (PDL) is the connective tissue located between the tooth root and alveolar bone. In a previous study, PDLs22 was isolated as a PDL-specific gene by using subtractive hybridization between cultured PDL fibroblasts and gingival fibroblasts. It was also suggested that PDLs22 plays important roles in the development, differentiation and maintenance of periodontal tissues. However, little is known about functional study of PDLs22 using recombinant protein in PDL fibroblast differentiation and periodontium formation. In this study, in order to produce the PDLs22 recombinat protein, PDLs22-expression vector were constructed and expressed its protein in various host cell and temperature conditions. The results were as follows: 1. PDLs22 protein was not strongly expressed in the induction system using pRSET-PDLs22 construct. 2. When the BL21(DE3) pLysS was used as a expression host, PDLS22 protein was strongly expressed in the induction system using pHCEIIBNd-PDLs22 construct. 3. The PDLs22 protein was recognized at a molecular weight of 28 kDa in western blots. 4. Almost of the expressed PDLs22 protein was not soluble and observed like as inclusion body. 5. The protein solubility was not improved after modification of induction time and temperature during PDLs22 protein production. In this study, the system for the PDLs22 protein production was connstructed. However, the results suggest that further studies will be needed to produce the considerable amount of PDLs22 recombinat protein, which can use for the periodontal regeneration.
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Western Blotting , Tejido Conectivo , Fibroblastos , Cuerpos de Inclusión , Peso Molecular , Ligamento Periodontal , Periodoncio , Regeneración , Solubilidad , Raíz del DienteRESUMEN
Protein microarray or protein chips is potentially powerful tools for analysis of protein-protein interactions. APin cDNA was previously identified and cloned from a rat odontoblast cDNA library. The purpose of this study was to investigate the APin-protein interactions during ameloblast differentiation. Protein microarray was carried with recombinant APin protein and MEF2, Aurora kinase A, BMPR-IB and EF-hand calcium binding protein were selected among 74 interacting proteins. Immortalized ameloblast cells (ALCs) were transfected with pCMV-APin construct and U6-APin siRNA construct. After transfection, the expression of the mRNAs for four proteins selected by protein micoarrays were assessed by RT-PCR. The results were as follows: 1. APin expression was increased and decreased markedly after its over-expression and inactivation, respectively. 2. Over-expression of the APin in the ALCs markedly down-regulated the expression of MEF2 and Aurora kinase A, whereas their expression remained unchanged by its inactivation. 3. Expression of BMPR-IB and EF-hand calcium binding protein were markedly increased by the overexpression of the APin in the ALCs, whereas expression of BMPR-IB remained unchanged and expression of EF-hand calcium binding protein was markedly decreased by its inactivation. These results suggest that APin plays an important role in ameloblast differentiation and mineralization by regulating the expression of MEF2, Aurora kinase A, BMPR-IB and EF-hand calcium binding protein.
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Animales , Ratas , Ameloblastos , Aurora Quinasa A , Calcio , Proteínas Portadoras , Células Clonales , ADN Complementario , Biblioteca de Genes , Odontoblastos , Análisis por Matrices de Proteínas , ARN Mensajero , ARN Interferente Pequeño , TransfecciónRESUMEN
Melatonin is the major hormone released from the pineal gland and regulates a variety of physiological and pathophysiological processes. According to the recent studies the melatonin plays an important role in regulation of bone growth. The purpose of this study was to determine whether melatonin promotes the cell differentiation and nodules formation in MDPC-23 pre-odontoblast cell line. MDPC-23 cells were cultured for up to 15 days in growth media containing differentiation medium with melatonin or without melatonin. Cultures were stained with Alizarin-S. The expression of the mRNAs for DSPP, OC, ALP and NFI-C were analyzed by RT-PCR. The results were as follows. Cultures containing melatonin at day 15 showed extensive mineralization as compared with control cultures. Melatonin increased the expression of DSPP and OC mRNAs in MDPC-23 cells in a concentration-dependent manner. However, melatonin did not changed ALP expression. Melatonin markedly decreased mRNA expression of NFI-C in early stage cultures as compared with control cultures. These results demonstrated that melatonin is capable of promoting MDPC-23 cells differentiation and mineralization and suggested that melatonin may play an important role in dentin formation.
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Desarrollo Óseo , Diferenciación Celular , Línea Celular , Dentina , Melatonina , Glándula Pineal , ARN MensajeroRESUMEN
OBJECTIVE: Periodontal ligament fibroblasts have an ectomesenchymal origin and are thought to play a crucial role for not only homeostasis of periodontal tissues but also bone remodeling, wound healing and regeneration of tissues. Recently, it has been reported that UNC-50 is not expressed in gingival fibroblasts but in PDL fibroblasts. The purpose of this study was to examine the expression of UNC-50 and osteocalcin in the periodontium after application of intermittent force. METHODS: Twelve rats had 40 grams of mesially-directed force applied at the upper molar for 1 hour/day. Four rats were sacrificed at 1, 3 and 5 days. Immunohistochemical localization of UNC-50 and osteocalcin antibody was carried out. The results showed apposition of new cellular cementum and a slight increase in periodontal space at the tension side. RESULTS: Strong UNC-50 expression was observed in the differentiating cementoblasts close to PDL fibroblasts in the tension side whereas it was barely expressed at the compression side. Expression was strong at day 3, and decreased at day 5. Osteocalcin immunoreactivity expression was strong in differentiating cementoblasts at the tension side. CONCLUSION: It can be suggested that UNC-50 is related to the differentiation of cementoblasts, and may be responsible for the molecular event in PDL cells under mechanical stress.