Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 2.631
Filtrar
1.
Braz Oral Res ; 33: e117, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31939498

RESUMO

The aim of this study was to evaluate the effect of mineral trioxide aggregate (MTA) and Brazilian propolis on the cell viability, mineralization, anti-inflammatory ability, and migration of human dental pulp cells (hDPCs). The cell viability was evaluated with CCK-8 kit after 1, 5, 7, and 9 days. The deposition of calcified matrix and the expression of osteogenesis-related genes were evaluated by Alizarin Red staining and real-time PCR after incubation in osteogenic medium for 21 days. The expression of inflammation-related genes in cells was determined after exposure to 1 µg/mL LPS for 3 h. Finally, the numbers of cells that migrated through the permeable membranes were compared during 15 h. Propolis and MTA significantly increased the viability of hDPCscompared to the control group on days 7 and 9. In the propolis group, significant enhancement of osteogenic potential and suppressed expression of IL-1ß and IL-6 was observed after LPS exposure compared to the MTA and control groups. The number of migration cells in the propolis group was similar to that of the control group, while MTA significantly promoted cell migration. Propolis showed comparable cell viability to that of MTA and exhibited significantly higher anti-inflammatory and mineralization promotion effects on hDPCs.


Assuntos
Compostos de Alumínio/farmacologia , Anti-Inflamatórios/farmacologia , Compostos de Cálcio/farmacologia , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Óxidos/farmacologia , Própole/farmacologia , Silicatos/farmacologia , Antraquinonas , Brasil , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Combinação de Medicamentos , Humanos , Interleucina-1beta/análise , Interleucina-6/análise , Odontoblastos/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Estatísticas não Paramétricas , Fator de Necrose Tumoral alfa/análise
2.
J Appl Oral Sci ; 28: e20190215, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31939521

RESUMO

OBJECTIVE: This study evaluated the angiogenesis-enhancing potential of a tricalcium silicate-based mineral trioxide aggregate (ProRoot MTA), Biodentine, and a novel bioceramic root canal sealer (Well-Root ST) in human dental pulp stem cells (hDPSCs), human periodontal ligament stem cells (hPLSCs), and human tooth germ stem cells (hTGSCs). METHODOLOGY: Dulbecco's modified Eagle's medium was conditioned for 24 h by exposure to ProRoot MTA, Biodentine, or Well-Root ST specimens (prepared according to the manufacturers' instructions). The cells were cultured in these conditioned media and their viability was assessed with 3-(4,5-dimethyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H tetrazolium (MTS) on days 1, 3, 7, 10, and 14. Angiogenic growth factors [platelet-derived growth factor (PDGF), basic fibroblast growth factor (FGF-2), and vascular endothelial growth factor (VEGF)] were assayed by sandwich enzyme-linked immunosorbent assay (ELISA) on days 1, 7, and 14. Human umbilical vein endothelial cell (HUVEC) migration assays were used to evaluate the vascular effects of the tested materials at 6-8 h. Statistical analyses included Kruskal-Wallis, Mann-Whitney U, and Friedman and Wilcoxon signed rank tests. RESULTS: None of tricalcium silicate-based materials were cytotoxic and all induced a similar release of angiogenic growth factors (PDGF, FGF-2, and VEGF) (p>0.05). The best cell viability was observed for hDPSCs (p<0.05) with all tricalcium silicate-based materials at day 14. Tube formation by HUVECs showed a significant increase with all tested materials (p<0.05). CONCLUSION: The tricalcium silicate-based materials showed potential for angiogenic stimulation of all stem cell types and significantly enhanced tube formation by HUVECs.


Assuntos
Indutores da Angiogênese/farmacologia , Compostos de Cálcio/farmacologia , Cerâmica/farmacologia , Materiais Restauradores do Canal Radicular/farmacologia , Silicatos/farmacologia , Células-Tronco/efeitos dos fármacos , Materiais Biocompatíveis/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Fator 2 de Crescimento de Fibroblastos/análise , Fator 2 de Crescimento de Fibroblastos/efeitos dos fármacos , Citometria de Fluxo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Teste de Materiais , Neovascularização Fisiológica/efeitos dos fármacos , Ligamento Periodontal/citologia , Ligamento Periodontal/efeitos dos fármacos , Fator de Crescimento Derivado de Plaquetas/análise , Fator de Crescimento Derivado de Plaquetas/efeitos dos fármacos , Reprodutibilidade dos Testes , Estatísticas não Paramétricas , Germe de Dente/citologia , Germe de Dente/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/análise , Fator A de Crescimento do Endotélio Vascular/efeitos dos fármacos
3.
J Appl Oral Sci ; 28: e20190023, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31800871

RESUMO

When exposure of the pulp to external environment occurs, reparative dentinogenesis can be induced by direct pulp capping to maintain pulp tissue vitality and function. These clinical situations require the use of materials that induce dentin repair and, subsequently, formation of a mineralized tissue. OBJECTIVE: This work aims to assess the effect of tricalcium silicate cements and mineral trioxide aggregate cements, including repairing dentin formation and inflammatory reactions over time after pulp exposure in Wistar rats. METHODOLOGY: These two biomaterials were compared with positive control groups (open cavity with pulp tissue exposure) and negative control groups (no intervention). The evaluations were performed in three stages; three, seven and twenty-one days, and consisted of an imaging (nuclear medicine) and histological evaluation (H&E staining, immunohistochemistry and Alizarin Red S). RESULTS: The therapeutic effect of these biomaterials was confirmed. Nuclear medicine evaluation demonstrated that the uptake of 99mTc-Hydroxymethylene diphosphonate (HMDP) showed no significant differences between the different experimental groups and the control, revealing the non-occurrence of differences in the phosphocalcium metabolism. The histological study demonstrated that in mineral trioxide aggregate therapies, the presence of moderate inflammatory infiltration was found after three days, decreasing during follow-ups. The formation of mineralized tissue was only verified at 21 days of follow-up. The tricalcium silicate therapies demonstrated the presence of a slight inflammatory infiltration on the third day, increasing throughout the follow-up. The formation of mineralized tissue was observed in the seventh follow-up day, increasing over time. CONCLUSIONS: The mineral trioxide aggregate (WhiteProRoot®MTA) and tricalcium silicate (Biodentine™) present slight and reversible inflammatory signs in the pulp tissue, with the formation of mineralized tissue. However, the exacerbated induction of mineralized tissue formation with the tricalcium silicate biomaterial may lead to the formation of pulp calcifications.


Assuntos
Compostos de Alumínio/farmacologia , Materiais Biocompatíveis/farmacologia , Compostos de Cálcio/farmacologia , Polpa Dentária/efeitos dos fármacos , Dentina/efeitos dos fármacos , Dentinogênese/efeitos dos fármacos , Óxidos/farmacologia , Silicatos/farmacologia , Animais , Polpa Dentária/patologia , Capeamento da Polpa Dentária/métodos , Exposição da Polpa Dentária/tratamento farmacológico , Exposição da Polpa Dentária/patologia , Combinação de Medicamentos , Proteínas da Matriz Extracelular/análise , Imuno-Histoquímica , Masculino , Imagem Molecular/métodos , Odontoblastos/efeitos dos fármacos , Fosfoproteínas/análise , Agentes de Capeamento da Polpa Dentária e Pulpectomia/farmacologia , Pulpite/tratamento farmacológico , Pulpite/patologia , Distribuição Aleatória , Ratos Wistar , Reprodutibilidade dos Testes , Sialoglicoproteínas/análise , Fatores de Tempo
4.
J Appl Oral Sci ; 28: e20190105, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31800873

RESUMO

Calcium aluminate cement (CAC) has been highlighted as a promising alternative for endodontic use aiming at periapical tissue repair. However, its effects on dental pulp cells have been poorly explored. OBJECTIVE: This study assessed the impact of calcium chloride (CaCl2) and bismuth oxide (Bi2O3) or zinc oxide (ZnO) additives on odontoblast cell response to CAC. METHODOLOGY: MDPC-23 cells were exposed for up to 14 d: 1) CAC with 2.8% CaCl2 and 25% ZnO (CACz); 2) CAC with 2.8% CaCl2 and 25% Bi2O3 (CACb); 3) CAC with 10% CaCl2 and 25% Bi2O3 (CACb+); or 4) mineral trioxide aggregate (MTA), placed on inserts. Non-exposed cultures served as control. Cell morphology, cell viability, gene expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), and dentin matrix protein 1 (DMP-1), ALP activity, and extracellular matrix mineralization were evaluated. Data were compared using ANOVA (α=5%). RESULTS: Lower cell density was detected only for MTA and CACb+ compared with Control, with areas showing reduced cell spreading. Cell viability was similar among groups at days one and three (p>0.05). CACb+ and MTA showed the lowest cell viability values at day seven (p>0.05). CACb and CACb+ promoted higher ALP and BSP expression compared with CACz (p<0.05); despite that, all cements supported ALP activity. Matrix mineralization were enhanced in CACb+ and MTA. CONCLUSION: In conclusion, CAC with Bi2O3, but not with ZnO, supported the expression of odontoblastic phenotype, but only the composition with 10% CaCl2 promoted mineralized matrix formation, rendering it suitable for dentin-pulp complex repair.


Assuntos
Compostos de Alumínio/química , Compostos de Alumínio/farmacologia , Compostos de Cálcio/química , Compostos de Cálcio/farmacologia , Cimentos Dentários/química , Cimentos Dentários/farmacologia , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Fosfatase Alcalina/análise , Fosfatase Alcalina/efeitos dos fármacos , Animais , Bismuto/química , Bismuto/farmacologia , Cloreto de Cálcio/química , Cloreto de Cálcio/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Combinação de Medicamentos , Expressão Gênica/efeitos dos fármacos , Teste de Materiais , Camundongos , Odontoblastos/efeitos dos fármacos , Óxidos/química , Óxidos/farmacologia , Reprodutibilidade dos Testes , Silicatos/química , Silicatos/farmacologia , Fatores de Tempo , Óxido de Zinco/química , Óxido de Zinco/farmacologia
5.
Braz Oral Res ; 33: e059, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31664357

RESUMO

We recently demonstrated that a co-culture system of human umbilical vein endothelial cells (HUVECs) and human dental pulp stem cells (hDPSCs) could enhance angiogenesis ability in vitro. However, whether tumor necrosis factor α (TNF-α) could promote blood vessel formation during pulp regeneration remained unknown. The aim of this study was to investigate the effects of TNF-α on the formation of endothelial tubules and vascular networks in a co-culture system of hDPSCs and HUVECs. hDPSCs were co-cultured with HUVECs at a ratio of 1:5. The Matrigel assay was performed to detect the total tubule branching lengths and numbers of branches, and the Cell-Counting Kit 8 assay was performed to examine the effect of TNF-α on cell proliferation. Real-time polymerase chain reactions and western blot were used to detect vascular endothelial growth factor (VEGF) mRNA and protein expression. The Matrigel assay showed significantly greater total branching lengths and numbers of branches formed in the experimental groups treated with different concentrations of TNF-α compared with the control group. The decomposition times of the tubule structures were also significantly prolonged (P < 0.05). Treatment with 50 ng/ml TNF-α did not significantly change the proliferation of co-cultured cells, but it significantly increased the VEGF mRNA and protein expression levels (p < 0.05). In addition, the migration abilities of HUVECs and hDPSCs increased after co-culture with TNF-α (p < 0.05). TNF-α enhanced angiogenic ability in vitro in the co-culture system of hDPSCs and HUVECs.


Assuntos
Indutores da Angiogênese/farmacologia , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologia , Adolescente , Adulto , Western Blotting , Contagem de Células , Ensaios de Migração Celular , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/fisiologia , Células Cultivadas , Colágeno , Polpa Dentária/fisiologia , Combinação de Medicamentos , Células Endoteliais da Veia Umbilical Humana/fisiologia , Humanos , Laminina , Neovascularização Fisiológica/fisiologia , Proteoglicanas , Reação em Cadeia da Polimerase em Tempo Real , Valores de Referência , Reprodutibilidade dos Testes , Fatores de Tempo , Fator A de Crescimento do Endotélio Vascular/análise , Fator A de Crescimento do Endotélio Vascular/efeitos dos fármacos , Adulto Jovem
6.
J Endod ; 45(11): 1332-1341, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31585735

RESUMO

INTRODUCTION: Leptin is secreted as a peptide hormone from adipose tissues. The aim of this study was to evaluate the effects of leptin on reparative dentin formation and angiogenesis in the pulp tissue of teeth in vivo. METHODS: Twenty-four 7-week-old male rats were anesthetized. Cavities were prepared in maxillary first molars. Pulp cappings were performed with collagen scaffold (Col) with a phosphate-buffered saline (PBS) vehicle (Col + PBS), leptin 1 µmol/L with Col (L1 + Col), or leptin 10 µmol/L with Col (L10 + Col). For the negative control group (no pulp capping), pulp capping was not performed. All cavities were sealed with resin-modified glass ionomer followed by a micro-computed tomographic scan, histologic examination, and immunohistochemical analysis. RESULTS: The volume of newly formed mineralized tissue in the leptin group was significantly (P < .01) higher than that in the control group based on micro-computed tomographic analysis. In histologic examination, hard tissue formation was rarely shown in the no pulp capping and Col + PBS groups. However, significantly (P < .01) larger amounts of newly mineralized tissue deposition were observed in the leptin groups. In immunohistochemical analysis, reparative dentin and new vessels formed in the pulp cavity of the leptin groups. Vascular endothelial growth factor, dentin sialoprotein, and dentin sialophosphoprotein were expressed around the newly formed mineralized tissue area. CONCLUSIONS: Leptin showed the ability to induce angiogenesis, odontogenic differentiation, and mineralization in exposed rat pulps. Leptin also exhibited favorable inflammatory responses in the pulp tissue. Not only osteodentin but also tubular dentin and new vessels were observed in the pulp cavity.


Assuntos
Polpa Dentária , Dentina Secundária , Leptina , Neovascularização Fisiológica , Odontoblastos , Animais , Diferenciação Celular , Polpa Dentária/efeitos dos fármacos , Capeamento da Polpa Dentária , Leptina/fisiologia , Masculino , Neovascularização Fisiológica/efeitos dos fármacos , Odontoblastos/efeitos dos fármacos , Ratos , Fator A de Crescimento do Endotélio Vascular
7.
Cell Prolif ; 52(6): e12675, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31553127

RESUMO

OBJECTIVES: To evaluate the regenerative potential of human dental pulp stem cells (hDPSCs) in an animal model of stress urinary incontinence (SUI). SUI, an involuntary leakage of urine, is due to physical stress involving an increase in bladder pressure and a damage of external urethral sphincter affecting muscles and nerves. Conventional therapies can only relieve the symptoms. Human DPSCs are characterized by peculiar stemness and immunomodulatory properties and might provide an alternative tool for SUI therapy. MATERIALS AND METHODS: In vitro phase: hDPSCs were induced towards the myogenic commitment following a 24 hours pre-conditioning with 5-aza-2'-deoxycytidine (5-Aza), then differentiation was evaluated. In vivo phase: pudendal nerve was transected in female rats to induce stress urinary incontinence; then, pre-differentiated hDPSCs were injected in the striated urethral sphincter. Four weeks later, urethral sphincter regeneration was assayed through histological, functional and immunohistochemical analyses. RESULTS: Human DPSCs were able to commit towards myogenic lineage in vitro and, four weeks after cell injection, hDPSCs engrafted in the external urethral sphincter whose thickness was almost recovered, committed towards myogenic lineage in vivo, promoted vascularization and an appreciable recovery of the continence. Moreover, hDPSCs were detected within the nerve, suggesting their participation in repair of transected nerve. CONCLUSIONS: These promising data and further investigations on immunomodulatory abilities of hDPSCs would allow to make them a potential tool for alternative therapies of SUI.


Assuntos
Polpa Dentária/efeitos dos fármacos , Células-Tronco/citologia , Uretra/efeitos dos fármacos , Incontinência Urinária por Estresse/tratamento farmacológico , Animais , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Polpa Dentária/citologia , Modelos Animais de Doenças , Feminino , Humanos , Ratos , Uretra/citologia
8.
Mater Sci Eng C Mater Biol Appl ; 104: 109955, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31500064

RESUMO

Calcium phosphate cement (CPC), functionalized with iron oxide nanoparticles (IONP), is of great promise to promote osteoinduction and new bone formation. In this work, the IONP powder was added into the CPC powder to fabricate CPC + IONP scaffolds and the effects of the novel composite on bone matrix formation and osteogenesis of human dental pulp stem cells (hDPSCs) were explored. A series of CPC + IONP magnetic scaffolds with different IONP contents (1%, 3% and 6%) were fabricated using 5% chitosan solution as the cement liquid. Western blotting and RT-PCR were used to analyze the signaling pathway. The IONP incorporation substantially enhanced the performance of CPC + IONP, with increases in both mechanical strength and cellular activities. The IONP addition greatly promoted the osteogenesis of hDPSCs, elevating the ALP activity, the expression of osteogenic marker genes and bone matrix formation with 1.5-2-fold increases. The 3% IONP incorporation showed the most enhancement among all groups. Activation of the extracellular signal-related kinases WNT/ß-catenin in DPSCs was observed, and this activation was attenuated by the WNT inhibitor DKK1. The results indicated that the osteogenic behavior of hDPSCs was likely driven by CPC + IONP via the WNT signaling pathway. In conclusion, incorporate IONP into CPC scaffold remarkably enhanced the spreading, osteogenic differentiation and bone mineral synthesis of stem cell. Therefore, this method had great potential for bone tissue engineering. The novel CPC + IONP composite scaffolds with stem cells are promising to provide an innovative strategy to enhance bone regenerative therapies.


Assuntos
Cimentos para Ossos/química , Cimentos para Ossos/farmacologia , Fosfatos de Cálcio/química , Compostos Férricos/química , Nanopartículas/química , Osteogênese/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Células-Tronco/efeitos dos fármacos , Regeneração Óssea/efeitos dos fármacos , Osso e Ossos/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Quitosana/química , Cimentos Dentários/química , Polpa Dentária/efeitos dos fármacos , Polpa Dentária/metabolismo , Humanos , Células-Tronco/metabolismo , Engenharia Tecidual/métodos , Tecidos Suporte/química , Proteínas Wnt/metabolismo , beta Catenina/metabolismo
9.
Shanghai Kou Qiang Yi Xue ; 28(3): 251-258, 2019 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-31489411

RESUMO

PURPOSE: To evaluate the effect of iRoot BP Plus as pulp-capping agents on the biological behaviors of stem cells from human exfoliated deciduous teeth (SHED) and human dental pulp stem cells (DPSC). METHODS: iRoot BP Plus and ProRoot MTA sample disks were prepared and the extractive solution was extracted from iRoot BP Plus and ProRoot MTA sample disks. The influence of iRoot BP Plus and MTA extracts on the SHED and DPSC proliferation capacity was detected by CCK-8 method at 1, 3, 5 and 7 days.The influence of iRoot BP Plus and MTA extract on the SHED and DPSC migration capacity was observed by Transwell chamber and scratch repair experiments. SHED and DPSC were respectively inoculated on the sample disks of iRoot BP Plus and MTA for culture. Phalloidin and DAPI (4',6-diamidino-2-phenylindole) were used for immunofluorescence staining on 1, 3 and 5 days respectively to observe cytoskeleton changes. SHED and DPSC underwent mineralization induction respectively in osteoblastic induction medium, the osteoblastic induction medium containing MTA extract and the osteoblastic induction medium containing iRoot BP Plus extract. Alkaline phosphatase (ALP) staining and quantitative analysis of ALP were performed at 7 and 14 days, Alizarin Red staining and semi-quantitative analysis of calcium salt deposition were performed at 21 days. SPSS 19.0 software package was used for statistical analysis of the data. RESULTS: iRoot BP Plus and MTA extracts could promote cell proliferation of SHED and DPSC. In cell migration and adhesion experiments, iRoot BP Plus and MTA both promoted migration and adhesion of SHED and DPSC, and iRoot BP Plus played a more significant role (P=0.000). After mineralization induction, the ALP activity of SHED and DPSC in iRoot BP Plus group was significantly greater than that of MTA. Alizarin red staining and semi-quantitative analysis of calcium salt deposition showed that both iRoot BP Plus and MTA could promote cell mineralization. Moreover, the ability of iRoot BP Plus to promote cell mineralization was significantly stronger than that of MTA (P=0.000). CONCLUSIONS: iRoot BP Plus and MTA has good biocompatibility and good osteogenetic differentiation ability, it can promote SHED and DPSC cell proliferation, adhesion, migration, and BP Plus has better affect of promoting iRoot SHED and DPSC adhesion, migration and distribution of differentiation than MTA, therefore iRoot BP Plus and MTA may be used as pulp capping agent both for deciduous and permanent teeth.


Assuntos
Polpa Dentária , Silicatos , Diferenciação Celular , Polpa Dentária/efeitos dos fármacos , Humanos , Silicatos/farmacologia , Células-Tronco , Dente Decíduo
10.
Rev. Asoc. Odontol. Argent ; 107(3): 110-115, jul.-sept. 2019.
Artigo em Espanhol | LILACS | ID: biblio-1048552

RESUMO

Este trabajo pretende actualizar los conocimientos acerca de los diversos materiales utilizados en las terapias pulpares (tanto en dientes primarios como en permanentes) que buscan una respuesta reparativa cada vez más conservadora, biológica y sustentable. Se trata de un recorrido por el uso de agentes como el agregado de trióxido mineral (MTA), el láser, Biodentine® y los concentrados de plasma rico en plaquetas, con sus características, sus posibles aplicaciones y su eficacia, evaluadas clínica y radiográficamente en múltiples trabajos de investigación, informes de casos, estudios comparativos (in vitro e in vivo) y ensayos experimentales en animales que documentan sus resultados (AU)


This literature review aims to update knowledge about the different materials used in pulp therapies (both in primary and permanent teeth) that seek for an increasingly conservative, biological and sustainable reparative response. It is a journey through the use of agents such as mineral trioxide aggregate (MTA), lasers, biodentine and platelet rich plasma concentrates, their properties, applications and efficacy, clinically and radiographically evaluated in multiple research papers, case reports, comparative studies (in vitro and in vivo) and experimental studies in animals that document their results


Assuntos
Humanos , Animais , Materiais Biocompatíveis/uso terapêutico , Polpa Dentária/efeitos dos fármacos , Plasma Rico em Plaquetas , Dente Decíduo , Dentição Permanente , Terapia a Laser
11.
Cell Prolif ; 52(6): e12680, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31454111

RESUMO

OBJECTIVES: The odontoblastic differentiation of dental pulp stem cells (DPSCs) contributes to tertiary dentin formation. Our previous study indicated that epiregulin (EREG) enhanced odontogenesis potential of dental pulp. Here, we explored the effects of EREG during DPSC odontoblastic differentiation. METHODS: The changes in EREG were detected during tertiary dentin formation. DPSCs were treated with recombinant human EREG (rhEREG), EREG receptor inhibitor gefitinib and short hairpin RNAs. The odontoblastic differentiation was assessed with ALP staining, ALP activity assay, alizarin red S staining and real-time RT-PCR of DSPP, OCN, RUNX2 and OSX. Western blot was conducted to examine the levels of p38 mitogen-activated protein kinase (p38 MAPK), c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase 1/2 (Erk1/2). The expression of EREG and odontoblastic differentiation-related markers was investigated in human dental pulp from teeth with deep caries and healthy teeth. RESULTS: Epiregulin was upregulated during tertiary dentin formation. rhEREG enhanced the odontoblastic differentiation of DPSCs following upregulated p38 MAPK and Erk1/2 phosphorylation, but not JNK, whereas depletion of EREG suppressed DPSC differentiation. Gefitinib decreased odontoblastic differentiation with decreased phosphorylation of p38 MAPK and Erk1/2. And suppression of p38 MAPK and Erk1/2 pathways attenuated DPSC differentiation. In human dental pulp tissue, EREG upregulation in deep caries correlates with odontoblastic differentiation enhancement. CONCLUSION: Epiregulin is released during tertiary dentin formation. And EREG enhanced DPSC odontoblastic differentiation via MAPK pathways.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Polpa Dentária/efeitos dos fármacos , Epirregulina/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Células-Tronco/citologia , Animais , Proliferação de Células/efeitos dos fármacos , Polpa Dentária/citologia , Proteínas da Matriz Extracelular/metabolismo , Masculino , Odontoblastos/citologia , Odontoblastos/efeitos dos fármacos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos
12.
J Appl Oral Sci ; 27: e20180550, 2019 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-31365709

RESUMO

PURPOSE: To compare, both qualitatively and quantitatively, the inflammatory cells, vascular density and IL-6 immunolabeled cells present in the pulp after pulpotomy with white MTA versus 15.5% ferric sulfate (FS). METHODOLOGY: Forty-eight mandibular first molars from 24 Wistar rats were divided into MTA or FS groups and subdivided according to the period after pulpotomy procedure (24, 48 and 72 hours). Four teeth (sound and untreated) were used as controls. Histological sections were obtained and assessed through the descriptive analysis of morphological aspects of pulp tissue and the quantification of inflammatory cells, vascular density and interleukin-6 (IL-6) expression. Data were statistically analyzed (p<0.05). RESULTS: The number of inflammatory cells was similar in both groups, being predominantly localized at the cervical radicular third. In the MTA group, increased inflammation was observed at 48 hours. Vascular density was similar in both groups and over time, being predominant in the medium radicular third. No correlation was found between the number of inflammatory cells and the vascular density. Pulp tissue was more organized in MTA-treated teeth. In both groups, a weak to moderate IL-6 expression was detected in odontoblasts and inflammatory cells. Comparing both groups, there was a greater IL-6 expression in the cervical radicular third of teeth treated with MTA at 24 hours and in the medium and apical thirds at 72 hours, while in the FS group a greater IL-6 expression was found in the apical third at 24 hours. CONCLUSION: The MTA group presented better histological features and greater IL-6 expression than the FS group. However, no difference was observed between the groups regarding the inflammatory status and vascularization, suggesting the usefulness of FS as a low-cost alternative to MTA.


Assuntos
Compostos de Alumínio/farmacologia , Compostos de Cálcio/farmacologia , Compostos Férricos/farmacologia , Inflamação/imunologia , Interleucina-6/análise , Óxidos/farmacologia , Pulpotomia/efeitos adversos , Silicatos/farmacologia , Animais , Polpa Dentária/efeitos dos fármacos , Polpa Dentária/patologia , Combinação de Medicamentos , Masculino , Ratos Wistar , Estatísticas não Paramétricas , Fatores de Tempo
13.
BMC Oral Health ; 19(1): 174, 2019 08 06.
Artigo em Inglês | MEDLINE | ID: mdl-31387578

RESUMO

BACKGROUND: This study evaluated the antibacterial efficiency and ability of propolis to promote regeneration of immature permanent non-vital dogs' teeth. METHODS: Ninety six immature permanent premolars teeth in 6 mongrel dogs were divided randomly into: experimental teeth (N = 72) and control teeth (N = 24). Periapical pathosis was induced in all experimental and positive control teeth. Experimental teeth were classified according to the used intra-canal medication into: group I (N = 36), propolis paste was used and group II (N = 36), triple antibiotic paste (TAP) was used. Bacteriologic samplings were collected before and after exposure to intra-canal medicaments. After the disinfection period (3 weeks), revascularization was induced in all experimental teeth. Each group was subdivided according to the root canal orifice plug into: subgroup A (N = 18), propolis paste was used and subgroup B (N = 18), mineral trioxide aggregates (MTA) was used. Each subgroup was further subdivided according to the evaluation period into 3 subdivisions (6 teeth each): subdivision 1; after 2 weeks, subdivision 2; after one month and subdivision 3; after 2 months. Positive control group had 12 teeth with induced untreated periapical pathosis. Negative control group had 12 untouched sound teeth. All teeth were evaluated with radiography and histology. The bacteriologic and radiographic data were analyzed using repeated measures ANOVA and post-hoc Tukey tests. The histologic data were analyzed using Kruskal-Wallis test, Mann-Whitney U test with Bonferroni's adjustment and Chi-square test. The significance level was set at P ≤ .05. RESULTS: There was no significant difference in the antibacterial effectiveness between TAP and propolis groups (P > .05). In all subdivisions, there was no significant difference between the experimental groups in terms of increase in root length and dentin thickness, decrease in apical closure, new hard tissue formation, vital tissue formation inside the pulp canal and apical closure scores (P > .05). CONCLUSION: Propolis can be comparable with TAP as a disinfection treatment option in regenerative endodontic. As a root canal orifice plug after revascularization of necrotic immature permanent teeth in dogs, propolis induces a progressive increase in root length and dentin thickness and a decrease in apical diameter similar to those of MTA.


Assuntos
Antibacterianos/administração & dosagem , Necrose da Polpa Dentária/tratamento farmacológico , Polpa Dentária/efeitos dos fármacos , Dentina/efeitos dos fármacos , Própole/administração & dosagem , Endodontia Regenerativa/métodos , Tratamento do Canal Radicular/métodos , Raiz Dentária/efeitos dos fármacos , Animais , Antibacterianos/uso terapêutico , Polpa Dentária/irrigação sanguínea , Polpa Dentária/fisiologia , Dentina/irrigação sanguínea , Dentina/fisiologia , Cães , Tecido Periapical/irrigação sanguínea , Tecido Periapical/efeitos dos fármacos , Tecido Periapical/fisiologia , Própole/uso terapêutico , Distribuição Aleatória , Irrigantes do Canal Radicular/uso terapêutico , Ápice Dentário/patologia , Raiz Dentária/irrigação sanguínea , Raiz Dentária/fisiologia , Resultado do Tratamento
14.
J Mol Histol ; 50(3): 273-283, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31049797

RESUMO

Recent studies have demonstrated that IGF-1 modulates the pluripotent differentiation of dental pulp stem cells (DPSCs). Although mTOR pathway activation has been showed as responsible for IGF-1 induced pluripotent differentiation, the mechanism that the IGF-1-mTOR pathway induces the neural differentiation of DPSCs is still unclear. In our research, we have demonstrated that 0-10 ng/mL IGF-1 had no obvious effect on the proliferation of DPSCs, but IGF-1 nonetheless enhances the neural differentiation of DPSCs in a dose-dependent manner. Simultaneously, we found that phosphorylated mTOR was up-regulated, which indicated the involvement of mTOR in the process. Rapamycin, an inhibitor of mTOR activity, can reverse the effect of DPSCs stimulated by IGF-1. Next, we studied the role of mTORC1 and mTORC2, two known mTOR complexes, in the neural differentiation of DPSCs. We found that inhibition of mTORC1 can severely restricts the neural differentiation of DPSCs. However, inhibition of mTORC2 has the opposite effect. This latter effect disappears when both rictor and mTOR are inhibited, showing that the mTORC2 effect is mTORC1 dependent. This study has expanded the role of mTOR in DPSCs neural differentiation regulated by IGF-1.


Assuntos
Diferenciação Celular/genética , Polpa Dentária/enzimologia , Fator de Crescimento Insulin-Like I/genética , Células-Tronco/efeitos dos fármacos , Adolescente , Adulto , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Polpa Dentária/crescimento & desenvolvimento , Feminino , Humanos , Fator de Crescimento Insulin-Like I/farmacologia , Masculino , Alvo Mecanístico do Complexo 1 de Rapamicina/antagonistas & inibidores , Alvo Mecanístico do Complexo 2 de Rapamicina/antagonistas & inibidores , Fosforilação/efeitos dos fármacos , Proteínas Recombinantes/farmacologia , Transdução de Sinais/efeitos dos fármacos , Sirolimo/farmacologia , Serina-Treonina Quinases TOR/antagonistas & inibidores , Serina-Treonina Quinases TOR/genética , Adulto Jovem
15.
J Endod ; 45(7): 882-889, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31133343

RESUMO

INTRODUCTION: This study aimed to compare the cytocompatibility and angiogenic potential of 2 antibiotics (clindamycin [CLIN] and minocycline [MINO]) at distinct concentrations on dental pulp stem cells (DPSCs) and human umbilical vein endothelial cells (HUVECs). METHODS: DPSCs and HUVECs were exposed to cell culture media modified with CLIN or MINO at concentrations ranging from 30 µg/mL-1000 µg/mL. Cell toxicity and proliferation were investigated using the lactate dehydrogenase and tetrazolium reduction assays, respectively. A capillarylike tube formation in vitro assay was conducted to determine the angiogenic potential associated with each antibiotic. Additionally, selected morphometric angiogenesis parameters were determined using dedicated software (WimTube; Onimagin Technologies SCA, Córdoba, Spain). All statistical analyses were performed using 1-way analysis of variance and the Tukey post hoc test (α= .05). RESULTS: The collected data showed that compared with the control (cell culture media, alpha-minimum essential medium Eagle) increasing the antibiotic concentration significantly decreased cell viability and proliferation of both DPSCs and HUVECs. In terms of angiogenic potential, when tested at 30 µg/mL and 50 µg/mL, CLIN significantly amplified tube formation when compared with MINO with angiogenesis parameters (ie, tube length and tube number) similar to the effect promoted by exogenous vascular endothelial growth factor (50 ng/mL). CONCLUSIONS: CLIN was less cytotoxic when compared with MINO at higher concentrations. Of note, CLIN did not hinder the proangiogenic activity induced by vascular endothelial growth factor to the same extent as MINO, suggesting that the replacement of MINO by CLIN might translate into positive implications in the overall regenerative outcome.


Assuntos
Antibacterianos , Clindamicina , Minociclina , Antibacterianos/farmacologia , Antibacterianos/toxicidade , Proliferação de Células , Células Cultivadas , Clindamicina/farmacologia , Clindamicina/toxicidade , Polpa Dentária/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana , Humanos , Minociclina/farmacologia , Minociclina/toxicidade , Neovascularização Fisiológica , Espanha , Fator A de Crescimento do Endotélio Vascular
16.
Mol Immunol ; 111: 11-18, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30952010

RESUMO

Endodontic infection is a widespread oral problem. DNA methylation is a key epigenetic modification that plays important roles in various inflammatory responses, but its role in dental pulp inflammation is poorly understood. In this study, we assessed the expression of DNA methyltransferases (DNMTs) in human dental pulp cells (hDPCs) during lipopolysaccharide (LPS)-induced inflammation and found that DNMT3B mRNA expression was reduced and DNMT1 mRNA and protein levels decreased significantly. Pretreatment with the DNMT inhibitor 5-Aza-2'-deoxycytidine (5-Aza-CdR) significantly enhanced the expression of the inflammatory cytokines IL-6 and IL-8 in LPS-stimulated hDPCs, indicating that DNA methylation may play a role in hDPC inflammation. Studies have reported that some microRNAs (miRNAs) are involved in dental pulp infection. DNA methylation can modulate the inflammatory response by regulating miRNA expression, but this phenomenon has not yet been reported in pulp inflammation. The present study used next-generation sequencing to examine the effect of 5-Aza-CdR on the miRNA expression profile of LPS-treated hDPCs, and the results showed that 5-Aza-CdR pretreatment changed the miRNA expression pattern in hDPCs during inflammation. Among the changed miRNAs, miR-146a-5p, which is a pulp inflammation-related miRNA, demonstrated the most noticeably altered expression. miR-146a-5p could be induced by LPS in hDPCs, and 5-Aza-CdR preincubation or DNMT1 knockdown markedly increased its expression level. However, no significant difference was found in the methylation pattern of the MIR146A promoter with 5-Aza-CdR pretreatment or DNMT1 knockdown in LPS-stimulated hDPCs. These results indicate that DNA methylation may regulate the LPS-induced inflammatory response by changing the miRNA expression in hDPCs.


Assuntos
Metilação de DNA/genética , Polpa Dentária/metabolismo , Inflamação/induzido quimicamente , Inflamação/genética , Lipopolissacarídeos/farmacologia , MicroRNAs/genética , Azacitidina/farmacologia , Células Cultivadas , Citocinas/genética , DNA (Citosina-5-)-Metiltransferase 1/genética , DNA (Citosina-5-)-Metiltransferases/genética , Metilação de DNA/efeitos dos fármacos , Polpa Dentária/efeitos dos fármacos , Epigênese Genética/efeitos dos fármacos , Epigênese Genética/genética , Humanos , Regiões Promotoras Genéticas/efeitos dos fármacos , Regiões Promotoras Genéticas/genética , RNA Mensageiro/genética
17.
J Appl Oral Sci ; 27: e20180442, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30994776

RESUMO

OBJECTIVE: To assess pulp oxygen saturation levels (SaO2) in maxillary central incisors after dental bleaching. MATERIALS AND METHODS: 80 participants (160 teeth) were randomly allocated to four groups: G1 In-office bleaching with two applications of 35% hydrogen peroxide (HP) (20 minutes), followed by at-home bleaching with 10% carbamide peroxide (CP) (2 hours/day for 16 days); G2 - Same protocol as G1, plus desensitizing toothpaste; G3 - In-office bleaching with 35% HP and one application of placebo gel (20 minutes), followed by at-home bleaching with 10% CP (2 hours/day for 16 days); and G4 - Same protocol as G3, plus desensitizing toothpaste. Pulp SaO2 levels were measured before (T0) and immediately after (T1) in-office bleaching; on the 5th (T2), 8th (T3), 12th (T4), and 16th days of at-home bleaching (T5); and on the 7th (T6) and 30th (T7) days. Mean (SD) pulp SaO2 levels were compared within groups by generalized estimating equations (GEE) and Student's t-test (P<0.05). RESULTS: Mean pulp SaO2 at T0 was 84.29% in G1, 84.38% in G2, 84.79% in G3, and 85.83% in G4. At T1, these values decreased to 81.96%, 82.06%, 82.19%, and 81.15% in G1, G2, G3, and G4 respectively, with significant difference in G4 (P<0.05). During home bleaching, pulp SaO2 levels varied in all groups, with 86.55%, 86.60%, 85.71%, and 87.15% means at T7 for G1, G2, G3, and G4, respectively; G2 presented significant difference (P<0.05). CONCLUSIONS: Pulp SaO2 level in maxillary central incisors was similar at baseline, reducing immediately after in-office bleaching, regardless of using desensitizing toothpaste and increasing at 30 days after dental bleaching.


Assuntos
Polpa Dentária/metabolismo , Incisivo/metabolismo , Oxigênio/metabolismo , Clareadores Dentários/efeitos adversos , Clareamento Dental/efeitos adversos , Adolescente , Adulto , Peróxido de Carbamida/efeitos adversos , Polpa Dentária/efeitos dos fármacos , Dessensibilizantes Dentinários/uso terapêutico , Sensibilidade da Dentina/induzido quimicamente , Sensibilidade da Dentina/prevenção & controle , Feminino , Humanos , Peróxido de Hidrogênio/efeitos adversos , Incisivo/efeitos dos fármacos , Masculino , Oximetria/métodos , Valores de Referência , Fatores de Tempo , Clareamento Dental/métodos , Cremes Dentais/uso terapêutico , Resultado do Tratamento , Adulto Jovem
18.
Colloids Surf B Biointerfaces ; 179: 326-333, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-30981068

RESUMO

Carbamide peroxide is the popular home dental whitening agent. However, it has critical stability. Nanoparticles have been applied to develop products with advantages properties as better efficacy and stability increase. The aim of this study was the characterization of carbamide peroxide polymeric nanoparticles, their bleaching efficacy, effects on pulp damage and stability evaluation. Particle size demonstrated a spherical morphology and bimodal distribution (11 and 398 nm). Nanoparticles presented high entrapment efficiency (98.94%) and the zeta potential value was slightly positive (+10.26 mV). Regardless of the zeta potential, the steric effect may contribute to carbamide peroxide nanoparticle stabilization. The stability studies conducted at room temperature suggested that carbamide peroxide nanoparticles could maintain all the parameters evaluated (size, polydispersity index, zeta potential, entrapment efficiency, pH and content) for at least 90 days. Instability index was determined by dispersion analyzer (LUMiSizer ®), was 0.018, and the light transmission profile did not present sedimentation. Carbamide peroxide nanoparticles were able to prevent thermal degradation and photostability. Clinical efficacy of the whitening gels was obtained by color change in the spectrophotometer and the results showed that all the evaluated gels containing the nanoparticles (0, 1, 2 and 5% of real carbamide peroxide) were effective at bleaching after 2 h of home whitening treatment (during 30 days). After the treatment, the extracted teeth showed no in situ pulp damage by histological evaluation. The nanotechnology strategy of converting carbamide peroxide into polymeric nanoparticles revealed a new product with improved stability, a good approach for carbamide peroxide delivery.


Assuntos
Peróxido de Carbamida/farmacologia , Nanopartículas/química , Clareamento Dental , Polpa Dentária/efeitos dos fármacos , Nanopartículas/ultraestrutura , Temperatura Ambiente
19.
Clin Oral Investig ; 23(12): 4289-4299, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30864114

RESUMO

OBJECTIVES: To histologically evaluate the morphology of the newly formed mineralized tissue and of the adjacent cells, in intact human teeth subjected to mechanical pulp exposure and capping with a fast-setting mineral trioxide aggregate (RetroMTA). MATERIALS AND METHODS: Seven caries-free third molars from three adults were subjected to pulp exposure, direct capping with RetroMTA, and restoration with a composite resin. Seven months later, the teeth were clinically and radiographically evaluated, extracted, and subjected to histological processing and evaluation. RESULTS: All teeth were clinically and radiographically inconspicuous and showed no presence of severe inflammatory reactions. Bacteria were absent in all teeth. All cases exhibited some degree of mineralized tissue in the area of exposure to varying extent. This newly formed mineralized tissue was mostly atubular and did not display the features of regular dentine in any of the cases. No cells exhibiting the features of odontoblasts or odontoblast-like cells were observed. Instead, the cells exhibited a flat or cuboidal shape, resembling fibroblasts. CONCLUSIONS: When the exposed pulps were directly capped with RetroMTA, the new calcified hard tissue was not "regular dentine," and did not seem to be the product of genuine odontoblast differentiation. These results suggest that the formation of calcified tissues after direct pulp capping with RetroMTA may be more appropriately regarded as a reparative process than as a genuine regeneration response. CLINICAL RELEVANCE: This is the first histological study on humans showing that regular dentine was not regenerated when a bioactive pulp-capping material (RetroMTA) was placed over exposed pulp tissue. TRIAL REGISTRATION: NCT03631511.


Assuntos
Compostos de Alumínio/uso terapêutico , Compostos de Cálcio/uso terapêutico , Capeamento da Polpa Dentária/métodos , Polpa Dentária/efeitos dos fármacos , Agentes de Capeamento da Polpa Dentária e Pulpectomia/uso terapêutico , Silicatos/uso terapêutico , Adulto , Compostos de Alumínio/química , Compostos de Cálcio/química , Exposição da Polpa Dentária/terapia , Dentina Secundária/efeitos dos fármacos , Dentina Secundária/patologia , Combinação de Medicamentos , Humanos , Óxidos/química , Silicatos/química
20.
Int J Mol Sci ; 20(3)2019 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-30709061

RESUMO

Hyaline cartilage is a tissue of very low regenerative capacity because of its histology and limited nutrient supply. Cell-based therapies have been spotlighted in the regeneration of damaged cartilage. Dental pulp stem cells (DPSCs) are multipotent and are easily accessible for therapeutic purposes. In human gastrointestinal tracts, Enterococcus faecium is a naturally occurring commensal species of lactic acid bacteria. In this work, the human DPSCs were differentiated into chondrocytes using a chondrogenic differentiation medium with or without L-15 extract. We observed that chondrogenic differentiation improved in an E. faecium L-15 extract (L-15)-treated DPSC group via evaluation of chondrogenic-marker mRNA expression levels. In particular, we found that L-15 treatment promoted early-stage DPSC differentiation. Cells treated with L-15 were inhibited at later stages and were less likely to transform into hypertrophic chondrocytes. In L-15-treated groups, the total amount of cartilage extracellular matrix increased during the differentiation process. These results suggest that L-15 promotes chondrogenic differentiation, and that L-15 may be used for cartilage repair or cartilage health supplements. To our knowledge, this is the first report demonstrating the beneficial effect of L-15 treatment on chondrogenic differentiation.


Assuntos
Condrogênese , Meios de Cultura/farmacologia , Polpa Dentária/citologia , Enterococcus faecium/crescimento & desenvolvimento , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular , Sistema Livre de Células , Células Cultivadas , Meios de Cultura/química , Polpa Dentária/efeitos dos fármacos , Enterococcus faecium/metabolismo , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Marcadores Genéticos , Humanos , Células-Tronco/citologia , Células-Tronco/efeitos dos fármacos
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA