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 ï¬broblast 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 fármacosRESUMO
Abstract 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
Humanos , Materiais Restauradores do Canal Radicular/farmacologia , Células-Tronco/efeitos dos fármacos , Cerâmica/farmacologia , Silicatos/farmacologia , Compostos de Cálcio/farmacologia , Indutores da Angiogênese/farmacologia , Ligamento Periodontal/citologia , Ligamento Periodontal/efeitos dos fármacos , Germe de Dente/citologia , Germe de Dente/efeitos dos fármacos , Materiais Biocompatíveis/farmacologia , Teste de Materiais , Fator de Crescimento Derivado de Plaquetas/análise , Fator de Crescimento Derivado de Plaquetas/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Sobrevivência Celular/efeitos dos fármacos , Reprodutibilidade dos Testes , Fator 2 de Crescimento de Fibroblastos/análise , Fator 2 de Crescimento de Fibroblastos/efeitos dos fármacos , Estatísticas não Paramétricas , Neovascularização Fisiológica/efeitos dos fármacos , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/análise , Fator A de Crescimento do Endotélio Vascular/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Citometria de FluxoRESUMO
Dithiocarbamate propinebs are organometal fungicides that are widely used for the control of diseases in plants. In this study, pregnant female rats received 400 ppm propineb concentrations in 5 ml distilled water for 16 days of gestation, and then infant rats were obtained by cesarean section. In the histological analysis on the frontal sections, the use of propineb was found effective on odontoblast cell hyperplasia, cell infiltration in the dental papilla, and degeneration in the mesenchymal cells of the outer enamel. The expression of MMP-2 (Matrix Metalloproteinase-2) and VEGF (Endothelial cell growth factor) in the connective tissue was evaluated by immunohistochemistry. The drinking water given to the mothers in propineb tooth bud, enamel and dentin, resulted in morphological changes suggestive of a delay in formation, which cross the placental barrier and possibly affect the tooth development.
Los ditiocarbamatos (Propineb) son fungicidas organometálicos que son ampliamente utilizados para el control de enfermedades en las plantas. En este estudio, ratas hembras preñadas recibieron concentraciones de 4000 ppm de propineb en 5 ml de agua destilada durante 16 días de su gestación. Luego, las crías de las ratas fueron obtenidas mediante cesárea para su estudio estudio histológico. En el análisis histológico de las secciones frontales, el uso de propineb fue positivo para la hiperplasia de las células odontoblástica, infiltración de células en la papila dental, y la degeneración en las células mesenquimales del epitelio externo del esmalte. La expresión de MMP-2 (metaloproteinasa de la matriz 2) y VEGF (factor de crecimiento de células endoteliales) en el tejido conectivo se evaluó por inmunohistoquímica. El agua potable con propineb dada a las madres actuó sobre el brote dentario, esmalte y dentina; se tradujo en cambios morfológicos indicativos de un retraso en la formación. Por tanto, el propineb atraviesa la barrera placentaria y posiblemente afecten el desarrollo de los dientes.
Assuntos
Animais , Masculino , Feminino , Gravidez , Ratos , Fungicidas Industriais/toxicidade , Odontogênese/efeitos dos fármacos , Zineb/análogos & derivados , Esmalte Dentário/efeitos dos fármacos , Dentina/efeitos dos fármacos , Imuno-Histoquímica , Metaloproteinase 2 da Matriz/efeitos dos fármacos , Metaloproteinase 2 da Matriz/metabolismo , Germe de Dente/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/metabolismo , Zineb/toxicidadeRESUMO
Excessive fluoride ingestion has been identified as a risk factor for fluorosis and oxidative stress. The oxidative stress results from the loss of equilibrium between oxidative and antioxidative mechanisms that can produce kinase activation, mitochondrial disturbance and DNA fragmentation, resulting in apoptosis. Actually many people are exposed to no-adverted fluoride consumption in acute or chronic way. The aim of this study was to determine the effect of sodium fluoride on first molar germ in relation to its effect on antioxidative enzymes immunoexpression and apoptosis. Thirty first molar germs from 1-day-old Balb/c mice were cultured for 24 h with sodium fluoride (0 mM, 1 mM and 5 mM). Immunoexpression determination of CuZnSod, MnSod, catalase, Bax, Bid, caspase 8, caspase 9, caspase 3 and TUNEL assay were performed. Cellular disorganization in ameloblast and odontoblast-papilla zones was observed. CuZnSod and MnSod immunoexpression decrease in experimental groups. Caspase 8, caspase 3, Bax, Bid increase expression and more TUNEL positive cells in both experimental groups than control, suggest that apoptosis induced by fluoride is related to oxidative stress due to reduction of the enzymatic antioxidant.
Assuntos
Apoptose , Cariostáticos/toxicidade , Odontogênese/efeitos dos fármacos , Estresse Oxidativo , Fluoreto de Sódio/toxicidade , Germe de Dente/efeitos dos fármacos , Ameloblastos/metabolismo , Animais , Apoptose/fisiologia , Proteína Agonista de Morte Celular de Domínio Interatuante com BH3/biossíntese , Caspases/biossíntese , Catalase/biossíntese , Marcação In Situ das Extremidades Cortadas , Camundongos , Camundongos Endogâmicos BALB C , Estresse Oxidativo/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/biossíntese , Germe de Dente/enzimologia , Proteína X Associada a bcl-2/biossínteseRESUMO
OBJECTIVE: A polyclonal antibody was used to investigate the effects of ethanol ingestion before and during pregnancy, in the expression of EGF on dentinogenesis and amelogenesis of rat mandibular first molar. DESIGN: Ethanol was administered to drinking water (treated group) starting at concentrations of 1% and increasing weekly to 5, 10, 15, 20 and 25% (v/v). During week 7, these rats were mated and continued to receive the 25% alcoholic solution, up to delivery. The control group received tap water. On postnatal days 0, 4 and 9, two offspring of each litter were killed, their hemimandibles removed and prepared for paraffin processing and immunohistochemistry. RESULTS: At postnatal day 0 the EGF immunoreactivity of the inner enamel epithelium and presecretory ameloblasts was weak when compared to controls. At postnatal day 4 EGF immunoreactivity of the secretory ameloblasts and odontoblasts was only moderate compared to controls. At postnatal day 9 EGF staining of the ameloblasts was weak when compared to controls. CONCLUSIONS: These results suggest that, maternal alcoholism interferes with EGF expression during initial dentinogenesis and amelogenesis and in the secretion and maturation of the dentin and enamel, therefore, which may cause a reduction of dentin and enamel formation.
Assuntos
Fator de Crescimento Epidérmico/efeitos dos fármacos , Etanol/farmacologia , Transtornos do Espectro Alcoólico Fetal/metabolismo , Dente Molar/embriologia , Amelogênese/efeitos dos fármacos , Animais , Biometria , Dentinogênese/efeitos dos fármacos , Fator de Crescimento Epidérmico/metabolismo , Feminino , Transtornos do Espectro Alcoólico Fetal/fisiopatologia , Técnicas Imunoenzimáticas , Troca Materno-Fetal , Dente Molar/efeitos dos fármacos , Dente Molar/metabolismo , Gravidez , Ratos , Ratos Wistar , Germe de Dente/efeitos dos fármacos , Germe de Dente/embriologia , Germe de Dente/patologiaRESUMO
Vitamin A and its derivatives, retinoic acid, tretinoin and isotretinoin, are currently used in dermatological treatments. The administration of high doses of this vitamin provokes congenital malformations in mice: cleft palate, maxillary and mandibular hypoplasia and total or partial fusion of the maxillary incisors. This study compares the tooth germs of the first maxillary and mandibular molars of fetal mice submitted to isotretinoin during organogenesis. Twelve 60-day-old female Mus musculus were divided into two groups on the 7th day of pregnancy: treated group--1 mg isotretinoin per kg body weight, dissolved in vegetable oil, was administered from the 7th to the 13th day of pregnancy; control group--vegetable oil in equivalent volume was administered orally for the same period. On the 16th day of pregnancy, the females were sacrificed, the fetuses were removed and their heads amputated. After standard laboratory procedures, 6-micron thick serial slices were stained with hematoxylin and eosin for optical microscopy examination. The results showed that both groups had closed palates with no reminiscence of epithelial cells; however, the first molar germs of the isotretinoin-treated animals showed delayed development compared to the control animals.
Assuntos
Isotretinoína/toxicidade , Palato/efeitos dos fármacos , Palato/embriologia , Germe de Dente/efeitos dos fármacos , Germe de Dente/embriologia , Animais , Desenvolvimento Embrionário e Fetal/efeitos dos fármacos , Feminino , Camundongos , Dente Molar/efeitos dos fármacos , Dente Molar/embriologia , GravidezRESUMO
Utilizando a escala de cores Vita, dois avaliadores tomaram a cor do incisivo central, incisivo lateral e canino de 242 pacientes, sob iluminaçäo natural. Além da cor desses dentes, observou-se a existência ou näo da transparência incisal. Eliminando-se os pacientes cujos dentes apresentavam anomalias de cor, todos os dentes tinham uma cor correspondente no guia de cores da escala Vita. Houve dúvida entre os avaliadores apenas nas cores de baixa saturaçäo. Os dados obtidos na nossa amostra permitem as seguintes conclusöes: a) existe um padräo de matiz para os dentes do mesmo paciente: o incisivo central tem, na maioria das vezes, a mesma cor do incisivo lateral; o canino possui uma cor mais saturada. b) a cor amarela é a mais freqüente; com a idade, os dentes tornam-se mais saturados. c) a transparência incisal prevalece no incisivo central e no incisivo lateral, ao contrário do que acontece com o canino, em que prevalece a ausência de transparência incisal; com a idade, diminui a incidência da transparência incisal. d) na cor cinza a ausência de transparência incisal prevalece em todas as idades
Assuntos
Cor/normas , Estética Dentária , Germe de Dente/anatomia & histologia , Germe de Dente/efeitos dos fármacos , Dente Canino/anatomia & histologia , Incisivo/anatomia & histologiaRESUMO
Camundongos fêmeas prenhes foram injetadas no 14§ dia de gestaçäo com 200 µg/kg de peso corporal de ivermectina. Os filhotes foram sacrificados 1, 5 e 10 dias após o nascimento, e as cabeças processadas histologicamente para análise do palato e dos germes dentais. Os resultados näo mostraram diferenças no desenvolvimento, pois o palato estava fechado e os germes dentais dos animais tratados eram semelhantes aos dos animais controle. Desse modo, nas condiçöes experimentais deste trabalho, a ivermectina näo se mostrou tóxica às células, permitindo um desenvolvimento das estruturas analisadas com características compatíveis com as encontradas nos animais controle e com as descritas na literatura
Assuntos
Animais , Camundongos , Palato/efeitos dos fármacos , Germe de Dente/crescimento & desenvolvimento , Germe de Dente/efeitos dos fármacos , Ivermectina/farmacologia , PrenhezRESUMO
Literature reports appeared in the last years suggested that some type of dentigerous cyst (DC) in children would initiate by teh action of other factors than developmental and at different stages of dental sac growth. Periodontitis and pulp therapy with formocresol (FC) have been suggested as aeteological factors, both promoting DC in children by irritation of the underlaying dental sacs of premolar. The purpose of this study is to analyse the pathological changes in DC of patients aged 0 to 15 years, and to compare the findings according to the anatomical site of DC and the existence of previous FC therapy. Morphological changes observed in the epithelium and the connective wall did not show clear differences between both groups of DC. The histometric measurements of epithelial changes did not show significant statistical differences of various parameters in the DC walls. These facts would not support the hypotesis of DC initiation from FC effects.
Assuntos
Cisto Dentígero/induzido quimicamente , Formocresóis/efeitos adversos , Adolescente , Dente Pré-Molar , Criança , Pré-Escolar , Cisto Dentígero/patologia , Humanos , Lactente , Dente Molar , Pulpotomia/efeitos adversos , Germe de Dente/efeitos dos fármacos , Dente DecíduoRESUMO
Thirty six female mice were injected on the 12th day of the gestational period with 0.2 ml of distilled water (control group) or of an acqueous solution containing either 30 mg/Kg or 50 mg/Kg of body weight of cyclophosphamide (treated group). The animal were killed at 24, 48, 72, 96 and 120 hours after the injection and 3 days after birth. It was verified that cyclophosphamide interferes on the tooth germ development and that this effect is in directly ratio of the doses used.