ABSTRACT
Abstract Recent studies on functional tissue regeneration have focused on substances that favor cell proliferation and differentiation, including the bioactive phenolic compounds present in grape seed extract (GSE). The aim of this investigation was to evaluate the stimulatory potential of GSE in the functional activity of undifferentiated pulp cells and odontoblast-like cells. OD-21 and MDPC-23 cell lines were cultivated in odontogenic medium until subconfluence, seeded in 24-well culture plates in a concentration of 2x104/well and divided into: 1) OD-21 without GSE; 2) OD-21+10 µg/mL of GSE; 3) MDPC-23 without GSE; 4) MDPC-23+10 µg/mL of GSE. Cell proliferation, in situ detection of alkaline phosphatase (ALP) and total protein content were assessed after 3, 7 and 10 days, and mineralization was evaluated after 14 days. The data were analyzed by ANOVA statistical tests set at a 5% level of significance. Results revealed that cell proliferation increased after 10 days, and protein content, after 7 days of culture in MDPC-23 cells. In situ ALP staining intensity was higher in undifferentiated pulp cells and odontoblast-like cells after 7 and 10 days, respectively. A discrete increase in MDPC-23 mineralization after GSE treatment was observed despite OD-21 cells presenting a decrease in mineralized nodule deposits. Data suggest that GSE favors functional activity of differentiated cells more broadly than undifferentiated cells (OD-21). More studies with different concentrations of GSE must be conducted to confirm its benefits to cells regarding dentin regeneration.
Subject(s)
Animals , Mice , Dental Pulp/cytology , Dental Pulp/drug effects , Cell Proliferation/drug effects , Grape Seed Extract/pharmacology , Odontoblasts/drug effects , Reference Values , Time Factors , Cell Differentiation/drug effects , Cell Line , Cells, Cultured , Reproducibility of Results , Dentin/cytology , Dentin/drug effects , Odontogenesis/drug effectsABSTRACT
Abstract The aim of this study was to evaluate whether medium modification improves the odontogenic differentiation of human dental pulp stem cells (DPSC) in vitro and in vivo. DPSC isolated from human impacted third molar teeth were analysed for clusters of differentiation with flow cytometry. Odontogenic differentiation was stimulated by medium modification with the addition of bone morphogenetic protein 2 (BMP2). The expression of dentin sialophosphoprotein, dentin matrix protein 1, enamelysin/matrix metalloproteinase 20 and the phosphate-regulating gene with homologies to endopeptidases on the X chromosome of the cells were analysed with RT-PCR at 7, 14 and 21 days. Then, DPSC were transplanted on the back of immunocompromised mice via a hydroxyapatite tricalcium phosphate scaffold, and the structure of the formed tissue was investigated. The cells were identified as mesenchymal stem cells with a 98.3% CD73 and CD90 double-positive cell rate. The increase in mineralization capacity and expression of human enamel-dentin specific transcripts proportional to the culture period were determined after differentiation. Six weeks after transplantation, an osteo-dentin matrix was formed in the group in which odontogenic differentiation was stimulated, and the odontogenic characteristics of the matrix were confirmed by histological examination and RT-PCR analysis. Odontogenic differentiation of the isolated and characterized human DPSC was improved with medium modification by the addition of BMP2 in vitro and in vivo. The defined medium and applied technique have a potential use for forming reparative dentin in the future, but the effects of the method should be investigated in long-term studies.
Subject(s)
Humans , Animals , Adult , Mice , Young Adult , Stem Cells/cytology , Cell Differentiation/drug effects , Culture Media/chemistry , Dental Pulp/cytology , Bone Morphogenetic Protein 2/pharmacology , Phosphoproteins/analysis , Sialoglycoproteins/analysis , Time Factors , Cell Differentiation/physiology , Cells, Cultured , Reproducibility of Results , Extracellular Matrix Proteins/analysis , Actins/analysis , Reverse Transcriptase Polymerase Chain Reaction , Stem Cell Transplantation/methods , Cell Proliferation/drug effects , Cell Proliferation/physiology , Matrix Metalloproteinase 20/analysis , PHEX Phosphate Regulating Neutral Endopeptidase/analysis , Bone Morphogenetic Protein 2/chemistry , Flow Cytometry , Odontogenesis/drug effects , Odontogenesis/physiologyABSTRACT
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.
Subject(s)
Animals , Male , Female , Pregnancy , Rats , Fungicides, Industrial/toxicity , Odontogenesis/drug effects , Zineb/analogs & derivatives , Dental Enamel/drug effects , Dentin/drug effects , Immunohistochemistry , Matrix Metalloproteinase 2/drug effects , Matrix Metalloproteinase 2/metabolism , Tooth Germ/drug effects , Vascular Endothelial Growth Factor A/drug effects , Vascular Endothelial Growth Factor A/metabolism , Zineb/toxicityABSTRACT
Se hace un estudio de los efectos del consumo prolongado de etanol sobre las etapas tempranas del desarrollo dentario en ratones, se emplean para ello 6 ratonas albinas preñadas, divididas en un grupo control de 2 ratonas y un grupo experimental de 4 animales. Los 2 grupos se ubicaron en jaulas diferentes, disponían de agua y comida a voluntad. En el grupo experimental el auga contenía el 25 % de alcohol. Todos los animales fueron sacrificados a los 14 días de preñez se les extrajeron los fetos y se obtuvieron fragmentos de maxilar y mandíbula, que fueron procesados por la técnica de parafina y teñidos con HE.Los resultados corroboran la presencia de un alto número de reabsorciones embrionarias, así como se detectan anomalías visibles morfológicamente en la capa basaldel brote y los epitelios adamantinos del casquete. Las alteraciones más relevantes se presentan en el epitelio adamantino interno