RESUMO
Se describe el proceso de aislamiento y posterior cultivo de los odontoblastos humanos, identificándolos por sus características fenotípicas y funcionales, una vez identificados se procede la congelación de los mismos en nitrógeno líquido. Se preparan unos discos de cristal sobre los que se pincelan tres adhesivos dentinarios sobre los que se vierte una suspensión de odontoblastos en el medio de cultivo apropiado y se procede a su cultivo. Se estudian las características funcionales de las células en cuanto a síntesis de DNA, actividad de la fosfatasa alcalina, síntesis de colágeno y formación de matriz (AU)
This is a description of the isolation and cultivation of human odontoblasts, and their identification via phenotypic and functional characteristics. Once they have been identified they are frozen in liquid nitrogen. Glass discs are then prepared and three dentin adhesives are applied with a brush, and then a suspension of odontoblasts in a suitable cultivation is poured on and the culture is prepared. The functional characteristics of the cells are studied in terms of DNA synthesis, alkaline phosphate activity, collagen synthesis and matrix formation (AU)
Assuntos
Humanos , Odontoblastos/classificação , Retenção de Dentadura/métodos , Colagem Dentária , Odontoblastos/microbiologia , Colágeno/química , Fosfatase Alcalina/análise , Fosfatase Alcalina/síntese química , Espectrofotometria/classificação , Espectrofotometria/instrumentação , Timidina/uso terapêuticoRESUMO
Odontoblast-like cells derived from human tooth pulps were maintained in explant culture and grown either on glass coverslips only (used as control) or on glass coverslips coated with cyanoacrylate films. Ultrastructural and cyto-morphometric evidence showed that cells exposed to cyanoacrylate, in contrast to controls, display a significant decrease of rough endoplasmic reticulum and mitochondria. In addition, immunofluorescent staining and radioimmunoassays for type-I collagen suggested disturbances in production for the exposed cells. The use of anti-fibronectin antibodies with electron-microscopic immunoperoxidase-labelling demonstrated that the adherence of cells to cyanoacrylate can involve both adhesion plaques and fibronectin. These results therefore suggest that there were no apparent differences in the adhesion interaction of cells between glass and cyanoacrylate substrates.
