Aislamiento y cultivo de células osteoblásticas: Interés para la investigación en cirugía ortopédica y traumatología

Describimos un método de aislamiento y caracterización de células osteoblásticas procedentes de la superficie de trabéculas óseas de explantes obtenidos de metafisis de fémures y tibias de conejo y de cabezas femorales humanas de pacientes sometidos a cirugía protésica de cadera. Después de eliminar las células de la médula ósea de los explantes, estos se despositaron sobre dos tipos de mallas, de 80 y 100 µm., obteniéndose un cultivo en monocarpa que se mostró confluente a partir de los 20 días. La población celular aislada presentó un fenotipo osteoblástico utilizando métodos colorimétricos para determinación de la actividad de fosfatasa alcalina (FA) y morfológicos por medio de la observación con microscopía electrónica de barrido. Las células cultivadas exhibieron una morfología diferente según el grado de confluencia de cultivo, siendo en todo momento positivas las sucesivas determinaciones colorimétricas de FA realizadas a lo largo de la experiencia. Consideramos el método de gran interés en investigación en ortopédica, pues permite el estudio de comportamiento de estas células cuando están sometidas a cualquier tipo de estímulo externo, así como cuando se enfrentan a nuevas generaciones de biomateriales, etc.We describe a method of isolation and characterization of osteoblastic cells taken from the surface of bone trabeculae of explants obtained from femur and tibia metaphysis in rabbits and human femoral head in patients undergoing orthopaedic surgery. Afther removing the cells from the bone marrow of explants, they were placed over two mesh, 80 and 100 µmn, obtaining then a culture in monolayer which showed to be confluent after 20 days. The isolated cell population showed an osteoblastic phenotype using colourmetric methods to determinate the alkaline phosphatase (AF) activity and morphologic methods by means of scanning. The cultivated cell showed a different morphology according to the degree of confluence of the culture, being always positive the sucesive colourmetric determinations of AF along the experiment. We consider the method of a great interest for orthopaedic investigations, since it allows the study of the behaviour of these cells when they are subjet to different types of external stimulus as well as meet they the new generations of biomaterials

The antioxidant effect of a diet rich in Maillard reaction products is attenuated after consumption by healthy male adolescents. In vitro and in vivo comparative study

Presented in part at the COST Action 927-IMARS Congress, Naples, Italy, May 2006.[Background]: Maillard reaction products (MRPs) are widely consumed as a part of the human diet. A 2 week randomised two-period crossover trial to determine whether MRP intake affects the antioxidant defence system in male adolescents (11–14 years, n = 18) was carried out using two diets rich and poor in MRPs (brown diet, BD, and white diet, WD, respectively). Fasting blood samples were collected after the dietary intervention periods to measure oxidative status. The in vitro antioxidant activity of the diets was also assessed.[Results]: The BD had stronger in vitro antioxidant activity to scavenge free radicals and greater ability to reduce lipid peroxidation. However, in the in vivo assay, markers of oxidative damage (serum thiobarbituric acid-reactive substances and erythrocyte hydroperoxides) and antioxidant defence parameters (serum antioxidants and enzymatic activities of catalase, superoxide dismutase and glutathione peroxidase) were unchanged after the dietary treatments. Only treatment of biological samples with external oxidants revealed higher values of the antioxidant capacity after consumption of the MRP-rich diet.[Conclusion]: In spite of the higher antioxidant activity of the BD shown in vitro, consumption of diets rich in MRPs does not seem to modify oxidative status in healthy male adolescents. However, a protective effect against induced oxidation was shown.This work was supported by a project of the Spanish MEC.Peer reviewe