Acta bioquímica clínica latinoamericana
versión ISSN 0325-2957
SALAS, María Mercedes; DE LA CASA, María Luisa y LOPEZ, María Elena. Organic content of human and bovine dental pulp partially purified extracts. Acta bioquím. clín. latinoam. [online]. 2011, vol.45, n.2, pp. 297-304. ISSN 0325-2957.
The dental pulp contains connective tissue and is rich in proteins, mainly collagen. The aim of this study was to describe a purification method for organic compounds of human and bovine pulp tissues, either vital as necrotic, in order to validate the experimental bovine model. Extracts of 50 mg of the middle fraction of one root pulp teeth, 20 humans or 1 bovine, in 500 µL of cold 50 mM Tris - HCl pH 7.4, 1 mM phenyl methyl sulfonyl fluoride and 5 mM benzamidine hydrochloride were prepared. Extracts were homogenized and centrifuged at 10000 rpm. Supernatants were partially purified with 1% protamine sulphate, sediments re-suspended and dialyzed during 2 hours with a spare in 500 mM sodium acetate pH 6. Proteins, hydroxiproline and carbohydrates were determined, and the yield of the process was calculated according to the hydroxiproline content. Poliacrylamide gel electrophoresis was applied. A greater organic content was found in human than in bovine tissue, and in vital rather than in the necrotic one. Hydroxyproline/ proteins and yield evidenced that bovine extract would contain a larger proportion of collagen. Bands at 78 and 80 kDa in human pulp and at 74 and 76 kDa in the bovine tissue were observed. There was no difference between vital and necrotic tissues. The qualitative and quantitative evidences expressed in this work would validate the bovine research endodontic model.
Palabras llave : Dental human pulp; Dental bovine pulp; Proteins; Hydroxyproline; Carbohydrates.