TUNING FOR THE DETERMINATION OF RANKL AND OPG IN TEETH OF CAT BY IMMUNOCYTOCHEMISTRY

SACCOMANO, DM.; GAUNA AÑASCO, LG.; NEGRO, VB.; BOVIEZ, JD.; LOMBARDO, DM.; HERNÁNDEZ, SZ.

Tandil. Buenos Aires

Jornada; Primeras Jornadas de Ciencias Morfológicas de las Sierras de Tandil. Universidad del Centro de la Provincia de Buenos Aires (UNCPBA).; 2011

Universidad Nacional del Centro de la Provincia de Buenos Aires.

Several studies have shown that severe periodontal disease (PD) results in dental resorption in human and animal teeth. Although how these resorptions occur are not entirely clear, various signals exist at the molecular level that would trigger the responsible cells for dental destruction (osteoclasts / odontoclasts). Nuclear Receptor Activator Complex K (RANK), its ligand (RANKL) and osteoprotegerin (OPG) regulate the activation of cells that induce the formation or resorption of dental tissue, according to the stimulation they receive. No previous studies indicated that available antibodies could be used in teeth of cat, for that the aim of this paper is to report the development of the technique of immunocytochemistry (ICQ) for determination of RANKL and OPG in normal teeth and teeth with periodontal disease of cat. Two normal teeth and 5 with severe EP from 3 cats were analyzed. Teeth were fixed in buffered formaldehyde for at least 48 hours, decalcified with 10% formic acid and processed with routine technique (H/E). For ICQ rabbit anti goat secondary antibody (Biotin), rabbit anti goat ,primary OPG (N-20) and RANKL (N-19) primary polyclonal antibodies (Santa Cruz Biotechnology®, Inc) (1/100), Dako Streptavidin were used and revealed with DAB. Bone marrow (for RANKL) and rat small gut (for OPG) were used as positive control. Reaction was positive in the area of the periodontium. This study shows that these primary antibodies could be used in cat dental tissues with positive results.