RECEPTORES P2 MODULAN LA MIGRACIÓN Y DIFERENCIACIÓN CELULAR EN CULTIVOS PRIMARIOS DE OSTEOBLASTOS DE CALVARIA DE RATA NEONATA

AYALA PEÑA V.B.; LAIUPPA J.; SANTILLAN G.E.

buenos aires

Congreso; XXIX Reunión anual de la Asociación Argentina de Osteología y Metabolismo Mineral (AAOMM).; 2012

Bone cells are exposed to mechanical stimuli and injury and lysis during movements and trauma respectively. This favors ATP and UTP release to extracellular environment. Nucleotides can trigger different responses acting through plasma membrane P2 receptors (P2X ionotropic, and P2Y metabotropic). We have previously reported that ATP stimulates alkaline phosphatase activity in OBC after 7 days of treatment in osteogenic medium. In this work, we studied the participation of P2 receptors in the modulation of cell migration and differentiation. Cell migration was evaluated by ?wound? assay. Differentiation into osteoblast was analyzed by detection of mineralized nodules after Alizarin red staining, and Bone Sioaloprotein (BSP) gene expression by qRT-PCR. Cells were incubated with 1-100 μM of several nucleotides (UTP, UDP, ADP, ATPγS or ADPβS) for 1-22 days. Treatment of OBC with 10 M ADPβS seems to stimulate cell migration, whereas 100 M ATPS appears to show an inhibitory effect, respect to controls. OBC treated with 100 M ATP or UTP in osteogenic medium showed morphological changes from 7th day of treatment, these changes correlate with an increased BSP expression. In addition, under same conditions, we observed an increased calcium deposition around 22nd day, respect to controls. Our results suggest an important role for P2 receptors in bone cell migration and maturation. Extracellular nucleotides signaling would be involved in the regulation of cellular migration through P2Y1,12 y 13 receptors (responsive to ADP). Moreover, P2Y2 and/or P2Y4 receptors (ATP and UTP responsive) would regulate genes expression involved in osteoblast maturation and function.