EFFECT OF MELATONIN-TREATED TROPHOBLAST EXTRACELLULAR VESICLES ON PLACENTAL ENDOTHELIAL CELL MIGRATION

SZPILBARG N.; REPPETTI J.; SAR J.; DAMIANO A. E.; MARTINEZ N.

Congreso; REUNION ANUAL DE LA SOCIEDAD ARGENTINA DE FISIOLOGIA; 2023

EFFECT OF MELATONIN-TREATED TROPHOBLAST EXTRACELLULAR VESICLES ON PLACENTAL ENDOTHELIAL CELL MIGRATIONSzpilbarg N1, Reppetti J1, Sar J2, Damiano A1,3, Martínez N11.Laboratorio de Biología de la Reproducción. IFIBIO-Houssay (UBA-CONICET). Facultad de Medicina, Universidad de Buenos Aires. Buenos Aires, Argentina.2.Servicio de Obstetricia, Hospital Naval Cirujano Mayor Dr. Pedro Mallo, Ciudad de Buenos Aires, Argentina. 3.Cátedra de Biología Celular y Molecular. Departamento de Ciencias Biológicas, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina.Introduction:Placental angiogenesis is indispensable for successful gestation and it is regulated by trophoblast paracrine signaling. In this sense, placental extracellular vesicles (EVs) have been recognized as a major mediator of feto-maternal communication and have been involved in placental angiogenesis regulation. Trophoblast cells synthesize large amounts of melatonin and express its receptors. Melatonin acts in an autocrine and paracrine manner in this organ, prevents the injury produced by oxidative stress and regulates the expression of vascular endothelial growth factor (VEGF).Here, we investigated whether treatment of trophoblast with melatonin can change the content of released placental EVs and modify the biological functions of placental endothelial cells.Materials and methods:This study was approved by the Ethics Committee of Naval Hospital. Placentas were obtained from healthy-term pregnancies (n=3). Placental explants were cultured with and without melatonin (1-20 μM). Tissue viability was evaluated by the MTT assay. Placental EVs were isolated by differential centrifugation, filtration, and ultracentrifugation. EVs were characterized by DLS, NTA, TEM, and western blot for CD63 and HSP70. The EVs-VEGF expression was evaluated by western blot. EA.hy926 placental endothelial cells were treated with placental EVs for 24h and cell migration was assessed by wound healing assays.Results:The different concentrations of melatonin assayed did not alter trophoblast cell viability. VEGF expression was significantly decreased in the EVs cultured in the presence of melatonin (p