Ability of cationic and neutral hydrogels based on N-isopropylacrylamide to bind equine spermatozoa

EBEL BARRERA F; A C. LIAUDAT; RODRIGUEZ N; RIVAROLA C; BOSCH P

Congreso; Reunión Anual SAIC SAFE SAB SAP 2019; 2019

Ability of cationic and neutral hydrogels based on N-isopropylacrylamide to bind equine spermatozoaEbel F1, Liaudat AC1, Rodríguez N1, Rivarola C2, Bosch P11 Instituto de Biotecnología Ambiental y Salud (INBIAS) and 2 Instituto de Investigaciones en Tecnologías Energéticas y Materiales Avanzados (IITEMA), Universidad Nacional de Río Cuarto (UNRC) ? Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).PNIPAm (poli(N-isopropylacrylamide) based hydrogels are biocompatible materials extensively used in biomedicine. The aim of this study was to evaluate the ability of PNIPAm hydrogels to attach stallion spermatozoa. PNIPAm was co-polymerized with cationic 3-(acrylamido propyl) trimethylammonium chloride (APTA) or neutral N-[Tris (hydroxymethyl) methyl] acrylamide (HMA) monomers. Studied copolymers contained 5%APTA, 10%APTA, 15%APTA, 20%HMA and 20%HMA semi-interpenetrated with hyaluronic acid (20%HMA-HA, 1mg/mL). Each hydrogel was placed in a culture dish containing TALP medium and fresh stallion sperm suspension was added (1x106). The percentage of sperm attached to the surfaces was determined from the difference between the number of sperm initially added into the culture dish and the recovered non-bound cells after incubation (30 min at 37°C in 5% CO2). Sperm that were exposed to -20°C for 60 min to render them non-viable were incubated with 15%APTA and 20%HMA hydrogels-containing dishes, which served as negative controls. Data was analyzed by one way ANOVA; a P0.05). Few non-viable spermatozoa were observed attached to hydrogels (20%HMA: 16.1±8.3%; 15%APTA: 15.3±4.8%). In conclusion, cationic PNIPAm hydrogels could be an efficient support and binding substrate to adhere viable equine sperm, since stallion spermatozoa attaches more efficiently on them. It is expected that this strategy will allow the development of a technique for sperm selection, which isolates the cell subpopulation with high structural and functional quality characteristics, improving the efficiency of assisted reproduction techniques in horses.This work was supported by grants from UNRC and MinCyT, Córdoba