BULL SPERM SELECTION BY ATTACHMENT TO HYALURONIC ACID SEMI-INTERPENETRATED HYDROGLES

BLOIS D; A C. LIAUDAT; CAPELLA V; MORILLA G; RIVERO R; RIVAROLA C; BARBERO C; RODRIGUEZ N; BOSCH P

Congreso; Reunión Anual SAIC SAFE SAB SAP 2019; 2019

BULL SPERM SELECTION BY ATTACHMENT TO HYALURONIC ACID SEMI-INTERPENETRATED HYDROGLESBlois DA1, Liaudat AC1, Capella V1, Morilla G1, Rivero R2, Rivarola C2, Barbero C2, Rodríguez N1, Bosch P1.1 Instituto de Biotecnología Ambiental y Salud (INBIAS) y 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).Hyaluronic acid (HA) is a glycosaminoglycan present in oocyte-cumulus cell complexes. A population of motile mature spermatozoa with low incidence of genetic alterations preferentially interact with HA molecules. The aim of this work was to develop polymeric biomaterials containing HA for selection of bull spermatozoa based on their functional characteristics. Polymeric hydrogels composed of poli N-isopropylacrylamide-co-20% N-Tris (hydroxymethyl) methyl acrylamide semi-interpenetrated with HA (PNIPAM-HMA-HA) were synthesized and physical chemically characterized. The interaction and degree of frozen-thawed sperm/hydrogel surface binding was analyzed by phase contrast microscopy. Motility, viability, nuclear morphology, acrosome membrane integrity and plasma membrane functionality of attached/released and non-attached sperm populations were studied. Surfaces of hydrogels interpenetrated with HA were more hydrophilic according to angle of contact study; however presence of HA into the polymeric network was not associated with a change of the volume phase transition temperature. In addition, PNIPAM-HMA-HA hydrogels had higher swelling capacity than PNIPAM-HMA without HA. Fifty percent of frozen-thawed bull spermatozoa attached to PNIPAM-HMA-HA hydrogels and the 47% of them were released upon treatment with medium containing hyaluronidase. Selected sperm have acceptable characteristics of rectilinear motility (70% ± 2.58 %), high viability (58.7 ± 11.7 %), nuclear and cellular morphology and low percentage of acrosome reacted spermatozoa (23.3 ± 4.1 %). The plasma membrane integrity of the released population remained unchanged compared to initial sample. In conclusion, our results indicate that polymeric hydrogels semi-interpenetrated with HA could be useful to select high-quality sperm for use in assisted reproduction techniques.This work was supported by grants from UNRC and MinCyT, Córdoba