REAC technology as optimizer of stallion spermatozoa liquid storage.
BACKGROUND
REAC technology (acronym for Radio Electric Asymmetric Conveyor) is a technology platform for neuro and bio modulation. It has already proven to optimize the ions fluxes at the molecular level and the molecular mechanisms driving cellular asymmetry and polarization.


METHODS
This study was designed to verify whether this technology could extend spermatozoa life-span during liquid storage, while preserving their functions, DNA integrity and oxidative status. At 0, 24, 48, and 72 h. of storage at 4 °C, a battery of analyses was performed to assess spermatozoa viability, motility parameters, acrosome status, and DNA integrity during REAC treatment. Spermatozoa oxidative status was assessed by determining lipid peroxidation, the activity of superoxide dismutase (SOD), and the total antioxidant capacity.


RESULTS
During liquid storage REAC treated spermatozoa, while not showing an increased viability nor motility compared to untreated ones, had a higher acrosome (p > 0.001) and DNA integrity (p > 0.01). Moreover, the analysis of the oxidative status indicated that the mean activity of the intracellular superoxide dismutase (SOD) was significantly higher in REAC treated spermatozoa compared to untreated controls (p < 0.05), while the intracellular concentration of malondialdehyde (MDA), an end product of lipid peroxidation, at the end of the REAC treatment was higher in untreated controls (p > 0.05). The REAC efficacy on spermatozoa oxidative status was also evidenced by the higher trolox equivalent antioxidant capacity (TEAC) found in both the cellular extract (p < 0.05) and the storage media of REAC treated spermatozoa compared to untreated controls (p < 0.0001).


CONCLUSIONS
The present study demonstrated that REAC treatment during liquid storage preserves spermatozoa acrosome membrane and DNA integrity, likely due to the enhancement of sperm antioxidant defenses. These results open new perspective about the extending of spermatozoa functions in vitro and the clinical management of male infertility.
Department of Veterinary Medicine, University of Sassari, Viale Vienna 43/B, 07100, Sassari, Italy.
Fiammetta
F
Berlinguer
Department of Veterinary Medicine, University of Sassari, Viale Vienna 43/B, 07100, Sassari, Italy.
Valeria
V
Pasciu
Department of Veterinary Medicine, University of Sassari, Viale Vienna 43/B, 07100, Sassari, Italy.
Sara
S
Succu
AGRIS, Department of Research for Equine Reproduction, Ozieri, Sassari, Italy.
Ignazio
I
Cossu
Department of Veterinary Medicine, University of Sassari, Viale Vienna 43/B, 07100, Sassari, Italy.
Sabrina
S
Caggiu
Department of Veterinary Medicine, University of Sassari, Viale Vienna 43/B, 07100, Sassari, Italy.
Daniela
D
Addis
Departments of Regenerative Medicine, Rinaldi Fontani Institute, Viale Belfiore 43, 50144, Florence, Italy.
Alessandro
A
Castagna
Departments of Regenerative Medicine, Rinaldi Fontani Institute, Viale Belfiore 43, 50144, Florence, Italy.|Research Department, Rinaldi Fontani Foundation, Viale Belfiore 43, 50144, Florence, Italy.
Vania
V
Fontani
Departments of Regenerative Medicine, Rinaldi Fontani Institute, Viale Belfiore 43, 50144, Florence, Italy. srinaldi@irf.it.|Research Department, Rinaldi Fontani Foundation, Viale Belfiore 43, 50144, Florence, Italy. srinaldi@irf.it.
Salvatore
http://orcid.org/0000-0002-8961-6316
S
Rinaldi
Department of Veterinary Medicine, University of Sassari, Viale Vienna 43/B, 07100, Sassari, Italy.
Eraldo Sanna
ES
Passino
D000177:Acrosome / Q000378:metabolism; D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D002470:Cell Survival / Q000235:genetics; D020552:Comet Assay; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D004247:DNA / Q000235:genetics / Q000378:metabolism; D006736:Horses; D015227:Lipid Peroxidation; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D008856:Microscopy, Fluorescence; D055101:Semen Analysis / Q000379:methods / Q000662:veterinary*; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013081:Sperm Motility / Q000235:genetics; D013094:Spermatozoa / Q000166:cytology* / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism
D000975:Antioxidants; D008315:Malondialdehyde; D004247:DNA; D013482:Superoxide Dismutase