Effects of Sperm Processing Techniques Involving Centrifugation on Nitric Oxide, Reactive Oxygen Species Generation and Sperm Function~!2009-11-11~!2010-01-07~!2010-03-30~!

Abstract

This study was aimed at investigating the effects of sperm centrifugation on nitric oxide (NO) and reactive oxygen species (ROS) generation as well as sperm motility and viability. Human spermatozoa were centrifuged for 10 and 30 minutes (400 x g) in the presence or absence of the NOS inhibitor, N G -nitro-L-arginine methyl ester (L-NAME); ROS scavenger, N-(2-mercaptopropionyl)Glycine (MPG) or the combination of L-NAME + MPG. Total sperm motility was significantly decreased with 30 minutes of centrifugation whereas progressive motility and cell viability were significantly decreased with 10 and 30 minutes of centrifugation. These effects were reversed with the administration of MPG or L-NAME + MPG. Ten minutes centrifugation significantly elevated ROS and NO production (P < 0.05). Thirty minutes centrifugation elevated ROS generation (P < 0.01) whereas NO was attenuated. This study has demonstrated that 10 and 30 minutes of sperm centrifugation were detrimental to both sperm motility and viability, but generally 30 minutes centrifugation was more detrimental to sperm than 10 minutes. It has also demonstrated that 10 minutes centrifugation led to both NO and ROS elevation whereas 30 minutes centrifugation led to ROS elevation and NO attenuation. We therefore recommend that sperm separation techniques should avoid using centrifugation or prolonged centrifugation in assisted reproductive technologies.

Sperm DNA Fragmentation and Apoptosis Levels: A Comparison of the Swim up and the Density Gradient Centrifugation Methods for Sperm Preparation

“…In addition, repeated centrifugation during sperm preparation can induce iatrogenic DNA damage. This study supports the findings by Lampiao et al (2010) that centrifugation for 10 and 30 min damaged both the motility and the viability of sperm and that 30 min centrifugation was more damaging than 10 min. According to Marchesi et al (2010) both the DGC and the SU methods can improve sperm DNA integrity, but the SU method improved the quality of DNA more than the DGC method.…”

Alpha‐lipoic acid minimises reactive oxygen species‐induced damages during sperm processing

“…In semen, spermatozoon is protected from ROS by the antioxidants presented in the seminal fluid. Therefore, separation of spermatozoa from seminal antioxidant and their concomitant introduction to shearing forces during centrifugation further aggravate ROS production and induce DNA damage (Lampiao, Strijdom, & du Plessis, ; Li et al, ). In order to minimise the effect of ROS‐induced DNA damage, supplementation of sperm processing media with antioxidants is recommended (Chi et al, ; Michael et al, ).…”

Estrés oxidativo y el uso de antioxidantes en la producción in vitro de embriones mamíferos. Revisión

“…En los procesos de producción in vitro de embriones, el protocolo de preparación y capacitación del semen para la fertilización in vitro requiere el uso de centrifugación. Sin embargo, el tiempo y la fuerza centrífuga son factores que contribuyen a aumentar los niveles de ERO, causando daño oxidativo y afectando la función espermática (5,60) . Además, se observó que la centrifugación de espermatozoides sexados y no sexados por largos periodos de tiempo (45 min) a 700 xg causaba pérdida de la integridad de la membrana plasmática y fragmentación del ADN (61,62) .…”