The Consequence of Short Insemination Strategy on Sperm Biological Characteristics, Embryo Morphokinetics, and Clinical Outcomes in the IVF Program

Abstract

Overnight coincubation of the cumulus–oocyte complexes (COCs) with spermatozoa may be accompanied by overproduction of reactive oxygen species (ROS) and possible determinant effects on sperm cellular status in the in vitro fertilization (IVF) program. The COCs from 100 cases were divided into two groups of short (2 hr) and long (18 hr) inseminations. The malondialdehyde (MDA) level as by-product of ROS was assessed in the insemination medium. The sperm DNA integrity, the mitochondrial membrane potential (MMP), and acrosome reaction (AR) were evaluated using sperm chromatin dispersion (SCD), fluorescence stain of JC-1, and double staining, respectively. Normally, fertilized oocytes (n = 525) were assessed via time-lapse monitoring (TLM) for assessment of the time of fading (tPNF), 2 until 8 timing (t2-t8), s1 (t2-tPNF), s2 (t4-t3), s3 (t8-t5), and duration of cell cycles. Finally, the best embryos were transferred, and clinical outcomes were assessed. Higher rates of MDA concentration, DNA fragmentation, and AR and low rate of MMP were noticed in long compared to short insemination groups ( $p\le 0.0001$ ). The morphokinetic parameters showed five-cell stage (t5) and cell cycle 3 (cc3) that were significantly different between the groups ( $p=0.04$ and $p=0.03$ , respectively). A high level of ROS observed in the long insemination group might have a detrimental effect on sperm status. Although, similar embryo quality and clinical outcomes were noticed in two insemination groups, but the trend was toward the short insemination.