5 research outputs found
Combined Effect of Trolox and EDTA on Frozen-Thawed Sperm Quality
The freezing and thawing process not only is associated with serious damage to sperm such as damage to the plasma membrane and the acrosomal membrane but also changes the membrane permeability to some ions including calcium. Also, the generation of oxygen free radicals is increased during the freezing-thawing process. The purpose of this study was to evaluate of the effects of Trolox as an antioxidant and edetic acid (EDTA) as a calcium chelator on frozen-thawed (FT) sperm and compare these effects with those on fresh sperm. This study was done on these men of 25 healthy men, who referred to Shiraz Infertility Centerbetween2012 and2013. Normal samples were transferred to the ReproductivePhysiology Laboratory, Department of Physiology,Shiraz University of Medical Sciences, Shiraz. The samples were divided into two groups randomly: fresh and FT sperm groups. Each group was divided into five subgroups: control group, the solvent group (0.1%dimethyl sulfoxide [DMSO]), Trolox group (200μM), EDTA group (1.1mM), and Trolox+EDTA group. The percentages of motility, viability, and acrosome-reacted sperm were tested. The percentages of motility and viability in the FT sperm were lower than those in the fresh sperm. The progressive motility of the FT sperm was improved nonsignificantly with Trolox+EDTA. However, the effect of Trolox+EDTA on the progressive motility of the FT sperm was much more than that on the fresh sperm. The fewest acrosome-reacted sperm were observed in the EDTA-containingFT sperm. Antioxidant supplementation or omission of extracellular calcium may partly improve motility and also reduce acrosomal damage in FT sperm
The effects of laser assisted hatching on pregnancy rates
Background: For infertile women aged over 35 years, failure of the ZP
(zona pellucida) to rupture is believed to be associated with a
decreased implantation rate in in vitro fertilization (IVF) or intra
cytoplasmic sperm injection (ICSI). Objective: In this research, laser
assisted hatching (LAH) was offered to patients with advanced maternal
age to evaluate a possible benefit. Materials and Methods: Nine hundred
thirty two cycles of IVF/ICSI in females were analyzed. Women included
in this study were allocated in 4 groups. In group I and II, embryos
were cultured and transferred with and without LAH in women aged
≤ 35, whereas embryos of group III and IV were examined with and
without LAH in women aged ≤ 35. Laser manipulations were
performed using a suturn – Tm3 system using 2-3 pulses of 0.8
millisecond with 400 voltage duration. The size of the hole made in the
zona was measured to be 5-10 µm, depending on the zona thickness
of each individual embryo. Results: The performance of LAH
significantly increased clinical pregnancy rates in all patients. In
group I and II, the chemical (50.99% and 31.61% respectively), clinical
(50% and 30.69% respectively) and multiple pregnancies (22.27% and
5.94% respectively) significantly differ between these groups. In the
patients with advanced female age ≤ 35 the performance of LAH
significantly increased chemical (30.12%) and clinical pregnancy
(27.71%) rates compared to whom without LAH (18.96% and 16.37%
respectively). Conclusion: Our data demonstrate in the patients who
were less than 35 years old, multiple pregnancy rates were
significantly increased compared to other groups who aged over 35 years
old. In addition benefit of LAH in improving pregnancy rates after IVF
or ICSI in women of advanced age (≤ 35) was show
Effect of combined density gradient centrifugation on X- and Y- sperm separation and chromatin integrity
Background: It has been claimed that by using different washing
methods, the sperms can be separated according to size, motility,
density, chromosomal content and surface markings and charge. These
methods also reduce sperm chromatin deficiencies and screen the sperms
before applying in assisted reproduction techniques. Objective: This
study compared simple density gradient methods and a combined method
with albumin density gradient and PureSperm separation (alb/PureSperm)
for sex preselection by double fluorescence in situ hybridization
(FISH) versus chromomycin A3 staining to determine chromatin integrity.
Materials and Methods: 30 normal semen samples were prepared with
PureSperm, albumin gradients and alb/PureSperm. All samples were then
stained by FISH and chromomycin A3. The results were compared with SPSS
11.5 and the Kruskal-Wallis test. Results: The proportion of X-bearing
spermatozoa by PureSperm separation (47.58±5.67) and Y-bearing
spermatozoa by albumin gradient (46.13±3.83) methods were slightly
higher than in putative normal sperm samples (1:1), but there were no
significant differences in the X- or Y- bearing spermatozoa counts
among the three methods. Albumin gradient separation tended to
underestimate abnormal spermatozoa compared to PureSperm and combined
alb/PureSperm. Conclusion: Routine separation methods slightly enriched
X- or Y- bearing spermatozoa, but the differences were not significant
for clinical purposes. The combined alb/PureSperm method had no
advantages for assessing sex ratio or chromatin integrity compared to
simpler gradient methods