Sperm Lipid Peroxidation and Antioxidants Activities as Predictors of Pregnancy Outcomes after Density Gradient Centrifugation in Medical Assistance Procreation

Aims: The purpose of this study was to assess the Density Gradient Centrifugation (DGC) effectiveness in spermatozoa selection by analyzing the oxidative stress profile and in pregnancy prediction outcomes of In Vitro Fertilization (IVF). Study Design: This was a prospective study. Place and Duration of Study: Department of Biology, laboratory of Physiopathology Molecular Genetic and Biotechnology. Ain Chock Faculty of Sciences, Hassan II University, Casablanca. And Anfa Fertility Center Private Clinic of in vitro fertilization and Endoscopic Surgery, Casablanca. Methodology: We included 101 men patients subdivided into three groups. Group A with normal sperm parameters and a positive pregnancy; Group B with normal sperm parameters and a negative pregnancy; Group C with abnormal sperm parameters and a negative pregnancy. After DGC, lipid peroxidation (MDA) level, as well as the antioxidant enzyme activities of superoxide dismutase (SOD), glutathione reductase (GR) and catalase (CAT), were evaluated. Results: Patients with normal and abnormal sperm parameters showed that MDA level, and antioxidant enzymes activities increased significantly from the pellet, to 80 %, and from 80% to 40% fractions. In addition, lipid peroxidation and enzymes activities levels were significantly lower in patients with positive pregnancy than in patients with negative pregnancy and especially those with abnormal spermatic parameters. Conclusion: The DGC would select sperm not only motile and viable but with less oxidative stress. Moreover, the rate of lipid peroxidation and antioxidant enzyme activities could differentiate between patients with normal and abnormal spermatic parameters and between patients with positive and negative pregnancy outcome.

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Conventional in vitro fertilization (IVF) is as efficient as intracytoplasmic sperm injection (ICSI) in art cycles with low number of retrieved oocytes and normal sperm parameters

Fertility and Sterility ◽

10.1016/j.fertnstert.2007.07.388 ◽

2007 ◽

Vol 88 ◽

pp. S121

Author(s):

S.-H. Lee ◽

H. Song ◽

M.K. Kim ◽

M.K. Koong ◽

I.O. Song ◽

...

Keyword(s):

In Vitro Fertilization ◽

Intracytoplasmic Sperm Injection ◽

Sperm Parameters ◽

Vitro Fertilization ◽

Normal Sperm

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Effect of toluene on erythrocyte membrane stability under in vivo and in vitro conditions with assessment of oxidant/antioxidant status

Toxicology and Industrial Health ◽

10.1177/0748233709346758 ◽

2009 ◽

Vol 25 (8) ◽

pp. 545-550 ◽

Cited By ~ 15

Author(s):

Ismail Karabulut ◽

Z. Dicle Balkanci ◽

Bilge Pehlivanoglu ◽

Aysen Erdem ◽

Ersin Fadillioglu

Keyword(s):

Oxidative Stress ◽

Lipid Peroxidation ◽

Unsaturated Fatty Acids ◽

Osmotic Fragility ◽

Human Erythrocytes ◽

Antioxidant Enzyme Activities ◽

Oxidative Stress Parameters ◽

Stress Parameters

Toluene, an organic solvent used widely in the industry, is highly lipophilic and accumulates in the cell membrane impeding transport through it. Its metabolites cause oxygen radical formation that react with unsaturated fatty acids and proteins in erythrocytes leading to lipid peroxidation and protein breakdown. In this study, we aimed to investigate the membrane stabilizing and the oxidative stress—inducing effects of toluene in human erythrocytes. Measurements of osmotic fragility, mean corpuscular volume (MCV), oxidative stress parameters and antioxidant enzyme activities were performed simultaneously both in individuals exposed to toluene professionally (in vivo) and human erythrocytes treated with toluene (in vitro). To measure osmotic fragility, erythrocytes were placed in NaCl solutions at various concentrations (0.1% [blank], 0.38%, 0.40%, 0.42%, 0.44%, 0.46%, 0.48% and 1% [stock]). Percentage of haemolysis in each solution was calculated with respect to the 100% haemolysis in the blank solution. The erythrocyte packs prepared at the day of the above-mentioned measurements were kept at —80°C until the time for determination of malonyldialdehyde and protein carbonyl levels, and catalase (CAT) and glutathione peroxidase activities as indicators of oxidative stress. Toluene increased oxidative stress parameters significantly both in vivo and in vitro; it also caused a significant decrease in the activities of antioxidant enzymes. Osmotic fragility was altered only in the case of in vitro exposure. In conclusion, toluene exposure resulted in increased lipid peroxidation and protein damage both in vivo and in vitro. Although, it is natural to expect increased osmotic fragility due to oxidative properties of toluene, its membrane-stabilizing effect overcame the oxidative properties leading to decreased osmotic fragility or preventing its deterioration in vitro and in vivo toluene exposures, respectively, in the present study.

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Protective effects of casein-derived peptide VLPVPQK against hydrogen peroxide–induced dysfunction and cellular oxidative damage in rat osteoblastic cells

Human & Experimental Toxicology ◽

10.1177/0960327116678293 ◽

2017 ◽

Vol 36 (9) ◽

pp. 967-980 ◽

Cited By ~ 16

Author(s):

SB Mada ◽

S Reddi ◽

N Kumar ◽

S Kapila ◽

R Kapila

Keyword(s):

Oxidative Stress ◽

Hydrogen Peroxide ◽

Lipid Peroxidation ◽

Oxidative Damage ◽

Osteoblastic Cells ◽

Antioxidant Enzyme Activities ◽

Protective Agent ◽

Protective Effects ◽

Differentiation Markers

Oxidative stress inhibits osteoblast differentiation and function that lead to the development of osteoporosis. Casein-derived peptide VLPVPQK (PEP), a potent antioxidant, was isolated from β-casein of buffalo milk. We used an in vitro oxidative stress model induced by hydrogen peroxide (H2O2) in rat osteoblastic cells to investigate the protective effects of PEP against H2O2-induced dysfunction and oxidative damage. Cells were pretreated with PEP (50–200 ng/mL) for 2, 7 or 21 days followed by 0.3 mM H2O2 treatment for 24 h and then markers of osteogenic development, oxidative damage and apoptosis were examined. PEP significantly increased the viability and differentiation markers of osteoblast cells such as alkaline phosphatase and calcium mineralization. Moreover, PEP suppressed the production of reactive oxygen species (ROS), lipid peroxidation and ameliorated H2O2-induced reduction in glutathione, superoxide dismutase and catalase activities. In addition, PEP partially inhibited caspase-9 and-3 activities and reduced propidium iodide–positive cells. Altogether, our results demonstrated that PEP could protect rat osteoblast against H2O2-induced dysfunction and oxidative damage by reduction of ROS production, lipid peroxidation and increased antioxidant enzyme activities. Thus, our data suggest that PEP might be a valuable protective agent against oxidative stress–related diseases such as osteoporosis.

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Pregnancy from in vitro fertilization of human eggs after separation of motile spermatozoa by density gradient centrifugation**Supported in part by National Health and Medical Research Council grants to R. V. H. and A. L. and by Pharmacia (South Seas) Pty. Ltd., Melbourne, Victoria, Australia.

Fertility and Sterility ◽

10.1016/s0015-0282(16)49103-x ◽

1986 ◽

Vol 45 (1) ◽

pp. 93-96 ◽

Cited By ~ 52

Author(s):

Ross V. Hyne ◽

Alexander Stojanoff ◽

Gary N. Clarke ◽

Alexander Lopata ◽

W. Ian H. Johnston

Keyword(s):

In Vitro Fertilization ◽

Medical Research ◽

Density Gradient ◽

National Health ◽

Research Council ◽

Medical Research Council ◽

Density Gradient Centrifugation ◽

Gradient Centrifugation ◽

Vitro Fertilization

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Comparison of in vitro fertilization outcomes in ICSI cycles after human sperm preparation by density gradient centrifugation and direct micro swim-up without centrifugation

JBRA Assisted Reproduction ◽

10.5935/1518-0557.20170022 ◽

2017 ◽

Vol 21 (2) ◽

Cited By ~ 6

Author(s):

Simone Palini ◽

Silvia De Stefani ◽

Mariangela Primiterra ◽

Serena Benedetti ◽

Stefano Barone ◽

...

Keyword(s):

In Vitro Fertilization ◽

Density Gradient ◽

Density Gradient Centrifugation ◽

Gradient Centrifugation ◽

Human Sperm ◽

Vitro Fertilization ◽

Sperm Preparation

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The Effects of In Vitro Incubation of Asthenoteratozoospermic Semen after Density Gradient Centrifugation at Room Temperature and 37C on Sperm Parameters, Chromatin Quality and DNA Fragmentation in a Short Time Period

Journal of Reproduction & Infertility ◽

10.18502/jri.v21i4.4332 ◽

2020 ◽

Author(s):

Motahareh Karimi Zarchi ◽

Behnam Maleki ◽

Mahmood Dehghani Ashkezari ◽

Leila Motamed Zadeh ◽

Azam Agha-Rahimi

Keyword(s):

Dna Fragmentation ◽

Density Gradient ◽

Room Temperature ◽

Density Gradient Centrifugation ◽

Sperm Quality ◽

Gradient Centrifugation ◽

Sperm Parameters ◽

Sperm Dna Fragmentation ◽

In Vitro Incubation

Background: Sperm quality is an important factor in assisted reproductive technology (ART) that affects the success rate of infertile couples treatment. In vitro incubation of sperm can influence its parameters and DNA integrity. The present study focused on the effect of different incubation temperatures sperm parameters on asthenoteratozoospermia semen prepared with density gradient centrifugation at different times. Methods: Twenty-seven samples were collected and prepared. Then, the suspension was divided into two parts. One part was incubated at room temperature (RT), and another was incubated at 37°C. Immediately and after 2 hr (2H) and 4 hr (4H), sper-matozoa were evaluated regarding motility, viability, morphology, sperm protamine deficiency, chromatin and DNA fragmentation. Statistical analysis was performed using paired t-test and repeated measures. The p<0.05 was considered statistically significant. Results: Our results showed that following 2 and 4 hr of incubation at RT, sperm progressive motility and viability decreased significantly. Sperm DNA fragmentation increased significantly following 2 and 4 hr of incubation at RT and 37°C. The Trend analysis confirmed that there were no significant differences between sperm parameters and DNA fragmentation after different times at RT and 37°C. Conclusion: Incubation of sperm at RT in comparison to 37°C didn’t preserve sperm parameters and DNA efficiently. Therefore, IVF, ICSI and IUI procedure should be performed in the soonest possible time after sperm preparation.

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Practical use of percoll density gradient centrifugation on sperm sex determination in commercial dairy farm in Thailand

Indian Journal of Animal Research ◽

10.18805/ijar.8427 ◽

2016 ◽

Author(s):

Kornkanok Promthep ◽

Saranya Satitmanwiwat ◽

Narisorn Kitiyanant ◽

Pisit Tantiwattanakul ◽

Kanya Jirajaroenrat ◽

...

Keyword(s):

In Vitro Fertilization ◽

Density Gradient ◽

Density Gradient Centrifugation ◽

Gradient Centrifugation ◽

Female Offspring ◽

Vitro Fertilization ◽

Dairy Farmer ◽

Percoll Density Gradient Centrifugation ◽

Significant Difference

Our experiment demonstrated an extension of Percoll® Density Gradient Centrifugation to use as a selection tool for sperm sexing developed for dairy farmer. We develop a simple protocol for bovine sperm sexing and used for artificial insemination (AI) and in vitro fertilization (IVF) for embryo transfer (ET). The quality of sperm after centrifugation in each layer were significantly different (P <0.01). There was no significant difference between percentage of live sperm after centrifugation and fresh sperm before centrifugation (P >0.05). The phenotype and swimming capacity of sperms in each layer were significantly different (P < 0.01). It was found that the most appropriate sperm in the layer after centrifugation and after quinacrine staining was between 65-70% Percoll®. X-bearing spermatozoa at this layer was 60.75% and motility was 95.86±0.46%. After insemination, the pregnancy ratio in the experiment was 40% which is similar to the percentage of normal AI. The number of female offspring in the experiment was 71.4% in selected sperm vs 50% in non-selected sperm AI. In the In Vitro fertilization experiment, sperm from our 7 layer Percoll® gradient concentration can fertilize and produce blastocyst embryo successfully. The simple 7 layer Percoll® gradient concentration protocol is appropriate for dairy farmer to do as a pre-AI step to increase their female offspring in their herd.

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Fertilization capacity of cryopreserved Iberian ibex epididymal sperm in a heterologous in vitro fertilization assay

Zygote ◽

10.1017/s0967199413000518 ◽

2013 ◽

Vol 23 (1) ◽

pp. 136-144 ◽

Cited By ~ 3

Author(s):

J. López-Saucedo ◽

J. Santiago-Moreno ◽

R. Fierro ◽

D. Izquierdo ◽

M.A. Coloma ◽

...

Keyword(s):

Density Gradient Centrifugation ◽

Gradient Centrifugation ◽

Blastocyst Development ◽

Vitro Fertilization ◽

Air Atmosphere ◽

Epididymal Spermatozoa ◽

Mountain Ungulates ◽

Fertilization Capacity ◽

Oviductal Fluid

SummaryIn vitro fertilization (IVF) can be used to assess the fertilization capacity of sperm. Heterologous IVF may be useful when assessing that of wild animals as it is often difficult to obtain adequate numbers of naturally corresponding oocytes. The aim of the present study was to assess the fertilization capacity of frozen–thawed ibex epididymal spermatozoa via heterologous IVF involving the oocytes of prepubertal domestic goats. The effect on fertilization and embryo development of adding oestrous sheep serum (ESS) to the fertilization medium was also examined. Cumulus–oocyte complexes (COCs) were matured in TCM-199 for 24–27 h at 38.5°C in a 5% CO2 in air atmosphere. Frozen–thawed epididymal spermatozoa were selected by density gradient centrifugation. After maturation, the oocytes were co-incubated with spermatozoa in synthetic oviductal fluid (SOF) with different concentrations of ESS: SOF-C (0%), SOF-2 (2%) and SOF-20 (20%). At 17 h post-insemination (hpi), zygotes with one female and one male pronucleus (2PN) were categorised as normal; zygotes with 3PN were recorded as polyspermic, and oocytes with 1PN as asynchronous. Cleavage and blastocyst development were assessed at 48 and 168 hpi respectively. The percentage of zygotes with 2PN was higher in the SOF-2 than in the SOF-20 treatment group (27.7% versus 2.9% P < 0.05). The percentage of blastocysts formed with the SOF-C, SOF-2 and SOF-20 treatments were 1.1%, 7.5% and 0% respectively. These results show that the presence of 2% ESS achieves better results than the use of no serum or the standard 20% concentration. Heterologous IVF may be an effective method for predicting the fertilization capacity of ibex spermatozoa, and therefore perhaps that of other wild mountain ungulates.

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