Bio-Functional Sperm Parameters: Does Age Matter?

The evaluation of biofunctional sperm parameters can explain some cases of idiopathic male infertility. Among these, sperm DNA fragmentation (fDNA) is the most studied biofunctional sperm parameter. Mitochondrial membrane potential (MMP) correlates positively with sperm motility, the evaluation of sperm apoptosis by flow cytometry allows us to identify a population of spermatozoa not recognizable at the optical microscopy and finally, lipid peroxidation (LP) and mitochondrial superoxide levels measurements are rational oxidative stress indices. Male age seems to affect sperm concentration and sperm fDNA. For these reasons, this study was undertaken to evaluate the correlation, if any, between male age and biofunctional sperm parameters evaluating their possible impact on fDNA. To accomplish this, MMP, degree of chromatin compactness, sperm apoptosis/vitality, fDNA, LP, and mitochondrial superoxide levels were evaluated by flow cytometry in a cohort of 874 men. A significant negative correlation was found between age and the percentage of alive spermatozoa (r = -0.75, p < 0.05). The percentage of spermatozoa with low MMP (L-MMP) correlated positively with the percentage of spermatozoa with abnormal chromatin compactness (r = 0.24, p < 0.05). Spermatozoa with abnormal chromatin compactness and L-MMP correlated negatively with the percentage of alive spermatozoa (r = 0.83, p < 0.05) and positively with spermatozoa with PS externalization (r = 0.13, p < 0.01). The percentage of alive spermatozoa correlated negatively with both the percentage of spermatozoa with PS externalization (r = 0.24, p < 0.01) and of the spermatozoa with fDNA (r = 0.10, p < 0.05). Spermatozoa with PS externalization correlated positively with the percentage of spermatozoa with fDNA (r = 0.09, p < 0.05). Spermatozoa with LP correlated positively with the percentage of spermatozoa with increased mitochondrial superoxide (r = 0.11, p < 0.01) In conclusion, these findings in a large number of men suggest that age, mitochondrial damage, and alteration of chromatin compactness could activate the apoptotic cascade which could result in an increased fDNA rate.

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The effects of Pb on sperm parameters and sperm DNA fragmentation of men investigated for infertility

Journal of Basic and Clinical Physiology and Pharmacology ◽

10.1515/jbcpp-2019-0239 ◽

2020 ◽

Vol 31 (4) ◽

Author(s):

G.U.S. Wijesekara ◽

D.M.S. Fernando ◽

S. Wijeratne

Keyword(s):

Dna Fragmentation ◽

Seminal Plasma ◽

Sperm Concentration ◽

Sperm Parameters ◽

World Health ◽

Sperm Dna Fragmentation ◽

Progressive Motility ◽

Sperm Dna ◽

The Mean ◽

Pb Concentration

AbstractBackgroundLead (Pb) is one of the metals most prevalent in the environment and is known to cause infertility and deoxyribonucleic acid (DNA) fragmentation. This study aimed to determine the association between seminal plasma Pb and sperm DNA fragmentation in men investigated for infertility.MethodsMale partners (n = 300) of couples investigated for infertility were recruited after informed consent was obtained. Sperm parameters were assessed according to the World Health Organization (WHO) guidelines. Seminal plasma Pb was estimated by atomic absorption spectrophotometry after digestion with nitric acid.ResultsIn Pb-positive and -negative groups the sperm parameters and sperm DNA fragmentation were compared using independent sample t-test and the Mann-Whitney U-test, respectively. The mean [standard deviation (SD)] age and duration of infertility were 34.8 (5.34) years and 45.7 (35.09) months, respectively, and the mean Pb concentration was 15.7 μg/dL. In Pb positives compared to Pb negatives the means (SD) of sperm count, progressive motility viability and normal morphology were lower (p > 0.05) but the DNA fragmentation was significantly higher 39.80% (25.08) than Pb negatives 22.65% (11.30). Seminal plasma Pb concentration and sperm DNA fragmentation had a positive correlation (r = 0.38, p = 0.03). A negative correlation was observed between sperm DNA fragmentation and sperm concentration, progressive motility, total motility and viability. When the DNA fragmentation was ≥30% sperm concentration and viability decreased (p < 0.05).ConclusionsPb in seminal plasma had a significant effect on sperm DNA fragmentation but not with other sperm parameters.

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The Effect of COVID-19 Infection on Sperm Quality and Male Fertility

Jentashapir Journal of Cellular and Molecular Biology ◽

10.5812/jjcmb.115390 ◽

2021 ◽

Vol In Press (In Press) ◽

Author(s):

Hamid Piroozmanesh ◽

Ebrahim Cheraghi ◽

Leila Naserpoor ◽

Masoumeh Aghashahi ◽

Rahil Jannatifar

Keyword(s):

Sperm Concentration ◽

Reproductive Hormones ◽

Sperm Parameters ◽

World Health ◽

Control Group ◽

Dna Integrity ◽

Sperm Dna Fragmentation ◽

Rt Pcr ◽

Sperm Dna Integrity ◽

Sperm Dna

Background: Coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) may lead to the infertility of men. Objectives: This study aimed to investigate the impact of COVID-19 infection on sperm parameters and reproductive hormones in fertile men. Methods: A total of 100 males were selected and divided into two groups: (1) patients in convalescence (patients suffering from COVID-19 infection in pharyngeal swab in accordance with reverse-transcription polymerase chain reaction [RT-PCR] or antibodies); (2) negative control group (without antibodies). Semen and blood samples were gathered from all subjects. In the native semen, immunoglobulin (Ig) M and IgG antibodies in the blood were confirmed, and COVID-19 was detected via RT-PCR. To this end, based on the World Health Organization (WHO) guidelines, semen analysis, total antioxidant capacity (TAC), and sperm DNA integrity were assessed. Results: Results demonstrated that sperm concentration, motility, sperm viability, and TAC significantly reduced in fertile males with virus infection. In comparison with the control group, sperm DNA integrity was significantly increased (P < 0.05). Data indicated that the semen volume was not significantly correlated with COVID-19, and there was a significantly negative correlation between sperm concentration, sperm total motility, sperm vitality, sperm normal forms, and TAC with COVID-19. Sperm DNA fragmentation index had a significant and positive correlation with COVID-19 (P < 0.05). In addition, reproductive hormones significantly reduced in fertile males with COVID-19 infection (P < 0.05). Conclusions: COVID-19 infection has a negative influence on sperm parameters and reproductive hormones in fertile males.

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P–042 Impact of semen parameters, sperm DNA fragmentation and sperm aneuploidy in male infertility

Human Reproduction ◽

10.1093/humrep/deab130.041 ◽

2021 ◽

Vol 36 (Supplement_1) ◽

Author(s):

M Maia ◽

C Almeida ◽

M Cunha ◽

A Gonçalves ◽

S S Soares ◽

...

Keyword(s):

Male Infertility ◽

Dna Fragmentation ◽

Sperm Concentration ◽

Semen Parameters ◽

Sperm Dna Fragmentation ◽

Male Age ◽

Fertility Status ◽

Sperm Aneuploidy ◽

Sperm Dna ◽

Aneuploidy Testing

Abstract Study question Should sperm aneuploidies and sperm DNA fragmentation (sDNAfrag) be included as valid tests in the routine investigation of male infertility? Summary answer Sperm DNA fragmentation was associated with male age, oligozoospermia (OZ), oligoteratozoospermia (OT), astenoteratozoospermia (AT) and oligoastenoteratozoospermia (OAT). Sperm aneuploidies were associated with OT and OAT. What is known already Semen parameters assist male infertility diagnosis and treatment, but sDNAfrag and aneuploidy analysis could add useful information, as abnormal values compromise fertility. To include these tests in the routine diagnosis it should be determined if behave as informative parameter and add information regarding the fertility status. For that, further studies comparing these tests to semen parameters are needed, since previous results are not consensual. Additionally, standardization of a sDNAfrag cut-off is needed, as different sample sizes and techniques originate distinct results. Also, until a standardization of the protocol is missing, a cut-off value should be defined for each laboratory. Study design, size, duration A retrospective and prospective investigation was performed, within a 12 years period (April 2007-December 2019). A total of 835 infertile males with a normal karyotype (46,XY) were included. Karyotyping and evaluation of sDNAfrag and sperm aneuploidies were made at a public Genetic unit. All normozoospermic (NZ) patients with a born child and patients whose infertility treatments were done due to female factors were selected from our database and used as controls (60 individuals). Participants/materials, setting, methods Semen analysis followed WHO–2010 guidelines. sDNAfrag was evaluated using the TUNEL assay. Sperm aneuploidies were detected using FISH (chromosomes 13, 18, 21, X, Y). Several tests were applied: correlations for linear associations between numerical variables, ANOVA for comparisons between means, Dunn-test for post-hoc comparisons. To determine the sDNAfrag cut-off value, the area under the ROC curve, sensitivity and specificity, were calculated, with the Youden-Index used to find a threshold that maximizes both sensitivity and specificity. Main results and the role of chance Regarding male age, it was observed a positive correlation with sperm concentration, a negative correlation with sperm vitality (VT) and hypoosmolality, and a positive correlation with sDNAfrag. Regarding sDNAfrag, it was observed negative correlation with sperm concentration, total progressive motility (TPM), morphology, VT and hypoosmolality. Regarding sperm aneuploidies, both total sperm aneuploidy and total sperm disomy exhibited a negative association with sperm concentration, TPM and morphology. It was also investigated whose groups of individuals could be indicated for sDNAfrag or sperm aneuploidy testing. The NZ group evidenced significant lower sDNAfrag, total sperm aneuploidy and total sperm disomy in relation to the non-NZ group. In the NZ group, sDNAfrag was significantly lower in relation to the OZ, OT, AT and OAT groups. The NZ group presented significant lower percentages of sperm aneuploidy in relation to the OT and OAT groups, and significant lower percentages of sperm disomy in relation to the OAT group. Additionally, sDNAfrag was positively correlated with total sperm aneuploidy and total sperm disomy. From the present large population, ROC curve analysis allowed estimating a cut-off value of 18.8% for the TUNEL-assay (sDNAfrag), with 0.658 of area under the curve, 53.9% sensitivity and 76.7% specificity. Limitations, reasons for caution Although presenting a high number of cases and strict controls, the present study was unable to include as controls healthy men with proven fertility. Additionally, the present study did not take into account life-style factors and male associated pathologies besides infertility. Wider implications of the findings: Semen parameters were shown to be negatively correlated with sDNAfrag and sperm aneuploidies. As sDNAfrag testing and sperm aneuploidy testing were associated with semen abnormalities and male age, it is suggested their inclusion in the routine evaluation of infertile men, thus adding important complementary information about the fertility status. Trial registration number Not Appliable

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O-207 Which infertile patients mostly deserve to have a sperm DNA fragmentation index done? Findings from a cross-sectional study

Human Reproduction ◽

10.1093/humrep/deab128.018 ◽

2021 ◽

Vol 36 (Supplement_1) ◽

Author(s):

E Pozzi ◽

L Boeri ◽

L Candela ◽

D Cignoli ◽

G Colandrea ◽

...

Keyword(s):

Logistic Regression ◽

Regression Analysis ◽

Dna Fragmentation ◽

Logistic Regression Analysis ◽

Roc Curve ◽

Sperm Concentration ◽

Testicular Volume ◽

Sperm Dna Fragmentation ◽

Sperm Dna ◽

Increased Risk

Abstract Study question Current scientific guidelines do not clearly suggest which patients would benefit the most from a sperm DNA fragmentation (SDF) test. Summary answer We aimed to investigate potential predictive factors for altered SDF in a homogenous cohort of white-European men presenting for primary couple’s infertility. What is known already High SDF has been associated with reduced fertilization rates, reduced chances of natural conception and an increased risk of early pregnancy loss. Study design, size, duration Data from 478 consecutive men with normal or altered SDF were analysed. Infertility was defined according to the WHO criteria. Semen analysis, SDF (according to SCSA) and serum hormones were measured in every patient. Health significant comorbidities were scored with the Charlson Comorbidity Index (CCI). Altered SDF was considered with a threshold of > 30%. Participants/materials, setting, methods Descriptive statistics compared the overall characteristics of patients with normal SDF and altered SDF. Logistic regression analysis tested potential predictors of altered SDF. ROC curve was used to test the accuracy of the model in predicting SDF alteration Main results and the role of chance Of 478 patients, 253 (57.7%) had altered SDF. Median (IQR) age and BMI of the whole cohort were 38 (35-42) years and 25.1 (23.3-27.1) kg/m2 respectively. Patients with altered SDF were older (median (IQR) age: 39 (36-43) vs. 37 (34-38) years, p < 0.0001), had lower sperm concentration (5 (1.1–18) vs. 17 x106/mL (6–38.8), p < 0.0001), testicular volume (15.1 (12 –20) vs. 16.8 (12 – 25) Prader, p = 0.0005), and total motile sperm count (TMSC) (1.8 (0.21–10.71) vs. 11.8x106 (2–37.26), p < 0.0001). Conversely, men with altered SDF had higher FSH (6.1 (3.85–9.7) vs. 4.8 (3.85 – 7.9) mIU/mL, p < 0.0001) and prolactin levels (9.8 (7.43–14.04) vs. 8.3 (6.6–11.3) pg/mL, p = 0.0004) than those with normal SDF. At multivariable logistic regression analysis, patients’ age >35 years (OR: 2.45, p = 0.0009), FSH > 8.0 mIU/mL (OR: 2.23, p < 0.0001) and lower TMSC (OR: 2.04, p = 0.002) were identified as indipendent predictors of altered SDF, after adjusting for testicular volume and CCI≥1. ROC curve (Figure 1) revealed that the model has a good predictive ability to identify patients with SDF alteration (AUC: 0.72, 95%CI: 0.67 - 0.77). Limitations, reasons for caution It is a retrospective analysis at a single, tertiary-referral academic centre, thus raising the possibility of selection biases. In spite of this, all patients have been consistently analysed over time with a rigorous follow-up, thus limiting potential heterogeneity in terms of data reporting Wider implications of the findings Primary infertile men older than 35 years, with high serum FSH and low TMSC at baseline are the ones who mostly deserve a SDF test over their diagnostic work-up and that would potentially benefit the most of certain treatments to improve SDF value, thus increasing chances of conceiving. Trial registration number Not applicable

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From Spermiogram to Bio-Functional Sperm Parameters: When and Why Request Them?

Journal of Clinical Medicine ◽

10.3390/jcm9020406 ◽

2020 ◽

Vol 9 (2) ◽

pp. 406 ◽

Cited By ~ 4

Author(s):

Rosita A. Condorelli ◽

Aldo E. Calogero ◽

Giorgio I. Russo ◽

Sandro La Vignera

Keyword(s):

Normal Form ◽

Sperm Motility ◽

Sperm Count ◽

Semen Analysis ◽

Sperm Concentration ◽

Sperm Parameters ◽

Sperm Parameter ◽

Progressive Motility ◽

Semen Volume ◽

Chromatin Compactness

The aim of this experimental study was to evaluate whether infertile patients may benefit from the evaluation of bio-functional sperm parameters in addition to the conventional semen analysis. To accomplish this, we evaluated the correlation between conventional and bio-functional sperm parameters based on their percentile distribution in search of a potential threshold of these latter that associates with conventional sperm parameter abnormalities. The study was conducted on 577 unselected patients with infertility lasting at least 12 months. We identified cut-off values according to the median of the population for mitochondrial membrane potential (MMP), number of alive spermatozoa, and chromatin abnormality. High MMP (HMMP) (≥46.25%) was associated with sperm concentration, sperm count, progressive motility, and normal form. Low MMP (LMMP) (≥36.5%) was found to be associated with semen volume, sperm concentration, total sperm count, progressive motility, total motility, and normal form. The number of alive spermatozoa (≥71.7%) was associated with sperm concentration and progressive motility whereas abnormal chromatin compactness (≥21.10%) was associated with sperm concentration, total sperm count, and progressive motility. The data would suggest that, for every increase in the percentile category of sperm concentration, the risk of finding an HMMP≤46.25 is reduced by 0.4 and by 0.66 for a total sperm count. This risk is also reduced by 0.60 for every increase in the percentile category of sperm progressive motility and by 0.71 for total sperm motility. Each increment of percentile category of the following sperm parameter was followed by a decrease in the risk of finding an LMMP≤36.5: sperm concentration 1.66, total sperm count 1.28, sperm progressive motility 1.27, total sperm motility 1.76, and normal form 1.73. Lastly, the data showed that, for every increase in the percentile category of total sperm count, the risk of finding an abnormal chromatin compactness ≤21.10 is reduced by 1.25 (1.04–1.51, p < 0.05) and an increase of total sperm motility is associated with a reduced risk by 1.44 (1.12–1.85, p < 0.05). Results suggest a correlation between bio-functional and conventional sperm parameters that impact the sperm fertilizing potential. Therefore, the evaluation of bio-functional sperm parameters by flow cytometry may be useful to explain some cases of idiopathic male infertility.

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Relationship between Testicular Volume and Conventional or Nonconventional Sperm Parameters

International Journal of Endocrinology ◽

10.1155/2013/145792 ◽

2013 ◽

Vol 2013 ◽

pp. 1-6 ◽

Cited By ~ 37

Author(s):

Rosita Condorelli ◽

Aldo E. Calogero ◽

Sandro La Vignera

Keyword(s):

Flow Cytometry ◽

Testicular Volume ◽

Endocrine Function ◽

Sperm Parameters ◽

Serum Concentrations ◽

Ultrasound Evaluation ◽

Serum Hormone ◽

Chromatin Compactness ◽

Tv 12 ◽

First Time

Background. Reduced testicular volume (TV) (<12 cm3) is associated with lower testicular function. Several studies explored the conventional sperm parameters (concentration, motility, and morphology) and the endocrine function (gonadotropins and testosterone serum concentrations) in the patients with reduction of TV. No other parameters have been examined.Aim. This study aims at evaluating some biofunctional sperm parameters by flow cytometry in the semen of men with reduced TV compared with that of subjects with normal TV.Methods. 78 patients without primary scrotal disease were submitted to ultrasound evaluation of the testis. They were divided into two groups according to testicular volume: A Group, including 40 patients with normal testicular volume (TV > 15 cm3) and B Group, including 38 patients with reduced testicular volume (TV ≤ 12 cm3). All patients underwent serum hormone concentration, conventional and biofunctional (flow cytometry) sperm parameters evaluation.Results. With regard to biofunctional sperm parameters, all values (mitochondrial membrane potential, phosphatidylserine externalization, chromatin compactness, and DNA fragmentation) were strongly negatively correlated with testicular volume ().Conclusions. This study for the first time in the literature states that the biofunctional sperm parameters worsen and with near linear correlation, with decreasing testicular volume.

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313 ASSESSMENT OF SPERM DNA FRAGMENTATION IN CANINE EJACULATES USING THE Sperm-Halomax® KIT: PRELIMINARY RESULTS

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab313 ◽

2010 ◽

Vol 22 (1) ◽

pp. 312 ◽

Cited By ~ 6

Author(s):

M. Hidalgo ◽

M. R. Murabito ◽

M. J. Gálvez ◽

S. Demyda ◽

L. J. De Luca ◽

...

Keyword(s):

Dna Fragmentation ◽

Sperm Concentration ◽

Semen Parameters ◽

Sperm Chromatin ◽

Sperm Dna Fragmentation ◽

Fragmentation Index ◽

Sperm Dna ◽

Commercial Kit ◽

Sperm Dna Fragmentation Index ◽

Dna Fragmentation Index

Recently, a new procedure for the analysis of sperm DNA fragmentation has been developed for humans and different mammalian species, using a commercial kit based on the sperm chromatin dispersion (SCD) test; however, a descriptive study in canine semen has not been performed. The aim of this work was to assess the sperm DNA fragmentation in canine ejaculates using the SCD test and 2 different staining techniques. For this purpose, ejaculates were collectedby digital manipulation from4 healthy dogs of different breeds (1 German Pointer, 2 Spanish Greyhounds, and 1 Crossbreed). After collection, the sperm-rich fraction of the ejaculates from 3 dogs were pooled each time (n = 4) and then extended in Dulbecco’s phosphate buffered saline. All the pooled semen samples presented physiological values concerning routine semen parameters (motility, morphology, and sperm concentration). The sperm DNA fragmentation was assessed using the Sperm-Halomax® commercial kit specifically developed for canine semen (Halotech DNA SL, Madrid, Spain). Two semen aliquots of the diluted pooled semen samples were processed on each pre-treated slide provided in the kit following the manufacturer’s instructions. The last step was the staining technique. We stained each slide with 2 different staining procedures. The first half of the slide was stained with propidium iodide (Sigma-Aldrich, St. Louis, MO, USA) mixed in a proportion 1 : 1 with an antifading solution. The second half of the slide was stained for 15 min in Wright solution (1.01383.0500, Merck, Whitehouse Station, NJ, USA) 1 :1 in Phosphate Buffer pH 6.88 (1.07294.1000, Merck). The stained slides were observed using fluorescence and light microscopy, respectively. Five hundred sperm per slide were counted. Spermatozoa with fragmented DNA showed a large and spotty halo of chromatin dispersion. Unfragmented sperm only showed a small and compact halo. Statistical analyses were performed using the Statistical Package for Social Science version 12.0 (SPSS Inc., Chicago, IL, USA). The sperm DNA fragmentation index was compared between Wright and fluorescence staining methods by ANOVA. Results were expressed as mean ± standard error of the mean. The first report of the sperm DNA fragmentation index in canine ejaculates was 2.26 ± 0.53% for Wright staining and 1.99 ± 0.10% for fluorescence technique. No differences were found between staining procedures. In conclusion, it was possible to assess the sperm DNA fragmentation of canine ejaculates using 2 different staining procedures, expecting that continuous research could be useful in defining the role of DNA fragmentation SCD test in canine semen evaluation and cryopreservation.

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Utility and Predictive Value of Human Standard Semen Parameters and Sperm DNA Dispersion for Fertility Potential

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph16112004 ◽

2019 ◽

Vol 16 (11) ◽

pp. 2004 ◽

Cited By ~ 5

Author(s):

Kamil Gill ◽

Joanna Jakubik ◽

Aleksandra Rosiak-Gill ◽

Michał Kups ◽

Mariusz Lukaszuk ◽

...

Keyword(s):

Predictive Value ◽

Male Fertility ◽

Operating Characteristic ◽

Sperm Concentration ◽

Semen Parameters ◽

Sperm Dna Fragmentation ◽

Predictive Values ◽

Sperm Dna ◽

Semen Characteristics ◽

Fertility Potential

Because the assessment of sperm DNA fragmentation (SDF) plays a key role in male fertility, our study was designed to find the relationships between SDF and standard semen parameters. The receiver operating characteristic (ROC) curve showed that 18% SDF is a prognostic parameter for discriminating between men with normal and abnormal standard semen parameters (n = 667). Men with > 18% SDF had significantly lower quality semen, a higher prevalence of abnormal semen characteristics, and a higher odds ratio for abnormal semen parameters compared to men with ≤ 18% SDF. An ROC analysis provided predictive values for age and semen parameters to distinguish between men with SDF > 18% and men with ≤ 18% SDF. SDF was positively correlated with male age and teratozoospermia index but negatively with sperm concentration, total number of spermatozoa, sperm morphology, progressive motility, and vitality. Our study shows that 18% SDF has a predictive value for distinguishing between men with normal and abnormal semen characteristics. Men with >18% SDF have a higher risk for abnormal semen parameters, while age and obtained semen parameters have a predictive value for SDF. There is a relationship between SDF and conventional sperm characteristics, and thus, SDF can be incorporated into male fertility assessment.

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Environment and Male Fertility: Effects of Benzo-α-Pyrene and Resveratrol on Human Sperm Function In Vitro

Journal of Clinical Medicine ◽

10.3390/jcm8040561 ◽

2019 ◽

Vol 8 (4) ◽

pp. 561 ◽

Cited By ~ 6

Author(s):

Angela Alamo ◽

Rosita A. Condorelli ◽

Laura M. Mongioì ◽

Rossella Cannarella ◽

Filippo Giacone ◽

...

Keyword(s):

Protective Effect ◽

Sperm Motility ◽

Dose Response ◽

Superoxide Anion ◽

Male Fertility ◽

Negative Impact ◽

Sperm Parameters ◽

Mitochondrial Superoxide ◽

Chromatin Compactness

Lifestyle, cigarette smoking and environmental pollution have a negative impact on male fertility. Therefore, the aim of this study was to evaluate the in-vitro effects of benzo-α-pyrene (BaP) and aryl hydrocarbon receptor (AHR) agonists on motility and bio-functional sperm parameters. We further assessed whether resveratrol (RES), an AHR antagonist and antioxidant molecule, had any protective effect. To accomplish this, 30 normozoospermic, healthy, non-smoker men not exposed to BaP were enrolled. Spermatozoa of 15 men were incubated with increasing concentrations of BaP to evaluate its effect and to establish its dose response. Then, spermatozoa of the 15 other men were incubated with BaP (15 µM/mL), chosen according to the dose-response and/or RES to evaluate its antagonistic effects. The effects of both substances were evaluated after 3 h of incubation on total and progressive sperm motility and on the following bio-functional sperm parameters evaluated by flow cytometry: Degree of chromatin compactness, viability, phosphatidylserine externalization (PS), late apoptosis, mitochondrial membrane potential (MMP), DNA fragmentation, degree of lipoperoxidation (LP), and concentrations of mitochondrial superoxide anion. Benzo-α-pyrene decreased total and progressive sperm motility, impaired chromatin compactness, and increased sperm lipoperoxidation and mitochondrial superoxide anion levels. All these effects were statistically significant at the lowest concentration tested (15 µM/mL) and they were confirmed at the concentration of 45 µM/mL. In turn, RES was able to counteract the detrimental effects of BaP on sperm motility, abnormal chromatin compactness, lipid peroxidation, and mitochondrial superoxide. This study showed that BaP alters sperm motility and bio-functional sperm parameters and that RES exerts a protective effect on BaP-induced sperm damage.

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