35,575 research outputs found
In Vitro Effect of Gold or Silver Nanoparticles on Meiotic and Postmeiotic Fractions of Rat Germinal Cells
No full textThe cytotoxicity of Au or Ag nanoparticles (NPs) on rat germinal cells was investigated in vitro. Rat germ cells separated by the STAPUT method in two different populations, pachytene spermatocytes (F5) and round spermatids (F3), were incubated (37°C, 60'-120') with 60 μM, 125 μM, 250 μM and 500 μM of Au/Ag-NPs. Cell viability was assessed with the Eosin Y test. Au or Ag-NPs were investigated with FEG-STEM/EDS and TEM. A dose-dependent effect on the viability of F3 and F5 populations was observed after incubation with Au-NPs or Ag-NPs (P<0.001). A significant decrease in cell viability was observed in F5 fractions compared to F3 fractions (P<0.05), except for the samples treated with 60μM of Au-NPs, and the decrease was also significant in all the samples treated with Ag-NPs compared to those incubated with Au-NPs (P< 0.05). Au-NPs were localized in spermatocytes and spermatids whereas Ag-NPs were undetectable. In conclusion, Au- NPs and Ag-NPs seem to exert a negative effect on rat germ cells, particularly on spermatocytes, that appeared significantly more compromised than spermatids. Further research is needed, mainly to carefully explore the possible genotoxicity of these NPs on germinal cells. © Collodel et al
Immunolocalization of aquaporin 7 in human sperm and its relationship with semen parameters
No full textAquaporins (AQPs) are a family of 13 small hydrophobic trans-membrane proteins expressed in numerous tissues and cells. Some AQPs work as strict water channels, others are permeable to a range of substances, including glycerol. In the male reproductive system their localization in testis, efferent ducts, epididymis, and spermatozoa has been described. We studied the distribution of AQP7 in ejaculated human sperm and the relationship between AQP7 labeling and sperm characteristics. Semen samples from 33 men were examined by light and transmission electron microscopy (TEM). TEM data were quantified using a mathematical formula that calculates a fertility index (FI) and the percentages of sperm apoptosis, immaturity, and necrosis. Immunocytochemistry with a polyclonal antibody anti-AQP7 was performed on the sperm samples. Normal sperm were labeled in the pericentriolar area, midpiece, equatorial segment, and weakly in the tail (grade 1). Abnormal sperm showed a diffuse low intensity of fluorescence evident in the cytoplasmic residues, coiled tails, in the entire head, and acrosome (grade 2). A high number of motile sperm obtained by swim up were labeled in a dotted manner in the mitochondria. A significant positive correlation was found between the spermatozoa with AQP7 grade 1 labeling and the percentage of normal form (P<0.008), progressive motility and FI (P<0.005); a negative correlation was noted with the percentages of cytoplasmic residues (P<0.010) and immaturity (P<0.006) and coiled tails (P<0.012). The link between AQP7 distribution and sperm morphology and the particular dotted labeling in swim up selected motile sperm are novel and deserve additional studies. © Copyright ©2012 Informa Healthcare USA, Inc
Semen characteristics and malondialdehyde levels in men with different reproductive problems
No full textOxidative stress is an important factor in the aetiology of poor sperm function, inducing morphological alterations and oxidative damage to DNA, membrane and proteins. Previous studies reported that malondialdehyde (MDA), the end product in membrane lipid peroxidation, correlates with abnormal seminal parameters (Atig et al., 2012; Benedetti et al. 2012). In this study MDA was dosed in semen of men who had consulted our centre to know their seminal status. They had a normal 46,XY karyotype evaluated by conventional cytogenetic analysis, no history of radiotherapy and chemotherapy. They were subjected to microbiological analysis of semen specimen and urethral fluid to verify the presence of genitourinary infections, to clinical and physical examinations and scrotal Eco-color Doppler to investigate the presence of varicocele or other anatomical problems. MDA levels in seminal plasma was determined by spectrofluorometry. Semen samples were examined by light microscopy according WHO guidelines (2010) and transmission electron microscopy (TEM). TEM data were quantified with a mathematical formula furnishing a fertility index and the percentage of sperm apoptosis, immaturity and necrosis (Collodel and Moretti 2008). On the basis of the presence of reproductive problems men were divided in three groups: men with genitourinary infections (group I), men with varicocele (group II) and individuals without varicocele and infections (group III); in each group we evaluated semen quality (concentration, motility, percentage of apoptosis, necrosis, immaturity and fertility index) and MDA levels in seminal plasma. Sperm concentration was comprised between 10 centile and 25 centile in groups II and III, between 25 centile and 50 centile in group I. Progressive sperm motility appeared similar in all examined groups (around 5 percentile). TEM analysis indicated that all the groups had fertility index lower than controls. Regarding sperm pathologies, the percentage of apoptosis was higher in all groups compared to control (group I: 7.3%, group II: 7.8%, group III: 8.5% vs 4.05%), the immaturity percentage was significantly elevated in group II (70.34%) compared to control (48.83%) and the other groups (group I: 49.3%; group III: 58.6%; p<0.001) and necrosis in group I (57.53%) compared to control (32.13%) and group II and III (respectively 24.99%, 24.95%; p<0.001). MDA concentration in group I was significantly higher than in group II (4.05 vs 1.70; p<0.001) and group III that showed level significantly lower than group II (0.62 vs 1.70; p<0.001). A negative correlation was observed between MDA levels and progressive sperm motility in group I (r=-0.58). Regarding sperm pathologies evaluated by TEM, MDA concentrations were positively correlated with necrosis in groups I and II (group I: r=0.68; group II: r= 0.61). These preliminary data indicate that MDA levels were significantly elevated in patients with genitourinary infections (group I) compared those measured in group II and group III. Moreover, group I had significantly higher levels of necrosis but this alteration was correlated with MDA also in groups II and III. Necrosis is a pathology characterised by disrupted chromatin, disassembled axonemal and periaxonemal structures, swollen mitochondria and broken plasma membrane. Also other authors reported that MDA and sperm decondensation are significantly linked (Montjean et al. 2010), and that MDA negatively correlated with sperm motility (Zribi et al., 2011). The role of MDA in semen of men with reproductive problems deserve further attention
Cytokines release and oxidative status in semen samples from rabbits treated with bacterial lipopolysaccharide
No full textThis study was aimed to evaluate the effects of a lipopolysaccharide- (LPS) induced inflammation on cytokines release and oxidative status of semen samples from buck rabbits at different times after treatment. Semen analysis was performed by optical microscopy and sperm motility evaluation by the computer-assisted sperm analyzer. The presence of activated macrophages and apoptotic/necrotic sperm was evaluated by fluorescent microscopy. A panel of cytokines, interleukin (IL)-6, IL-8, IL-1 beta, and tumor necrosis factor-alpha, were detected and quantified in seminal plasma using the Bio-Plex Cytokine assay. Reactive oxygen metabolite and thiobarbituric acid reactive substance determinations were carried out by spectrophotometry and tocopherol analysis by high performance liquid chromatography. The sperm motility and track speed were reduced in LPS-treated rabbits. The activated macrophages in LPS-treated buck rabbits significantly increased from 0.50 x 10(6)/mL (baseline) to 27 x 10(6)/mL on Day 21; successively, there was a progressive reduction. Apoptotic and necrotic sperm in LPS rabbits followed more or less the same trend. The reactive oxygen metabolite levels in semen from LPS-treated rabbits showed higher values compared with those evaluated in controls, particularly during the lag time, Days 1 to 3. The sperm thiobarbituric acid reactive substances highlighted a peak in LPS-treated rabbits compared with those of controls on Day 1 after LPS treatment, and the different T isoforms (alpha, delta, and gamma+beta) showed a similar trend with a significant decrease on Day 1 after injection and a recovery on Days 30 to 56. Until Days 3 to 21 from the treatment, higher levels of IL-1 beta and tumor necrosis factor-alpha were detected in seminal plasma of LPS-treated rabbits. Interleukin-6 showed a peak on Day 3 after LPS treatment, and on Day 7, the value was similar to the control group. In conclusion, this study confirms that the buck rabbit is a good model for mimicking and understanding the inflammation mechanisms, which may induce male infertility, in particular that a systemic inflammatory status causes alterations to the sperm cells through a shift in the balance between the oxidant and antioxidant systems.
Are F2-isoprostanes a better marker of semen lipid peroxidation than MDA in reproductive pathologies with inflammatory basis?
No full textMany male reproductive pathologies and a part of undiagnosed infertility share an oxidative stress (OS) etiology with high reactive oxygen species and cytokine concentrations. The lack of reliable biomarkers to quantify oxidative injury is a crucial problem in the field of male infertility. In this observational study, IL-1 beta and the OS markers malondialdehyde (MDA) and F-2-isoprostanes (F-2-IsoPs) were quantified in seminal plasma of 46 infertile patients with varicocele, genitourinary infections, idiopathic infertility, and 11 fertile men. Semen analysis was performed following WHO guidelines, IL-1 beta was determined by ELISA, MDA was quantified by HPLC, and F-2-IsoPs by GC/NICI-MS analysis. F-2-IsoPs were immunolocalized in spermatozoa of fertile and infertile subjects. Results indicated that F-2-IsoP, MDA, and IL-1 beta seminal levels positively correlated pairwise (p < 0.001) and showed negative correlations with sperm parameters (p < 0.001). Then, the studied population was grouped following the cause of infertility and the variables were compared between the different groups and a control sample. Seminal IL-1 beta, F-2-IsoPs, and MDA were significantly higher in varicocele (p < 0.001, for MDA p < 0.01) and genitourinary infections (p < 0.001, for IL-1 beta p < 0.01) groups than those observed in fertile subjects. F-2-IsoPs seemed to discriminate more accurately than MDA the different conditions, in particular idiopathic infertility. ROC curves demonstrated that the three analyzed indices were able to discriminate fertile and infertile patients. The immunofluorescence studies showed a low presence of F-2-IsoPs in spermatozoa of fertile men and an evident labeling in the tail, and cytoplasmic residues of spermatozoa from infertile patients. In conclusion, this data confirmed that F-2-IsoP level is a suitable marker of OS in seminal plasma, even more accurate than MDA and can be proposed for measuring OS in the clinical setting. © 2025 The Author
In Vitro Effects of Charged and Zwitterionic Liposomes on Human Spermatozoa and Supplementation with Liposomes and Chlorogenic Acid during Sperm Freezing
No full textSemen handling and cryopreservation induce oxidative stress that should be minimized. In this study, human semen was supplemented during cryopreservation with formulations of handmade liposomes and chlorogenic acid (CGA), an antioxidant compound. Zwitterionic (ZL), anionic (AL), and cationic (CL) liposomes were synthesized and characterized. Three aliquots of swim-up-selected sperm were incubated with ZL, AL, and CL (1:10,000), respectively. The percentages of sperm with progressive motility, high mitochondrial membrane potential (MMP; JC-1), double-stranded DNA (dsDNA acridine orange), and acrosome integrity (Pisum sativum agglutinin) were assessed. Then, human semen was frozen using both 1:10,000 ZL and CGA as follows: freezing medium/empty ZL (EL), freezing medium/empty ZL/CGA in the medium (CGA + EL), freezing medium/CGA loaded ZL (CGA), freezing medium (CTR). The same sperm endpoints were evaluated. ZL were the most tolerated and used for semen cryopreservation protocols. All the supplemented samples showed better endpoints versus CTR (p < 0.001). In particular, spermatozoa from the CGA and CGA + EL A samples showed increased motility, dsDNA, and acrosome integrity versus CTR and EL (p < 0.001; motility EL vs. CGA + EL p < 0.05). ZL and CGA can improve post-thaw sperm quality, acting on both cold shock effect management and oxidative stress. These findings open new perspectives on human and animal reproduction
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