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The synthesis of iron oxide nanoparticles (IONPs)–antiubiquitin antibodies (Abs) complex for depletion of dead/damaged spermatozoa from buffalo semen was done. The IONPs synthesized were round in shape with size of 12.09 ± 0.91 nm. At the end of the two‐step functionalization, that is, silanization and pegylation of bare IONPs and bioconjugation of functionalized IOPNs, particles with the sizes of 19.15 ± 1.46, 20.72 ± 0.95, and 73.01 ± 7.56 nm, respectively, were obtained. Twenty‐four semen samples from four bulls with mean individual progressive motility (%) and sperm concentration (million/mL) of 77.1 ± 0.9 and 1,321.2 ± 84.7, respectively, were divided into Group I (control), and treatment groups viz. Groups II, III, and IV; with each group containing 150 ± 25 million dead/damaged spermatozoa. The IONPs–Abs complex was added at the ratio of 1:1 (0.5 μg/mL), 1:2 (1.0 μg/mL), and 1:4 (2.0 μg/mL), respectively, in the Groups II, III, and IV. The mean efficiency (%) of nanopurification was estimated to be greater in nanopurified semen with the increasing doses of the IONPs–Abs complex. A reduction of 29.3 ± 6.4%, 48.4 ± 5.3%, and 55.4 ± 4.4% in dead/damaged spermatozoa following nanopurification in Groups II, III, and IV, respectively, was observed. The study shows that in‐house synthesized IONPs–Abs complex can be successfully used to deplete dead/damaged spermatozoa from buffalo semen with improvement in quality. [ABSTRACT FROM AUTHOR]

Since ensuring stability of the nanoparticles in aqueous-phase is essential for biomedical applications, this study evaluated the effect of ultra-sonication followed by peptization on iron-oxide nanoparticles (IONPs) aqueous-phase stability. The IONPs synthesized via two different chemical co-precipitation methods were ultra-sonicated for −5, −10 and 20-min and evaluated using Dynamic light scattering at Day-1, -2, -7 and -15 of the synthesis. The Experiment II-IONPs had lesser hydrodynamic size with better aqueous-phase stability over the time. The decrease in the average size and increase in homogeneity of solution was better following ultra-sonication for 20-min. These IONPs were peptized with perchloric acid to evaluate the effect on stability and longevity. The results showed large proportion of IONPs less than 100 nm even on day-15 post-peptization. Moreover, transmission electron microscopy revealed average size of 11.23 nm with round shape. The study showed positive influence of peptization on the aqueous-phase stability of the IONPs. [ABSTRACT FROM AUTHOR]

Despite recent advances in technique of spermatozoa cryopreservation, there are still ejaculates present that fail to meet strict quality standard; mainly due to detrimental effect of imbalance of free radicals. The omnipresence of dead/defective spermatozoa in ejaculates of eutherian species is a major source of excessive free radicals. Though sperm-selection techniques, as well as addition of antioxidants addressed the problem to a certain extent, the major source of free radicals in the semen remained, causing much damage. This study attempts to remove dead/damaged spermatozoa using negative fertility-marker. The effect is unraveled by Hypo-osmotic (HOS), and fluorescein-conjugated Pisum sativum agglutinin (FITC-PSA) assay, further confirmed by Ca2+-regulating mechanisms and depolarization of sperm membrane potential, reduction in concentration of free radicals and finally by in vitro fertility assay. The study involved functionalization of iron oxide nanoparticles (IONPs) with silane followed by bio-conjugation with anti -ubiquitin antibodies. The nano-purification of semen using anti -ubiquitin conjugated iron oxide nanoparticles (IONPs) (antibody concentrations 0.5, 1.0 and 2.0 μg/ml) was attempted. The efficiency of nano-purification was 18.1%–43.8% in the study. The results revealed greater (P ≤ 0.05) spermatozoa population with intact plasma membrane, acrosome integrity, high mitochondrial membrane potential and pattern-F (least intracellular Ca2+), evidence of low lipid peroxidation and higher total antioxidant capacity in nano-purified groups. More number of spermatozoa were bound to zona pellucida of matured oocytes from nano-depleted than non-depleted group. The findings demonstrate antibody concentration of 1.0 μg/ml bio-conjugated with IONPs as most efficient in enriching the ejaculate with functional spermatozoa with the highest percentage of zona binding. • Nano-purification improves semen quality in buffalo. • Removal of dead and damaged spermatozoa reduces the oxidative stress level in semen. • Nano-purification improves the fertility of semen. [ABSTRACT FROM AUTHOR]