Oxidative Stress and Acrosomal Status of Human Spermatozoa Subjected to Hydrophobic Carbon Soot Treatments.

The fourth industrial revolution extensively reshapes the reality we are living in by blurring the boundaries of physical, digital and biological worlds. A good example is the previously unthinkable incursion of nanoscale waste materials, such as soot, into the technologies for assisted reproduction. Although the rapeseed oil soot may efficiently enhance the progressive motility of human spermatozoa, it is yet unknown whether this material induces undesirable oxidative stress and premature acrosome reaction, endangering the sperm-oocyte fusion and blastocyst formation. In an attempt to clarify this issue, we reveal that the three-hour incubation of human semen mixed with three main types of soot does not cause oxidative stress and spontaneous acrosome reaction of the sperm. These unique findings are attributed to synchronous elimination and stabilization of the oxidants via hydrogen bonding to the acidic groups of the soot (i.e., C=O and/or C-O-C) and electron donation by its basic chemical sites (i.e., C-OH and/or COOH). Moreover, the soot nanoparticles are electrostatically attracted by discrete positively charged areas on the sperm head, increasing its negative charge and in some cases interfering the acrosome reaction. Such novel mechanistic insights emphasize the credibility of rapeseed oil soot to confidently shift from the purely diagnostic and therapeutic phases in reproductive medicine to research dealing with the effect of carbon nanomaterials on the embryo development and implantation.

Cryopreservation of human semen by inherently-controlled icing probability: Or how the surface profile of superhydrophobic carbon soot coatings and the sperm volume affect the outcome of slow freezing?

The restoration of initial functionality of human spermatozoa subjected to cryopreservation is challenging, because the deleterious intracellular icing and the occurrence of osmotic shocks due to prolonged exposure to increased concentrations of intracellular solutes are oppositely dependent on the cooling rate. This longstanding problem could be overcome if using superhydrophobic soot coatings delaying the heat transfer rate, reducing the ice formation probability and triggering balanced and timely dehydration of the cells, but the effect of their surface profile and sperm volume on the success rate of slow freezing is unclear. Here, we show for the first time that the two-factor freezing injury is entirely avoidable by tailoring the solid-to-gas voids (pores) fraction in the soot, leading to increased nucleation free energy barrier, presumable incipiency of ice crystals with controllable shape and size and hence, fully (100 %) recovered post-thaw sperm motility. It is demonstrated that the reason for such a unique scientific result is the selection of soot coatings with appropriate morphochemical features, hypothetically (not directly proven yet) inducing equilibrium among the solution composition and ice crystals formation, retarding the undesirable compression of liquid-filled "slush ice" channels surrounding the cytoplasm and impeding the ice recrystallization. The novel insights introduced in this article open endless horizon for customizing and revolutionizing the technical protocols in cryobiology.

Manipulated sperm motility via soot nanoparticles-induced biochemical alterations in human seminal plasma.

Obtaining spermatozoa with progressive motility, via postejaculatory activation with pharmacological agents such as theophylline and pentoxifylline, is crucial for the success rate of assisted reproduction in couples with severe male factor infertility. Regrettably, the possibility of premature acrosome reactions and impared oocyte function questions the practical applicability of phosphodiesterase inhibitors. The rapid development of nanotechnologies promotes the use of hydrophobic rapeseed oil soot as a non-cytotoxic biomaterial for sperm motility activation, but the scarcity of knowledge regarding the interactions of soot with components from the seminal plasma hinders the eventual commercialization of this cutting-edge approach. Aiming to eliminate this shortcoming, the current study shows for the first time how the soot nanomaterials alter the biochemistry of human seminal plasma. Upon 270 min incubation with soot nanoparticles, the activity of AST, ALT, CK, LDH and GGT enzymes in the seminal plasma of ten patients changes inversely to the registered sperm motility (i.e., lower enzyme activity, higher sperm motility and vice versa). This phenomenon is primarily related to termination of the enzymes-substrate binding or extraction of enzymes from the gametes via chemical bonding with the soot. These novel mechanisms depend on the physicochemical features of used carbon nanomaterials, revealing opportunities for predictable tuning of the sperm reproductive potential.

Hydrophobic soot nanoparticles as a non-cytotoxic motility activator of human spermatozoa.

Sperm cryopreservation is vital in combating the human infertility, but regrettably, the toxicity of cryoprotectants and the occurrence of intracellular icing, osmotic shocks or shrinkage of the cells below a given threshold volume greatly affects the success rate of this technique. Using the virtue of nanotechnologies and depositing water-repellent soot nanoparticles on the inner walls of cryovials may outline new directions in the development of cryobiology, but doubts related to the soot's venomosity question its practical implementability. The scientific content of this article eliminates the existing apprehensions by analyzing the cytotoxicity of three types of rapeseed oil soot, differing in morphology, surface chemistry and zeta potential, towards human spermatozoa. Upon intermittent evaluations of the sperm motility within 270 min of incubation in vials comprising carbon nanoparticles, we reveal that this soot category is non-cytotoxic or at worst, faintly toxic to the gametes provided by twenty individuals. Enhanced progressive sperm motility is observed at ∼50-60% of patients following the soot treatments, which is attributed to electrostatic repulsions and biochemical alterations in the seminal plasma. These fascinating results open new horizons for incorporation of the rapeseed oil soot as a tool for functional preparation and activation of human spermatozoa preceding in vitro fertilization.

On the Development of Ultradurable Extremely Water-Repellent and Oleophobic Soot-Based Fabrics with Direct Relevance to Sperm Cryopreservation.

Nowadays, the tremendous progress of nanotechnologies and materials science facilitates the fabrication of universal and multifunctional superhydrophobic surfaces on a large scale. Yet, integrating icephobic and anti-bioadhesive properties in an individual water-repellent functional coating, for addressing the difficulties faced by cryobiologists, aircraft, and seacraft manufacturers, is quite tricky but feasible if using nonpolar soot nanoparticles, whose fragility, however, impedes their industrial applicability. Here, we advance the current state-of-the-art to an extent, permitting the introduction of economically affordable and ultradurable non-wettable soot-based coatings. The deposition of rapeseed oil soot, cyanoacrylate glue and fluorine compounds onto different fabrics confers the latter with superior tolerance to harsh mechanical and thermal interventions [e.g., scratching, blade scraping, liquid nitrogen immersion (T ∼ -196 °C), torsion and water jetting], while in the meantime retaining water repellency and oleophobicity. The as-prepared soot fabrics can stick continuously to the selected host surface and favor the recovery of ∼60% of the initial motility of human spermatozoa subjected to cryopreservation or being detached and utilized as standalone non-wettable membranes. Our invention may be considered as the first fundamental stage of safely (without any health concerns) transferring the soot in reproductive medicine and developing enhanced cryogenic and antibacterial medical devices.

When condensed matter physics meets biology: Does superhydrophobicity benefiting the cryopreservation of human spermatozoa?

With the increasing demand in regenerative and reproductive medicine for successful conservation of living matter, the need of reliable platform in cell banking seems inevitable. Whilst the cells storage at cryogenic temperatures is a well-developed method, far less is known about the efficiency of nanotechnology in cryogenics. The primary objective of this study is to represent the first of its kind experimental results related to cryopreservation of human spermatozoa by means of superhydrophobic carbon soot coatings. The inclusion of soot-based water repellent interface during the freezing and thawing of human semen minimizes the solid-liquid interfacial area, retards the heat transfer rate and promotes the recovery of up to 80% of initial motility of post-thaw sperm cells. Our discoveries reveal a fundamentally new and exciting direction of development of cryopreservation technologies in the battle against painful biopsies and repetitive surgeries.

Superhydrophobic Soot Coated Quartz Crystal Microbalances: A Novel Platform for Human Spermatozoa Quality Assessment.

The functionality of human spermatozoa is a key factor for the success rate of natural human reproduction, but unfortunately the infertility progressively increases due to multifarious environmental factors. Such disquieting statistics requires the employment of sophisticated computer-assisted methods for semen quality analysis, whose precision, however, is unreliable in cases of patients with low sperm concentrations. In this study, we report a novel quartz crystal microbalance (QCM) based biosensor for in-situ quality assessment of male gametes, comprising a superhydrophobic soot coating as an interface sensing material. The soot deposition on the surface of a 5 MHz QCM eliminates the noise that normally arises upon immersion of the uncoated sensor in the test liquid environment, allowing the detection of human spermatozoa down to 1000⁻100,000 units/mL (1⁻100 ppb). Furthermore, the soot coated QCM delimitates in a highly repeatable way the immotile and motile sperm cells by inducing fundamentally distinct responses in respect to sensor sensitivity and signal trends. The obtained results reveal the strong potential of the superhydrophobic QCM for future inclusion in diverse laboratory analyses closely related to the in vitro fertilization procedures, with a final aim of gaining practical approaches for diagnoses and selection of male gametes.