Adsorption of DNA and Aptamers to Sodium Urate Crystals and Inhibition of Crystal Growth.

An elevated level of blood uric acid (UA) can cause the formation of kidney stones, gout, and other diseases. We recently isolated a few DNA aptamers that can selectively bind to UA. In this work, we investigated the adsorption of a UA aptamer and random sequence DNA onto sodium urate crystals. Both DNA strands adsorbed similarly to urate crystals. In addition, both the UA aptamer and random DNA can inhibit the growth of urate crystals, suggesting a nonspecific adsorption mechanism rather than specific aptamer binding. In the presence of 500 nM DNA, the growth of needle-like sodium urate crystals was inhibited, and the crystals appeared granular after 6 h. To understand the mechanism of DNA adsorption, a few chemicals were added to desorb DNA. DNA bases contributed more to the adsorption than the phosphate backbone. Surfactants induced significant DNA desorption. Finally, DNA could also be adsorbed onto real UA kidney stones. This study provides essential insights into the interactions between DNA oligonucleotides and urate crystals, including the inhibition of growth and interface effects of DNA on sodium urate crystals.

Locomotion of bovine spermatozoa during the transition from individual cells to bundles.

Various locomotion strategies employed by microorganisms are observed in complex biological environments. Spermatozoa assemble into bundles to improve their swimming efficiency compared to individual cells. However, the dynamic mechanisms for the formation of sperm bundles have not been fully characterized. In this study, we numerically and experimentally investigate the locomotion of spermatozoa during the transition from individual cells to bundles of two cells. Three consecutive dynamic behaviors are found across the course of the transition: hydrodynamic attraction/repulsion, alignment, and synchronization. The hydrodynamic attraction/repulsion depends on the relative orientation and distance between spermatozoa as well as their flagellar wave patterns and phase shift. Once the heads are attached, we find a stable equilibrium of the rotational hydrodynamics resulting in the alignment of the heads. The synchronization results from the combined influence of hydrodynamic and mechanical cell-to-cell interactions. Additionally, we find that the flagellar beat is regulated by the interactions during the bundle formation, whereby spermatozoa can synchronize their beats to enhance their swimming velocity.

Flagellar Propulsion of Sperm Cells Against a Time-Periodic Interaction Force.

Sperm cells undergo complex interactions with external environments, such as a solid-boundary, fluid flow, as well as other cells before arriving at the fertilization site. The interaction with the oviductal epithelium, as a site of sperm storage, is one type of cell-to-cell interaction that serves as a selection mechanism. Abnormal sperm cells with poor swimming performance, the major cause of male infertility, are filtered out by this selection mechanism. In this study, collinear bundles, consisting of two sperm cells, generate propulsive thrusts along opposite directions and allow to observe the influence of cell-to-cell interaction on flagellar wave-patterns. The developed elasto-hydrodynamic model demonstrates that steric and adhesive forces lead to highly symmetrical wave-pattern and reduce the bending amplitude of the propagating wave. It is measured that the free cells exhibit a mean flagellar curvature of 6.4 ± 3.5 rad mm-1 and a bending amplitude of 13.8 ± 2.8 rad mm-1 . After forming the collinear bundle, the mean flagellar curvature and bending amplitude are decreased to 1.8 ± 1.1 and 9.6 ± 1.4 rad mm-1 , respectively. This study presents consistent theoretical and experimental results important for understanding the adaptive behavior of sperm cells to the external time-periodic force encountered during sperm-egg interaction.

Bundle formation of sperm: Influence of environmental factors.

Cooperative behaviour of sperm is one of the mechanisms that plays a role in sperm competition. It has been observed in several species that spermatozoa interact with each other to form agglomerates or bundles. In this study, we investigate the effect of physical and biochemical factors that will most likely promote bundle formation in bull sperm. These factors include fluid viscosity, swim-up process, post-thaw incubation time and media additives which promote capacitation. While viscosity does not seem to influence the degree of sperm bundling, swim-up, post-thaw migration time and suppressed capacitation increase the occurrence of sperm bundles. This leads to the conclusion that sperm bundling is a result of hydrodynamic and adhesive interactions between the cells which occurs frequently during prolonged incubation times.

Drug-Loaded IRONSperm clusters: modeling, wireless actuation, and ultrasound imaging.

Individual biohybrid microrobots have the potential to perform biomedicalin vivotasks such as remote-controlled drug and cell delivery and minimally invasive surgery. This work demonstrates the formation of biohybrid sperm-templated clusters under the influence of an external magnetic field and essential functionalities for wireless actuation and drug delivery. Ferromagnetic nanoparticles are electrostatically assembled around dead sperm cells, and the resulting nanoparticle-coated cells are magnetically assembled into three-dimensional biohybrid clusters. The aim of this clustering is threefold: First, to enable rolling locomotion on a nearby solid boundary using a rotating magnetic field; second, to allow for noninvasive localization; third, to load the cells inside the cluster with drugs for targeted therapy. A magneto-hydrodynamic model captures the rotational response of the clusters in a viscous fluid, and predicts an upper bound for their step-out frequency, which is independent of their volume or aspect ratio. Below the step-out frequency, the rolling velocity of the clusters increases nonlinearly with their perimeter and actuation frequency. During rolling locomotion, the clusters are localized using ultrasound images at a relatively large distance, which makes these biohybrid clusters promising for deep-tissue applications. Finally, we show that the estimated drug load scales with the number of cells in the cluster and can be retained for more than 10 h. The aggregation of microrobots enables them to collectively roll in a predictable way in response to an external rotating magnetic field, and enhances ultrasound detectability and drug loading capacity compared to the individual microrobots. The favorable features of biohybrid microrobot clusters place emphasis on the importance of the investigation and development of collective microrobots and their potential forin vivoapplications.

Impact of Segmented Magnetization on the Flagellar Propulsion of Sperm-Templated Microrobots.

Technical design features for improving the way a passive elastic filament produces propulsive thrust can be understood by analyzing the deformation of sperm-templated microrobots with segmented magnetization. Magnetic nanoparticles are electrostatically self-assembled on bovine sperm cells with nonuniform surface charge, producing different categories of sperm-templated microrobots. Depending on the amount and location of the nanoparticles on each cellular segment, magnetoelastic and viscous forces determine the wave pattern of each category during flagellar motion. Passively propagating waves are induced along the length of these microrobots using external rotating magnetic fields and the resultant wave patterns are measured. The response of the microrobots to the external field reveals distinct flow fields, propulsive thrust, and frequency responses during flagellar propulsion. This work allows predictions for optimizing the design and propulsion of flexible magnetic microrobots with segmented magnetization.

IRONSperm: Sperm-templated soft magnetic microrobots.

We develop biohybrid magnetic microrobots by electrostatic self-assembly of nonmotile sperm cells and magnetic nanoparticles. Incorporating a biological entity into microrobots entails many functional advantages beyond shape templating, such as the facile uptake of chemotherapeutic agents to achieve targeted drug delivery. We present a single-step electrostatic self-assembly technique to fabricate IRONSperms, soft magnetic microswimmers that emulate the motion of motile sperm cells. Our experiments and theoretical predictions show that the swimming speed of IRONSperms exceeds 0.2 body length/s (6.8 ± 4.1 µm/s) at an actuation frequency of 8 Hz and precision angle of 45°. We demonstrate that the nanoparticle coating increases the acoustic impedance of the sperm cells and enables localization of clusters of IRONSperm using ultrasound feedback. We also confirm the biocompatibility and drug loading ability of these microrobots, and their promise as biocompatible, controllable, and detectable biohybrid tools for in vivo targeted therapy.

Sperm-Driven Micromotors Moving in Oviduct Fluid and Viscoelastic Media.

Biohybrid micromotors propelled by motile cells are fascinating entities for autonomous biomedical operations on the microscale. Their operation under physiological conditions, including highly viscous environments, is an essential prerequisite to be translated to in vivo settings. In this work, a sperm-driven microswimmer, referred to as a spermbot, is demonstrated to operate in oviduct fluid in vitro. The viscoelastic properties of bovine oviduct fluid (BOF), one of the fluids that sperm cells encounter on their way to the oocyte, are first characterized using passive microrheology. This allows to design an artificial oviduct fluid to match the rheological properties of oviduct fluid for further experiments. Sperm motion is analyzed and it is confirmed that kinetic parameters match in real and artificial oviduct fluids, respectively. It is demonstrated that sperm cells can efficiently couple to magnetic microtubes and propel them forward in media of different viscosities and in BOF. The flagellar beat pattern of coupled as well as of free sperm cells is investigated, revealing an alteration on the regular flagellar beat, presenting an on-off behavior caused by the additional load of the microtube. Finally, a new microcap design is proposed to improve the overall performance of the spermbot in complex biofluids.

Gelatin Microcartridges for Onboard Activation and Antioxidant Protection of Sperm.

Sperm-driven micromotors are promising devices for developing autonomous, noninvasive diagnostic and therapeutic tools, specifically in reproductive biology and medicine. In this article, we present template-based microstructures fabricated from gelatin for sperm capture, activation, and antioxidant protection. The cartridge-like structures with one closed end-capture single bovine sperm cells and are propelled by the sperm's flagellum. We demonstrate the pH-dependent release of heparin from the gelatin microcartridges to induce capacitation, a crucial maturation process prior to fertilization. Further, the gelatin cartridges exhibit a protection against oxidative stress, one of the main causes for sperm damage. Finally, we investigate the stability and degradability of the microstructures in various physiological environments, such as endometrial cell culture, different pH, and in the presence of proteases. Overall, we prove that these gelatin spermbots display great potential for the development of noninvasive theranostic tools in reproductive biology and medicine for the protection and activation of sperm, especially useful for studying sperm migration. We present their beneficial features which comprise not only biocompatibility and biodegradability but also pH response, loading stability, and antioxidant protection.

The motility-based swim-up technique separates bull sperm based on differences in metabolic rates and tail length.

Swim-up is a sperm purification method that is being used daily in andrology labs around the world as a simple step for in vitro sperm selection. This method accumulates the most motile sperm in the upper fraction and leaves sperm with low or no motility in the lower fraction. However, the underlying reasons are not fully understood. In this article, we compare metabolic rate, motility and sperm tail length of bovine sperm cells of the upper and lower fraction. The metabolic assay platform reveals oxygen consumption rates and extracellular acidification rates simultaneously and thereby delivers the metabolic rates in real time. Our study confirms that the upper fraction of bull sperm has not only improved motility compared to the cells in the lower fraction but also shows higher metabolic rates and longer flagella. This pattern was consistent across media of two different levels of viscosity. We conclude that the motility-based separation of the swim-up technique is also reflected in underlying metabolic differences. Metabolic assays could serve as additional or alternative, label-free method to evaluate sperm quality.

Nano-and Micromotors Designed for Cancer Therapy.

Research on nano- and micromotors has evolved into a frequently cited research area with innovative technology envisioned for one of current humanities' most deadly problems: cancer. The development of cancer targeting drug delivery strategies involving nano-and micromotors has been a vibrant field of study over the past few years. This review aims at categorizing recent significant results, classifying them according to the employed propulsion mechanisms starting from chemically driven micromotors, to field driven and biohybrid approaches. In concluding remarks of section 2, we give an insight into shape changing micromotors that are envisioned to have a significant contribution. Finally, we critically discuss which important aspects still have to be addressed and which challenges still lie ahead of us.

Sperm-Particle Interactions and Their Prospects for Charge Mapping.

In this article, a procedure to investigate sperm charge distribution by electrostatic sperm-particle interactions is demonstrated. Differently chargedparticles are fabricated and their attachment distribution on the bovine sperm membrane is investigated. The sperm-particle attachment sites are observed using bright field and cryo-scanning electron microscopy combined with energy-dispersive X-ray analysis. The findings suggest that the charge distribution of the sperm membrane is not uniform, and although the overall net charge of the sperm cell is negative, positively charged areas are especially found on the sperm heads. The newly developed method is used to investigate the dynamic charge distribution of the sperm cell membrane upon maturation induced by heparin, as a representation of the multitude of changes during the development of a sperm.

Characterization of Flagellar Propulsion of Soft Microrobotic Sperm in a Viscous Heterogeneous Medium.

Several microorganisms swim by a beating flagellum more rapidly in solutions with gel-like structure than they do in low-viscosity mediums. In this work, we aim to model and investigate this behavior in low Reynolds numbers viscous heterogeneous medium using soft microrobotic sperm samples. The microrobots are actuated using external magnetic fields and the influence of immersed obstacles on the flagellar propulsion is investigated. We use the resistive-force theory to predict the deformation of the beating flagellum, and the method of regularized Stokeslets for computing Stokes flows around the microrobot and the immersed obstacles. Our analysis and experiments show that obstacles in the medium improves the propulsion even when the Sperm number is not optimal (S p ≠ 2.1). Experimental results also show propulsion enhancement for concentration range of 0-5% at relatively low actuation frequencies owing to the pressure gradient created by obstacles in close proximity to the beating flagellum. At relatively high actuation frequency, speed reduction is observed with the concentration of the obstacles.

Sperm-Hybrid Micromotor for Targeted Drug Delivery.

A sperm-driven micromotor is presented as a targeted drug delivery system, which is appealing to potentially treat diseases in the female reproductive tract. This system is demonstrated to be an efficient drug delivery vehicle by first loading a motile sperm cell with an anticancer drug (doxorubicin hydrochloride), guiding it magnetically, to an in vitro cultured tumor spheroid, and finally freeing the sperm cell to deliver the drug locally. The sperm release mechanism is designed to liberate the sperm when the biohybrid micromotor hits the tumor walls, allowing it to swim into the tumor and deliver the drug through the sperm-cancer cell membrane fusion. In our experiments, the sperm cells exhibited a high drug encapsulation capability and drug carrying stability, conveniently minimizing  toxic side effects and unwanted drug accumulation in healthy tissues. Overall, sperm cells are excellent candidates to operate in physiological environments, as they neither express pathogenic proteins nor proliferate to form undesirable colonies, unlike other cells or microorganisms. This sperm-hybrid micromotor is a biocompatible platform with potential application in gynecological healthcare, treating or detecting cancer or other diseases in the female reproductive system.

Spermatozoa as Functional Components of Robotic Microswimmers.

In recent years, the combination of synthetic micro- and nanomaterials with spermatozoa as functional components has led to the development of tubular and helical spermbots - microrobotic devices with potential applications in the biomedical and nanotechnological field. Here, the initial advances in this field are discussed and the use of spermatozoa as functional parts in microdevices elaborated. Besides the potential uses of these hybrid robotic microswimmers, the obstacles along the way are discussed, with suggestions for solutions of the encountered challenges also given.

Dynamic Polymeric Microtubes for the Remote-Controlled Capture, Guidance, and Release of Sperm Cells.

Remote-controlled release of single sperm cells is demonstrated by the use of polymeric microtubes that unfold upon temperature increase to 38 °C. Thermoresponsive, ferromagnetic multilayers are tailored to catch sperm cells and remotely control them by external magnetic fields. These polymeric spermbots are propelled by the sperm flagella. When the temperature is increased, the tubes unfold and the cell is set free.

Sperm dynamics in tubular confinement.

An on-chip system that mimics tubular microenvironments is presented for the study of spermatozoa motion in confinement. Using rolled up transparent silicon oxide/dioxide microtubes, the influence of tube diameter on the velocity, directionality, and linearity of spermatozoa is investigated. Tubular microenvironments of diameters 20-45 µm facilitate sperm migration through channels.

Effect of surfactants on the performance of tubular and spherical micromotors - a comparative study.

The development of artificial micromotors is one of the greatest challenges of modern nanotechnology. Even though many kinds of motors have been published in recent times, systematic studies on the influence of components of the fuel solution are widely missing. Therefore, the autonomous movement of Pt-microtubes and Pt-covered silica particles is comparatively observed in the presence and absence of surfactants in the medium. One representative of each of the three main surfactant classes - anionic (sodium dodecyl sulfate, SDS), cationic (benzalkonium chloride, BACl) and non-ionic (Triton X) - has been chosen and studied.

Propulsion Mechanism of Catalytic Microjet Engines.

We describe the propulsion mechanism of the catalytic microjet engines that are fabricated using rolled-up nanotech. Microjets have recently shown numerous potential applications in nanorobotics but currently there is a lack of an accurate theoretical model that describes the origin of the motion as well as the mechanism of self-propulsion. The geometric asymmetry of a tubular microjet leads to the development of a capillary force, which tends to propel a bubble toward the larger opening of the tube. Because of this motion in an asymmetric tube, there emerges a momentum transfer to the fluid. In order to compensate this momentum transfer, a jet force acting on the tube occurs. This force, which is counterbalanced by the linear drag force, enables tube velocities of the order of 100 µm/s. This mechanism provides a fundamental explanation for the development of driving forces that are acting on bubbles in tubular microjets.

Spermbots: potential impact for drug delivery and assisted reproductive technologies.

Micromotors and nanomotors are an emerging research field that aims at achieving locomotion on the microscale for a variety of applications such as drug delivery, single cell manipulation, microsensors and lab-on-a-chip devices, just to point out a few. The enthusiastic development of hybrid micromotors harnessing biological power sources for physiologically compatible nano/microdevices has recently brought a lot of attention to the international research community that is looking for a solution for the actuation and locomotion on the microscale. This article describes the potential of sperm-driven micro-bio-robots in the biomedical field such as drug delivery or single cell manipulation. Herein, a specific potential of the sperm-driven micro-bio-robot is described that might have impact on the development of assisted reproductive technologies.