Spermbots and Their Applications in Assisted Reproduction: Current Progress and Future Perspectives.

Sperm quality is declining dramatically during the past decades. Male infertility has been a serious health and social problem. The sperm cell driven biohybrid nanorobot opens a new era for automated and precise assisted reproduction. Therefore, it is urgent and necessary to conduct an updated review and perspective from the viewpoints of the researchers and clinicians in the field of reproductive medicine. In the present review, we first update the current classification, design, control and applications of various spermbots. Then, by a comprehensive summary of the functional features of sperm cells, the journey of sperms to the oocyte, and sperm-related dysfunctions, we provide a systematic guidance to further improve the design of spermbots. Focusing on the translation of spermbots into clinical practice, we point out that the main challenges are biocompatibility, effectiveness, and ethical issues. Considering the special requirements of assisted reproduction, we also propose the three laws for the clinical usage of spermbots: good genetics, gentle operation and no contamination. Finally, a three-step roadmap is proposed to achieve the goal of clinical translation. We believe that spermbot-based treatments can be validated and approved for in vitro clinical usage in the near future. However, multi-center and multi-disciplinary collaborations are needed to further promote the translation of spermbots into in vivo clinical applications.

G-Quadruplex-Programmed Versatile Nanorobot Combined with Chemotherapy and Gene Therapy for Synergistic Targeted Therapy.

Developing synergistic targeted therapeutics to improve treatment efficacy while reducing side effects has proven promising for anticancer therapies, but how to conveniently modulate multidrug cooperation remains a challenge. Here, a novel synergistic strategy using a G-quadruplex-programmed versatile nanorobot (G4VN) containing two subunits of DNAzyme (DzG4) and ligand-drug conjugates (LDCs) is proposed to precisely target tumors and then execute both gene silencing and chemotherapy. As the core module of this nanorobot, a well-designed G4 responding to a high level of K+ in tumor microenvironment smartly kills three birds with one stone, which makes two TfR aptamers proximate to improve their efficiency of targeting tumor cells, and in situ activates a split 10-23 DNAzyme to downregulate target mRNA expression, meanwhile promotes the cell uptake of a GSH-responsive LDCs to enhance drug efficacy. Such a design enables a potently synergistic anticancer therapy with low side effects in vivo, showing great promise for broad applications in precision disease treatment.

Magnetically actuated sonodynamic nanorobot collectives for potentiated ovarian cancer therapy.

Ovarian cancer presents a substantial challenge due to its high mortality and recurrence rates among gynecological tumors. Existing clinical chemotherapy treatments are notably limited by drug resistance and systemic toxic side effects caused by off target drugs. Sonodynamic therapy (SDT) has emerged as a promising approach in cancer treatment, motivating researchers to explore synergistic combinations with other therapies for enhanced efficacy. In this study, we developed magnetic sonodynamic nanorobot (Fe3O4@SiO2-Ce6, FSC) by applying a SiO2 coating onto Fe3O4 nanoparticle, followed by coupling with the sonosensitizer Ce6. The magnetic FSC nanorobot collectives could gather at fixed point and actively move to target site regulated by magnetic field. In vitro experiments revealed that the magnetic FSC nanorobot collectives enabled directional navigation to the tumor cell area under guidance. Furthermore, under low-intensity ultrasonic stimulation, FSC nanorobot collectives mediated sonodynamic therapy exhibited remarkable anti-tumor performance. These findings suggest that magnetically actuated sonodynamic nanorobot collectives hold promising potential for application in target cancer therapy.

An FOF1-ATPase motor-embedded chromatophore as a nanorobot for overcoming biological barriers and targeting acidic tumor sites.

Despite the booming progress of anticancer nanomedicines in the past two decades, precise tumor-targetability and sufficient tumor-accumulation are less successful and still require further research. To tackle this challenge, herein we present a biomolecular motor (FOF1-ATPase)-embedded chromatophore as nanorobot to efficiently overcome biological barriers, and thoroughly investigate its chemotactic motility, tumor-accumulation ability and endocytosis. Chromatophores embedded with FOF1-ATPase motors were firstly extracted from Thermus thermophilus, then their properties were fully characterized. Specifically, two microfluidic platforms (laminar flow microchip and tumor microenvironment (TME) microchip) were designed and developed to fully investigate the motility, tumor-accumulation ability and endocytosis of the chromatophore nanorobot (CN). The results from the laminar flow microchip indicated that the obtained CN possessed the strongly positive chemotaxis towards protons. And the TME microchip experiments verified that the CN had a desirable tumor-accumulation ability. Cellular uptake experiments demonstrated that the CN efficiently promoted the endocytosis of the fluorescence DiO into the HT-29 cells. And the in vivo studies revealed that the intravenously administered CN exhibited vigorous tumor-targetability and accumulation ability as well as highly efficient antitumor efficacy. All the results suggested that FOF1-ATPase motors-embedded CN could be promising nanomachines with powerful self-propulsion for overcoming physiological barriers and tumor-targeted drug delivery. STATEMENT OF SIGNIFICANCE: In this study, we demonstrated that FOF1-ATPase-embedded chromatophore nanorobots exhibit a strong proton chemotaxis, which not only plays a key role in tumor-targetability and accumulation, but also promotes tumor tissue penetration and internalization. The results of in vitro and in vivo studies indicated that drug-loaded chromatophore nanorobots are capable to simultaneously accomplish tumor-targeting, accumulation, penetration and internalization for enhanced tumor therapy. Our study provides a fundamental basis for further study on FOF1-ATPase-embedded chromatophore as tumor-targeting drug delivery systems that have promising clinical applications. It offers a new and more efficient delivery vehicle for cancer related therapeutics.

AcousticRobots: Smart acoustically powered micro-/nanoswimmers for precise biomedical applications.

Although nanotechnology has evolutionarily progressed in biomedical field over the past decades, achieving satisfactory therapeutic effects remains difficult with limited delivery efficiency. Ultrasound could provide a deep penetration and maneuverable actuation to efficiently power micro-/nanoswimmers with little harm, offering an emerging and fascinating alternative to the active delivery platform. Recent advances in novel fabrication, controllable concepts like intelligent swarm and the integration of hybrid propulsions have promoted its function and potential for medical applications. In this review, we will summarize the mechanisms and types of ultrasonically propelled micro/nanorobots (termed here as "AcousticRobots"), including the interactions between AcousticRobots and acoustic field, practical design considerations (e.g., component, size, shape), the synthetic methods, surface modification, controllable behaviors, and the advantages when combined with other propulsion approaches. The representative biomedical applications of functional AcousticRobots are also highlighted, including drug delivery, invasive surgery, eradication on the surrounding bio-environment, cell manipulation, detection, and imaging, etc. We conclude by discussing the challenges and outlook of AcousticRobots in biomedical applications.

Application of micro/nanorobot in medicine.

The development of micro/nanorobots and their application in medical treatment holds the promise of revolutionizing disease diagnosis and treatment. In comparison to conventional diagnostic and treatment methods, micro/nanorobots exhibit immense potential due to their small size and the ability to penetrate deep tissues. However, the transition of this technology from the laboratory to clinical applications presents significant challenges. This paper provides a comprehensive review of the research progress in micro/nanorobotics, encompassing biosensors, diagnostics, targeted drug delivery, and minimally invasive surgery. It also addresses the key issues and challenges facing this technology. The fusion of micro/nanorobots with medical treatments is poised to have a profound impact on the future of medicine.

Design and Control of the Magnetically Actuated Micro/Nanorobot Swarm toward Biomedical Applications.

Recently, magnetically actuated micro/nanorobots hold extensive promises in biomedical applications due to their advantages of noninvasiveness, fuel-free operation, and programmable nature. While effectively promised in various fields such as targeted delivery, most past investigations are mainly displayed in magnetic control of individual micro/nanorobots. Facing practical medical use, the micro/nanorobots are required for the development of swarm control in a closed-loop control manner. This review outlines the recent developments in magnetic micro/nanorobot swarms, including their actuating fundamentals, designs, controls, and biomedical applications. The fundamental principles and interactions involved in the formation of magnetic micro/nanorobot swarms are discussed first. The recent advances in the design of artificial and biohybrid micro/nanorobot swarms, along with the control devices and methods used for swarm manipulation, are presented. Furthermore, biomedical applications that have the potential to achieve clinical application are introduced, such as imaging-guided therapy, targeted delivery, embolization, and biofilm eradication. By addressing the potential challenges discussed toward the end of this review, magnetic micro/nanorobot swarms hold promise for clinical treatments in the future.

Nanorobotic artificial blood components and its therapeutic applications: A minireview.

Numerous scientific and medical domains have been revolutionized by nanotechnology, opening up unprecedented opportunities for healthcare applications. Among these developments, the creation of nanorobots for artificial blood components is a novel field of research that seeks to overcome the constraints of conventional pharmacological therapy. This review article provides a comprehensive overview of the nanorobotic artificial blood components and their therapeutic uses. The article begins by outlining the core concepts of nanotechnology and nanorobotic systems, emphasizing their design and control methods. It then delves into various types of nanorobotic artificial blood components, such as oxygen transporters (artificial RBCs), clotting agents (artificial platelets), and immune modulators (artificial WBCs). It goes into detail about their properties, functioning, and capabilities, which allow them to replicate the physiological activities of actual blood components. The article also assesses the clinical uses of artificial blood components in a variety of medical circumstances. It highlights their potential value in the management of certain blood-related diseases.

Magnetically Manipulated Optoelectronic Hybrid Microrobots for Optically Targeted Non-Genetic Neuromodulation.

Optically controlled neuromodulation is a promising approach for basic research of neural circuits and the clinical treatment of neurological diseases. However, developing a non-invasive and well-controllable system to deliver accurate and effective neural stimulation is challenging. Micro/nanorobots have shown great potential in various biomedical applications because of their precise controllability. Here, a magnetically-manipulated optoelectronic hybrid microrobot (MOHR) is presented for optically targeted non-genetic neuromodulation. By integrating the magnetic component into the metal-insulator-semiconductor junction design, the MOHR has excellent magnetic controllability and optoelectronic properties. The MOHR displays a variety of magnetic manipulation modes that enables precise and efficient navigation in different biofluids. Furthermore, the MOHR could achieve precision neuromodulation at the single-cell level because of its accurate targeting ability. This neuromodulation is achieved by the MOHR's photoelectric response to visible light irradiation, which enhances the excitability of the targeted cells. Finally, it is shown that the well-controllable MOHRs effectively restore neuronal activity in neurons damaged by ß-amyloid, a pathogenic agent of Alzheimer's disease. By coupling precise controllability with efficient optoelectronic properties, the hybrid microrobot system is a promising strategy for targeted on-demand optical neuromodulation.

Dual-Functional Laser-Guided Magnetic Nanorobot Collectives against Gravity for On-Demand Thermo-Chemotherapy of Peritoneal Metastasis.

Combining hyperthermic intraperitoneal chemotherapy with cytoreductive surgery is the main treatment modality for peritoneal metastatic (PM) carcinoma despite the off-target effects of chemotherapy drugs and the ineluctable side effects of total abdominal heating. Herein, a laser-integrated magnetic actuation system that actively delivers doxorubicin (DOX)-grafted magnetic nanorobot collectives to the tumor site in model mice for local hyperthermia and chemotherapy is reported. With intraluminal movements controlled by a torque-force hybrid magnetic field, these magnetic nanorobots gather at a fixed point coinciding with the position of the localization laser, moving upward against gravity over a long distance and targeting tumor sites under ultrasound imaging guidance. Because aggregation enhances the photothermal effect, controlled local DOX release is achieved under near-infrared laser irradiation. The targeted on-demand photothermal therapy of multiple PM carcinomas while minimizing off-target tissue damage is demonstrated. Additionally, a localization/treatment dual-functional laser-integrated magnetic actuation system is developed and validated in vivo, offering a potentially clinically feasible drug delivery strategy for targeting PM and other intraluminal tumors.

Untethered Micro/Nanorobots for Remote Sensing: Toward Intelligent Platform.

Untethered micro/nanorobots that can wirelessly control their motion and deformation state have gained enormous interest in remote sensing applications due to their unique motion characteristics in various media and diverse functionalities. Researchers are developing micro/nanorobots as innovative tools to improve sensing performance and miniaturize sensing systems, enabling in situ detection of substances that traditional sensing methods struggle to achieve. Over the past decade of development, significant research progress has been made in designing sensing strategies based on micro/nanorobots, employing various coordinated control and sensing approaches. This review summarizes the latest developments on micro/nanorobots for remote sensing applications by utilizing the self-generated signals of the robots, robot behavior, microrobotic manipulation, and robot-environment interactions. Providing recent studies and relevant applications in remote sensing, we also discuss the challenges and future perspectives facing micro/nanorobots-based intelligent sensing platforms to achieve sensing in complex environments, translating lab research achievements into widespread real applications.

Micro/nanorobots for remediation of water resources and aquatic life.

Nowadays, global water scarcity is becoming a pressing issue, and the discharge of various pollutants leads to the biological pollution of water bodies, which further leads to the poisoning of living organisms. Consequently, traditional water treatment methods are proving inadequate in addressing the growing demands of various industries. As an effective and eco-friendly water treatment method, micro/nanorobots is making significant advancements. Based on researches conducted between 2019 and 2023 in the field of water pollution using micro/nanorobots, this paper comprehensively reviews the development of micro/nanorobots in water pollution control from multiple perspectives, including propulsion methods, decontamination mechanisms, experimental techniques, and water monitoring. Furthermore, this paper highlights current challenges and provides insights into the future development of the industry, providing guidance on biological water pollution control.

A Spatially Programmable DNA Nanorobot Arm to Modulate Anisotropic Gold Nanoparticle Assembly by Enzymatic Excision.

Programmable assembly of gold nanoparticle superstructures with precise spatial arrangement has drawn much attention for their unique characteristics in plasmonics and biomedicine. Bio-inspired methods have already provided programmable, molecular approaches to direct AuNP assemblies using biopolymers. The existing methods, however, predominantly use DNA as scaffolds to directly guide the AuNP interactions to produce intended superstructures. New paradigms for regulating AuNP assembly will greatly enrich the toolbox for DNA-directed AuNP manipulation and fabrication. Here, we developed a strategy of using a spatially programmable enzymatic nanorobot arm to modulate anisotropic DNA surface modifications and assembly of AuNPs. Through spatial controls of the proximity of the reactants, the locations of the modifications were precisely regulated. We demonstrated the control of the modifications on a single 15 nm AuNP, as well as on a rectangular DNA origami platform, to direct unique anisotropic AuNP assemblies. This method adds an alternative enzymatic manipulation to DNA-directed AuNP superstructure assembly.

Enhanced Inhibition of Amyloid Formation by Heat Shock Protein 90 Immobilized on Nanoparticles.

As the population ages, an epidemic of neurodegenerative diseases with devastating social consequences is looming. To address the pathologies leading to amyloid-related dementia, novel therapeutic strategies must be developed for the treatment or prevention of neural protein-folding disorders. Nanotechnology will be crucial to this scenario, especially in the design of nanoscale systems carrying therapeutic compounds that can navigate the nervous system and identify amyloid to treat it in situ. In this line, we have recently designed a highly simplified and versatile nanorobot consisting of a protein coating based on the heat shock protein 90 (Hsp90) chaperone that not only propels nanoparticles using ATP but also endows them with the extraordinary ability to fold and restore the activity of heat-denatured proteins. Here, we assess the effectiveness of these nanosystems in inhibiting/reducing the aggregation of amyloidogenic proteins. Using Raman spectroscopy, we qualitatively and quantitatively analyze amyloid by identifying and semi-quantifying the Amide I band. Our findings indicate that the coupling of Hsp90 to nanoparticles results in a more potent inhibition of amyloid formation when compared to the soluble protein. We propose that this enhanced performance may be attributed to enhanced release-capture cycles of amyloid precursor oligomers by Hsp90 molecules nearby on the nanosurface. Intelligent biocompatible coatings, like the one described here, that enhance the diffusivity and self-propulsion of nanoparticles while enabling them to carry out critical functions such as environmental scanning, identification, and amyloid prevention, present an exceptional opportunity for the development of advanced nanodevices in biomedical applications. This approach, which combined active biomolecules with synthetic materials, is poised to reveal remarkable prospects in the field of nanomedicine and biotechnology.

Untethered Small-Scale Machines for Microrobotic Manipulation: From Individual and Multiple to Collective Machines.

Untethered small-scale machines (USSMs) that can actively adjust their motion, deformation, and collective states in response to external stimuli have gained enormous interest in various manipulation, sensing, and biomedical applications. Because they can be efficiently operated in confined and tortuous environments, USSMs are capable of conducting wireless microrobotic manipulation tasks that tethered machines find hard to achieve. Over the past decade of development, significant research progress has been achieved in designing USSM-based manipulation strategies, which are enabled by investigating machine-object, machine-environment, and machine-machine interactions. This review summarizes the latest developments in USSMs for microrobotic manipulation by utilizing individual machines, coordinating multiple machines, and inducing collective behaviors. Providing recent studies and relevant applications in microrobotic and biomedical areas, we also discuss the challenges and future perspectives facing USSMs-based intelligent manipulation systems to achieve manipulation in complex environments with imaging-guided processes and increasing autonomy levels.

Nanorobot-Mediated Synchronized Neuron Activation.

Interactions between active materials lead to collective behavior and even intelligence beyond the capability of individuals. Such behaviors are prevalent in nature and can be observed in animal colonies, providing these species with diverse capacities for communication and cooperation. In artificial systems, however, collective intelligence systems interacting with biological entities remains unexplored. Herein, we describe black (B)-TiO2@N/Au nanorobots interacting through photocatalytic pure water splitting-induced electrophoresis that exhibit periodic swarming oscillations under programmed near-infrared light. The periodic chemical-electric field generated by the oscillating B-TiO2@N/Au nanorobot swarm leads to local neuron activation in vitro. The field oscillations and neurotransmission from synchronized neurons further trigger the resonance oscillation of neuron populations without synaptic contact (about 2 mm spacing), in different ways from normal neuron oscillation requiring direct contact. We envision that the oscillating nanorobot swarm platforms will shed light on contactless communication of neurons and offer tools to explore interactions between neurons.

Active Micro/Nanoparticles in Colloidal Microswarms.

Colloidal microswarms have attracted increasing attention in the last decade due to their unique capabilities in various complex tasks. Thousands or even millions of tiny active agents are gathered with distinctive features and emerging behaviors, demonstrating fascinating equilibrium and non-equilibrium collective states. In recent studies, with the development of materials design, remote control strategies, and the understanding of pair interactions between building blocks, microswarms have shown advantages in manipulation and targeted delivery tasks with high adaptability and on-demand pattern transformation. This review focuses on the recent progress in active micro/nanoparticles (MNPs) in colloidal microswarms under the input of an external field, including the response of MNPs to external fields, MNP-MNP interactions, and MNP-environment interactions. A fundamental understanding of how building blocks behave in a collective system provides the foundation for designing microswarm systems with autonomy and intelligence, aiming for practical application in diverse environments. It is envisioned that colloidal microswarms will significantly impact active delivery and manipulation applications on small scales.

Wake-Riding Effect-Inspired Opto-Hydrodynamic Diatombot for Non-Invasive Trapping and Removal of Nano-Biothreats.

Contamination of nano-biothreats, such as viruses, mycoplasmas, and pathogenic bacteria, is widespread in cell cultures and greatly threatens many cell-based bio-analysis and biomanufacturing. However, non-invasive trapping and removal of such biothreats during cell culturing, particularly many precious cells, is of great challenge. Here, inspired by the wake-riding effect, a biocompatible opto-hydrodynamic diatombot (OHD) based on optical trapping navigated rotational diatom (Phaeodactylum tricornutum Bohlin) for non-invasive trapping and removal of nano-biothreats is reported. Combining the opto-hydrodynamic effect and optical trapping, this rotational OHD enables the trapping of bio-targets down to sub-100 nm. Different nano-biothreats, such as adenoviruses, pathogenic bacteria, and mycoplasmas, are first demonstrated to be effectively trapped and removed by the OHD, without affecting culturing cells including precious cells such as hippocampal neurons. The removal efficiency is greatly enhanced via reconfigurable OHD array construction. Importantly, these OHDs show remarkable antibacterial capability, and further facilitate targeted gene delivery. This OHD serves as a smart micro-robotic platform for effective trapping and active removal of nano-biothreats in bio-microenvironments, and especially for cell culturing of many precious cells, with great promises for benefiting cell-based bio-analysis and biomanufacturing.

A Dual-Biomineralized Yeast Micro-/Nanorobot with Self-Driving Penetration for Gastritis Therapy and Motility Recovery.

Micro-/nanorobots have attracted great interest in the field of drug delivery and treatment, while preparations for biocompatible robots are extremely challenging. Here, a self-driving yeast micro-/nanorobot (Cur@CaY-robot) is designed via dual biomineralization and acid catalysis of calcium carbonate (CaCO3). Inner nano-CaCO3 inside yeast cells (CaY) is biomineralized through cell respiration and provides nanoscaffolds for highly encapsulating curcumin (Cur). Meanwhile, the CaCO3 crystals outside yeast cells (outer-CaCO3) through uniaxial growth offer an asymmetric power source for self-propelled motility. The Cur@CaY-robot displays an efficient motion in gastric acid, with the potential for deep penetration to the thick gastric mucus, which significantly improves the accumulation of drug agents in the stomach wall tissue for robust gastritis therapy. More importantly, Ca2+ cations released from the Cur@CaY-robot also synergistically repair the gastric motility of gastritis mice. Such yeast micro-/nanorobots exhibit desirable biocompatibility and biodegradability with a good loading capacity for drugs. This work provides an idea for the design of micro-/nanorobots through an environmentally friendly biosynthesis strategy for active drug delivery and precise therapy.