Bacterial outer membrane vesicle nanorobot.

Autonomous nanorobots represent an advanced tool for precision therapy to improve therapeutic efficacy. However, current nanorobotic designs primarily rely on inorganic materials with compromised biocompatibility and limited biological functions. Here, we introduce enzyme-powered bacterial outer membrane vesicle (OMV) nanorobots. The immobilized urease on the OMV membrane catalyzes the decomposition of bioavailable urea, generating effective propulsion for nanorobots. This OMV nanorobot preserves the unique features of OMVs, including intrinsic biocompatibility, immunogenicity, versatile surface bioengineering for desired biofunctionalities, capability of cargo loading and protection. We present OMV-based nanorobots designed for effective tumor therapy by leveraging the membrane properties of OMVs. These involve surface bioengineering of robotic body with cell-penetrating peptide for tumor targeting and penetration, which is further enhanced by active propulsion of nanorobots. Additionally, OMV nanorobots can effectively safeguard the loaded gene silencing tool, small interfering RNA (siRNA), from enzymatic degradation. Through systematic in vitro and in vivo studies using a rodent model, we demonstrate that these OMV nanorobots substantially enhanced siRNA delivery and immune stimulation, resulting in the utmost effectiveness in tumor suppression when juxtaposed with static groups, particularly evident in the orthotopic bladder tumor model. This OMV nanorobot opens an inspiring avenue to design advanced medical robots with expanded versatility and adaptability, broadening their operation scope in practical biomedical domains.

Heterogeneous Sensor-Carrier Microswarms for Collaborative Precise Drug Delivery toward Unknown Targets with Localized Acidosis.

Micro/nanorobots hold the potential to revolutionize biomedicine by executing diverse tasks in hard-to-reach biological environments. Nevertheless, achieving precise drug delivery to unknown disease sites using swarming micro/nanorobots remains a significant challenge. Here we develop a heterogeneous swarm comprising sensing microrobots (sensor-bots) and drug-carrying microrobots (carrier-bots) with collaborative tasking capabilities for precise drug delivery toward unknown sites. Leveraging robust interspecific hydrodynamic interactions, the sensor-bots and carrier-bots spontaneously synchronize and self-organize into stable heterogeneous microswarms. Given that the sensor-bots can create real-time pH maps employing pH-responsive structural-color changes and the doxorubicin-loaded carrier-bots exhibit selective adhesion to acidic targets via pH-responsive charge reversal, the sensor-carrier microswarm, when exploring unknown environments, can detect and localize uncharted acidic targets, guide itself to cover the area, and finally deploy therapeutic carrier-bots precisely there. This versatile platform holds promise for treating diseases with localized acidosis and inspires future theranostic microsystems with expandability, task flexibility, and high efficiency.

Swarms of Enzyme-Powered Nanomotors Enhance the Diffusion of Macromolecules in Viscous Media.

Over the past decades, the development of nanoparticles (NPs) to increase the efficiency of clinical treatments has been subject of intense research. Yet, most NPs have been reported to possess low efficacy as their actuation is hindered by biological barriers. For instance, synovial fluid (SF) present in the joints is mainly composed of hyaluronic acid (HA). These viscous media pose a challenge for many applications in nanomedicine, as passive NPs tend to become trapped in complex networks, which reduces their ability to reach the target location. This problem can be addressed by using active NPs (nanomotors, NMs) that are self-propelled by enzymatic reactions, although the development of enzyme-powered NMs, capable of navigating these viscous environments, remains a considerable challenge. Here, the synergistic effects of two NMs troops, namely hyaluronidase NMs (HyaNMs, Troop 1) and urease NMs (UrNMs, Troop 2) are demonstrated. Troop 1 interacts with the SF by reducing its viscosity, thus allowing Troop 2 to swim more easily through the SF. Through their collective motion, Troop 2 increases the diffusion of macromolecules. These results pave the way for more widespread use of enzyme-powered NMs, e.g., for treating joint injuries and improving therapeutic effectiveness compared with traditional methods.

Nanoarchitectonic Engineering of Thermal-Responsive Magnetic Nanorobot Collectives for Intracranial Aneurysm Therapy.

Stent-assisted coiling is a main treatment modality for intracranial aneurysms (IAs) in clinics, but critical challenges remain to be overcome, such as exogenous implant-induced stenosis and reliance on antiplatelet agents. Herein, an endovascular approach is reported for IA therapy without stent grafting or microcatheter shaping, enabled by active delivery of thrombin (Th) to target aneurysms using innovative phase-change material (PCM)-coated magnetite-thrombin (Fe3O4-Th@PCM) FTP nanorobots. The nanorobots are controlled by an integrated actuation system of dynamic torque-force hybrid magnetic fields. With robust intravascular navigation guided by real-time ultrasound imaging, nanorobotic collectives can effectively accumulate and retain in model aneurysms constructed in vivo, followed by controlled release of the encapsulated Th for rapid occlusion of the aneurysm upon melting the protective PCM (thermally responsive in a tunable manner) through focused magnetic hyperthermia. Complete and stable aneurysm embolization is confirmed by postoperative examination and 2-week postembolization follow-up using digital subtraction angiography (DSA), contrast-enhanced ultrasound (CEUS), and histological analysis. The safety of the embolization therapy is assessed through biocompatibility evaluation and histopathology assays. This strategy, seamlessly integrating secure drug packaging, agile magnetic actuation, and clinical interventional imaging, avoids possible exogenous implant rejection, circumvents cumbersome microcatheter shaping, and offers a promising option for IA therapy.

A Nanorobotics-Based Approach of Breast Cancer in the Nanotechnology Era.

We are living in an era of advanced nanoscience and nanotechnology. Numerous nanomaterials, culminating in nanorobots, have demonstrated ingenious applications in biomedicine, including breast cancer (BC) nano-theranostics. To solve the complicated problem of BC heterogeneity, non-targeted drug distribution, invasive diagnostics or surgery, resistance to classic onco-therapies and real-time monitoring of tumors, nanorobots are designed to perform multiple tasks at a small scale, even at the organelles or molecular level. Over the last few years, most nanorobots have been bioengineered as biomimetic and biocompatible nano(bio)structures, resembling different organisms and cells, such as urchin, spider, octopus, fish, spermatozoon, flagellar bacterium or helicoidal cyanobacterium. In this review, readers will be able to deepen their knowledge of the structure, behavior and role of several types of nanorobots, among other nanomaterials, in BC theranostics. We summarized here the characteristics of many functionalized nanodevices designed to counteract the main neoplastic hallmark features of BC, from sustaining proliferation and evading anti-growth signaling and resisting programmed cell death to inducing angiogenesis, activating invasion and metastasis, preventing genomic instability, avoiding immune destruction and deregulating autophagy. Most of these nanorobots function as targeted and self-propelled smart nano-carriers or nano-drug delivery systems (nano-DDSs), enhancing the efficiency and safety of chemo-, radio- or photodynamic therapy, or the current imagistic techniques used in BC diagnosis. Most of these nanorobots have been tested in vitro, using various BC cell lines, as well as in vivo, mainly based on mice models. We are still waiting for nanorobots that are low-cost, as well as for a wider transition of these favorable effects from laboratory to clinical practice.

Colloidal Self-Assembly: From Passive to Active Systems.

Self-assembly fundamentally implies the organization of small sub-units into large structures or patterns without the intervention of specific local interactions. This process is commonly observed in nature, occurring at various scales ranging from atomic/molecular assembly to the formation of complex biological structures. Colloidal particles may serve as micrometer-scale surrogates for studying assembly, particularly for the poorly understood kinetic and dynamic processes at the atomic scale. Recent advances in colloidal self-assembly have enabled the programmable creation of novel materials with tailored properties. We here provide an overview and comparison of both passive and active colloidal self-assembly, with a discussion on the energy landscape and interactions governing both types. In the realm of passive colloidal assembly, many impressive and important structures have been realized, including colloidal molecules, one-dimensional chains, two-dimensional lattices, and three-dimensional crystals. In contrast, active colloidal self-assembly, driven by optical, electric, chemical, or other fields, involves more intricate dynamic processes, offering more flexibility and potential new applications. A comparative analysis underscores the critical distinctions between passive and active colloidal assemblies, highlighting the unique collective behaviors emerging in active systems. These behaviors encompass collective motion, motility-induced phase segregation, and exotic properties arising from out-of-equilibrium thermodynamics. Through this comparison, we aim to identify the future opportunities in active assembly research, which may suggest new application domains.

Magnetically Driven Self-Degrading Zinc-Containing Cystine Microrobots for Treatment of Prostate Cancer.

Prostate cancer is the most commonly diagnosed tumor disease in men, and its treatment is still a big challenge in standard oncology therapy. Magnetically actuated microrobots represent the most promising technology in modern nanomedicine, offering the advantage of wireless guidance, effective cell penetration, and non-invasive actuation. Here, new biodegradable magnetically actuated zinc/cystine-based microrobots for in situ treatment of prostate cancer cells are reported. The microrobots are fabricated via metal-ion-mediated self-assembly of the amino acid cystine encapsulating superparamagnetic Fe3 O4 nanoparticles (NPs) during the synthesis, which allows their precise manipulation by a rotating magnetic field. Inside the cells, the typical enzymatic reducing environment favors the disassembly of the aminoacidic chemical structure due to the cleavage of cystine disulfide bonds and disruption of non-covalent interactions with the metal ions, as demonstrated by in vitro experiments with reduced nicotinamide adenine dinucleotide (NADH). In this way, the cystine microrobots served for site-specific delivery of Zn2+ ions responsible for tumor cell killing via a "Trojan horse effect". This work presents a new concept of cell internalization exploiting robotic systems' self-degradation, proposing a step forward in non-invasive cancer therapy.

Imaging-Navigated Surgery.

It is vitally important to guide or navigate therapeutic proceedings with a direct and visual approach in order to carefully undertake precision medical manipulations and efficiently evaluate the treatments. Imaging-navigated surgery is one of the common and prevailing technologies to realize this target, and more importantly it merges visualized medicine into next-generation theranostic paradigms in modern medicine. Endoscopes, surgical robots, and nanorobots are three major domains in terms of imaging-navigated surgery. The history of endoscopy has seen upgraded developments since the early 1800s. In contrast, surgical robots have been widely used and investigated in recent years, and they came into clinical uses only in the past decades. Nanorobots which closely depend on innovated and multifunctional biomaterials are still in their infancy. All these imaging-navigated technologies show similar and apparent advantages such as minimal invasiveness, minimized pain, positive prognosis, and relatively expected recovery, which have greatly improved surgery efficiency and patients' life quality. Therefore, the imaging-navigated surgery will be discussed in this chapter, and advanced clinical and preclinical medical applications will also be demonstrated for a diverse readers and comprehensive understanding.

Magnetic Biohybrid Robots as Efficient Drug Carrier to Generate Plant Cell Clones.

Micro/nanorobots represent a new generation of micromachines that can accomplish various tasks, such as loading and transporting specific targets or pharmaceuticals for a given application. Biohybrid robots consisting of biological cells (bacteria, sperm, and microalgae) combined with inorganic particles to control or propel their movement are of particular interest. The skeleton of these biohybrid robots can be used to load biomolecules. In this work, the authors create biohybrid robots based on tomato plants by coculturing ferromagnetic nanoparticles (Fe3 O4 ) with tomato callus cells. The tomato-based biohybrid robots (Tomato-Biobots) containing Fe3 O4 nanoparticles  are driven by a transversely rotating magnetic field. In addition, biohybrid robots are used to load vitamin C, to generate clones of tomato cells. It is shown that the presence of Fe3 O4  does not affect the growth of tomato callus. This study opens a wide range of possibilities for the use of biohybrid robots@Fe3 O4  to deliver conventional agrochemicals, including fertilizers, pesticides, and herbicides, and allows for a gradual and sustained release of nutrients and agrochemicals, leading to precise dosing that reduces the amount of agrochemicals used. This conceptually new type of micromachine with application to plants and agronomy shall find broad use in this field.

Plant-Actuated Micro-Nanorobotics Platforms: Structural Designs, Functional Prospects, and Biomedical Applications.

Plants are anatomically and physiologically different from humans and animals; however, there are several possibilities to utilize the unique structures and physiological systems of plants and adapt them to new emerging technologies through a strategic biomimetic approach. Moreover, plants provide safe and sustainable results that can potentially solve the problem of mass-producing practical materials with hazardous and toxic side effects, particularly in the biomedical field, which requires high biocompatibility. In this review, it is investigated how micro-nanostructures available in plants (e.g., nanoparticles, nanofibers and their composites, nanoporous materials, and natural micromotors) are adapted and utilized in the design of suitable materials for a micro-nanorobot platform. How plants' work on micro- and nanoscale systems (e.g., surface roughness, osmotically induced movements such as nastic and tropic, and energy conversion and harvesting) that are unique to plants, can provide functionality on the platform and become further prospective resources are examined. Furthermore, implementation across organisms and fields, which is promising for future practical applications of the plant-actuated micro-nanorobot platform, especially on biomedical applications, is discussed. Finally, the challenges following its implementation in the micro-nanorobot platform are also presented to provide advanced adaptation in the future.

Design, fabrication and application of magnetically actuated micro/nanorobots: a review.

Magnetically actuated micro/nanorobots are typical micro- and nanoscale artificial devices with favorable attributes of quick response, remote and contactless control, harmless human-machine interaction and high economic efficiency. Under external magnetic actuation strategies, they are capable of achieving elaborate manipulation and navigation in extreme biomedical environments. This review focuses on state-of-the-art progresses in design strategies, fabrication techniques and applications of magnetically actuated micro/nanorobots. Firstly, recent advances of various robot designs, including helical robots, surface walkers, ciliary robots, scaffold robots and biohybrid robots, are discussed separately. Secondly, the main progresses of common fabrication techniques are respectively introduced, and application achievements on these robots in targeted drug delivery, minimally invasive surgery and cell manipulation are also presented. Finally, a short summary is made, and the current challenges and future work for magnetically actuated micro/nanorobots are discussed.

Engineering Nanorobots for Tumor-Targeting Drug Delivery: From Dynamic Control to Stimuli-Responsive Strategy.

Nanotechnology has been widely applied to the fabrication of drug delivery systems in the past decades. Recently, with the progress made in microfabrication approaches, nanorobots are steadily becoming a promising means for tumor-targeting drug delivery. In general, nanorobots can be divided into two categories: nanomotors and stimuli-responsive nanorobots. Nanomotors are nanoscale systems with the ability to convert surrounding energies into mechanical motion, whereas stimuli-responsive nanorobots are featured with activatable capacity in response to various endogenous and exogenous stimulations. In this minireview, the dynamic control of nanomotors and the rational design of stimuli-responsive nanorobots are overviewed, with particular emphasis on their contribution to tumor-targeting therapy. Moreover, challenges and perspectives associated with the future development of nanorobots are presented.

Engineering a Second-Order DNA Logic-Gated Nanorobot to Sense and Release on Live Cell Membranes for Multiplexed Diagnosis and Synergistic Therapy.

Tumor biomarker-based theranostics have achieved broad interest and success in recent years. However, single biomarker-based recognition can cause false-positive feedback, including the on-target off-tumor phenomenon, in the absence of tumor-specific antigen. Multibiomarker-based recognition molecules often elicit nonspecific and undesired internalization when they bind to "bystander" cells. We report a universal DNA tetrahedral scaffold (DTS) that anchors on the cell membrane to load multiple aptamers and therapeutics for precise and effective theranostics. This DNA logic-gated nanorobot (DLGN) not only facilitates precise discrimination among five cell lines, but also triggers synergistic killing of effector aptamer-tethered synergistic drugs (EASDs) to target cancer cells. Logic-gated recognition integrated into aptamer-functionalized molecular machines will prompt fast tumor profiling, in situ capture and isolation, and safe delivery of precise medicine.

Chemotactic Micro- and Nanodevices.

Preparation of autonomous chemotactic micro- and nanomachines represents one of the most difficult challenges of modern materials science. To construct a device mimicking the behavior of many microorganisms, which evolved their chemotactic abilities during the millennia of evolution, places extreme demands on the imagination and abilities of researchers. However, with the chemotactic devices in hand, many novel and interesting applications of micromachines could be implemented. The introduction of an autonomous navigation, independent of the external control with electric, magnetic, or electromagnetic field is crucial for applications in environmental remediation and might be advantageous in medical applications. This Minireview summarizes the development in the field of chemotactic micro- and nanomachines, describes the trends in their construction, and compares the different approaches to their construction considering the areas of possible application of the devices.

Biomimetic DNA Nanotubes: Nanoscale Channel Design and Applications.

Biomacromolecular nanotubes play important physiological roles in transmembrane ion/molecule channeling, intracellular transport, and inter-cellular communications. While genetically encoded protein nanotubes are prevalent in vivo, the in vitro construction of biomimetic DNA nanotubes has attracted intense interest with the rise of structural DNA nanotechnology. The abiotic use of DNA assembly provides a powerful bottom-up approach for the rational construction of complex materials with arbitrary size and shape at the nanoscale. More specifically, a typical DNA nanotube can be assembled either with parallel-aligned DNA duplexes or by closing DNA tile lattices. These artificial DNA nanotubes can be tailored and site-specifically modified to realize biomimetic functions including ionic or molecular channeling, bioreactors, drug delivery, and biomolecular sensing. In this Minireview, we aim to summarize recent advances in design strategies, including the characterization and applications of biomimetic DNA nanotubes.

Magnetic Propulsion of Microswimmers with DNA-Based Flagellar Bundles.

We show that DNA-based self-assembly can serve as a general and flexible tool to construct artificial flagella of several micrometers in length and only tens of nanometers in diameter. By attaching the DNA flagella to biocompatible magnetic microparticles, we provide a proof of concept demonstration of hybrid structures that, when rotated in an external magnetic field, propel by means of a flagellar bundle, similar to self-propelling peritrichous bacteria. Our theoretical analysis predicts that flagellar bundles that possess a length-dependent bending stiffness should exhibit a superior swimming speed compared to swimmers with a single appendage. The DNA self-assembly method permits the realization of these improved flagellar bundles in good agreement with our quantitative model. DNA flagella with well-controlled shape could fundamentally increase the functionality of fully biocompatible nanorobots and extend the scope and complexity of active materials.

Magnetically Propelled Fish-Like Nanoswimmers.

The swimming locomotion of fish involves a complex interplay between a deformable body and induced flow in the surrounding fluid. While innovative robotic devices, inspired by physicomechanical designs evolved in fish, have been created for underwater propulsion of large swimmers, scaling such powerful locomotion into micro-/nanoscale propulsion remains challenging. Here, a magnetically propelled fish-like artificial nanoswimmer is demonstrated that emulates the body and caudal fin propulsion swimming mechanism displayed by fish. To mimic the deformable fish body for periodic shape changes, template-electrosynthesized multisegment nanowire swimmers are used to construct the artificial nanofishes (diameter 200 nm; length 4.8 µm). The resulting nanofish consists a gold segment as the head, two nickel segments as the body, and one gold segment as the caudal fin, with three flexible porous silver hinges linking each segment. Under an oscillating magnetic field, the propulsive nickel elements bend the body and caudal fin periodically to generate travelling-wave motions with speeds exceeding 30 µm s-1 . The propulsion dynamics is studied theoretically using the immersed boundary method. Such body-deformable nanofishes exhibit a high swimming efficiency and can serve as promising biomimetic nanorobotic devices for nanoscale biomedical applications.

Motion Control of Micro-/Nanomotors.

As we progress towards employing self-propelled micro-/nanomotors in envisioned applications such as cargo delivery, environmental remediation, and therapeutic treatments, precise control of the micro-/nanomotors direction and their speed is essential. In this Review, major emerging approaches utilized for the motion control of micro-/nanomotors have been discussed, together with the lastest publications describing these approaches. Future studies could incorporate investigations on micro-/nanomotors motion control in a real-world environment in which matrix complexity might disrupt successful manipulation of these small-scale devices.

Magnetically and Near-Infrared Light-Powered Supramolecular Nanotransporters for the Remote Control of Enzymatic Reactions.

Cancer is one of the primary causes of death worldwide. A high-precision analysis of biomolecular behaviors in cancer cells at the single-cell level and more effective cancer therapies are urgently required. Here, we describe the development of a magnetically- and near infrared light-triggered optical control method, based on nanorobotics, for the analyses of cellular functions. A new type of nanotransporters, composed of magnetic iron nanoparticles, carbon nanohorns, and liposomes, was synthesized for the spatiotemporal control of cellular functions in cells and mice. Our technology will help to create a new state-of-the-art tool for the comprehensive analysis of "real" biological molecular information at the single-cell level, and it may also help in the development of innovative cancer therapies.

A DNA origami nanorobot controlled by nucleic acid hybridization.

A prototype for a DNA origami nanorobot is designed, produced, and tested. The cylindrical nanorobot (diameter of 14 nm and length of 48 nm) with a switchable flap, is able to respond to an external stimulus and reacts by a physical switch from a disarmed to an armed configuration able to deliver a cellular compatible message. In the tested design the robot weapon is a nucleic acid fully contained in the inner of the tube and linked to a single point of the internal face of the flap. Upon actuation the nanorobot moves the flap extracting the nucleic acid that assembles into a hemin/G-quadruplex horseradish peroxidase mimicking DNAzyme catalyzing a colorimetric reaction or chemiluminescence generation. The actuation switch is triggered by an external nucleic acid (target) that interacts with a complementary nucleic acid that is beard externally by the nanorobot (probe). Hybridization of probe and target produces a localized structural change that results in flap opening. The flap movement is studied on a two-dimensional prototype origami using Förster resonance energy transfer and is shown to be triggered by a variety of targets, including natural RNAs. The nanorobot has potential for in vivo biosensing and intelligent delivery of biological activators.