Nanoparticle-modified microrobots for in vivo antibiotic delivery to treat acute bacterial pneumonia.

Bioinspired microrobots capable of actively moving in biological fluids have attracted considerable attention for biomedical applications because of their unique dynamic features that are otherwise difficult to achieve by their static counterparts. Here we use click chemistry to attach antibiotic-loaded neutrophil membrane-coated polymeric nanoparticles to natural microalgae, thus creating hybrid microrobots for the active delivery of antibiotics in the lungs in vivo. The microrobots show fast speed (>110 µm s-1) in simulated lung fluid and uniform distribution into deep lung tissues, low clearance by alveolar macrophages and superb tissue retention time (>2 days) after intratracheal administration to test animals. In a mouse model of acute Pseudomonas aeruginosa pneumonia, the microrobots effectively reduce bacterial burden and substantially lessen animal mortality, with negligible toxicity. Overall, these findings highlight the attractive functions of algae-nanoparticle hybrid microrobots for the active in vivo delivery of therapeutics to the lungs in intensive care unit settings.

Switching from Chemical to Electrical Micromotor Propulsion across a Gradient of Gastric Fluid via Magnetic Rolling.

To address and extend the finite lifetime of Mg-based micromotors due to the depletion of the engine (Mg-core), we examine electric fields, along with previously studied magnetic fields, to create a triple-engine hybrid micromotor for driving these micromotors. Electric fields are a facile energy source that is not limited in its operation time and can dynamically tune the micromotor mobility by simply changing the frequency and amplitude of the field. Moreover, the same electrical fields can be used for cell trapping and transport as well as drug delivery. However, the limitations of these propulsion mechanisms are the low pH (and high conductivity) environment required for Mg dissolution, while the electrical propulsion is quenched at these conditions as it requires low conductivity mediums. In order to translate the micromotor between these two extreme medium conditions, we use magnetic rolling as means of self-propulsion along with magnetic steering. Interestingly, electrical propulsion also necessitates at least the partial consumption of the Mg, resulting in a sufficient geometrical asymmetry of the micromotor. We have successfully demonstrated the rapid propulsion switching capability of the micromotor, from chemical to electrical motions, via magnetic rolling within a microfluidic device with the concentration gradient of the simulated gastric fluid. Such triple-engine micromotor propulsion holds considerable promise for in vitro studies mimicking gastric conditions and performing various bioassay tasks.

Smart Materials for Microrobots.

Over the past 15 years, the field of microrobotics has exploded with many research groups from around the globe contributing to numerous innovations that have led to exciting new capabilities and important applications, ranging from in vivo drug delivery, to intracellular biosensing, environmental remediation, and nanoscale fabrication. Smart responsive materials have had a profound impact on the field of microrobotics and have imparted small-scale robots with new functionalities and distinct capabilities. We have identified four large categories where the majority of future efforts must be allocated to push the frontiers of microrobots and where smart materials can have a major impact on such future advances. These four areas are the propulsion and biocompatibility of microrobots, the cooperation between individual units and human operators, and finally, the intelligence of microrobots. In this Review, we look critically at the latest developments in these four categories and discuss how smart materials contribute to the progress in the exciting field of microrobotics and will set the stage for the next generation of intelligent and programmable microrobots.

Electrical Propulsion and Cargo Transport of Microbowl Shaped Janus Particles.

Herein the effective electrical propulsion, cargo trapping, and transport capabilities of microbowl-shaped Janus particles (JPs) are demonstrated and evaluated. These active JPs are made by deposition of Au and Ti layers onto sacrificial spherical polystyrene particles, followed by oxidation of the Ti to TiO2 . In contrast to the commonly studied spherical JP, the dual broken symmetry of both geometrical and electrical properties of the microbowl renders a strong dependence of its mobility and cargo loading on the order of the layering of Au and TiO2 . Specifically, an opposite direction of motion is obtained for interchanged layers of Au and TiO2 , using only electrical propulsion as the sole mechanism of motion. The concave side of the microbowl exhibits a negative dielectrophoretic trap of large size wherein trapped cargo is protected from hydrodynamic shearing, leading to an enhanced cargo loading capacity compared to that obtained using common spherical JP. Such enhanced cargo capability of the microbowl along with the ease of engineering it by interchanging the order of the layers are very attractive for future in vitro biological and biomedical applications.

Physical Disruption of Solid Tumors by Immunostimulatory Microrobots Enhances Antitumor Immunity.

The combination of immunotherapy with other forms of treatment is an emerging strategy for boosting antitumor responses. By combining multiple modes of action, these combinatorial therapies can improve clinical outcomes through unique synergisms. Here, a microrobot-based strategy that integrates tumor tissue disruption with biological stimulation is shown for cancer immunotherapy. The microrobot is fabricated by loading bacterial outer membrane vesicles onto a self-propelling micromotor, which can react with water to generate a propulsion force. When administered intratumorally to a solid tumor, the disruption of the local tumor tissue coupled with the delivery of an immunostimulatory payload leads to complete tumor regression. Additionally, treatment of the primary tumor results in the simultaneous education of the host immune system, enabling it to control the growth of distant tumors. Overall, this work introduces a distinct application of microrobots in cancer immunotherapy and offers an attractive strategy for amplifying cancer treatment efficacy when combined with conventional therapies.

A Microstirring Pill Enhances Bioavailability of Orally Administered Drugs.

Majority of drugs are administered orally, yet their efficient absorption is often difficult to achieve, with a low dose fraction reaching the blood compartment. Here, a microstirring pill technology is reported with built-in mixing capability for oral drug delivery that greatly enhances bioavailability of its therapeutic payload. Embedding microscopic stirrers into a pill matrix enables faster disintegration and dissolution, leading to improved release profiles of three widely used model drugs, aspirin, levodopa, and acetaminophen, without compromising their loading. Unlike recently developed drug-carrying nanomotors, drug molecules are not associated with the microstirrers, and hence there is no limitation on the loading capacity. These embedded microstirrers are fabricated through the asymmetric coating of titanium dioxide thin film onto magnesium microparticles. In vitro tests illustrate that the embedded microstirrers lead to substantial enhancement of local fluid transport. In vivo studies using murine and porcine models demonstrate that the localized stirring capability of microstirrers leads to enhanced bioavailability of drug payloads. Such improvements are of considerable importance in clinical scenarios where fast absorption and high bioavailability of therapeutics are critical. The encouraging results obtained in porcine model suggest that the microstirring pill technology has translational potential and can be developed toward practical biomedical applications.

ACE2 Receptor-Modified Algae-Based Microrobot for Removal of SARS-CoV-2 in Wastewater.

The coronavirus SARS-CoV-2 can survive in wastewater for several days with a potential risk of waterborne human transmission, hence posing challenges in containing the virus and reducing its spread. Herein, we report on an active biohybrid microrobot system that offers highly efficient capture and removal of target virus from various aquatic media. The algae-based microrobot is fabricated by using click chemistry to functionalize microalgae with angiotensin-converting enzyme 2 (ACE2) receptor against the SARS-CoV-2 spike protein. The resulting ACE2-algae-robot displays fast (>100 µm/s) and long-lasting (>24 h) self-propulsion in diverse aquatic media including drinking water and river water, obviating the need for external fuels. Such movement of the ACE2-algae-robot offers effective "on-the-fly" removal of SARS-CoV-2 spike proteins and SARS-CoV-2 pseudovirus. Specifically, the active biohybrid microrobot results in 95% removal of viral spike protein and 89% removal of pseudovirus, significantly exceeding the control groups such as static ACE2-algae and bare algae. These results suggest considerable promise of biologically functionalized algae toward the removal of viruses and other environmental threats from wastewater.

Swimmers Heal on the Move Following Catastrophic Damage.

Herein, we describe the development of 2D self-healing small-scale swimmers capable of autonomous propulsion and "on-the-fly" structural recovery in large containers. Incorporation of magnetic Nd2Fe14B microparticles in specialized printed strips results in rapid reorientation and reattachment of the moving tail to its complementary broken static piece to restore the original swimmer structure and propulsion behavior. The swimmers display functional recovery independent of user input. Measurements of the magnetic hysteresis and fields were used to assess the behavior of the healing mechanism in real swimming situations. Damage position and multiple magnetic strip patterns have been examined and their influence upon the recovery efficiency has been compared. Owing to its versatility, fast response, and simplicity the new self-healing strategy represents an important step toward the development of new "on-the-fly" repairing strategies for small-scale swimmers and robots.

Enzyme-powered Janus platelet cell robots for active and targeted drug delivery.

Transforming natural cells into functional biocompatible robots capable of active movement is expected to enhance the functions of the cells and revolutionize the development of synthetic micromotors. However, present cell-based micromotor systems commonly require the propulsion capabilities of rigid motors, external fields, or harsh conditions, which may compromise biocompatibility and require complex actuation equipment. Here, we report on an endogenous enzyme-powered Janus platelet micromotor (JPL-motor) system prepared by immobilizing urease asymmetrically onto the surface of natural platelet cells. This Janus distribution of urease on platelet cells enables uneven decomposition of urea in biofluids to generate enhanced chemophoretic motion. The cell surface engineering with urease has negligible impact on the functional surface proteins of platelets, and hence, the resulting JPL-motors preserve the intrinsic biofunctionalities of platelets, including effective targeting of cancer cells and bacteria. The efficient propulsion of JPL-motors in the presence of the urea fuel greatly enhances their binding efficiency with these biological targets and improves their therapeutic efficacy when loaded with model anticancer or antibiotic drugs. Overall, asymmetric enzyme immobilization on the platelet surface leads to a biogenic microrobotic system capable of autonomous movement using biological fuel. The ability to impart self-propulsion onto biological cells, such as platelets, and to load these cellular robots with a variety of functional components holds considerable promise for developing multifunctional cell-based micromotors for a variety of biomedical applications.

Multicompartment Tubular Micromotors Toward Enhanced Localized Active Delivery.

A tubular micromotor with spatially resolved compartments is presented toward efficient site-specific cargo delivery, with a back-end zinc (Zn) propellant engine segment and an upfront cargo-loaded gelatin segment further protected by a pH-responsive cap. The multicompartment micromotors display strong gastric-powered propulsion with tunable lifetime depending on the Zn segment length. Such propulsion significantly enhances the motor distribution and retention in the gastric tissues, by pushing and impinging the front-end cargo segment onto the stomach wall. Once the micromotor penetrates the gastric mucosa (pH ≥ 6.0), its pH-responsive cap dissolves, promoting the autonomous localized cargo release. The fabrication process, physicochemical properties, and propulsion behavior are systematically tested and discussed. Using a mouse model, the multicompartment motors, loaded with a model cargo, demonstrate a homogeneous cargo distribution along with approximately four-fold enhanced retention in the gastric lining compared to monocompartment motors, while showing no apparent toxicity. Therapeutic payloads can also be loaded into the pH-responsive cap, in addition to the gelatin-based compartment, leading to concurrent delivery and sequential release of dual cargos toward combinatorial therapy. Overall, this multicompartment micromotor system provides unique features and advantages that will further advance the development of synthetic micromotors for active transport and localized delivery of biomedical cargos.

Small-Scale Propellers Deliver Miniature Versions of Themselves.

Small-scale actuators and propellers have benefited from advances in materials and manufacturing to become more lifelike. Inspired by animal species, multi-generational chemically powered artificial propellers that carry small versions of themselves and deliver them "on-the-fly" are described. The released replicas are capable of autonomous propulsion and propelling immediately after detachment. Release occurs without human involvement and relies solely on sacrificial layers separating the carriers and replicas. These layers are composed of transient natural polymers, which dissolve under the swimming conditions to release the confined replicas. Judicious selection of the responsive transient materials, layer thickness, and solution conditions (e.g., pH), leads to programmable delivery of the replicas. Finally, the ability of the same carrier propellers to carry and transport multiple generations of propellers and deliver them at predetermined times is demonstrated.

Multigear Bubble Propulsion of Transient Micromotors.

Transient, chemically powered micromotors are promising biocompatible engines for microrobots. We propose a framework to investigate in detail the dynamics and the underlying mechanisms of bubble propulsion for transient chemically powered micromotors. Our observations on the variations of the micromotor active material and geometry over its lifetime, from initial activation to the final inactive state, indicate different bubble growth and ejection mechanisms that occur stochastically, resulting in time-varying micromotor velocity. We identify three processes of bubble growth and ejection, and in analogy with macroscopic multigear machines, we call each process a gear. Gear 1 refers to bubbles that grow on the micromotor surface before detachment while in Gear 2 bubbles hop out of the micromotor. Gear 3 is similar in nature to Gear 2, but the bubbles are too small to contribute to micromotor motion. We study the characteristics of these gears in terms of bubble size and ejection time, and how they contribute to micromotor displacement. The ability to tailor the shell polarity and hence the bubble growth and ejection and the surrounding fluid flow is demonstrated. Such understanding of the complex multigear bubble propulsion of transient chemical micromotors should guide their future design principles and serve for fine tuning the performance of these micromotors.

Onion-like Multifunctional Microtrap Vehicles for Attraction-Trapping-Destruction of Biological Threats.

A multifunctional motile microtrap is developed that is capable of autonomously attracting, trapping, and destroying pathogens by controlled chemoattractant and therapeutic agent release. The onion-inspired multi-layer structure contains a magnesium engine core and inner chemoattractant and therapeutic layers. Upon chemical propulsion, the magnesium core is depleted, resulting in a hollow structure that exposes the inner layers and serves as structural trap. The sequential dissolution and autonomous release of the chemoattractant and killing agents result in long-range chemotactic attraction, trapping, and destruction of motile pathogens. The dissolved chemoattractant (l-serine) significantly increases the accumulation and capture of motile pathogens (E. coli) within the microtrap structure, while the internal release of silver ions (Ag+ ) leads to lysis of the pathogen accumulated within the microtrap cavity.

Micromotors for Active Delivery of Minerals toward the Treatment of Iron Deficiency Anemia.

As the most common nutritional disorder, iron deficiency represents a major public health problem with broad impacts on physical and mental development. However, treatment is often compromised by low iron bioavailability and undesired side effects. Here, we report on the development of active mineral delivery vehicles using Mg-based micromotors, which can autonomously propel in gastrointestinal fluids, aiding in the dynamic delivery of minerals. Iron and selenium are combined as a model mineral payload in the micromotor platform. We demonstrate the ability of our mineral-loaded micromotors to replenish iron and selenium stores in an anemic mouse model after 30 days of treatment, normalizing hematological parameters such as red blood count, hemoglobin, and hematocrit. Additionally, the micromotor platform exhibits no toxicity after the treatment regimen. This proof-of-concept study indicates that micromotor-based active delivery of mineral supplements represents an attractive approach toward alleviating nutritional deficiencies.

A dynamic thermoregulatory material inspired by squid skin.

Effective thermal management is critical for the operation of many modern technologies, such as electronic circuits, smart clothing, and building environment control systems. By leveraging the static infrared-reflecting design of the space blanket and drawing inspiration from the dynamic color-changing ability of squid skin, we have developed a composite material with tunable thermoregulatory properties. Our material demonstrates an on/off switching ratio of ~25 for the transmittance, regulates a heat flux of ~36 W/m2 with an estimated mechanical power input of ~3 W/m2, and features a dynamic environmental setpoint temperature window of ~8 °C. Moreover, the composite can manage one fourth of the metabolic heat flux expected for a sedentary individual and can also modulate localized changes in a wearer's body temperature by nearly 10-fold. Due to such functionality and associated figures of merit, our material may substantially reduce building energy consumption upon widespread deployment and adoption.

A Macrophage-Magnesium Hybrid Biomotor: Fabrication and Characterization.

Magnesium (Mg)-based micromotors are combined with live macrophage (MΦ) cells to create a unique MΦ-Mg biohybrid motor system. The resulting biomotors possess rapid propulsion ability stemming from the Mg micromotors and the biological functions provided by the live MΦ cell. To prepare the biohybrid motors, Mg microparticles coated with titanium dioxide and poly(l-lysine) (PLL) layers are incubated with live MΦs at low temperature. The formation of such biohybrid motors depends on the relative size of the MΦs and Mg particles, with the MΦ swallowing up Mg particles smaller than 5 µm. The experimental results and numerical simulations demonstrate that the motion of MΦ-Mg motors is determined by the size of the Mg micromotor core and the position of the MΦ during the attachment process. The MΦ-Mg motors also perform biological functions related to free MΦs such as endotoxin neutralization. Cell membrane staining and toxin neutralization studies confirm that the MΦs maintain their viability and functionality (e.g., endotoxin neutralization) after binding to the Mg micromotors. This new MΦ-Mg motor design can be expanded to different types of living cells to fulfill diverse biological tasks.

Biomimetic Micromotor Enables Active Delivery of Antigens for Oral Vaccination.

Vaccination represents one of the most effective means of preventing infectious disease. In order to maximize the utility of vaccines, highly potent formulations that are easy to administer and promote high patient compliance are desired. In the present work, a biomimetic self-propelling micromotor formulation is developed for use as an oral antivirulence vaccine. The propulsion is provided by a magnesium-based core, and a biomimetic cell membrane coating is used to detain and neutralize a toxic antigenic payload. The resulting motor toxoids leverage their propulsion properties in order to more effectively elicit mucosal immune responses. After demonstrating the successful fabrication of the motor toxoids, their uptake properties are shown in vitro. When delivered to mice via an oral route, it is then confirmed that the propulsion greatly improves retention and uptake of the antigenic material in the small intestine in vivo. Ultimately, this translates into markedly elevated generation of antibody titers against a model toxin. This work provides a proof-of-concept highlighting the benefits of active oral delivery for vaccine development, opening the door for a new set of applications, in which biomimetic motor technology can provide significant benefits.

Cell-Like Micromotors.

In the past decade, versatile micro- and nanosized machines have emerged as active agents for large-scale detoxification, sensing, microfabrication, and many other promising applications. Micromachines have also been envisioned as the next advancement in dynamic therapy with numerous proof-of-concept studies in drug delivery, microsurgery, and detoxification. However, the practical use of synthetic micromotors in the body requires the development of fully biocompatible designs facilitating micromotor movement in biological fluids of diverse composition and displaying desired functions in specific locations. The combination of the efficient movement of synthetic micromotors with the biological functions of natural cells has resulted in cell-like micromotors with expanded therapeutic and toxin-removing capabilities toward different biological applications. Thus, these biocompatible and biomimetic cell-like micromotors can provide efficient movement in complex biofluids and mimic the functionalities of natural cells. This Account highlights a variety of recent proof-of-concept examples of cell-like micromotors, based on different designs and actuation mechanisms, which perform diverse in vivo tasks. The cell-like micromotors are divided into two groups: (i) cell membrane-coated micromotors, which use natural cell membranes derived from red blood cells, platelets, or a combination of different cells to cloak and functionalize synthetic motors, and (ii) cell-based micromotors, which directly use entire cells such as blood cells, spermatozoa, and bacteria as the micromotor engine. Cell-like micromotors, composed of different cellular components and actuated by different mechanisms, have shown unique advantages for operation in complex biofluids such as blood. Due to the inherent biocompatibility of cell-derived materials, these cell-like micromotors do not provoke an immune response while utilizing useful secondary functions of the blood cells such as strong ability to soak up foreign agents or bind toxins. Additionally, the utilization of autonomously motile cells (e.g., bacteria) allows for built-in chemotactic motion, which eliminates the need for harmful fuels or complex actuation equipment. Furthermore, a broad range of cells, both passive and motile, can be incorporated into micromachine designs constituting a large library of functional components depending on the limits of the desired application. The coupling of cellular and artificial components has led to active biohybrid swimming microsystems with greatly enhanced capabilities and functionalities compared to the individual biological or synthetic components. These characteristics have positioned these cell-like micromotors as promising biomimetic dynamic tools for potential actuation in vivo. Finally, the key challenges and limitations of cell-like micromotors are discussed in the context of expanded future clinical uses and translation to human trials.

Micromotor Pills as a Dynamic Oral Delivery Platform.

Tremendous progress has been made during the past decade toward the design of nano/micromotors with high biocompatibility, multifunctionality, and efficient propulsion in biological fluids, which collectively have led to the initial investigation of in vivo biomedical applications of these synthetic motors. Despite these recent advances in micromotor designs and mechanistic research, significant effort is needed to develop appropriate formulations of micromotors to facilitate their in vivo administration and thus to better test their in vivo applicability. Herein, we present a micromotor pill and demonstrate its attractive use as a platform for in vivo oral delivery of active micromotors. The micromotor pill is comprised of active Mg-based micromotors dispersed uniformly in the pill matrix, containing inactive (lactose/maltose) excipients and other disintegration-aiding (cellulose/starch) additives. Our in vivo studies using a mouse model show that the micromotor pill platform effectively protects and carries the active micromotors to the stomach, enabling their release in a concentrated manner. The micromotor encapsulation and the inactive excipient materials have no effects on the motion of the released micromotors. The released cargo-loaded micromotors propel in gastric fluid, retaining the high-performance characteristics of in vitro micromotors while providing higher cargo retention onto the stomach lining compared to orally administrated free micromotors and passive microparticles. Furthermore, the micromotor pills and the loaded micromotors retain the same characteristics and propulsion behavior after extended storage in harsh conditions. These results illustrate that combining the advantages of traditional pills with the efficient movement of micromotors offer an appealing route for administrating micromotors for potential in vivo biomedical applications.

Chemical/Light-Powered Hybrid Micromotors with "On-the-Fly" Optical Brakes.

Hybrid micromotors capable of both chemically powered propulsion and fuel-free light-driven actuation and offering built-in optical brakes for chemical propulsion are described. The new hybrid micromotors are designed by combining photocatalytic TiO2 and catalytic Pt surfaces into a Janus microparticle. The chemical reactions on the different surfaces of the Janus particle hybrid micromotor can be tailored by using chemical or light stimuli that generate counteracting propulsion forces on the catalytic Pt and photocatalytic TiO2 sides. Such modulation of the surface chemistry on a single micromotor leads to switchable propulsion modes and reversal of the direction of motion that reflect the tuning of the local ion concentration and hence the dominant propulsion force. An intermediate Au layer (under the Pt surface) plays an important role in determining the propulsion mechanism and operation of the hybrid motor. The built-in optical braking system allows "on-the-fly" control of the chemical propulsion through a photocatalytic reaction on the TiO2 side to counterbalance the chemical propulsion force generated on the Pt side. The adaptive dual operation of these chemical/light hybrid micromotors, associated with such control of the surface chemistry, holds considerable promise for designing smart nanomachines that autonomously reconfigure their propulsion mode for various on-demand operations.