Light-Responsive Hydrogel Microcrawlers, Powered and Steered with Spatially Homogeneous Illumination.

Sub-millimeter untethered locomoting robots hold promise to radically change multiple areas of human activity such as microfabrication/assembly or health care. To overcome the associated hurdles of such a degree of robot miniaturization, radically new approaches are being adopted, often relying on soft actuating polymeric materials. Here, we present light-driven, crawling microrobots that locomote by a single degree of freedom actuation of their light-responsive tail section. The direction of locomotion is dictated by the robot body design and independent of the spatial modulation of the light stimuli, allowing simultaneous multidirectional motion of multiple robots. Moreover, we present a method for steering such robots by reversibly deforming their front section, using ultraviolet (UV) light as a trigger. The deformation dictates the robot locomotion, performing right- or left-hand turning when the UV is turned on or off respectively. The robots' motion and navigation are not coupled to the position of the light sources, which enables simultaneous locomotion of multiple robots, steering of robots and brings about flexibility with the methods to deliver the light to the place of robot operation.

A New Class of Single-Material, Non-Reciprocal Microactuators.

A crucial component in designing soft actuating structures with controllable shape changes is programming internal, mismatching stresses. In this work, a new paradigm for achieving anisotropic dynamics between isotropic end-states-yielding a non-reciprocal shrinking/swelling response over a full actuation cycle-in a microscale actuator made of a single material, purely through microscale design is demonstrated. Anisotropic dynamics is achieved by incorporating micro-sized pores into certain segments of the structures; by arranging porous and non-porous segments (specifically, struts) into a 2D hexagonally-shaped microscopic poly(N-isopropyl acrylamide) hydrogel particle, the rate of isotropic shrinking/swelling in the structure is locally modulated, generating global anisotropic, non-reciprocal, dynamics. A simple mathematical model is introduced that reveals the physics that underlies these dynamics. This design has the potential to be used as a foundational tool for inducing non-reciprocal actuation cycles with a single material structure, and enables new possibilities in producing customized soft actuators and modular anisotropic metamaterials for a range of real-world applications, such as artificial cilia.

Friction-directed self-assembly of Janus lithographic microgels into anisotropic 2D structures.

We present a method for creating ordered 2D structures with material anisotropy from self-assembling micro-sized hydrogel particles (microgels). Microgel platelets of polygonal shapes (hexagon, square, and rhombus), obtained by a continuous scalable lithographic process, are suspended in an aqueous environment and sediment on an inclined plane. As a consequence of gravitational pull, they slide over the plane. Each half of the microgel is composed of a different type of hydrogel [poly(N-isopropylacrylamide) (PNIPAM), and poly(ethylene glycol) diacrylate (PEGDA), respectively] which exhibit different frictional coefficients when sheared over a substrate. Hence the microgels self-orientate as they slide, and the side with the lower frictional coefficient positions in the direction of sliding. The self-oriented microgels concentrate at the bottom of the tilted plane. Here they form densely packed structures with translational as well as orientational order.

Photoresponsive Hydrogel Microcrawlers Exploit Friction Hysteresis to Crawl by Reciprocal Actuation.

Mimicking the locomotive abilities of living organisms on the microscale, where the downsizing of rigid parts and circuitry presents inherent problems, is a complex feat. In nature, many soft-bodied organisms (inchworm, leech) have evolved simple, yet efficient locomotion strategies in which reciprocal actuation cycles synchronize with spatiotemporal modulation of friction between their bodies and environment. We developed microscopic (∼100 µm) hydrogel crawlers that move in aqueous environment through spatiotemporal modulation of the friction between their bodies and the substrate. Thermo-responsive poly-n-isopropyl acrylamide hydrogels loaded with gold nanoparticles shrink locally and reversibly when heated photothermally with laser light. The out-of-equilibrium collapse and reswelling of the hydrogel is responsible for asymmetric changes in the friction between the actuating section of the crawler and the substrate. This friction hysteresis, together with off-centered irradiation, results in directional motion of the crawler. We developed a model that predicts the order of magnitude of the crawler motion (within 50%) and agrees with the observed experimental trends. Crawler trajectories can be controlled enabling applications of the crawler as micromanipulator that can push small cargo along a surface.

Arbitrarily-shaped microgels composed of chemically unmodified biopolymers.

Biohydrogels, composed of naturally occurring biopolymers are typically preferred over their synthetic analogues in bioapplications thanks to their biocompatibility, bioactivity, mechanical or degradation properties. Shaping biohydrogels on the single-cell length scales (micrometers) is a key ability needed to create bioequivalent artificial cell/tissue constructs and cannot be achieved with current methods. This work introduces a method for photolithographic synthesis of arbitrarily shaped microgels composed purely of a biopolymer of choice. The biopolymer is mixed with a sacrificial photocrosslinkable polymer, and the mixture is photocrosslinked in a lithographic process, yielding anisotropic microgels with the biopolymer entrapped in the network. Subsequent ionic or covalent biopolymer crosslinking followed by template cleavage yields a microgel composed purely of a biopolymer with the 3D shape dictated by the photocrosslinking process. Method feasibility is demonstrated with two model polysaccharide biopolymers (alginate, chitosan) using suitable crosslinking methods. Next, alginate microgels were used as microtaggants on a pharmaceutical oral solid dose formulation to prevent its counterfeiting. Since the alginate is approved as an additive in the food and pharmaceutical industries, the presented tagging system can be implemented in practical use much easier than systems comprising synthetic polymers.

Biodegradable Microparticles for Simultaneous Detection of Counterfeit and Deteriorated Edible Products.

In an era of globalized trade relations where food and pharmaceutical products cross borders effortlessly, consumers face counterfeit and deteriorated products at elevated rates. This paper presents multifunctional, biodegradable hydrogel microparticles that can provide information on the authenticity and the potential deterioration of the tagged food or pharmaceutical formulations. These microparticles integrate spatially patterned authenticity code with two sensors-the first one detects possible presence of pathogenic microbes through monitoring pH while the second one identifies products stored above optimal temperatures via optical monitoring of the microparticle degradation. Particles are synthesized from a biocompatible polymer and a photoinitiator, dextran modified with 2-hydroxyethylmethacrylate and riboflavin, respectively, using a continuous, high throughput method stop-flow lithography. The proposed synthesis approach also enables crosslinking with visible light bringing about additional flexibility to flow lithography. Model liquid and solid food and pharmaceutical products are successfully labeled with microparticles and the functionality of the sensors in aqueous solutions is demonstrated.

Designing the nanobiointerface of fluorescent nanodiamonds: highly selective targeting of glioma cancer cells.

Core-shell nanoparticles based on fluorescent nanodiamonds coated with a biocompatible N-(2-hydroxypropyl)methacrylamide copolymer shell were developed for background-free near-infrared imaging of cancer cells. The particles showed excellent colloidal stability in buffers and culture media. After conjugation with a cyclic RGD peptide they selectively targeted integrin αvß3 receptors on glioblastoma cells with high internalization efficacy.

Plasmonic nanodiamonds: targeted core-shell type nanoparticles for cancer cell thermoablation.

Targeted biocompatible nanostructures with controlled plasmonic and morphological parameters are promising materials for cancer treatment based on selective thermal ablation of cells. Here, core-shell plasmonic nanodiamonds consisting of a silica-encapsulated diamond nanocrystal coated in a gold shell are designed and synthesized. The architecture of particles is analyzed and confirmed in detail using electron tomography. The particles are biocompatibilized using a PEG polymer terminated with bioorthogonally reactive alkyne groups. Azide-modified transferrin is attached to these particles, and their high colloidal stability and successful targeting to cancer cells overexpressing the transferrin receptor are demonstrated. The particles are nontoxic to the cells and they are readily internalized upon binding to the transferrin receptor. The high plasmonic cross section of the particles in the near-infrared region is utilized to quantitatively ablate the cancer cells with a short, one-minute irradiation by a pulse 750-nm laser.

Unambiguous observation of shape effects on cellular fate of nanoparticles.

Cellular fate of nanoparticles is vital to application of nanoparticles to cell imaging, bio-sensing, drug delivery, suppression of drug resistance, gene delivery, and cytotoxicity analysis. However, the current studies on cellular fate of nanoparticles have been controversial due to complications of interplay between many possible factors. By well-controlled experiments, we demonstrated unambiguously that the morphology of nanoparticles independently determined their cellular fate. We found that nanoparticles with sharp shapes, regardless of their surface chemistry, size, or composition, could pierce the membranes of endosomes that carried them into the cells and escape to the cytoplasm, which in turn significantly reduced the cellular excretion rate of the nanoparticles. Such features of sharp-shaped nanoparticles are essential for drug delivery, gene delivery, subcellular targeting, and long-term tracking. This work opens up a controllable, purely geometrical and hence safe, degree of freedom for manipulating nanoparticle-cell interaction, with numerous applications in medicine, bio-imaging, and bio-sensing.

Fluorescent nanodiamonds embedded in biocompatible translucent shells.

High pressure high temperature (HPHT) nanodiamonds (NDs) represent extremely promising materials for construction of fluorescent nanoprobes and nanosensors. However, some properties of bare NDs limit their direct use in these applications: they precipitate in biological solutions, only a limited set of bio-orthogonal conjugation techniques is available and the accessible material is greatly polydisperse in shape. In this work, we encapsulate bright 30-nm fluorescent nanodiamonds (FNDs) in 10-20-nm thick translucent (i.e., not altering FND fluorescence) silica shells, yielding monodisperse near-spherical particles of mean diameter 66 nm. High yield modification of the shells with PEG chains stabilizes the particles in ionic solutions, making them applicable in biological environments. We further modify the opposite ends of PEG chains with fluorescent dyes or vectoring peptide using click chemistry. High conversion of this bio-orthogonal coupling yielded circa 2000 dye or peptide molecules on a single FND. We demonstrate the superior properties of these particles by in vitro interaction with human prostate cancer cells: while bare nanodiamonds strongly aggregate in the buffer and adsorb onto the cell membrane, the shell encapsulated NDs do not adsorb nonspecifically and they penetrate inside the cells.

Fluorescent Nanodiamonds with Bioorthogonally Reactive Protein-Resistant Polymeric Coatings.

The novel synthesis of a polymeric interface grown from the surface of bright fluorescent nanodiamonds is reported. The polymer enables bioorthogonal attachment of various molecules by click chemistry; the particles are resistant to nonspecific protein adsorption and show outstanding colloidal stability in buffers and biological media. The coating fully preserves the unique optical properties of the nitrogen-vacancy centers that are crucial for bioimaging and sensoric applications.

Gadolinium- and manganite-based contrast agents with fluorescent probes for both magnetic resonance and fluorescence imaging of pancreatic islets: a comparative study.

Three magnetic resonance (MR)/fluorescence imaging probes were tested for visualization, cellular distribution, and survival of labeled pancreatic islets in vitro and following transplantation. As T(1) contrast agents (CAs), gadolinium(III) complexes linked to ß-cyclodextrin (Gd-F-ßCD) or bound to titanium dioxide (TiO2 @RhdGd) were tested. As a T(2) CA, perovskite manganite nanoparticles (LSMO@siF@si) were examined. Fluorescein or rhodamine was incorporated as a fluorescent marker in all probes. Islets labeled with gadolinium(III) CAs were visible as hyperintense spots on MR in vitro, but detection in vivo was inconclusive. Islets labeled with LSMO@siF@si CA were clearly visible as hypointense spots or areas on MR scans in vitro as well as in vivo. All CAs were detected inside the islet cells by fluorescence. Although the vitality and function of the labeled islets was not impaired by any of the tested CAs, results indicate that LSMO@siF@si CA is a superior marker for islet labeling, as it provides better contrast enhancement within a shorter scan time.

Boosting nanodiamond fluorescence: towards development of brighter probes.

A novel approach for preparation of ultra-bright fluorescent nanodiamonds (fNDs) was developed and the thermal and kinetic optimum of NV center formation was identified. Combined with a new oxidation method, this approach enabled preparation of particles that were roughly one order of magnitude brighter than particles prepared with commonly used procedures.

Phosphonate-titanium dioxide assemblies: platform for multimodal diagnostic-therapeutic nanoprobes.

Multimodal imaging-therapeutic nanoprobe TiO(2)@RhdGd was prepared and successfully used for in vitro and in vivo cell tracking as well as for killing of cancer cells in vitro. TiO(2) nanoparticles were used as a core for phosphonic acid modified functionalities, responsible for contrast in MRI and optical imaging. The probe shows high (1)H relaxivity and relaxivity density values. Presence of fluorescent dye allows for visualization by means of fluorescence microscopy. The applicability of the probe was studied, using mesenchymal stem cells, cancer HeLa cells, and T-lymphocytes. The probe did not exhibit toxicity in any of these systems. Labeled cells were successfully visualized in vitro by means of fluorescence microscopy and MRI. Furthermore, it was shown that the probe TiO(2)@RhdGd can be changed into a cancer cell killer upon UV light irradiation. The above stated results represent a valuable proof of a principle showing applicability of the probe design for diagnosis and therapy.