3D Bioprinted In Vitro Metastatic Models via Reconstruction of Tumor Microenvironments

The development of 3D in vitro models capable of recapitulating native tumor microenvironments could improve the translatability of potential anticancer drugs and treatments. Here, 3D bioprinting techniques are used to build tumor constructs via precise placement of living cells, functional biomaterials, and programmable release capsules. This enables the spatiotemporal control of signaling molecular gradients, thereby dynamically modulating cellular behaviors at a local level. Vascularized tumor models are created to mimic key steps of cancer dissemination (invasion, intravasation, and angiogenesis), based on guided migration of tumor cells and endothelial cells in the context of stromal cells and growth factors. The utility of the metastatic models for drug screening is demonstrated by evaluating the anticancer efficacy of immunotoxins. These 3D vascularized tumor tissues provide a proof‐of‐concept platform to i) fundamentally explore the molecular mechanisms of tumor progression and metastasis, and ii) preclinically identify therapeutic agents and screen anticancer drugs.     

Modelling of Protein‐Protein Interactions of Bone Morphogenetic Protein‐2 (BMP‐2) by 3D‐Rapid Prototyping

Recently we were able to apply the technique of 3D‐Rapid Prototyping (3D‐RP) to the construction of highly accurate three‐dimensional plastic models of biomolecules [Laub, M. et al. (2001), Materialwiss. Werkstofftech. 32, 926]. These models are derived from x‐ray crystallographic data and therefore represent exact replicas of the depicted molecules. Due to their accuracy these models should be suitable for the modelling of protein‐protein‐interactions. In a first study using 3D‐Rapid Prototyping models of bone morphogenetic protein‐2 (BMP‐2) we were able to identify a novel structural motive on the concave side of this protein which we termed anthelix since a left‐handed helix (radius ca. 0.8–1 nm, pitch 8–9 nm) can be fitted into this groove. Based on these structural findings we identified a 15mer polypeptide (KNMTPYRSPPPYVPP) from the Brookhaven database as a potential physiological ligand. Molecular docking studies using a geometric recognition approach confirmed the anthelix as a possible binding site for this peptide. However in affinity chromatography experiments no binding between BMP‐2 and the immobilized peptide was observed. As the question arose whether 3D‐Rapid Prototyping is in general suitable for modelling protein‐protein interaction we used dimeric BMP‐2 to study exemplary monomer‐dimer interaction. Molecular docking studies using the monomeric BMP‐2 subunits predicted a structure which is nearly identical to that found in dimeric BMP‐2 (root mean square deviation < 1 Å) proving the suitability of geometric docking. 3D‐RP‐BMP‐2‐monomers (size 140 mm × 75 mm × 65; magnification ca. 22 × 10⁶ fold) constructed from dimeric BMP‐2 could be assembled by hand yielding a structure highly homologous to dimeric BMP‐2. Differences between the 3D‐Rapid Prototyping model of dimeric BMP‐2 and the assembled monomers arose in several gaps at the interface between the two monomers which are not visible in the dimeric structure. These gaps can be explained by the way the solvent‐accessible molecular surface is generated. During this process an exterior probe sphere is rolled over the spherical atoms of the molecule. Distances between the monomers smaller than the diameter of this sphere are bridged thus resulting in a coherent surface. We conclude that 3D‐Rapid Prototyping is in general eligible for the modelling of protein‐protein‐interaction though there are further efforts needed to increase our understanding of this process.     

Rationally Designed 3D Hydrogels Model Invasive Lung Diseases Enabling High‐Content Drug Screening

Cell behavior is highly dependent upon microenvironment. Thus, to identify drugs targeting metastatic cancer, screens need to be performed in tissue mimetic substrates that allow cell invasion and matrix remodeling. A novel biomimetic 3D hydrogel platform that enables quantitative analysis of cell invasion and viability at the individual cell level is developed using automated data acquisition methods with an invasive lung disease (lymphangioleiomyomatosis, LAM) characterized by hyperactive mammalian target of rapamycin complex 1 (mTORC1) signaling as a model. To test the lung‐mimetic hydrogel platform, a kinase inhibitor screen is performed using tuberous sclerosis complex 2 (TSC2) hypomorphic cells, identifying Cdk2 inhibition as a putative LAM therapeutic. The 3D hydrogels mimic the native niche, enable multiple modes of invasion, and delineate phenotypic differences between healthy and diseased cells, all of which are critical to effective drug screens of highly invasive diseases including lung cancer.     

A 3D‐Structured Sustainable Solar‐Driven Steam Generator Using Super‐Black Nylon Flocking Materials

Solar‐driven evaporation is a promising way of using abundant solar energy for desalinating polluted water or seawater, which addresses the challenge of global fresh water scarcity. Cost‐effectiveness and durability are key factors for practical solar‐driven evaporation technology. The present cutting‐edge techniques mostly rely on costly and complex fabricated nanomaterials, such as metallic nanoparticles, nanotubes, nanoporous hydrogels, graphene, and graphene derivatives. Herein, a black nylon fiber (BNF) flocking board with a vertically aligned array prepared via a convenient electrostatic flocking technique is reported, presenting an extremely high solar absorbance (99.6%), a water self‐supply capability, and a unique salt self‐dissolution capability for seawater desalination. Through a carefully designed 3D structure, a plug‐in‐type BNF flocking board steam generator realizes a high evaporation rate of 2.09 kg m^?2 h^?1 under 1 kW m^?2 solar illumination, well beyond its corresponding upper limit of 1.50 kg m^?2 h^?1 (assuming 100% solar energy is being used for evaporation latent heat). With the advantages of high‐efficiency fabrication, cost‐effectiveness, high evaporation rate, and high endurance in seawater desalination, this 3D design provides a new strategy to build up an economic, sustainable, and rapid solar‐driven steam generation system.     

3D‐Printed Scanning‐Probe Microscopes with Integrated Optical Actuation and Read‐Out

Scanning‐probe microscopy (SPM) is the method of choice for high‐resolution imaging of surfaces in science and industry. However, SPM systems are still considered as rather complex and costly scientific instruments, realized by delicate combinations of microscopic cantilevers, nanoscopic tips, and macroscopic read‐out units that require high‐precision alignment prior to use. This study introduces a concept of ultra‐compact SPM engines that combine cantilevers, tips, and a wide variety of actuator and read‐out elements into one single monolithic structure. The devices are fabricated by multiphoton laser lithography as it is a particularly flexible and accurate additive nanofabrication technique. The resulting SPM engines are operated by optical actuation and read‐out without manual alignment of individual components. The viability of the concept is demonstrated in a series of experiments that range from atomic‐force microscopy engines offering atomic step height resolution, their operation in fluids, and to 3D printed scanning near‐field optical microscopy. The presented approach is amenable to wafer‐scale mass fabrication of SPM arrays and capable to unlock a wide range of novel applications that are inaccessible by current approaches to build SPMs.     

3D‐based hp‐adaptive first‐order shell finite element for modelling and analysis of complex structures—Part 2: Application to structural analysis

The paper presents a 3D‐based adaptive first‐order shell finite element to be applied to hierarchical modelling and adaptive analysis of complex structures. The main feature of the element is that it is equipped with 3D degrees of freedom, while its mechanical model corresponds to classical first‐order shell theory. Other useful features of the element are its modelling and adaptive capabilities. The element is assigned to hierarchical modelling and hpq‐adaptive analysis of shell parts of complex structures consisting of solid, thick‐ and thin‐shell parts, as well as of transition zones, where h, p and q denote the mesh density parameter and the longitudinal and transverse orders of approximation, respectively. The proposed hp‐adaptive first‐order shell element can be joined with 3D‐based hpq‐adaptive hierarchical shell elements or 3D hpp‐adaptive solid elements by means of the family of 3D‐based hpq/hp‐ or hpp/hp‐adaptive transition elements. The main objective of the first part of our research, presented in the first part of the paper, was to provide non‐standard information on the original parts of the element algorithm. Here we describe the second part of the research, devoted to the methodology and results of the application of the element to various plate and shell problems. The main objective of this part is to verify algorithms of the element and to show its usefulness in modelling and adaptive analysis of shell and plate parts of complex structures. In order to do that, there is a presentation of the results of a comparative analysis of model plate and shell problems using the classical and our elements, and equidistributed and integrated Legendre shape functions. For the plate problem a comparison of the results obtained from the adaptive and non‐adaptive analysis is also included. Additionally, some advantages of the application of our element are shown through a comparative analysis of p‐convergence of the thin plate problem and an adaptive analysis of the exemplary complex structure. Copyright © 2006 John Wiley & Sons, Ltd.     

A Cut‐and‐Paste Approach to 3D Graphene‐Oxide‐Based Architectures

Properly cut sheets can be converted into complex 3D structures by three basic operations including folding, bending, and pasting to render new functions. Folding and bending are extensively employed in crumpling, origami, and pop‐up fabrications for 3D structures. Pasting joins different parts of a material together, and can create new geometries that are fundamentally unattainable by folding and bending. However, it has been much less explored, likely due to limited choice of weldable thin film materials and residue‐free glues. Here it is shown that graphene oxide (GO) paper is one such suitable material. Stacked GO sheets can be readily loosened up and even redispersed in water, which upon drying, restack to form solid structures. Therefore, water can be utilized to heal local damage, glue separated pieces, and release internal stress in bent GO papers to fix their shapes. Complex and dynamic 3D GO architectures can thus be fabricated by a cut‐and‐paste approach, which is also applicable to GO‐based hybrid with carbon nanotubes or clay sheets.     

3D‐based hp‐adaptive first‐order shell finite element for modelling and analysis of complex structures—Part 1: The model and the approximation

The paper presents a 3D‐based adaptive first‐order shell finite element to be applied to hierarchical modelling and adaptive analysis of complex structures. The main feature of the element is that it is equipped with 3D degrees of freedom, while its mechanical model corresponds to classical first‐order shell theory. Other useful features of the element are its modelling and adaptive capabilities. The element is assigned to hierarchical modelling and hpq‐adaptive analysis of shell parts of complex structures consisting of solid, thick‐ and thin‐shell parts, as well as of transition zones, where h, p and q denote the mesh density parameter and the longitudinal and transverse orders of approximation, respectively. The proposed hp‐adaptive first‐order shell element can be joined with 3D‐based hpq‐adaptive hierarchical shell elements or 3D hpp‐adaptive solid elements by means of the family of 3D‐based hpq/hp‐ or hpp/hp‐adaptive transition elements. The main objective of the first part of our research, presented in this paper, is to provide non‐standard information on the original parts of the element algorithm. In order to do that, we present the definition of shape functions necessary for p‐adaptivity, as well as the procedure for imposing constraints corresponding to the lack of elongation of the straight lines perpendicular to the shell mid‐surface, which is the procedure necessary for q‐adaptivity. The 3D version of constrained approximation presented next is the basis for h‐adaptivity of the element. The second part of our research, devoted to methodology and results of the numerical research on application of the element to various plate and shell problems, are described in the second part of this paper. Copyright © 2006 John Wiley & Sons, Ltd.     

A 3D Real‐Scale,Biomimetic, and Biohybrid Model of the Blood‐Brain Barrier Fabricated through Two‐Photon Lithography

The investigation of the crossing of exogenous substances through the blood‐brain barrier (BBB) is object of intensive research in biomedicine, and one of the main obstacles for reliable in vitro evaluations is represented by the difficulties at the base of developing realistic models of the barrier, which could resemble as most accurately as possible the in vivo environment. Here, for the first time, a 1:1 scale, biomimetic, and biohybrid BBB model is proposed. Microtubes inspired to the brain capillaries were fabricated through two‐photon lithography and used as scaffolds for the co‐culturing of endothelial‐like bEnd.3 and U87 glioblastoma cells. The constructs show the maturation of tight junctions, good performances in terms of hindering dextran diffusion through the barrier, and a satisfactory trans‐endothelial electrical resistance. Moreover, a mathematical model is developed, which assists in both the design of the 3D microfluidic chip and its characterization. Overall, these results show the effective formation of a bioinspired cellular barrier based on microtubes reproducing brain microcapillaries to scale. This system will be exploited as a realistic in vitro model for the investigation of BBB crossing of nanomaterials and drugs, envisaging therapeutic and diagnostic applications for several brain pathologies, including brain cancer.     

3D‐Printed Biomimetic Super‐Hydrophobic Structure for Microdroplet Manipulation and Oil/Water Separation

Biomimetic functional surfaces are attracting increasing attention for various technological applications, especially the superhydrophobic surfaces inspired by plant leaves. However, the replication of the complex hierarchical microstructures is limited by the traditional fabrication techniques. In this paper, superhydrophobic micro‐scale artificial hairs with eggbeater heads inspired by Salvinia molesta leaf was fabricated by the Immersed surface accumulation three dimensional (3D) printing process. Multi‐walled carbon nanotubes were added to the photocurable resins to enhance the surface roughness and mechanical strength of the microstructures. The 3D printed eggbeater surface reveals interesting properties in terms of superhydrophobilicity and petal effect. The results show that a hydrophilic material can macroscopically behave as hydrophobic if a surface has proper microstructured features. The controllable adhesive force (from 23 μN to 55 μN) can be easily tuned with different number of eggbeater arms for potential applications such as micro hand for droplet manipulation. Furthermore, a new energy‐efficient oil/water separation solution based on our biomimetic structures was demonstrated. The results show that the 3D‐printed eggbeater structure could have numerous applications, including water droplet manipulation, 3D cell culture, micro reactor, oil spill clean‐up, and oil/water separation.     

Immunogold FIB‐SEM: Combining Volumetric Ultrastructure Visualization with 3D Biomolecular Analysis to Dissect Cell–Environment Interactions

Volumetric imaging techniques capable of correlating structural and functional information with nanoscale resolution are necessary to broaden the insight into cellular processes within complex biological systems. The recent emergence of focused ion beam scanning electron microscopy (FIB‐SEM) has provided unparalleled insight through the volumetric investigation of ultrastructure; however, it does not provide biomolecular information at equivalent resolution. Here, immunogold FIB‐SEM, which combines antigen labeling with in situ FIB‐SEM imaging, is developed in order to spatially map ultrastructural and biomolecular information simultaneously. This method is applied to investigate two different cell–material systems: the localization of histone epigenetic modifications in neural stem cells cultured on microstructured substrates and the distribution of nuclear pore complexes in myoblasts differentiated on a soft hydrogel surface. Immunogold FIB‐SEM offers the potential for broad applicability to correlate structure and function with nanoscale resolution when addressing questions across cell biology, biomaterials, and regenerative medicine.     

Keyhole‐3D phase contrast magnetic resonance angiography: A time‐resolved reconstruction method

In this study, we studied the keyhole imaging technique to 3D Phase‐contrast magnetic resonance angiography (PC MRA) to improve its temporal resolution. Previously, our research group has already studied the 2D PC MRA combined with keyhole technique, and evaluated the applicability. For keyhole‐3D PC MRA, the keyhole factor was used from 12.5% to 50% of the full k‐space. With keyhole factors above 50%, the images were similar to the original image and the vessels in the brain were well observed. We believe the keyhole‐3D PC MRA will give some advantages for improving the temporal resolution of MR systems.