Magnetic self-assembly of 3D multicellular microscaffolds: A biomimetic brain tumor-on-a-chip for drug delivery and selectivity testing.

In recent years, the need for highly predictive brain cancer models to test new anticancer compounds and experimental therapeutic approaches has significantly increased. Realistic in vitro brain tumor-on-a-chip platforms would allow a more accurate selection of valid candidate drugs and nanomedicines, therefore alleviating the economic and ethical issues of unsuccessful studies in vivo. Here, we present a multi-functional self-assembled brain tumor-on-a-chip model characterized by 3D glioma cultures interfaced both to nonmalignant brain cells of the peritumoral niche and to a 3D-real-scale blood-brain barrier (BBB) microfluidic system. This platform allowed us to screen multiple features, such as BBB crossing capabilities, apoptotic efficacy against GBM cells, and side effects on nonmalignant brain cells of a promising anticancer drug, nutlin-3a, which is fundamental for the treatment of brain cancer.

Direct laser writing of liquid crystal elastomers oriented by a horizontal electric field.

Background: The ability to fabricate components capable of performing actuation in a reliable and controlled manner is one of the main research topics in the field of microelectromechanical systems (MEMS). However, the development of these technologies can be limited in many cases by 2D lithographic techniques employed in the fabrication process. Direct Laser Writing (DLW), a 3D microprinting technique based on two-photon polymerization, can offer novel solutions to prepare, both rapidly and reliably, 3D nano- and microstructures of arbitrary complexity. In addition, the use of functional materials in the printing process can result in the fabrication of smart and responsive devices. Methods: In this study, we present a novel methodology for the printing of 3D actuating microelements comprising Liquid Crystal Elastomers (LCEs) obtained by DLW. The alignment of the mesogens was performed using a static electric field (1.7 V/µm) generated by indium-tin oxide (ITO) electrodes patterned directly on the printing substrates. Results: When exposed to a temperature higher than 50°C, the printed microstructures actuated rapidly and reversibly of about 8% in the direction perpendicular to the director. Conclusions: A novel methodology was developed that allows the printing of directional actuators comprising LCEs via DLW. To impart the necessary alignment of the mesogens, a static electric field was applied before the printing process by making use of flat ITO electrodes present on the printing substrates. The resulting microelements showed a reversible change in shape when heated higher than 50 °C.

A 3D Biohybrid Real-Scale Model of the Brain Cancer Microenvironment for Advanced In Vitro Testing.

The modeling of the pathological microenvironment of the central nervous system (CNS) represents a disrupting approach for drug screening for advanced therapies against tumors and neuronal disorders. The in vitro investigations of the crossing and diffusion of drugs through the blood-brain barrier (BBB) are still not completely reliable, due to technological limits in the replication of 3D microstructures that can faithfully mimic the in vivo scenario. Here, an innovative 1:1 scale 3D-printed realistic biohybrid model of the brain tumor microenvironment, with both luminal and parenchyma compartments, is presented. The dynamically controllable microfluidic device, fabricated through two-photon lithography, enables the triple co-culture of hCMEC/D3 cells, forming the internal biohybrid endothelium of the capillaries, of astrocytes, and of magnetically-driven spheroids of U87 glioblastoma cells. Tumor spheroids are obtained from culturing glioblas-toma cells inside 3D microcages loaded with superparamagnetic iron oxide nanoparticles (SPIONs). The system proves to be capable in hindering dextran diffusion through the bioinspired BBB, while allowing chemotherapy-loaded nanocarriers to cross it. The proper formation of the selective barrier and the good performance of the anti-tumor treatment demonstrate that the proposed device can be successfully exploited as a realistic in vitro model for high-throughput drug screening in CNS diseases.

A protein-coated micro-sucker patch inspired by octopus for adhesion in wet conditions.

In medical robotics, micromanipulation becomes particularly challenging in the presence of blood and secretions. Nature offers many examples of adhesion strategies, which can be divided into two macro-categories: morphological adjustments and chemical adaptations. This paper analyzes how two successful specializations from different marine animals can converge into a single biomedical device usable in moist environments. Taking inspiration from the morphology of the octopus sucker and the chemistry of mussel secretions, we developed a protein-coated octopus-inspired micro-sucker device that retains in moist conditions about half of the adhesion it shows in dry environments. From a robotic perspective, this study emphasizes the advantages of taking inspiration from specialized natural solutions to optimize standard robotic designs.

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.

Air Trapping Mechanism in Artificial Salvinia-Like Micro-Hairs Fabricated via Direct Laser Lithography.

Salvinia leaves represent an extraordinary example of how nature found a strategy for the long term retainment of air, and thus oxygen, on a surface, the so-called 'Salvinia effect', thanks to the peculiar three-dimensional and hierarchical shape of the hairs covering the leaves. Here, starting from the natural model, we have microfabricated hairs inspired by those present on the Salvinia molesta leaves, by means of direct laser lithography. Artificial hairs, like their natural counterpart, are composed of a stalk and a crown-like head, and have been reproduced in the microscale since this ensures, if using a proper design, an air-retaining behavior even if the bulk structural material is hydrophilic. We have investigated the capability of air retainment inside the heads of the hairs that can last up to 100 h, demonstrating the stability of the phenomenon. For a given dimension of the head, the greater the number of filaments, the greater the amount of air that can be trapped inside the heads since the increase in the number of solid⁻air interfaces able to pin the liquid phase. For this reason, such type of pattern could be used for the fabrication of surfaces for controlled gas retainment and gas release in liquid phases. The range of applications would be quite large, including industrial, medical, and biological fields.

Three-Dimensional Soft Material Micropatterning via Direct Laser Lithography of Flexible Molds.

Three-dimensionally micropatterned surfaces are attracting increasing interest in soft robotics owing to the potential of mimicking natural morphologies at the micro/nanoscale. We employ direct laser lithography to fabricate molds with complex three-dimensional (3D) micrometric features, in a positive photoresist on flexible substrates, to pattern curved macroscopic soft surfaces with shapes not achievable with standard methods (e.g., reentrant angles). We present several 3D intricate microstructures in poly(dimethylsiloxane) (PDMS) and show a soft cylinder patterned with 3D microstructures with one molding process. Finally, we deform PDMS-based 3D architectures and show soft microgripping capability, indicating the potentiality of this approach for future application in soft robotics.

3D Micropatterned Surface Inspired by Salvinia molesta via Direct Laser Lithography.

Biomimetic functional surfaces are attracting increasing attention for their relevant technological applications. Despite these efforts, inherent limitations of microfabrication techniques prevent the replication of complex hierarchical microstructures. Using a 3D laser lithography technique, we fabricated a 3D patterned surface bioinspired to Salvinia molesta leaves. The artificial hairs, with crownlike heads, were reproduced by scaling down (ca. 100 times smaller) the dimensions of natural features, so that microscale hairs with submicrometric resolution were attained. The micropatterned surface, in analogy with the natural model, shows interesting properties in terms of hydrophobicity and air retention when submerged by water, even if realized with a hydrophilic material. Furthermore, we successfully demonstrated the capability to promote localized condensation of water droplets from moisture in the atmosphere.