Direct laser writing-enabled 3D printing strategies for microfluidic applications.

Over the past decade, additive manufacturing-or "three-dimensional (3D) printing"-has attracted increasing attention in the Lab on a Chip community as a pathway to achieve sophisticated system architectures that are difficult or infeasible to fabricate via conventional means. One particularly promising 3D manufacturing technology is "direct laser writing (DLW)", which leverages two-photon (or multi-photon) polymerization (2PP) phenomena to enable high geometric versatility, print speeds, and precision at length scales down to the 100 nm range. Although researchers have demonstrated the potential of using DLW for microfluidic applications ranging from organ on a chip and drug delivery to micro/nanoparticle processing and soft microrobotics, such scenarios present unique challenges for DLW. Specifically, microfluidic systems typically require macro-to-micro fluidic interfaces (e.g., inlet and outlet ports) to facilitate fluidic loading, control, and retrieval operations; however, DLW-based 3D printing relies on a micron-to-submicron-sized 2PP volume element (i.e., "voxel") that is poorly suited for manufacturing these larger-scale fluidic interfaces. In this Tutorial Review, we highlight and discuss the four most prominent strategies that researchers have developed to circumvent this trade-off and realize macro-to-micro interfaces for DLW-enabled microfluidic components and systems. In addition, we consider the possibility that-with the advent of next-generation commercial DLW printers equipped with new dynamic voxel tuning, print field, and laser power capabilities-the overall utility of DLW strategies for Lab on a Chip fields may soon expand dramatically.

Correction: Direct laser writing-enabled 3D printing strategies for microfluidic applications.

Correction for 'Direct laser writing-enabled 3D printing strategies for microfluidic applications' by Olivia M. Young et al., Lab Chip, 2024, DOI: https://doi.org/10.1039/D3LC00743J.

FABRICATION OF MULTILUMEN MICROFLUIDIC TUBING FOR EX SITU DIRECT LASER WRITING.

Among the numerous additive manufacturing or "three-dimensional (3D) printing" techniques, two-photon Direct Laser Writing (DLW) is distinctively suited for applications that demand high geometric versatility with micron-to-submicron-scale feature resolutions. Recently, "ex situ DLW (esDLW)" has emerged as a powerful approach for printing 3D microfluidic structures directly atop meso/macroscale fluidic tubing that can be manipulated by hand; however, difficulties in creating custom esDLW-compatible multilumen tubing at such scales has hindered progress. To address this impediment, here we introduce a novel methodology for fabricating submillimeter multilumen tubing for esDLW 3D printing. Preliminary fabrication results demonstrate the utility of the presented strategy for resolving 743 µm-in-diameter tubing with three lumens-each with an inner diameter (ID) of 80 µm. Experimental results not only revealed independent flow of discrete fluorescently labelled fluids through each of the three lumens, but also effective esDLW-printing of a demonstrative 3D "MEMS" microstructure atop the tubing. These results suggest that the presented approach could offer a promising pathway to enable geometrically sophisticated microfluidic systems to be 3D printed with input and/or output ports fully sealed to multiple, distinct lumens of fluidic tubing for emerging applications in fields ranging from drug delivery and medical diagnostics to soft surgical robotics.

TOWARD CONTROLLED-RELEASE DRUG DELIVERY MICROCARRIERS ENABLED BY DIRECT LASER WRITING 3D PRINTING.

Controlled-release, and especially long-acting, drug delivery systems hold promise for improving treatments for numerous medical conditions. Previously, we reported an additive manufacturing or "three-dimensional (3D) printing" approach for fabricating liquid-core-shell-cap microcarriers comprising standard photoresists. Here we explore the potential to extend this strategy to achieve microcarriers comprising biodegradable materials as a new pathway to controlled-release drug delivery options. Specifically, we investigate the use of "Two-Photon Direct Laser Writing (DLW)" as a means to 3D print microcarriers composed of: (i) a bottle-shaped "shell" with an orifice, (ii) an aqueous liquid "core", and (iii) a biodegradable "cap". The cap, which is DLW-printed directly onto the shell's orifice, is designed to degrade over time in the body-e.g., with degradation time proportional to cap thickness-to ultimately facilitate release of the liquid core at desired time points. Fabrication results based on the use of a biodegradable poly(ethylene glycol) diacrylate (PEGDA) photomaterial for the cap revealed that shell designs incorporating microfluidic obstruction structures appeared to limit undesired entry of the liquid-phase PEGDA into the shell (i.e., directly preceding cap printing), thereby resulting in improved retention of the liquid core after completion of the cap printing process. These results mark an important first step toward evaluating the utility of the presented DLW 3D printing strategy for possible drug delivery applications.

GEOMETRIC DETERMINANTS OF CELL VIABILITY FOR 3D-PRINTED HOLLOW MICRONEEDLE ARRAY-MEDIATED DELIVERY.

A wide range of emerging biomedical applications and clinical interventions rely on the ability to deliver living cells via hollow, high-aspect-ratio microneedles. Recently, microneedle arrays (MNA) have gained increasing interest due to inherent benefits for drug delivery; however, studies exploring the potential to harness such advantages for cell delivery have been impeded due to the difficulties in manufacturing high-aspect-ratio MNAs suitable for delivering mammalian cells. To bypass these challenges, here we leverage and extend our previously reported hybrid additive manufacturing (or "three-dimensional (3D) printing) strategy-i.e., the combined the "Vat Photopolymerization (VPP)" technique, "Liquid Crystal Display (LCD)" 3D printing with "Two-Photon Direct Laser Writing (DLW)"-to 3D print hollow MNAs that are suitable for cell delivery investigations. Specifically, we 3D printed four sets of 650 µm-tall MNAs corresponding to needle-specific inner diameters (IDs) of 25 µm, 50 µm, 75 µm, and 100 µm, and then examined the effects of these MNAs on the post-delivery viability of both dendritic cells (DCs) and HEK293 cells. Experimental results revealed that the 25 µm-ID case led to a statistically significant reduction in post-MNA-delivery cell viability for both cell types; however, MNAs with needle-specific IDs ≥ 50 µm were statistically indistinguishable from one another as well as conventional 32G single needles, thereby providing an important benchmark for MNA-mediated cell delivery.

Differences in the mechanical properties of intestinal tissue based on preservation freezing duration and temperature.

In this study, tissue samples were stress tested to determine if freezing duration and temperature alter their mechanical properties. Tissue samples taken from the small intestine of pigs were assigned to 5 groups: fresh tissue, -28.9 °C for 7 days, -62.2 °C for 7 days, -28.9 °C for 30 days, and -62.2 °C for 30 days. Tissue was stored in PBS for the assigned temperature and duration until testing occurred with the exception of fresh tissue which was tested at sample collection. Before testing, samples were thawed in a room temperature bath, and the thickness was measured. Samples were then mounted in a biaxial test system using four anchoring rakes. Each sample was pulled to a strain of 0.2 with the corresponding forces recorded. This cycle of relaxation to 0.2 strain was repeated 5 times per sample. The thickness and force values were used to find the first Piola-Kirchhoff stress experienced at 0.2 strain and the strain energy. The average stress values in the circumferential direction were: fresh tissue: 22.3 ± 9.85 kPa; -28.9 °C for 7 days: 37.8 ± 14.1 kPa; -62.2 °C for 7 days: 46.5 ± 19.0 kPa; -28.9 °C for 30 days: 46.4 ± 22.7 kPa; -62.2 °C for 30 days: 40.1 ± 19.5 kPa. The stress and strain energy values of frozen tissue were statistically higher than the fresh tissue, although no statistical difference was found by varying duration or temperature. Based on this result, we determined that freezing tissue at any of the tested temperatures or durations increases the stiffness of the thawed tissue. This possibly occurs due to the directional formation of ice, which increases ion concentrations and glycosaminoglycan (GAG) interactions near collagen fibrils.

3D-Printed Microinjection Needle Arrays via a Hybrid DLP-Direct Laser Writing Strategy.

Microinjection protocols are ubiquitous throughout biomedical fields, with hollow microneedle arrays (MNAs) offering distinctive benefits in both research and clinical settings. Unfortunately, manufacturing-associated barriers remain a critical impediment to emerging applications that demand high-density arrays of hollow, high-aspect-ratio microneedles. To address such challenges, here, a hybrid additive manufacturing approach that combines digital light processing (DLP) 3D printing with "ex situ direct laser writing (esDLW)" is presented to enable new classes of MNAs for fluidic microinjections. Experimental results for esDLW-based 3D printing of arrays of high-aspect-ratio microneedles-with 30 µm inner diameters, 50 µm outer diameters, and 550 µm heights, and arrayed with 100 µm needle-to-needle spacing-directly onto DLP-printed capillaries reveal uncompromised fluidic integrity at the MNA-capillary interface during microfluidic cyclic burst-pressure testing for input pressures in excess of 250 kPa (n = 100 cycles). Ex vivo experiments perform using excised mouse brains reveal that the MNAs not only physically withstand penetration into and retraction from brain tissue but also yield effective and distributed microinjection of surrogate fluids and nanoparticle suspensions directly into the brains. In combination, the results suggest that the presented strategy for fabricating high-aspect-ratio, high-density, hollow MNAs could hold unique promise for biomedical microinjection applications.

A Factorial Approach for Optimizing the Design Parameters of a Tissue Attachment Mechanism for Drug Delivery.

Biological macromolecule drugs or biologics are not suited for commonly preferred oral delivery due to their intrinsic instability and physical, chemical, or immunological barriers to the gastrointestinal tract. Ingestible capsule robots (ICR) have become a versatile platform, including use for drug delivery applications for various gastrointestinal pathologies with future potential for systemic drug delivery. In this work, a tissue attachment mechanism (TAM) for a drug delivery ICR is introduced that can facilitate a non-invasive systemic delivery of unaltered biologics via direct injection through the insensate layers of the small intestine. The main prerequisite for achieving systemic drug delivery via this device is to have a strong tissue attachment of the TAM. This study aimed to optimize the attachment success rate for drug delivery and characterize attachment duration in vivo. A fractional factorial approach was used in vivo to identify and optimize factors that most influence attachment of the TAM to maximize attachment rate. Multiple in vivo optimization levels were performed using the small intestine of anesthetized pigs, and an attachment success rate of 92% was achieved. Optimal TAMs were surgically placed in vivo to determine the duration of attachment following anesthetization and surgery recovery. The average in vivo attachment duration was 32.2±9.4 hours. This work establishes a device for consistent and reliable attachment duration, making the TAM a suitable candidate for a 24-hour systemic drug delivery platform.

A Novel Capsule-Delivered Enteric Drug-Injection Device for Delivery of Systemic Biologics: A Pilot Study in a Porcine Model.

Innovative swallowable capsule technologies such as drug-loaded, dissolvable microneedles, mucoadhesive patches, and various microdevices present unique drug-carrying capabilities to overcome challenges regarding oral delivery of biologics. Here, we report a swallowable capsule for intestinal drug delivery (SCIDD) with the potential of directly injecting biological therapeutics into the insensate small intestine wall. The design, optimization, and validation of the SCIDD's primary subsystems were performed both ex-vivo and in-vivo. The assembled capsule was further tested in vivo to validate the actuation sequence and showed a 70% (n = 17) success rate in an animal model. Additionally, a drug delivery study indicated systemic uptake of adalimumab via SCIDD compared with luminal delivery in the small intestine. The pilot study presented here establishes that the novel platform could be used to orally deliver systemic biologics.

Computational optimization of a novel atraumatic catheter for local drug delivery in coronary atherosclerotic plaques.

Early identification and treatment of high-risk plaques before they rupture, and precipitate adverse events constitute a major challenge in cardiology today. Computational simulations are a time- and cost-effective way to study the performance, and to optimize a system. The main objective of this work is to optimize the flow of a novel atraumatic local drug delivery catheter for the treatment of coronary atherosclerosis. The mixing and spreading effectiveness of a drug fluid was analyzed utilizing computational fluid dynamics (CFD) in a coronary artery model. The optimum infusion flow of the nanoparticle-carrying drug fluid was found by maximizing the drug volume fraction and minimizing drug velocity at the artery wall, while maintaining acceptable wall shear stress (WSS). Drug velocities between 15 m/s and 20 m/s are optimum for local drug delivery. The resulting parameters from this study will be used to fabricate customized prototypes for future in-vivo experiments.