Dieter Spiehl

Fused filament fabrication (FFF) is a widely used additive manufacturing method in which the process forces within the hotend play an important role in terms of print quality and speed, particularly in high-throughput applications. This work reports on the influence of filament diameter, melting zone length, and nozzle set temperature on the process forces and the maximum achievable volumetric flow rate. Experimental measurements were carried out using a test rig that integrates a load cell to capture the resulting forces, complemented by non-isothermal numerical simulations. The results show that increasing the melting zone length reduces process forces and increases the attainable volumetric flow rate at high feed rates, as the filament has more time to melt. However, the effect depends strongly on filament diameter. For a diameter of 2.85 mm, extending the melting zone leads to a monotonic increase in the maximum achievable flow rate across the entire investigated range. For a diameter of 1.75 mm, an optimum is observed at an intermediate melting zone length, beyond which additional flow resistance outweighs the benefit of improved melting and thus reduces the attainable flow rate. When normalizing for the maximum transferable extruder force, the smaller filament diameter consistently yields superior throughput performance. The simulations reproduce the experimentally observed trends well and support the interpretation that throughput is limited by the competition between heat-transfer-controlled melting and viscous pressure losses.

3D-bioprinting is a promising technique to mimic the complex anatomy of natural tissues, as it comprises a precise and gentle way of placing bioinks containing cells and hydrogel. Although hydrogels expose an ideal growth environment due to their extracellular matrix (ECM)-like properties, high water amount and tissue like microstructure, they lack mechanical strength and possess a diffusion limit of a couple of hundred micrometers. Integration of electrospun fibers could hereby benefit in multiple ways, for instance by controlling mechanical characteristics, cell orientation, direction of diffusion and anisotropic swelling behavior. The aim of this study was to create an advanced ECM-biomimicking scaffold material for tissue engineering, which offers enhanced diffusion properties. PCL bulk membranes were successfully electrospun and fragmented using a cryo cutting technique. Subsequently, these short single fibers (500 µm), swelling properties (20%–60% of control), and higher mechanical strength, while its viscosity (5–30 mPas*s) and gelation kinetics (28 °C) remained almost unaffected. The diffusion tests indicate a high level of size selectivity, which can be utilized for targeted biomolecule transport in the future. Finally, applying 3D-bioprinting technology (drop-on-demand vs. microextrusion) a print setting dependent post-dispensing orientation of the fibers could be induced, which ultimately paves way for the fabrication of metamaterials with anisotropic material properties. As expected, the fiber-filled bioink was found to be non-cytotoxic in cell culture trials using HUVECs and HepG2 (>80% viability). In summary, microfiber integration holds great promise for 3D-bioprinting of tissue percursors with advanced metamaterial properties and thus offers high applicability in various fields of research, such as in-vitro tissue models, tissue engineered implants or cultivated meat.

To combat the growing threat of antibiotic resistance, environmental testing for antibiotic contamination is gaining an increasing role. This study aims to develop an easy-to-use assay for the detection of the fluoroquinolone antibiotic levofloxacin. Levofloxacin is used in human and veterinary medicine and has been detected in wastewater and river water. An RNA aptamer against levofloxacin was selected using RNA Capture-SELEX. The 73 nt long aptamer folds into three stems with a central three-way junction. It binds levofloxacin with a K_d of 6 µM and discriminates the closely related compound ciprofloxacin. Furthermore, the selection process was analyzed using a next-generation sequencing approach to better understand the sequence evolution throughout the selection. The aptamer was used as a bioreceptor for the development of a lateral flow assay. The biosensor exploited the innate characteristic of RNA Capture-SELEX to select aptamers that displace a complementary DNA oligonucleotide upon ligand binding. The lateral flow assay achieved a limit of visual detection of 100 µM. While the sensitivity of this assay constrains its immediate use in environmental testing, the present study can serve as a template for the selection of RNA aptamer-based biosensors.

The filament drive is a key part of the extrusion assembly of a Fused Filament Fabrication printer. This investigation examines the maximum feed force and the slip of different driving rollers using a filament made of polylactic-acid (PLA) on a test stand. The test stand systematically varies the main feed process parameters: feed velocity, pinch force between the rollers, and feed force. The maximum feed force has a characteristic dependency on the pinch force combined with a feed-velocity-dependency, which is influenced by the outer diameter of the driving roller. The slip was found to increase linearly with the feed force. The slip decreases with increasing pinch force and is nearly constant for pinch forces above 77 N—172 N, depending on the driving roller tooth geometry and outer diameter. A model derived from contact mechanics was used for phenomenological modeling of the slip in relation to pinch force and feed velocity. An exponential ansatz provided good modeling of the slip at a constant pinch force. The model of the slip combined with the extrusion forces in the liquefier can be used to estimate the material flow in the future, thus leading to increased precision of the parts in a magnitude of systems.

Since the pandemic outbreak of Covid-19 in December 2019, several lateral flow assay (LFA) devices were developed to enable the constant monitoring of regional and global infection processes. Additionally, innumerable lateral flow test devices are frequently used for determination of different clinical parameters, food safety, and environmental factors. Since common LFAs rely on non-biodegradable nitrocellulose membranes, we focused on their replacement by cellulose-composed, biodegradable papers. We report the development of cellulose paper-based lateral flow immunoassays using a carbohydrate-binding module-fused to detection antibodies. Studies regarding the protein binding capacity and potential protein wash-off effects on cellulose paper demonstrated a 2.7-fold protein binding capacity of CBM-fused antibody fragments compared to the sole antibody fragment. Furthermore, this strategy improved the spatial retention of CBM-fused detection antibodies to the test area, which resulted in an enhanced sensitivity and improved overall LFA-performance compared to the naked detection antibody. CBM-assisted antibodies were validated by implementation into two model lateral flow test devices (pregnancy detection and the detection of SARS-CoV-2 specific antibodies). The CBM-assisted pregnancy LFA demonstrated sensitive detection of human gonadotropin (hCG) in synthetic urine and the CBM-assisted Covid-19 antibody LFA was able to detect SARS-CoV-2 specific antibodies present in serum. Our findings pave the way to the more frequent use of cellulose-based papers instead of nitrocellulose in LFA devices and thus potentially improve the sustainability in the field of POC diagnostics.