Biology subjects

Caffrey, T. M.

Publications and source records attributed to Caffrey, T. M..

5 recordsLinked to original sources

bioRxiv2026

Strategies to improve neurite outgrowth from primary neurons ingelatin methacrylate hydrogels polymerized with visible lightexposure inside a microscale 3D model

Gelatin methacrylate (GelMA) hydrogels offer many advantageous properties such as excellent biocompatibility, tunable stiffness, and rapid fabrication, making them attractive materials for neural systems-on-a-chip devices used to study axonal outgrowth in three-dimensional (3D) environments. In this study, we evaluate properties of GelMA hydrogels crosslinked with visible blue light and application of these hydrogels for encapsulation of primary dorsal root ganglion (DRG) explants inside a custom microscale device. We selected low GelMA polymer concentrations of 3 % and 6 % w/v for hydrogel preparation to create soft materials and demonstrate the effect of degree of functionalization (DoF) on stiffness of GelMA in a blue light-induced crosslinking system. Additionally, a decrease in crosslinking efficiency and stiffness of GelMA was observed upon reconstitution of the polymer in DMEM cell culture media compared to PBS. To evaluate the axonal outgrowth, DRG explants were encapsulated in various GelMA formulations and cultured for 7 days. Following incubation, the explants were fixed and immunochemically labelled with anti-beta-III-tubulin antibody. We further describe an application of custom-developed semi-automated image analysis tool together with linear mixed effects modeling to evaluate axonal outgrowth across different GelMA formulations. Among the formulations tested in our study, 3 % GelMA 95 % DoF hydrogels supported the longest neurite outgrowth suggesting that matrix properties influenced neurite extension.

bioengineering↗

bioRxiv2026

Effects of MRI on an Injectable Hydrogel with Magnetically Alignable Microstructures for Oriented Cell Growth

Injectable biomaterials with aligned microstructures play a critical role in tissue engineering and drug-delivery applications where control over the position and orientation of cells and nano/micron-scale architectures enhance intervention efficacy. Patients are often subject to MRI scans; for patient safety and treatment efficacy, we investigated the effects of MRI on a biomaterial treatment consisting of aligned magnetic microstructures being developed for guiding cell growth. Under MRI exposure, potential movement of aligned structures could be detrimental to nearby cells, and potential MRI-induced heating could adversely affect traumatized tissue. In this work, the alignment state and heat conduction of such a treatment were studied using a 9.4 T preclinical MRI. The treatment comprises short magnetic rod-shaped polycaprolactone fibers (rods) with embedded magnetic nanoparticles in a surrounding hydrogel (gelatin methacrylate), with rod alignment observed before and after a 45-minute MRI scan. No change in rod alignment state was observed, and no heat generation was measured. A theoretical framework was developed which supports the experimental observation that the biomaterial is stable under MRI. This work can be extended to other biomaterial systems with aligned architectures used in tissue engineering applications such as spinal cord, muscle and tendon.

bioengineering↗

bioRxiv2025

Engineering a Multilayer Microfluidic Airway-On-A-Chip with Tunable GelMA Hydrogel for Physiologically Relevant Aerosol Exposure Studies

Climate change-driven increases in forest fires pose a major global health risk due to exposure to smoke containing hazardous gases and fine particulates, emphasizing the need for physiologically relevant in vitro airway models for studying smoke-induced responses. Microfluidic lung-on-a-chip technologies provide a strong foundation for in vitro airway modeling and ongoing developments are expanding their ability to incorporate multicellular organization, extracellular matrix complexity, and physiologically relevant exposure methods. This work presents the optimization and integration of a photopolymerizable gelatin methacrylate (GelMA)-based hydrogel into a microfluidic airway-on-a-chip that models the human small conducting airways and supports controlled aerosol exposure to wood smoke. The GelMA hydrogel was optimized to support fibroblast encapsulation, endothelial, and epithelial adhesion and robust mechanical stability. The device combines the hydrogel with a compartmentalized microchannel layout, and sacrificial molding to create a 3D organotypic airway culture featuring a multilayer architecture, 3D stromal matrix, and a perfusable vasculature-like lumen. Coupling the platform with a custom aerosol exposure system enables precise, biomimetic exposure to whole wood smoke. Proof-of-concept studies using transforming growth factor beta1 (TGF-{beta}1) and whole wood smoke elicited expected inflammatory and fibrotic responses, validating the platforms physiological relevance for inhalation studies and investigating smoke-induced airway remodeling and inflammation.

bioengineering↗

bioRxiv2025

Optical Properties of Gelatin Methacrylate and Implications for In Situ Cross-Linking in Spinal Cord Lesions and EngineeredHydrogel Systems

Understanding how light interacts with biomaterials is critical not only for therapeutic delivery in vivo but also for enabling emerging light-based fabrication methods such as volumetric bioprinting. In particular, optical characterization of photosensitive hydrogels like Gelatin Methacrylate (GelMA) provides foundational data for additive manufacturing of complex, tissue-engineered constructs with spatially controlled architecture. GelMA is a promising candidate for use as an injectable hydrogel for spinal cord injury (SCI) repair. This is primarily due to its highly tunable properties, allowing it to mimic tissue. Additionally, GelMA can be optically crosslinked. Optical simulations of this material within the spinal cord can inform whether the cross-linking of this material is feasible and uniform within a particular injury with a chosen exposure paradigm. These simulations require the optical properties of the tissues and materials of interest. However, the optical properties of GelMA have not been studied. The optical properties of GelMA were measured using a double integrating sphere setup and modelled with inverse adding doubling. We measured the absorption coefficient ({micro}a [mm-1]), reduced scattering coefficient ({micro}s[mm-1]), and scattering anisotropy (g [unitless]) for GelMA hydrogels, and examined the effect on these properties when magnetically alignable microstructures were suspended in the hydrogel. We conducted Monte Carlo simulations using these optical properties to determine whether the GelMA hydrogels would be cross-linked under different illumination conditions within a representative spinal cord injury geometry. The depth of hydrogel cross-linking for this representative spinal cord injury geometry was experimentally validated. Overall, it was found that GelMA without rods has an {micro}a = 0.516 {+/-} 0.025 mm-1, with insignificant scattering at 450 nm, GelMA with rods has an {micro}a = 0.588 {+/-} 0.018 mm-1, {micro}s = 0.196 {+/-} 0.024 mm-1, and g = 0.906 {+/-} 0.014 at 450 nm. Additionally, it was observed that in fluid-filled lesions, external illumination is often sufficient to achieve uniform gel curing. In contrast, fibrotic lesions require an intraspinal approach to adequately deliver light throughout the lesion, although the dose distribution will be nonuniform. Finally, experimental results confirmed the simulation findings, showing that when exposing GelMA to 63.29 mW/cm2 of 450 nm light, curing is uniform within the first 4 mm of gel and worsens with depth. These findings support the conclusion that larger volumes of photosensitive GelMA can be feasibly cured within the porcine spinal cord, but an intraspinal exposure will likely be required within human spinal tissue. These findings not only support the feasibility of in situ photopolymerization for SCI repair, but also provide critical optical parameters and validation approaches relevant to bioprinting systems.

bioengineering↗

bioRxiv2022

Tau depletion in human neurons mitigates A beta-driven toxicity

BackgroundAlzheimers disease (AD) is an age-related neurodegenerative condition and the most common type of dementia, characterised by pathological accumulation of extracellular plaques and intracellular neurofibrillary tangles that mainly consist of amyloid-{beta} (A{beta}) and hyperphosphorylated tau aggregates, respectively. Previous studies in mouse models with a targeted knock-out of the microtubule-associated protein tau (Mapt) gene demonstrated that A{beta}-driven toxicity is tau-dependent. However, human cellular models with chronic tau lowering remain unstudied. MethodsIn this study, we generated stable tau-depleted human induced pluripotent stem cell (iPSC) isogenic panels from two healthy individuals using CRISPR-Cas9 technology. We then differentiated these iPSCs into cortical neurons in vitro in co-culture with primary rat cortical astrocytes before conducting electrophysiological and imaging experiments for a wide range of disease-relevant phenotypes. Both AD brain derived and recombinant A{beta} were used in this study to elicit toxic responses from the iPSC- derived cortical neurons. ResultsWe showed that tau depletion in human iPSC-derived cortical neurons caused considerable reductions in neuronal activity without affecting synaptic density. We also observed neurite outgrowth impairments in two of the tau-depleted lines used. We found axonal transport of mitochondria, mitochondrial function, and cortical neuron differentiation propensity remained unaffected regardless of tau expression levels. Finally, tau depletion protected neurons from adverse effects mitigating the impact of exogenous A{beta}-induced hyperactivity, deficits in retrograde axonal transport of mitochondria, and neurodegeneration. ConclusionsOur study established stable human iPSC isogenic panels with chronic tau depletion from two healthy individuals. Cortical neurons derived from these iPSC lines showed that tau is essential in A{beta}-driven hyperactivity, axonal transport deficits, and neurodegeneration, consistent with studies conducted in Mapt-/- mouse models. These findings highlight the protective effects of chronic tau lowering strategies in AD pathogenesis and reinforce the potential in clinical settings. The tau-depleted human iPSC models can now be applied at scale to investigate the involvement of tau in disease-relevant pathways and cell types.

neuroscience↗