Biology subjects

Rawson, F. J.

Publications and source records attributed to Rawson, F. J..

4 recordsLinked to original sources

bioRxiv2026

FluoVolt Staining Induces Photodamage During Live-Cell Voltage Imaging

Fluorescent voltage-sensitive dyes (VSDs) enable non-invasive, high-throughput optical measurement of membrane potential in living cells, but the analytical reliability of such measurements depends critically on whether the dye and associated imaging conditions perturb the system under study. Here, we systematically characterise the photophysical performance and cell-perturbing effects of FluoVolt, a widely adopted VSD, across cancer cell lines (GIN31 glioblastoma and SK-MEL-30 melanoma) and primary human macrophages. Photobleaching kinetics were strongly cell-type-dependent, with SK-MEL-30 cells exhibiting complete fluorescence loss within 400 seconds under standard widefield conditions. FluoVolt staining combined with laser excitation caused an approximately 2.5-fold increase in cell detachment relative to unstained controls, and dual-wavelength excitation (488 + 405 nm) reduced GIN31 cell viability by approximately 17.5%. Critically, morphological changes, a transition from elongated to amoeboid-like phenotypes, were detected under staining conditions alone, prior to any laser exposure, indicating baseline dye-induced perturbation independent of phototoxicity. Halving dye concentration and loading time significantly attenuated these effects while preserving measurable fluorescence signal. These findings identify FluoVolt staining and excitation as previously uncharacterised sources of systematic measurement artefact and provide practical, actionable guidance for protocol design, control selection, and data interpretation in optical membrane potential imaging.

biophysics↗

bioRxiv2025

Bioelectronic Modulation of Glioblastoma via Wireless Carbon Nanotube Porin Interfaces

The membrane potential (Vmem) and faradaic charge transfer, resulting from altered charge distribution due to ion channels, play a crucial role in cellular bioelectricity. Disruption of Vmem can activate pathways associated with cancer proliferation. Manipulating ion channels may therefore present an effective strategy for treating cancers that fail to respond to conventional therapies. One approach to target these channels, is to manipulate the membrane charge which involves the use of wireless bipolar electrodes such as carbon nanotube porins (CNTPs), which could be inserted into cell membranes to mimic these channels. By utilizing membrane dyes, we observed alterations in Vmem induced by CNTPs and externally applied electric fields. Analyses of cellular behaviors and processes indicated that Vmem is more receptive to stimuli in invasive cancers, while it leads to increased metabolism in less invasive cancers, with notable changes in the cell cycle occurring at approximately 48 hours post-treatment in GBM cell lines. This work shows that CNTPs and electric fields can be used to modulate Vmem and alter cancer cell processes, supporting their potential therapeutic capability.

biophysics↗

bioRxiv2022

Electric Field Responsive Nanotransducers for Glioblastoma

Electric field therapies such as Tumor Treating Fields (TTFields) have emerged as a bioelectronic treatment for isocitrate dehydrogenase wild-type and IDH mutant grade 4 astrocytoma Glioblastoma (GBM). TTFields rely on alternating current (AC) electric fields (EF) leading to the disruption of dipole alignment and induced dielectrophoresis during cytokinesis. Although TTFields have a favourable side effect profile, particularly compared to cytotoxic chemotherapy, survival benefits remain limited ([~] 4.9 months) after an extensive treatment regime (20 hours/day for 18 months). The cost of the technology also limits its clinical adoption worldwide. Therefore, the discovery of new technology that can enhance survival benefit and improve the cost per added quality of life year per patient, of these TTFields will be of great benefit to cancer treatment and decrease healthcare costs worldwide. In this work, we report the role of electrically conductive gold (GNPs), dielectric silica oxide (SiO2), and semiconductor zinc oxide (ZnO) nanoparticles (NPs) as transducers for enhancing EF mediated anticancer effects on patient derived GBM cells. Physicochemical properties of these NPs were analyzed using spectroscopic, electron microscopy, and light-scattering techniques. In vitro TTFields studies indicated an enhanced reduction in the metabolic activity of patient-derived Glioma INvasive marginal (GIN 28) and Glioma contrast enhanced core (GCE 28) GBM cells in groups treated with NPs vs. control groups, irrespective of NPs dielectric properties. Our results indicate the inorganic NPs used in this work enhance the intracellular EF effects by virtue of bipolar dielectrophoretic and electrophoretic effects. This work presents preliminary evidence which could help to improve future EF applications for bioelectronic medicine. Furthermore, the merits of spherical morphology, excellent colloidal stability, and low toxicity, make these NPs ideal for future studies for elucidating the detailed mechanism and efficacy upon their delivery in GBM preclinical models.

cancer biology↗

bioRxiv2022

Bio-inspired artificial printed bioelectronic cardio-3D-cellular constructs

Bioelectronics is a growing field where novel smart materials are required to interface biology with electronic components. Conductive hydrogels have recently emerged as a promising material for biosensing/actuating applications as they can provide a wet, nanostructured and electrically conductive environment, minimising the mismatch between biological and electronic systems. In this work, we propose a strategy to develop conductive bioinks compatible with the freeform reversible embedding of suspended hydrogels (FRESH) extrusion bioprinting method. These bioinks are based on decellularized extracellular matrix (dECM), extracted from three different tissues (small intestine submucosa, liver and bone) and were characterised. 3D structures were manufactured containing human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs), exhibiting cell viabilities >80%. Multi-walled carbon nanotubes (MWCNTs) were selected as an additional component of the bioinks. The addition of the MWCNTs enhanced the conductive features of the hydrogels and the morphology of the dECM fibres. Electrical stimulation (ES) through alternating currents was applied to hPSC-CMs encapsulated in 3D structures manufactured with the previous material and our results indicated two main findings: (1) in the absence of external ES, the conductive properties of the materials can improve the contractile behaviour of the hPSC-CMs and (2) this effect is significantly enhanced under the application of external ES. Genetic markers analysed showed a trend towards a more mature state of the cells evaluated by the TNNI3/TNNI1 ratio, with upregulated SERCA2 and RYR2 calcium handling proteins when compared to controls and downregulation of calcium channels involved in the generation of pacemaking currents (CACNA1H). These results demonstrate the potential of our strategy to manufacture conductive hydrogels in complex geometries for bioactuating purposes. However, further development of the 3D bioprinting techniques is required to achieve higher control over the nano- and microarchitectures of the structures to improve their biomimicry.

bioengineering↗