RESUMEN

BACKGROUND: Silicon-based micro-pillar substrates (MPS), as three-dimensional cell culture platforms with vertically aligned micro-patterned scaffolding structures, are known to facilitate high-quality growth and morphology of dorsal root ganglion (DRG) sensory neurons, promote neurite outgrowth and enhance neurite alignment. However, the electrophysiological aspects of DRG neurons cultured on silicon MPSs have not been thoroughly investigated, which is of greatest importance to ensure that such substrates do not disrupt neuronal homeostasis and function before their widespread adoption in diverse biomedical applications. NEW METHOD: We conducted whole-cell patch-clamp recordings to explore the electrophysiological properties of DRG neurons cultured on MPS arrays, utilizing a custom-made upright patch-clamp setup. RESULTS: Our findings revealed that DRG neurons exhibited similar electrophysiological responses on patterned MPS samples when compared to the control planar glass surfaces. Notably, there were no significant differences observed in the action potential parameters or firing patterns of action potentials between neurons grown on either substrate. COMPARISON WITH EXISTING METHODS: In the current study we for the first time confirmed that successful electrophysiological recordings can be obtained from the cells grown on MPS. CONCLUSION: Our results imply that, despite the potential alterations caused by the cumulative trauma of tissue harvest and cell dissociation, essential functional cell properties of DRG neurons appear to be relatively maintained on MPS surfaces. Therefore, vertically aligned silicon MPSs could be considered as a potentially effective three-dimensional system for supporting a controlled cellular environment in culture.

Asunto(s)

Ganglios Espinales , Técnicas de Placa-Clamp , Silicio , Ganglios Espinales/fisiología , Ganglios Espinales/citología , Animales , Técnicas de Placa-Clamp/instrumentación , Técnicas de Placa-Clamp/métodos , Células Cultivadas , Potenciales de Acción/fisiología , Neuronas/fisiología , Neuronas/citología , Ratas Sprague-Dawley , Ratas , Técnicas de Cultivo Tridimensional de Células/métodos , Técnicas de Cultivo Tridimensional de Células/instrumentación , Fenómenos Electrofisiológicos/fisiología

RESUMEN

Biofilms are complex three-dimensional microbial communities that adhere to a variety of surfaces and interact with their surroundings. Because of the dynamic nature of biofilm formation, establishing a uniform technique for quantifying and monitoring biofilm volume, shape, and features in real-time is challenging. Herein, we describe a noninvasive electrochemical impedance approach for real-time monitoring of dental plaque-derived multispecies biofilm growth on a range of substrates. A working equation relating electrochemical impedance to live biofilm volume has been developed that is applicable to all three surfaces examined, including glass, dental filling resin, and Ca2+-releasing resin composites. Impedance changes of 2.5, 35, 50, and 65% correlated to biofilm volumes of 0.10 ± 0.01, 16.9 ± 2.2, 29.7 ± 2.3, and 38.6 ± 2.8 µm3/µm2, respectively. We discovered that glass, dental filling resin, and Ca2+-releasing dental composites required approximately 3.5, 4.5, and 6 days, respectively, to achieve a 50% change in impedance. The local pH change at the biofilm-substrate interfaces also monitored with potentiometry pH microsensor, and pH change varied according to biofilm volume. This impedance-based technique can be a useful analytical method for monitoring the growth of biofilms on a variety of substrates in real-time. Therefore, this technique may be beneficial for examining antibacterial properties of novel biomaterials.