Laser light can be used during endodontic procedures to sterilize the root canal by destroying bacteria. Previous in-vitro studies that investigated the mechanism of the destruction of bacteria inhabiting the root canal by 1,064-nm Nd:YAG and 808-nm diode laser light used substrates that absorb light in the near-infrared (NIR) spectrum. These substrates heat the bacterial microenvironment, which possibly contributes to cell death. To determine the direct effect of laser light on the bacterial sample in the absence of detrimental heating, a sapphire substrate, which is virtually transparent in NIR spectrum, was inoculated with bacterial samples and subjected to laser irradiation at 1,064 nm (1.5 W, 15 Hz) and at 808 nm (1.5 W, 20 Hz). Enterococcus faecalis, Escherichia coli, and Porphyromonas gingivalis bacteria were used. E. faecalis and E. coli were largely unaffected by laser light. The viability of P. gingivalis, a pigmented bacterium, was directly affected by both NIR wavelengths (a 57% decrease of viability at 1,064 nm and a 31% decrease at 808 nm). Our results indicate that the primary mediator of cell death appears to be the interaction between NIR laser light and the bacterial microenvironment, most likely in the form of heating. Our research suggests that when optimizing the efficacy of laser-assisted endodontic sterilization of the root canal, the optical characteristics of the bacterial microenvironment play a key role, as nonpigmented bacteria appear to be virtually transparent at 808 nm and 1,064 nm.
Our aim was to evaluate thermal damage to endodontic pathogen Enterococcus faecalis (E. faecalis) caused by sub-second laser-generated heat pulses by determining the parameters for the thermal damage survival curve (TDSC). A novel experimental method for thermal pulsing of bacteria in the millisecond range was developed. After cultivation, E. faecalis was inoculated on anodized aluminum substrate and heated with a pulsed Nd:YAG laser. Viability was assessed with both plate count and flow cytometry methods. An E. faecalis TDSC for single-pulse millisecond range heating times was derived from the Arrhenius equation. Results gained from single-pulse heating viability measurements were used to predict the bactericidal effect of multiple sequential pulses (pulse train), and compared to experimental measurements. The thermal damage model was then applied to determine the relationship between laser fluence, pulse width, and the viability decrease of E. faecalis in a simulated root canal disinfection procedure. The application of the model to calculate the required lethal laser fluence levels on dentin during endodontic laser procedures seems to indicate that for endodontic procedures, the sub-millisecond pulsed Nd:YAG lasers are more effective in comparison with continuous-mode diode lasers and will cause less undesirable bulk heating of the tooth and surrounding tissues. The results of the study can be applied to create a model for predicting the impact of sub-second temperature increase on viability of bacteria on various surfaces and calculate required fluences and pulse widths to achieve the aforementioned effects with laser pulses.
The plasticity of astrocytes is fundamental for their principal function, maintaining homeostasis of the central nervous system throughout life, and is associated with diverse exposomal challenges. Here, we used cultured astrocytes to investigate at subcellular level basic cell processes under controlled environmental conditions. We compared astroglial functional and signaling plasticity in standard serum-containing growth medium, a condition mimicking pathologic conditions, and in medium without serum, favoring the acquisition of arborized morphology. Using optoÀ/electrophysiologic techniques, we examined cell viability, expression of astroglial markers, vesicle dynamics, and cytosolic Ca 2+ and cAMP signaling. The results revealed altered vesicle dynamics in arborized astrocytes that was associated with increased resting [Ca 2 + ] i and increased subcellular heterogeneity in [Ca 2+ ] i , whereas [cAMP] i subcellular dynamics remained stable in both cultures, indicating that cAMP signaling is less prone to plastic remodeling than Ca 2+ signaling, possibly also in in vivo contexts. K E Y W O R D S astrocyte, Ca 2 + , cAMP, confocal microscopy, electrophysiology, vesicles 1 | INTRODUCTION Astrocytes are morphologically and functionally heterogeneous glial cells in the central nervous system. Immunolabeling of glial fibrillary acidic protein (GFAP) reveals major processes with finer cellular parts remaining unstained (Connor & Berkowitz, 1985) making astrocytes appear as stellate cells (Wolfes et al., 2017; Wolfes & Dean, 2018).Advanced visualization techniques revealed that astrocytes exhibit a more complex, spongioform structure (Benediktsson et al., 2005;Bushong et al., 2004). The morphological complexity of astrocytes arguably correlates with their extended homeostatic roles. Astroglia assist neuro-and synaptogenesis, provide substrates to neurons, regulate blood flow and the blood-brain barrier, control uptake and recycling of neurotransmitters, and produce and secrete various neurotrophic factors to regulate memory formation (Araque et al., 1999;
Cover Illustration: Live arborized astrocyte in culture (NB+), containing numerous dextran‐laden endocytotic vesicles (green), revealed by confocal microscopy. Arborized astrocytes display increased resting levels of intracellular Ca2+ concentration ([Ca2+]i) and [cAMP]i, increased subcellular heterogeneity in [Ca2+]i, but not in [cAMP]i, and attenuated vesicle mobility. (See Pirnat, S., et al, https://doi.org/10.1002/glia.24076.)image
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