Structural and permeability sensitivity of cells to low intensity ultrasound: Infrared and fluorescence evidence in vitro

Domenici, Fabio; Giliberti, Claudia; Bedini, Angelico; Palomba, Raffaele; Udroiu, Ion; Giambattista, Lucia Di; Pozzi, Deleana; Morrone, Stefania; Bordi, F.; Castellano, A. Congiu

doi:10.1016/j.ultras.2013.12.003

Cited by 17 publications

(20 citation statements)

References 39 publications

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“…Moreover, the treatments did not show any significant dependence of the studied effects on the US wave duty cycle. These observations are consistent with the low US absorption at 1 MHz; on the contrary, thermal effects and duty cycle could affect cells and tissues exposed to higher frequencies, being the US attenuation coefficient proportional to the frequency 23 , 31 .…”

Section: Resultssupporting

confidence: 83%

Differential effects on membrane permeability and viability of human keratinocyte cells undergoing very low intensity megasonic fields

Domenici

Brasili

Giantulli

et al. 2017

Sci Rep

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Among different therapeutic applications of Ultrasound (US), transient membrane sonoporation (SP) - a temporary, non-lethal porosity, mechanically induced in cell membranes through US exposure - represents a compelling opportunity towards an efficient and safe drug delivery. Nevertheless, progresses in this field have been limited by an insufficient understanding of the potential cytotoxic effects of US related to the failure of the cellular repair and to the possible activation of inflammatory pathway. In this framework we studied the in vitro effects of very low-intensity US on a human keratinocyte cell line, which represents an ideal model system of skin protective barrier cells which are the first to be involved during medical US treatments. Bioeffects linked to US application at 1 MHz varying the exposure parameters were investigated by fluorescence microscopy and fluorescence activated cell sorting. Our results indicate that keratinocytes undergoing low US doses can uptake drug model molecules with size and efficiency which depend on exposure parameters. According to sub-cavitation SP models, we have identified the range of doses triggering transient membrane SP, actually with negligible biological damage. By increasing US doses we observed a reduced cells viability and an inflammatory gene overexpression enlightening novel healthy relevant strategies.

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Section: Resultssupporting

confidence: 83%

Differential effects on membrane permeability and viability of human keratinocyte cells undergoing very low intensity megasonic fields

Domenici

Brasili

Giantulli

et al. 2017

Sci Rep

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“…Focusing on air-filled elastomeric PVA-shelled MBs, we demonstrated here that such agents when dispersed in water medium are found to become strongly stable over time in terms of morphology and structure, as compared to lipid-shelled MBs (months vs. hours). The system showed a good echogenicity in a range of frequencies centred at around 10 MHz, complementary to that of MBs with a lipid shell and useful to minimise the biological risks related to high-resolution imaging [11,20,57,58]. This should facilitate their use as UCAs as well as in derivatisation protocols for drug delivery and theranostics [1,7,18].…”

Section: Discussionmentioning

confidence: 99%

Long-term physical evolution of an elastomeric ultrasound contrast microbubble

Domenici

Brasili

Oddo

et al. 2019

Journal of Colloid and Interface Science

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Hypothesis One of the main assets of crosslinked polymer-shelled microbubbles (MBs) as ultrasound-active theranostic agents is the robustness of the shells, combined with the chemical versatility in modifying the surface with ligands and/or drugs. Despite the long shelf-life, subtle modifications occur in the MB shells involving shifts in acoustic, mechanical and structural properties. Experiments We carried out a long-term morphological and acoustic evolution analysis on elastomeric polyvinyl-alcohol (PVA)-shelled MBs, a novel platform accomplishing good acoustic and surface performances in one agent. Confocal laser scanning microscopy, acoustic spectroscopy and AFM nanomechanics were integrated to understand the mechanism of PVA MBs ageing. The changes in the MB acoustic properties were framed in terms of shell thickness and viscoelasticity using a linearised oscillation theory, and compared to MB morphology and to nanomechanical analysis. Findings We enlightened a novel, intriguing ageing time evolution of the PVA MBs with double behaviour with respect to a crossover time of ∼50 days. Before, significant changes occur in MB stiffness and shell thickness, mainly due to a massive release of entangled PVA chains. Then, the MB resonance frequency increases together with shell thickening and softening. Our benchmark study is of general interest for emerging viscoelastomeric bubbles towards personalised medicine.

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“…(19,20) Still, regarding the cellular percentage reduction in some groups, other authors corroborate with our findings, showing that the acoustic cavitation is intrinsically related to the TUS bioeffects and by generating numerous compressions and rarefaction can become unstable and trigger changes in the cellular size And membrane compliance, with even cellular implosions due to the increase of temperature and intracellular pressure, resulting in the decrease of viable cells irreversibly. (10,21,22) Already, specifically in in vitro fibroblasts, Oliveira et al, (7) reported that the statistical significance was only found in the group irradiated with ultrasound with analog dose (0.5 W/cm 2 ) and pulse rate of 10% (p=0.003). In this group, cell viability increased from 24 to 48 hours, and from 48 to 72 hours.…”

Section: Discussionmentioning

confidence: 99%

“…(6) In this sense, several authors report the efficiency of low intensity ultrasound in the percentage increase of viable cells cultured in vitro, with acceleration of proliferation and, (7)(8)(9) with the continuous, relevant therapeutic effects both in vivo and in vitro. (1,10) Furthermore, in the continuous mode, effects on the expression of cartilage specific mRNA in chondrocyte culture, (11) hypertrophy of muscle fibers, (1) or with pulsed modulation in biosynthetic activity and expression of integrins (12) and stimulation of anti-inflammatory systems in synovial membrane cells. (13) It is worth mentioning that although the effects of ultrasound have already been well described, the ideal dose-effect relationship still needs to be better elucidated and delimited.…”

Section: Introductionmentioning

confidence: 99%

Effect of therapeutic ultrasound of low intensity pulsed and continuous in fibroblastic cells in vitro

Oliveira¹,

Pires-Oliveira²,

Pacheco‐Soares³

et al. 2020

mtprehabjournal

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Introduction: The ultrasound is widely used clinical practice focused on tissue repair, because it is a secure resource, minimally-invasive and low investment. However, despite the effects of ultrasound have been well described, the ideal relationship between dose and effect, still needs to be better elucidated and bounded. Objective: In this way, the objective of this study was to evaluate the influence of irradiation pulsed ultrasound and continuous in the viability of fibroblastic cells L929 in vitro. Method: The cells were distributed in TPP plates 12 wells in the concentration of 5x10 5 cells/µL and subdivided into the following groups: G1: control (not received radiation), G2: irradiated to 0.5 W/cm 2-30% and G3: irradiated to 0.5 W/cm 2-100%. Then received ultrasonic irradiation with intervals of 24, 48 and 72 hours, and after 24 hours of each irradiation was performed MTT cytotoxicity cell. Results: Among the three groups analyzed, only the G2 group showed a significant difference between the time 48 and 72 hours (p=0.05). In other times, despite variation in the percentage of viable cells, were not significant. When compared to peers at post-test, it was possible to observe a difference of 29% of viable cells between the groups G1 and G2 (p=0.05). However, the biggest difference was observed between the groups G2 and G3 (p=0.05). Conclusion: In this way, it can be concluded that irradiation pulsed ultrasound showed higher proliferation of fibroblasts L929 in time 48 hours, whereas in group irradiated in continuous mode, there was no statistically significant difference between the times.

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Structural and permeability sensitivity of cells to low intensity ultrasound: Infrared and fluorescence evidence in vitro

Cited by 17 publications

References 39 publications

Differential effects on membrane permeability and viability of human keratinocyte cells undergoing very low intensity megasonic fields

Differential effects on membrane permeability and viability of human keratinocyte cells undergoing very low intensity megasonic fields

Long-term physical evolution of an elastomeric ultrasound contrast microbubble

Effect of therapeutic ultrasound of low intensity pulsed and continuous in fibroblastic cells in vitro

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