Microbubble Sonodestruction Rate as a Metric to Evaluate Sonoporation Efficiency

Tamošiūnas, Mindaugas; Jurkonis, Rytis; Mir, Lluis M.; Lukoševičius, Arūnas; Venslauskas, Mindaugas Saulius; Šatkauskas, Saulius

doi:10.7863/jum.2012.31.12.1993

Cited by 16 publications

(19 citation statements)

References 31 publications

(44 reference statements)

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“…The viability dynamics of CHO cells after Ca 2+ delivery via SP are very similar to those obtained in our previous works on BLM delivery [26,27]. Similarly, we have attained efficient Ca 2+ delivery via SP only after combined US-MB employment, as we have observed no significant changes in cell viability after 6 d due to the application of US alone, except at 800 kPa using 20 mM Ca 2+ concentration.…”

Section: Discussionsupporting

confidence: 86%

“…Therefore, broadband noise is considered to be an unambiguous confirmation of MB collapse and, respectively, IC [25,28,29]. IC has been substantiated to be involved in SP according to high correlation of IC based metrics, MB sonodestruction rate [26] and inertial cavitation dose [21], with the efficacy of therapeutic agent sonotransfer [23,24,26,30] and cell viability [21,23,26]. Therefore, passively registered acoustic emissions from MBs, induced by US, were analyzed in order to acquire valuable information about cavitation pattern.…”

Section: Discussionmentioning

confidence: 99%

“…The role of SC in SP becomes dominant when MBs are positioned near or bound to the cell surface; thereby, the distance between the cell and nearby MBs is reduced [31,32]. Since the impact of microstreaming is significantly downscaled with the increment in cell-MB separation, SC intensity is reduced and, therefore, IC prevails [22][23][24]26,27,30]. The assumption is reinforced by effective SP, obtained at SC regime, conducted on monolayers, where cells are tightly embedded and come into close contact with adjacent MBs [33,34].…”

Section: Discussionmentioning

confidence: 99%

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Sudden Cell Death Induced by Ca2+ Delivery via Microbubble Cavitation

et al. 2021

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Intracellular calcium ion delivery via sonoporation has been validated to be a substitute for classical chemotherapy. However, the mechanism behind calcium sonoporation remains unclear to this day. To elucidate the role of calcium in the process of sonoporation, we aimed to investigate the influence of different calcium concentration on cell membrane permeabilization and cell viability after sonoporation. In this study, we present experimental evidence that extracellular calcium plays a major role in cell membrane molecular transport after applying ultrasound pulses. Ultrasound-microbubble cavitation in the presence of different calcium concentration affects fundamental cell bio-physio-chemical conditions: cell membrane integrity, metabolic activity, and colony formation. Corresponding vital characteristics were evaluated using three independent viability tests: propidium iodide assay (20 min–3 h), MTT assay (48 h), and cell clonogenic assay (6 d). The results indicate instant cell death, as the level of cell viability was determined to be similar within a 20 min–48 h–6 d period. Inertial cavitation activities have been determined to be directly involved in calcium delivery via sonoporation according to high correlation (R2 > 0.85, p < 0.01) of inertial cavitation dose with change in either cell membrane permeabilization, metabolic activity, and colony formation efficiency. In general, calcium delivery via sonoporation induces rapid cell death, occurring within 20 min after treatment, that is the result of ultrasound mediated microbubble cavitation.

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Section: Discussionsupporting

confidence: 86%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Sudden Cell Death Induced by Ca2+ Delivery via Microbubble Cavitation

et al. 2021

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“…USMB therapy would benefit from simultaneous (real-time) cavitation monitoring with a single transducer of a clinical US system. This solution would allow for monitoring of bioeffects ( Chen et al, 2003 ; Hallow et al, 2006 ; Tamosiunas et al, 2012 ; Maciulevicius et al, 2015 ), while using standardized US settings. Currently, simultaneous USMB therapy and cavitation monitoring is not yet available on clinical US systems, although our work and the work by Keller et al show it is technically feasible ( Keller et al, 2021 ).…”

Section: Discussionmentioning

confidence: 99%

Ultrasound-Mediated Drug Delivery With a Clinical Ultrasound System: In Vitro Evaluation

et al. 2021

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Chemotherapy efficacy is often reduced by insufficient drug uptake in tumor cells. The combination of ultrasound and microbubbles (USMB) has been shown to improve drug delivery and to enhance the efficacy of several drugs in vitro and in vivo, through effects collectively known as sonopermeation. However, clinical translation of USMB therapy is hampered by the large variety of (non-clinical) US set-ups and US parameters that are used in these studies, which are not easily translated to clinical practice. In order to facilitate clinical translation, the aim of this study was to prove that USMB therapy using a clinical ultrasound system (Philips iU22) in combination with clinically approved microbubbles (SonoVue) leads to efficient in vitro sonopermeation. To this end, we measured the efficacy of USMB therapy for different US probes (S5-1, C5-1 and C9-4) and US parameters in FaDu cells. The US probe with the lowest central frequency (i.e. 1.6 MHz for S5-1) showed the highest USMB-induced intracellular uptake of the fluorescent dye SYTOX™ Green (SG). These SG uptake levels were comparable to or even higher than those obtained with a custom-built US system with optimized US parameters. Moreover, USMB therapy with both the clinical and the custom-built US system increased the cytotoxicity of the hydrophilic drug bleomycin. Our results demonstrate that a clinical US system can be used to perform USMB therapy as efficiently as a single-element transducer set-up with optimized US parameters. Therefore, future trials could be based on these clinical US systems, including validated US parameters, in order to accelerate successful translation of USMB therapy.

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“…1 However, sonoporation can also induce the death of the treated cells. [2][3][4] Therefore, it is crucial to evaluate cell viability when performing sonotransfer experiments. A few membrane integrity-based cell viability assays are compatible with sonoporation experiments; these use propidium iodide, 1,5,6 ethidium bromide (EB), 7,8 or trypan blue (TB) [7][8][9] fluorescent dyes.…”

Section: Introductionmentioning

confidence: 99%

FRET-based method for evaluation of the efficiency of reversible and irreversible sonoporation

et al. 2017

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It is widely known that not all of the treated cells survive after introduction of exogenous molecules via any physical method. Therefore, it is important to develop methods that would allow simultaneous evaluation of both molecular delivery efficiency and cell viability. This study presents Förster resonance energy transfer (FRET)-based method that allows molecular transfer and cell viability evaluation in a single measurement by employing two common fluorescent dyes, namely, ethidium bromide and trypan blue. The method has been validated using cell sonoporation. The FRET-based method allows the efficiency evaluation of both reversible and irreversible sonoporation in a single experiment. Therefore, this method could be used to reduce time, labor, and material cost while improving the accuracy of evaluations.

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Microbubble Sonodestruction Rate as a Metric to Evaluate Sonoporation Efficiency

Abstract: These results show that the microbubble sonodestruction rate can be used to predict the percentage of reversible and irreversible sonoporation.

Cited by 16 publications

References 31 publications

Sudden Cell Death Induced by Ca2+ Delivery via Microbubble Cavitation

Sudden Cell Death Induced by Ca2+ Delivery via Microbubble Cavitation

Ultrasound-Mediated Drug Delivery With a Clinical Ultrasound System: In Vitro Evaluation

FRET-based method for evaluation of the efficiency of reversible and irreversible sonoporation

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