Enhancement of ultrasound-induced apoptosis and cell lysis by echo-contrast agents

Feril, Loreto B.; Kondo, Takashi; Zhao, Qing‐Li; Ogawa, Ryohei; Tachibana, Katsuro; Kudo, Nobuki; Fujimoto, Shinichi; Nakamura, Shinobu

doi:10.1016/s0301-5629(02)00700-7

Cited by 161 publications

(115 citation statements)

References 33 publications

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“…The existence of the additional killed cells was puzzling and conceivably could indicate the operation of the two cell death pathways (i. e., apoptosis, as well as necrosis). Ultrasonic cavitation has most often been associated with cell rapid necrosis (i. e., immediate mechanical cell membrane disruption at the time of exposure); however, apoptosis recently has been reported in vitro (Feril et al 2003) and in vivo (Vykhodtseva et al 2006) for ultrasound aided by contrast agents. The purpose of this study was to better characterize the Evans blue stained cardiomyocytes with regard to the type of cell death seen: apoptosis vs. necrosis.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, ultrasound exposure has been shown to induce apoptosis. In vitro exposure of cells induced apoptosis in various cell lines (Takeuchi et al 2006;Kinoshita et al 2007), particularly with added contrast agents (Feril et al 2003). In vivo, ultrasound exposure of rabbit brain with contrast agent gas bodies in the circulation has been shown to induce apoptosis in brain cells (Vykhodtseva et al 2006).…”

Section: Introductionmentioning

confidence: 99%

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Evans Blue Staining of Cardiomyocytes Induced by Myocardial Contrast Echocardiography in Rats: Evidence for Necrosis Instead of Apoptosis

Miller

Peng

Dou

et al. 2007

Ultrasound in Medicine & Biology

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High Mechanical Index (MI) echocardiography with contrast agent has been shown to induce Evans blue staining of cardiomyocytes, seen one day after exposure, in addition to contraction band necrosis, seen immediately after exposure. This research examined the roles of necrosis vs. apoptosis in these bioeffects. Myocardial contrast echocardiography at high MI with 1:4 ECG triggering was performed in anesthetized rats at 1.5 MHz. Histologically observable cell injury was accumulated by infusing a high dose of 50 μl/kg Definity® via tail vein for 5 min at the start of 10 min of scanning. Evans blue dye or propidium iodide was injected as an indicator of cardiomyocyte plasma membrane integrity. Histological sections were stained using the TUNEL method for labeling nuclei with DNA degradation (e.g. apoptosis). Evans blue fluorescent cells were counted on frozen sections or on hematoxylin-stained and TUNEL labeled paraffin sections. In addition, transmission electron microscopy was used to assess potential apoptotic nuclei. Hypercontraction and propidium iodide staining were observed immediately after imaging-exposure. Although TUNEL positive cells were evident after 4 h, these also had indications of contraction band necrosis and features of apoptosis were not confirmed by electron microscopy. Inflammatory cell infiltration was evident after 24 h. A second, more subtle injury was recognized by Evans blue staining, with minimal inflammatory cell infiltration at the morphologically intact stained cells after 24 h. Apoptosis was not detected by the TUNEL method in the cardiomyocytes stained with Evans blue at 24 h. However, Evans blue stained cell numbers declined after 48 h, with continued inflammatory cell infiltration. The initial insult for Evans blue stained cardiomyocytes apparently induced partial permeability of the plasma membrane, which led to gradual degeneration (but not apoptosis) and necrosis after 24-48 h.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Evans Blue Staining of Cardiomyocytes Induced by Myocardial Contrast Echocardiography in Rats: Evidence for Necrosis Instead of Apoptosis

Miller

Peng

Dou

et al. 2007

Ultrasound in Medicine & Biology

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“…46,47 Cell viability following USinduced membrane poration in the absence of CA remains largely overlooked and as such, cell viability is generally reported in terms of the combined effect of US with CA rather than the effects of US "alone." 48 Additionally, commercially available CAs appear to have varying threshold levels before inducing cell death. 49 Cell death mechanisms (apoptosis, necrosis, autolysis, autophagy, and oncosis) display considerable overlap, 50 adding further complexity.…”

Section: A Microfluidic Device Characterisationmentioning

confidence: 99%

Contrast agent-free sonoporation: The use of an ultrasonic standing wave microfluidic system for the delivery of pharmaceutical agents

et al. 2011

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Sonoporation is a useful biophysical mechanism for facilitating the transmembrane delivery of therapeutic agents from the extracellular to the intracellular milieu. Conventionally, sonoporation is carried out in the presence of ultrasound contrast agents, which are known to greatly enhance transient poration of biological cell membranes. However, in vivo contrast agents have been observed to induce capillary rupture and haemorrhage due to endothelial cell damage and to greatly increase the potential for cell lysis in vitro. Here, we demonstrate sonoporation of cardiac myoblasts in the absence of contrast agent (CA-free sonoporation) using a low-cost ultrasound-microfluidic device. Within this device an ultrasonic standing wave was generated, allowing control over the position of the cells and the strength of the acoustic radiation forces. Real-time single-cell analysis and retrospective postsonication analysis of insonated cardiac myoblasts showed that CA-free sonoporation induced transmembrane transfer of fluorescent probes (CMFDA and FITC-dextran) and that different mechanisms potentially contribute to membrane poration in the presence of an ultrasonic wave. Additionally, to the best of our knowledge, we have shown for the first time that sonoporation induces increased cell cytotoxicity as a consequence of CA-free ultrasound-facilitated uptake of pharmaceutical agents (doxorubicin, luteolin, and apigenin). The US-microfluidic device designed here provides an in vitro alternative to expensive and controversial in vivo models used for early stage drug discovery, and drug delivery programs and toxicity measurements.

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“…Cavitation is the formation of vapor cavities in the cell solution. Cavitation may be inertial or stable [11][12][13][14][15]. The former occurs at lower intensity: the bubbles oscillate about some equilibrium size for many acoustic cycles.…”

Section: Introductionmentioning

confidence: 99%

Study of cellular response induced by low intensity ultrasound frequency sweep pattern on myelomonocytic lymphoma U937 cells

et al. 2016

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Purpose This study will analyze the mechanical effects (immediate lysis) and biological effects (cell survival, apoptosis, cell cycle) on U937 cells subjected to different sonication conditions with increasing and decreasing frequencies and burst rate (number of burst of a repeating signal in a specific time unit), in order to determine the best conditions of sonication to produce high mortality, apoptosis and inhibition of hyperproliferation. Method Cells are been stressed by pulse wave ultrasounds with increasing and decreasing frequencies between 400 and 620 kHz, at burst rates of 0.5, 10, 50 Hz and 50 % duty cycle (percentage of one period in which a signal is active), ultrasound intensities (spatial average-temporal peak) 0.045 and 0.09 W/cm 2 . The sonication durations were 90 and 180 s. Results The decreasing mode was found to be better than the increasing mode for 10 and 50 Hz burst rates, while at 0.5 Hz the increasing mode gave better results for the time of 180 s. For 10 Hz burst rate, decreasing frequency, 180 s, 0.09 W/cm 2 , 20 % survival rate was found; after 6-h incubation, cells showed 13 % of early apoptosis and 11 % of late apoptosis. For these conditions of sonication, the hyperproliferation of cells was inhibited. Conclusion Survival rate decreases for increasing intensity and duration with each burst rate. The best performance is decreasing mode in a range between 620 and 400 kHz, duty cycle 50 %, burst rate 10 Hz. In these conditions after 180 s duration, the average survival rate is 20 %, the survived cells manifest apoptosis after 6-h incubation and hyperproliferation is prevented. The results seem to lead toward a non-invasive and effective purging of leukemic cells.Keywords Low intensity ultrasound Á Cell lysis Á Hyperproliferation apoptosis Á Cell cycle Sommario Scopo Questo studio analizzerà gli effetti meccanici (lisi valutata immediatamente dopo la sonicazione) e gli effetti biologici (sopravvivenza delle cellule, apoptosis e lisi cellulare) su cellule U937 soggette a differenti condizioni di sonicazione con frequenze crescenti e decrescenti, a burst rate (numero di impulsi di un segnale che si ripete in una specifica unità di tempo) variabili, al fine di determinare le migliori condizioni di sonicazione per produrre alta mortalità, apoptosi e inibizione dell'iperproliferazione cellulare. Metodi Le cellule sono state sollecitate con ultrasuoni ad onda pulsata con frequenze crescenti e decrescenti fra 400 e 620 kHz a burst rate di 0.5 Hz, 10 Hz, 50 Hz, con un ciclo di lavoro (frazione di tempo in cui il segnale è attivo in proporzione al tempo totale) del 50 %, intensità (picco spaziale medio-temporale) 0.045 W/cm 2 and 0.09 W/cm 2 . I tempi di sonicazione erano di 90 s e 180 s. Risultati La modalità a frequenza decrescente risultava essere migliore di quella crescente per burst rate 10 Hz e 50 Hz, mentre a 0.5 Hz la modalità a frequenza crescente risultava migliore per un tempo di 180 s. Con un burst rate di 10 Hz, a frequenza decrescente fra 620 e 400 kHz, esposizione per ...

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Enhancement of ultrasound-induced apoptosis and cell lysis by echo-contrast agents

Cited by 161 publications

References 33 publications

Evans Blue Staining of Cardiomyocytes Induced by Myocardial Contrast Echocardiography in Rats: Evidence for Necrosis Instead of Apoptosis

Evans Blue Staining of Cardiomyocytes Induced by Myocardial Contrast Echocardiography in Rats: Evidence for Necrosis Instead of Apoptosis

Contrast agent-free sonoporation: The use of an ultrasonic standing wave microfluidic system for the delivery of pharmaceutical agents

Study of cellular response induced by low intensity ultrasound frequency sweep pattern on myelomonocytic lymphoma U937 cells

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