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Conference Paper: How sonoporation disrupts cellular structural integrity: morphological and cytoskeletal observations

TitleHow sonoporation disrupts cellular structural integrity: morphological and cytoskeletal observations
Authors
Issue Date2013
PublisherISTU2013.
Citation
The 13th International Symposium on Therapeutic Ultrasound (ISTU 2013), Shanghai, China, 12-16 May 2013. In 13th ISTU Final Programme, 2013, p. 149 How to Cite?
AbstractOBJECTIVES: In considering sonoporation for drug delivery applications, it is essential to understand how living cells respond to this puncturing force. Here we seek to investigate the effects of sonoporation on cellular structural integrity. We hypothesize that the membrane morphology and cytoskeletal behavior of sonoporated cells under recovery would inherently differ from that of normal viable cells. METHODS: A customized and calibrated exposure platform was developed for this work, and the ZR-75-30 breast carcinoma cells were used as the cell model. The cells were exposed to either single or multiple pulses of 1 MHz ultrasound (pulse length: 30 or 100 cycles; PRF: 1kHz; duration: up to 60s) with 0.45 MPa spatial-averaged peak negative pressure and in the presence of lipid-shelled microbubbles. Confocal microscopy was used to examine insitu the structural integrity of sonoporated cells (identified as ones with exogenous fluorescent marker internalization). For investigations on membrane morphology, FM 4-64 was used as the membrane dye (red), and calcein was used as the sonoporation marker (green); for studies on cytoskeletal behavior, CellLight (green) and propidium iodide (red) were used to respectively label actin filaments and sonoporated cells. Observation started from before exposure to up to 2 h after exposure, and confocal images were acquired at real-time frame rates. Cellular structural features and their temporal kinetics were quantitatively analyzed to assess the consistency of trends amongst a group of cells. RESULTS: Sonoporated cells exhibited membrane shrinkage (decreased by 61% in a cell’s cross-sectional area) and intracellular lipid accumulation (381% increase compared to control) over a 2 h period. The morphological repression of sonoporated cells was also found to correspond with post-sonoporation cytoskeletal processes: actin depolymerization was observed as soon as pores were induced on the membrane. These results show that cellular structural integrity is indeed disrupted over the course of sonoporation. CONCLUSIONS: Our investigation shows that the biophysical impact of sonoporation is by no means limited to the induction of membrane pores: e.g. structural integrity is concomitantly affected in the process. This prompts the need for further fundamental studies to unravel the complex sequence of biological events involved in sonoporation.
DescriptionPosters: no. 1
Control ID: 1672429
Persistent Identifierhttp://hdl.handle.net/10722/189834

 

DC FieldValueLanguage
dc.contributor.authorChen, Xen_US
dc.contributor.authorHu, Yen_US
dc.contributor.authorLeow, RSen_US
dc.contributor.authorYu, ACHen_US
dc.contributor.authorWan, JMFen_US
dc.date.accessioned2013-09-17T15:00:47Z-
dc.date.available2013-09-17T15:00:47Z-
dc.date.issued2013en_US
dc.identifier.citationThe 13th International Symposium on Therapeutic Ultrasound (ISTU 2013), Shanghai, China, 12-16 May 2013. In 13th ISTU Final Programme, 2013, p. 149en_US
dc.identifier.urihttp://hdl.handle.net/10722/189834-
dc.descriptionPosters: no. 1-
dc.descriptionControl ID: 1672429-
dc.description.abstractOBJECTIVES: In considering sonoporation for drug delivery applications, it is essential to understand how living cells respond to this puncturing force. Here we seek to investigate the effects of sonoporation on cellular structural integrity. We hypothesize that the membrane morphology and cytoskeletal behavior of sonoporated cells under recovery would inherently differ from that of normal viable cells. METHODS: A customized and calibrated exposure platform was developed for this work, and the ZR-75-30 breast carcinoma cells were used as the cell model. The cells were exposed to either single or multiple pulses of 1 MHz ultrasound (pulse length: 30 or 100 cycles; PRF: 1kHz; duration: up to 60s) with 0.45 MPa spatial-averaged peak negative pressure and in the presence of lipid-shelled microbubbles. Confocal microscopy was used to examine insitu the structural integrity of sonoporated cells (identified as ones with exogenous fluorescent marker internalization). For investigations on membrane morphology, FM 4-64 was used as the membrane dye (red), and calcein was used as the sonoporation marker (green); for studies on cytoskeletal behavior, CellLight (green) and propidium iodide (red) were used to respectively label actin filaments and sonoporated cells. Observation started from before exposure to up to 2 h after exposure, and confocal images were acquired at real-time frame rates. Cellular structural features and their temporal kinetics were quantitatively analyzed to assess the consistency of trends amongst a group of cells. RESULTS: Sonoporated cells exhibited membrane shrinkage (decreased by 61% in a cell’s cross-sectional area) and intracellular lipid accumulation (381% increase compared to control) over a 2 h period. The morphological repression of sonoporated cells was also found to correspond with post-sonoporation cytoskeletal processes: actin depolymerization was observed as soon as pores were induced on the membrane. These results show that cellular structural integrity is indeed disrupted over the course of sonoporation. CONCLUSIONS: Our investigation shows that the biophysical impact of sonoporation is by no means limited to the induction of membrane pores: e.g. structural integrity is concomitantly affected in the process. This prompts the need for further fundamental studies to unravel the complex sequence of biological events involved in sonoporation.-
dc.languageengen_US
dc.publisherISTU2013.-
dc.relation.ispartof13th International Symposium on Therapeutic Ultrasound Final Programmeen_US
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.titleHow sonoporation disrupts cellular structural integrity: morphological and cytoskeletal observationsen_US
dc.typeConference_Paperen_US
dc.identifier.emailYu, ACH: alfred.yu@hku.hken_US
dc.identifier.emailWan, JMF: jmfwan@hku.hken_US
dc.identifier.authorityYu, ACH=rp00657en_US
dc.identifier.authorityWan, JMF=rp00798en_US
dc.description.naturepostprint-
dc.identifier.hkuros222139en_US
dc.identifier.spage149-
dc.identifier.epage149-
dc.publisher.placeChina-

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