Saving cells from ultrasound-induced apoptosis: quantification of cell death and uptake following sonication and effects of targeted calcium chelation.
Applications of ultrasound for noninvasive drug and gene delivery have been limited by associated cell death as a result of sonication. In this study, we sought to quantify the distribution of cellular bioeffects caused by low-frequency ultrasound (24 kHz) and test the hypothesis that Ca(2+) chelation after sonication can shift this distribution by saving cells from death by apoptosis. Using flow cytometry, we quantitatively categorized sonicated cells among four populations: (i) cells that appear largely unaffected, (ii) cells reversibly permeabilized, (iii) cells rendered nonviable during sonication and (iv) cells that appear to be viable shortly after sonication, but later undergo apoptosis and die. By monitoring cells for 6 h after ultrasound exposure, we found that up to 15% of intact cells fell into this final category. Those apoptotic cells initially had the highest levels of uptake of a marker compound, calcein; also had highly elevated levels of intracellular Ca(2+); and contained an estimated plasma membrane wound radius of 100-300 nm. Finally, we showed that chelation of intracellular Ca(2+) after sonication reduced apoptosis by up to 44%, thereby providing a strategy to save cells. We conclude that cells can be saved from ultrasound-induced death by appropriate selection of ultrasound conditions and Ca(2+) chelation after sonication.
Hutcheson
J D
JD
School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA 30332-0100, USA.
Schlicher
R K
RK
Hicks
H K
HK
Prausnitz
M R
MR
D017209:Apoptosis; D002118:Calcium / Q000378:metabolism*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002614:Chelating Agents / Q000494:pharmacology*; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D011467:Prostate / Q000000981:diagnostic imaging*; D013010:Sonication; D014463:Ultrasonography
D002614:Chelating Agents; D002118:Calcium
Annu Rev Biomed Eng. 2007;9:415-47
17651012
Ultrasound Med Biol. 1993;19(2):147-65
8516961
Nature. 1998 Oct 15;395(6703):645-8
9790183
Ultrasound Med Biol. 2003 Aug;29(8):1211-22
12946524
Ultrasound Med Biol. 1994;20(3):205-18
8059482
Ultrasound Med Biol. 2001 Jun;27(6):841-50
11516544
Ultrasound Med Biol. 2004 May;30(5):683-92
15183235
Cytometry. 1998 Jan 1;31(1):1-9
9450519
Annu Rev Biomed Eng. 2004;6:229-48
15255769
Pharm Res. 2007 Jun;24(6):1047-57
17387604
Curr Pharm Biotechnol. 2007 Dec;8(6):355-61
18289044
Eur J Echocardiogr. 2004 Aug;5(4):245-56
15219539
Ultrasound Med Biol. 2001 Jan;27(1):111-7
11295277
Cancer Lett. 2005 Apr 28;221(2):145-52
15808400
Curr Protoc Immunol. 2004 Dec;Chapter 5:Unit 5.5
18432929
Cytometry. 1992;13(7):739-49
1451604
J Acoust Soc Am. 2001 Jul;110(1):588-96
11508983
Drug Discov Today. 2003 Mar 15;8(6):259-66
12623240
Ultrasound Med Biol. 2001 Aug;27(8):1107-13
11527597
Exp Hematol. 2002 Nov;30(11):1293-301
12423682
Ultrasound Med Biol. 2010 Apr;36(4):677-92
20350691
J Acoust Soc Am. 2004 Apr;115(4):1818-25
15101659
Expert Opin Drug Deliv. 2004 Nov;1(1):37-56
16296719
Science. 2004 Mar 19;303(5665):1818-22
15031496
Biophys J. 2001 Feb;80(2):755-64
11159443
Cytometry. 1992;13(8):795-808
1333943
Ultrasound Med Biol. 1988;14(1):43-50
3347962
J Cell Biol. 1997 Oct 6;139(1):63-74
9314529
Biophys J. 2008 Nov 1;95(9):4124-38
18676653
Ultrason Sonochem. 2004 Sep;11(6):349-63
15302020
Nat Rev Mol Cell Biol. 2005 Jun;6(6):499-505
15928713
Am J Physiol Heart Circ Physiol. 2006 Oct;291(4):H1595-601
16632548
Pharm Res. 2001 Nov;18(11):1514-20
11758757
Adv Drug Deliv Rev. 2008 Jun 30;60(10):1153-66
18486268
Ultrasound Med Biol. 2009 Mar;35(3):494-506
19010589
Biophys J. 2007 Sep 15;93(6):L29-31
17631537
J Biol Chem. 2001 Sep 7;276(36):34148-55
11435443
J Am Coll Cardiol. 2005 Feb 1;45(3):329-35
15680708
Ultrasound Med Biol. 2006 Jun;32(6):915-24
16785013
Int J Radiat Biol. 2004 Feb;80(2):165-75
15164798
Mol Imaging Biol. 2003 Jan-Feb;5(1):2-14
14499155
Science. 2001 Jul 6;293(5527):58-9
11441170
Annu Rev Cell Dev Biol. 2003;19:697-731
14570587
Med Res Rev. 2002 Mar;22(2):204-23
11857639
Cell Calcium. 1998 Feb-Mar;23(2-3):173-80
9601613
Cancer Res. 2000 Feb 15;60(4):1014-20
10706118
J Acoust Soc Am. 2001 Jul;110(1):597-606
11508985