Cultured Chinese hamster ovary cells attached to thin Mylar sheets were exposed to 3.3-MHz ultrasound in the presence of ultrasound contrast agent. The ultrasound beam was directed upward at the exposure chamber with the monolayer on the inside of the upper acoustic window. Cell membrane damage was detected by the firefly enzyme assay for released ATP and the subharmonic, fundamental, and second harmonic scattered signals were recorded. ATP release increased monotonically with increasing pressure amplitude above apparent thresholds of 0.28 MPa for 1-s continuous and 0.56 MPa for 100-s pulsed (10-μs pulses, 1-ms PRP) exposures with 5% The subharmonic signal and, to a lesser extent, the second harmonic signal both increased with the cell membrane damage, which suggests that these signals have predictive value for bioeffects. If the monolayer was positioned on the front window of the exposure chamber, cell membrane damage was greatly reduced, which confirms the protective influence of this configuration of monolayers reported in the literature. The effect decreased both at high (50%) or low (0.5%) concentrations of The strong nonlinear scattering of ultrasound by contrast agent gas bodies appears to provide useful indicators of gas body activity including cavitational bioeffects.
REFERENCES
1.
N.
deJong
, “Improvements in ultrasound contrast agents
,” IEEE Eng. Med. Biol. Mag.
15
, 72
–82
(1996
).2.
J. A.
Marsh
, C. S.
Hall
, M. S.
Hughes
, J.
Mobley
, J. G.
Miller
, and G. H.
Brandenburger
, “Broadband through-transmission signal loss measurements of suspensions at concentrations approaching in vivo doses
,” J. Acoust. Soc. Am.
101
, 1155
–1161
(1997
).3.
B.
Schrope
, V. L.
Newhouse
, and V.
Uhlendorf
, “Simulated capillary blood flow measurement using a nonlinear ultrasonic contrast agent
,” Ultrason. Imaging
14
, 134
–158
(1992
).4.
P. H.
Chang
, K. K.
Shung
, S.
Wu
, and H. B.
Levene
, “Second harmonic imaging and harmonic Doppler measurements with
” IEEE Trans. Ultrason. Ferroelectr. Freq. Control
42
, 1020
–1027
(1995
).5.
P. H.
Chang
, K. K.
Shung
, and H. B.
Levene
, “Quantitative measurements of second harmonic doppler using ultrasound contrast agents
,” Ultrasound Med. Biol.
22
, 1206
–1214
(1996
).6.
P. D.
Krishna
and V. L.
Newhouse
, “Second harmonic characteristics of the ultrasound contrast agents and FS069
,” Ultrasound Med. Biol.
23
, 453
–459
(1997
).7.
B. F. Vandenberg and H. E. Melton, “Acoustic lability of albumin microspheres,” J. Am. Soc. Echocardiogr. 7, 582–589 (1994).
8.
D. L.
Miller
and R. M.
Thomas
, “Ultrasound contrast agents nucleate inertial cavitation in vitro
,” Ultrasound Med. Biol.
21
, 1059
–1065
(1995
).9.
V. Uhlendorf and C. Hoffmann, “Nonlinear acoustical response of coated microbubbles in diagnostic ultrasound,” 1994 Ultrasonics Symposium, 1559–1562 (1994).
10.
D. L.
Miller
, “Ultrasonic detection of resonant cavitation bubbles in a flow tube by their second harmonic emissions
,” Ultrasonics
19
, 217
–224
(1981
).11.
D. R.
Gross
, D. L.
Miller
, and A. R.
Williams
, “A search for ultrasonic cavitation within the canine cardiovascular system
,” Ultrasound Med. Biol.
11
, 85
–97
(1985
).12.
K. I.
Morton
, G. R.
Ter Haar
, I. G.
Stratford
, and C. R.
Hill
, “The role of cavitation in the interaction of ultrasound with V79 Chinese hamster cells in vitro
,” Br. J. Cancer
45
, 147
–150
(1982
).13.
K. I.
Morton
, G. R.
Ter Haar
, I. G.
Stratford
, and C. R.
Hill
, “Subharmonic emission as an indicator of ultrasonically-induced biological damage
,” Ultrasound Med. Biol.
9
, 629
–633
(1983
).14.
P. D.
Edmonds
and P.
Ross
, “Acoustic emission as a measure of exposure of suspended cells in vitro
,” Ultrasound Med. Biol.
12
, 297
–305
(1986
).15.
P. D. Lele, “Cavitation and its effects on organized mammalian tissue,” in Ultrasound: its Applications in Medicine and Biology, edited by F. J. Fry (Elsevier Scientific, New York, 1978), Pt. II, App. I, pp. 737–742.
16.
E. C.
Everbach
, I. R. S.
Makin
, M.
Azadniv
, and R. S.
Meltzer
, “Correlation of ultrasound-induced hemolysis with cavitation detector output in vitro
,” Ultrasound Med. Biol.
23
, 619
–624
(1997
).17.
D. L.
Miller
and A. R.
Williams
, “Bubble cycling as the explanation of the promotion of ultrasonic cavitation in a rotating tube exposure system
,” Ultrasound Med. Biol.
15
, 641
–648
(1989
).18.
E. P.
Armour
and P. M.
Corry
, “Cytotoxic effects of ultrasound in vitro dependence on gas content, frequency, radical scavengers, and attachment
,” Radiat. Res.
89
, 369
–380
(1982
).19.
A. R.
Williams
and D. L.
Miller
, “Photometric detection of ATP release from human erythrocytes exposed to ultrasonically activated gas-filled pores
,” Ultrasound Med. Biol.
6
, 251
–256
(1980
).20.
D. L.
Miller
, R. A.
Gies
, and W. B.
Chrisler
, “Ultrasonically induced hemolysis at high cell and gas body concentrations in a thin-disc exposure chamber
,” Ultrasound Med. Biol.
23
, 625
–633
(1997
).21.
J. L.
Moore
and W. T.
Coakley
, “Ultrasonic treatment of Chinese hamster cells at high intensities and long exposure times
,” Br. J. Radiol.
50
, 46
–50
(1977
).22.
S. L.
Kerr
, D. W.
Gregory
, M.
Shammari
, D. J.
Watmough
, and D. N.
Wheatley
, “Differing effects of ultrasound-irradiation on suspension and monolayer cultured HeLa cells investigated by scanning electron microscopy
,” Ultrasound Med. Biol.
15
, 397
–401
(1989
).23.
E. G. Loch, A. B. Fischer, and E. Kuwert, “Effect of diagnostic and therapeutic intensities of ultrasonics on normal and malignant human cells in vitro,” Am. J. Obstet. Gynecol. 110, 457–460 (1971).
24.
M. J.
Pickworth
, P. P.
Dendy
, P. R.
Twentyman
, and T. G.
Leighton
, “Studies of the cavitational effects of clinical ultrasound by sonoluminescence: 4. The effect of therapeutic ultrasound on cells in monolayer culture in a standing wave field
,” Phys. Med. Biol.
34
, 1553
–1560
(1989
).25.
T. G. Leighton, The Acoustic Bubble (Academic, New York, 1994).
26.
C. C.
Church
, “The effects of an elastic solid surface layer on the radial pulsations of gas bubbles
,” J. Acoust. Soc. Am.
97
, 1510
–1521
(1995
).27.
A.
Eller
and H. G.
Flynn
, “Generation of subharmonics of order one-half by bubbles in a sound field
,” J. Acoust. Soc. Am.
46
, 722
–727
(1969
).28.
AIUM, Bioeffects and Safety of Diagnostic Ultrasound (American Institute of Ultrasound in Medicine, Rockville MD, 1993).
29.
WFUMB Symposium on Safety of Ultrasound in Medicine, Ultrasound Med. Biol. (in press).
30.
AIUM, Standard for Real-Time Display of Thermal and Mechanical Acoustic Output Indices on Diagnostic Ultrasound Equipment (American Institute of Ultrasound in Medicine, Rockville, MD, 1992).
This content is only available via PDF.
© 1998 Acoustical Society of America.
1998
Acoustical Society of America
You do not currently have access to this content.
