Tumor Cytotoxicity In Vivo and Radical Formation In Vitro Depend on the Shock Wave-Induced Cavitation Dose

Abstract Huber, P. E. and Debus, J. Tumor Cytotoxicity In Vivo and Radical Formation In Vitro Depend on the Shock Wave-Induced Cavitation Dose. Radiat. Res. 156, 301–309 (2001). Local tumor therapy using focused ultrasonic waves may become an important treatment option. This technique exploits the ability of mechanical waves to induce thermal and nonthermal effects noninvasively. The cytotoxicity to cultured cells and biological tissues in vivo that results from exposure to ultrasonic shock waves is considered to be a nonthermal effect that is partly a consequence of ultrasound-induced cavitation. Cavitation is defined as the formation of bubbles during the negative wave cycle; their subsequent oscillation and/or violent implosion can affect surrounding structures. To investigate cavitational effects in cells and tissues, defined cavitation doses must be applied while ideally holding all other potential ultrasound parameters constant. The application of independent cavitation doses has been difficult and has yielded little knowledge about quantitative cavitation–tissue interactions. By using a special shock-wave pulse regimen and laser optical calibration in this study, we were able to control the cavitation dose independently of other physical parameters such as the pressure amplitudes, and averaged acoustic intensity. We treated Dunning prostate tumors (subline R3327-AT1) transplanted into Copenhagen rats with shock waves at three cavitation dose levels and then determined the tumor growth delay and the histopathological changes. All of the treated animals exhibited a significant tumor growth delay compared to the controls. Higher cavitation doses were associated with a greater delay in the growth of the tumor and more severe effects on tumor histopathology, such as hemorrhaging, tissue disruption, and necrosis. In vitro, the cavitation dose level correlated with the amount of radical formation. We concluded that the process of acoustic cavitation was responsible; higher cavitation doses caused greater effects in tumors both in vivo and in vitro. These findings may prove important in local tumor therapy and other applications of ultrasound such as ultrasound-mediated drug delivery.

[1]  J. Mclean,et al.  A cavitation and free radical dosimeter for ultrasound. , 1988, Ultrasound in medicine & biology.

[2]  W. Brendel,et al.  A model of extracorporeal shock wave action: tandem action of shock waves. , 1988, Ultrasound in medicine & biology.

[3]  J E Saunders,et al.  The influence of fluid properties and pulse amplitude on bubble dynamics in the field of a shock wave lithotripter. , 1993, Physics in medicine and biology.

[4]  J F Greenleaf,et al.  Ultrasound-mediated transfection of mammalian cells. , 1996, Human gene therapy.

[5]  J. Debus,et al.  Sonochemically induced radicals generated by pulsed high-energy ultrasound in vitro and in vivo. , 1999, Ultrasound in medicine & biology.

[6]  K Hynynen,et al.  Histologic effects of high intensity pulsed ultrasound exposure with subharmonic emission in rabbit brain in vivo. , 1995, Ultrasound in medicine & biology.

[7]  R. M. Thomas,et al.  Induction of base damage in DNA solutions by ultrasonic cavitation. , 1995, Free radical biology & medicine.

[8]  Douglas L. Miller,et al.  Transfection of a reporter plasmid into cultured cells by sonoporation in vitro. , 1997, Ultrasound in medicine & biology.

[9]  R A Stephenson,et al.  High energy shock waves suppress tumor growth in vitro and in vivo. , 1986, The Journal of urology.

[10]  M. Jordan,et al.  Biological effects of shock waves: kidney haemorrhage by shock waves in dogs--administration rate dependence. , 1988, Ultrasound in medicine & biology.

[11]  E. Carstensen,et al.  Bioeffects of positive and negative acoustic pressures in mice infused with microbubbles. , 2000, Ultrasound in medicine & biology.

[12]  E. Endl,et al.  Flow cytometric analysis of cell suspensions exposed to shock waves in the presence of the radical sensitive dye hydroethidine. , 1995, Ultrasound in medicine & biology.

[13]  K. Hynynen.  The threshold for thermally significant cavitation in dog's thigh muscle in vivo. , 1991, Ultrasound in medicine & biology.

[14]  W. J. Lorenz,et al.  Control of cavitation activity by different shockwave pulsing regimes. , 1999, Physics in medicine and biology.

[15]  P. Huber,et al.  In vitro and in vivo transfection of plasmid DNA in the Dunning prostate tumor R3327-AT1 is enhanced by focused ultrasound , 2000, Gene Therapy.

[16]  D. Lifshitz,et al.  Quantitation of shock wave cavitation damage in vitro. , 1997, Ultrasound in medicine & biology.

[17]  A. Lorenz,et al.  Synergistic interaction of ultrasonic shock waves and hyperthermia in the dunning prostate tumor r3327‐at1 , 1999, International journal of cancer.

[18]  Douglas L. Miller,et al.  In vivo transfection of melanoma cells by lithotripter shock waves. , 1998, Cancer research.

[19]  E. Schmiedt,et al.  EXTRACORPOREALLY INDUCED DESTRUCTION OF KIDNEY STONES BY SHOCK WAVES , 1980, The Lancet.

[20]  C. Church,et al.  A theoretical study of cavitation generated by an extracorporeal shock wave lithotripter. , 1988, The Journal of the Acoustical Society of America.

[21]  F Dunn,et al.  Selective clinical ultrasound signals mediate differential gene transfer and expression in two human prostate cancer cell lines: LnCap and PC-3. , 1997, Biochemical and biophysical research communications.

[22]  L. Filipczyński,et al.  Estimation of the temperature increase in the focus of a lithotripter for the case of high rate administration. , 1990, Ultrasound in medicine & biology.

[23]  S. Mitragotri,et al.  Ultrasound-mediated transdermal protein delivery , 1995, Science.

[24]  B. Chauffert,et al.  Cytotoxic effects of acoustic cavitation on HT-29 cells and a rat peritoneal carcinomatosis in vitro. , 1991, Cancer research.

[25]  W. J. Lorenz,et al.  In vivo detection of ultrasonically induced cavitation by a fibre-optic technique. , 1994, Ultrasound in medicine & biology.

[26]  W. J. Lorenz,et al.  Treatment of the Dunning prostate rat tumor R3327-AT1 with pulsed high energy ultrasound shock waves (PHEUS): growth delay and histomorphologic changes. , 1991, The Journal of urology.

[27]  M Delius,et al.  Shock wave permeabilization with ribosome inactivating proteins: a new approach to tumor therapy. , 1999, Cancer research.

[28]  R. M. Thomas,et al.  The role of cavitation in the induction of cellular DNA damage by ultrasound and lithotripter shock waves in vitro. , 1996, Ultrasound in medicine & biology.

[29]  G. Kuhnle,et al.  High-energy shock waves enhance hyperthermic response of tumors: effects on blood flow, energy metabolism, and tumor growth. , 1994, Journal of the National Cancer Institute.

[30]  F. Brümmer,et al.  Histopathology of shock wave treated tumor cell suspensions and multicell tumor spheroids. , 1989, Ultrasound in medicine & biology.

[31]  M C Ziskin,et al.  The sensitivity of biological tissue to ultrasound. , 1997, Ultrasound in Medicine and Biology.

[32]  M. Barda-Saad,et al.  Apoptosis induction of human myeloid leukemic cells by ultrasound exposure. , 2000, Cancer research.

[33]  W. J. Lorenz,et al.  A new method of quantitative cavitation assessment in the field of a lithotripter. , 1996, Ultrasound in medicine & biology.

[34]  Robert E. Apfel,et al.  7. Acoustic Cavitation , 1981 .

[35]  Taylor Murray,et al.  Cancer statistics, 1999 , 1999, CA: a cancer journal for clinicians.

[36]  P. Hofschneider,et al.  Shock wave permeabilization as a new gene transfer method , 1997, Gene Therapy.

[37]  K. Suslick,et al.  The Temperature of Cavitation , 1991, Science.

[38]  Barber,et al.  Spectrum of synchronous picosecond sonoluminescence. , 1992, Physical review letters.

[39]  W. Wieland,et al.  In vitro investigations on cellular damage induced by high energy shock waves. , 1992, Ultrasound in medicine & biology.

[40]  J. Debus,et al.  Influence of shock wave pressure amplitude and pulse repetition frequency on the lifespan, size and number of transient cavities in the field of an electromagnetic lithotripter. , 1998, Physics in medicine and biology.

[41]  M. Jordan,et al.  Biological effects of shock waves: cavitation by shock waves in piglet liver. , 1990, Ultrasound in medicine & biology.

[42]  J F Greenleaf,et al.  Artificial cavitation nuclei significantly enhance acoustically induced cell transfection. , 1998, Ultrasound in medicine & biology.

[43]  F. Lohr,et al.  Measurement of the proliferative activity of three different sublines of the Dunning rat prostate tumor R3327. , 1993, Strahlentherapie und Onkologie : Organ der Deutschen Rontgengesellschaft ... [et al].

[44]  J. Fowlkes,et al.  Cavitation threshold measurements for microsecond length pulses of ultrasound. , 1988, The Journal of the Acoustical Society of America.

[45]  J E Saunders,et al.  A review of the physical properties and biological effects of the high amplitude acoustic field used in extracorporeal lithotripsy. , 1993, Ultrasonics.

[46]  Lawrence A. Crum,et al.  Acoustic cavitation generated by an extracorporeal shockwave lithotripter , 1986 .

[47]  H. G. Flynn.  Generation of transient cavities in liquids by microsecond pulses of ultrasound , 1982 .

[48]  J. Debus,et al.  A comparison of shock wave and sinusoidal-focused ultrasound-induced localized transfection of HeLa cells. , 1999, Ultrasound in medicine & biology.