SAFETY EVALUATION OF MID-FREQUENCY SONO­THROMBOLYSIS: ANIMAL BRAIN EXPERIMENT

J. Shimizu 1, J. Kubota 1, T. Fukuda 2, A. Sasaki 3, T. Azuma 3, K. Sasaki 4, K. Shimizu 5, T. Oishi 6, S. Umemura 7, H. Furuhata 1
1 Medical Engineering Laboratory,
2 Division of Neuropathology, The Jikei University School of Medicine – Tokyo, Japan
3 Faculty of Engineering, The University of Tokyo – Tokyo, Japan
4 Department of Veterinary Medicine, Tokyo University of Agriculture and Technology – Tokyo, Japan
5 Department of Zoology, Faculty of Science, Okayama University of Science – Okayama, Japan
6 Department of Cellular and Molecular Biology, Primate Research Institute, Kyoto University – Inuyama, Japan
7 Graduate School of Biomedical Engineering, Tohoku University – Sendai, Japan

Objective: We are developing mid-frequency sonothrombolysis, however, hemorrhagic complication in TRUMBI trial is a fence at these frequency. We evaluated safety in primate with a developed 490 kHz continuous waveform (CW)-US probe and investigated 400 kHz burst waveform (BW)-US emission for rabbit.

Material and Methods: 1.We applied a developed probe to Macaca monkey brain via sonication of the MCA through a temporal window. Each three cynomolgus monkeys were maintained for 1 day and 7 days after sonication. And more two elder rhesus monkeys were sonicated under the alteplase (0.9 mg/kg) i.v., and maintained for 7 days. An automatic switching circuit operated a therapeutic US (T-beam) generator for thrombolysis (490 kHz; CW-US, Ispta 0.72 W/cm2) and diagnostic TC-CFI (D-beam; 2.5 MHz; Ispta 0.20 W/cm2). A 15-min protocol, comprising 4 repeats of a sequence of 120-s T-beam followed by 30-s D-beam and then 5-min T-beam deactivation monitoring with D-beam, was repeated 4 times. 2. The three male rabbits were made a 2.5 cm of craniotomy window. 10days after surgery, they received the 15-min of transcutaneous BW-US (400 kHz, DC 20%, Ispta 0.70 W/cm2) from the craniotomy window. All animal brains were estimated histologically.

Results: 1.None of the monkeys showed neuropathological damage after sonication. 2. In rabbit, amyloid precursor protein positive axons in white matter and alpha-B crystallin positive astrocytes in cortex were observed which was similar to a diffuse axonal injury.

Discussion: One of the hemorrhagic causes in the TRUMBI trial is high effective mechanical index (eMI) over 2.0. There was no brain damage in monkey brain by developed probe at eMI=0.42. According to McDannold, the eMI threshold of the disruption in rabbit BBB by BW-US is 1.38. In our rabbit study, the similar traumatic change appeared at the eMI=1.56 by BW-US. It suggests that high eMI at brain induce traumatic damage.

Key words: mechanical index, neuropathology, safety, sonothrombolysis.