High-Frequency Ultrasound Properties of Multicellular Spheroids During Heating

Abstract

High-frequency ultrasound monitoring is a possible method for real-time imaging of thermal therapy in tissues at microscopic resolution. The objective of this work was to measure changes in the ultrasound properties of V79 spheroids (grown from Chinese Hamster lung fibroblasts) exposed to heating. Spheroids are clonal aggregates of cells that provide a useful model for investigating the ultrasound properties of cells in the absence of connective tissue. Relative echo signal power and attenuation coefficients were measured over the frequency range 30 MHz to 70 MHz, from spheroids heated from 37°C to 50°C or 60°C. Echo signal power from the viable rim decreased during the first 5 min by a factor of 1.08 before the spheroid reached 50°C. For the next 25 min, echo signal power rose to a factor of 1.27 above the initial level, after which it remained relatively constant over the remainder of the 50°C heating period. At 60°C, echo signal from the viable rim remained relatively constant, although it appeared to have possibly decreased slightly over the duration of the heating period. Echo signal power from the necrotic core fell to a factor of 1.4 and 1.54 below the initial level at 50°C and 60°C, respectively. First-order chemical rate analysis applied to the echo signal power results in the viable rim at 50°C revealed a rate constant for the 5–15-min heating interval. Interpretation of the echo signal power results in terms of histological stains indicates that the rise in echo signal power at 50°C was due to a loss of cell cohesion, and the possible drop in echo signal power at 60°C was due to spheroid coagulation. Attenuation coefficients decreased by up to 1.54 dB mm −1 over a 30-min period at 60°C. The appearance of a real-time ultrasound image of lesion formation in cells is discussed.