Preclinical evaluation of novel therapies using models of cancer is an important tool in cancer research, where imaging can provide non-invasive tools to characterise the internal structure and function of tumours. The short propagation paths when imaging tumours and organs in small animals allow the use of high frequencies for both ultrasound and shear waves, providing the opportunity for high-resolution shear wave elastography and hence its use for studying the heterogeneity of tissue elasticity, where heterogeneity may be a predictor of tissue response. Here we demonstrate vibrational shear wave elastography using a mechanical actuator to produce high frequency (up to 1000 Hz) shear waves in preclinical tumours, an alternative to the majority of preclinical ultrasound SWE studies where an acoustic radiation force impulse is required to create a relatively low-frequency broad-band shear-wave pulse. We implement vibrational shear wave elastography with a high frequency (17.8 MHz) probe running a focused line-by-line ultrasound imaging sequence which as expected was found to offer improved detection of 1000 Hz shear waves over an ultrafast planar wave imaging sequence in a homogenous tissue-mimicking phantom. We test the vibrational shear wave elastography in an ex-vivo tumour xenograft, demonstrating the ability to detect shear waves up to 10 mm from the contactor position at 1000Hz. By reducing the kernel size used for shear wave speed estimation to 1mm we are able to produce shear wave speed images with spatial resolution of this order. Finally, we present VSWE data from xenograft tumours in vivo, demonstrating the feasibility of the technique in mice under isoflurane sedation. Mean shear wave speeds in the tumours are in good agreements with those reported by previous authors. Characterising the frequency dependence of shear wave speed demonstrates the potential to quantify the viscoelastic properties of tumours in-vivo.

Preclinical investigation of the biomechanical properties of tissues and their treatment-induced changes are essential to support drug-discovery, clinical translation of biomarkers of treatment response, and studies of mechanobiology. Here we describe the first use of preclinical 3D elastography to map the shear wave speed (cs), which is related to tissue stiffness, in vivo and demonstrate the ability of our novel 3D vibrational shear wave elastography (3D-VSWE) system to detect tumour response to a therapeutic challenge. We investigate the use of one or two vibrational sources at vibrational frequencies of 700, 1000 and 1200 Hz. The within-subject coefficients of variation of our system were found to be excellent for 700 and 1000 Hz and 5.4 and 6.2%, respectively. The relative change in cs measured with our 3D-VSWE upon treatment with an anti-vascular therapy ZD6126 in two tumour xenografts reflected changes in tumour necrosis. U-87 MG drug vs vehicle: Δcs = −24.7 ± 2.5 % vs 7.5 ± 7.1%, (p = 0.002) and MDA-MB-231 drug vs vehicle: Δcs = −12.3 ± 2.7 % vs 4.5 ± 4.7%, (p = 0.02). Our system enables rapid (<5 min were required for a scan length of 15 mm and three vibrational frequencies) 3D mapping of quantitative tumour viscoelastic properties in vivo, allowing exploration of regional heterogeneity within tumours and speedy recovery of animals from anaesthesia so that longitudinal studies (e.g., during tumour growth or following treatment) may be conducted frequently.