Individuals with diabetes at risk of developing diabetic kidney disease (DKD) are challenging to identify using currently available clinical methods. Prognostic accuracy and initiation of treatment could be improved by a quantification of the renal microvascular rarefaction and the increased vascular tortuosity during the development of DKD. Super-resolution ultrasound (SRUS) imaging is an in vivo technique capable of visualizing blood vessels at sizes below 75 µm. This preclinical study aimed to investigate the alterations in renal blood vessels’ density and tortuosity in a type 2 diabetes rat model, Zucker diabetic fatty (ZDF) rats, as a prediction of DKD. Lean age-matched Zucker rats were used as controls. A total of 36 rats were studied, subdivided into ages of 12, 22, and 40 weeks. Measured albuminuria indicated the early stage of DKD, and the SRUS was compared with the ex vivo micro-computed tomography (µCT) of the same kidneys. Assessed using the SRUS imaging, a significantly decreased cortical vascular density was detected in the ZDF rats from 22 weeks of age compared to the healthy controls, concomitant with a significantly increased albuminuria. Already by week 12, a trend towards a decreased cortical vascular density was found prior to the increased albuminuria. The quantified vascular density in µCT corresponded with the in vivo SRUS imaging, presenting a consistently lower vascular density in the ZDF rats. Regarding vessel tortuosity, an overall trend towards an increased tortuosity was present in the ZDF rats. SRUS shows promise for becoming an additional tool for monitoring and prognosing DKD. In the future, large-scale animal studies and human trials are needed for confirmation.

Background: Vascular imaging is essential for clinical practice, research, and the diagnosis and management of vascular diseases. Super-resolution ultrasound (SRUS) imaging is an emerging high-resolution imaging technique with broad applications in soft tissue vascular imaging. However, the impact of biological and clinical variables on its imaging accuracy is currently unknown. This study investigates these factors in an animal model and compares SRUS with contrast-enhanced µCT. Methods: Kidney scans from 29 Zucker rats (Zucker Diabetic Fatty and Zucker Lean) were retrospectively analysed. The left kidney was imaged in vivo using SRUS during microbubble infusion, then filled with Microfil and excised for ex vivo µCT. SRUS parameters and clinical variables were analysed, and SRUS scans were co-registered with µCT to compare vascular density measurements. Results: Mean arterial blood pressure and anaesthesia time showed significant linear relationships with SRUS microbubble detection and vascular track reconstruction. The anaesthesia time was also strongly correlated with vascular density measurement. Visualisation and velocity estimations of renal arteries were limited with SRUS. Ultrasound signal attenuation had significant impacts, particularly in cortical far-field imaging. Despite differences between kidney regions, the vascular density distribution did not differ considerably between SRUS and µCT datasets for whole-kidney imaging. Conclusions: This study outlines key factors SRUS users must consider for optimal technique use. Careful region selection and control of clinical variables ensure more reliable and comparable images. Further research is necessary to translate these findings from a rat model into clinical application.

Introduction Tumour vascularity is an important biomarker of tumour growth and therapeutic response. Current in vivo imaging methods have a limited ability to detect individual tumour vessels due to the small vessel size and low flow rates commonly observed in murine oncology models. This paper presents three pilot studies which together aim to assess the performance of a developing imaging method, Super-resolution Ultrasound using Erythrocytes (SURE), for subcutaneous tumour imaging. Methods Across three experiments, we qualitatively compared the vascular imaging sensitivity and clinical applicability of SURE imaging with colour Doppler imaging and ex vivo immunohistochemistry. Two murine oncology models were investigated: the LNCaP (lymph node carcinoma of the prostate) xenograft model (n = 3) and the B16-F10 melanoma model (n = 7). Optimal SURE post-processing parameters for subcutaneous tumour imaging were characterised. Results SURE demonstrated enhanced visualisation of tumour microvasculature relative to conventional colour Doppler imaging. SURE successfully visualised tumour microvasculature, detecting vessels approximately 30 μm in diameter while enabling haemodynamic characterisation. Optimal post-processing involved a trade-off between vascular imaging sensitivity, final signal-to-noise ratio, and image artefact prevalence. Vascular density measurements obtained using SURE did not differ significantly from those obtained by immunohistochemistry. Conclusions SURE represents a significant advance in in vivo microvascular imaging. While post-processing time, image artefacts, and limited quantitative analysis currently limit preclinical application, the technique demonstrates strong potential for tumour vascular characterisation in both preclinical research and clinical oncology.

Background/Objectives: Diabetic kidney disease (DKD) is a significant concern for global healthcare, particularly in individuals with diabetes. The Zucker rat strain is a commonly used model of type 2 diabetes, despite awareness that this animal can develop hydronephrosis. In this study, we present novel imaging data evaluating the accuracy of this animal model in replicating the vascular aspects of human DKD while examining the impact of hydronephrosis on its validity as a disease model. Methods: This study reused data from a population of male Zucker Diabetic Fatty (ZDF; n = 22) rats and Zucker Lean (ZL) rats (n = 22) aged 12 to approximately 40 weeks. Vascular casting was performed to enable visualisation of the renal vasculature. Anatomical regional volumes and vascular density data were obtained from μCT scans using image thresholding and manual analysis. The effects of hydronephrosis were evaluated using renal functional parameters and histological examination. Results: A significantly lower cortical vascular density, as well as lower total renal vascular density, was seen in ZDF rats compared to ZL rats, independent of age. We identified that hydronephrosis affected 92% of ZDF rats and 69% of ZL rats. Hydronephrosis cavity size was significantly correlated with the degree of hyperglycaemia and rate of diuresis but had no other detected impact on renal function, vascularity, or tissue histological architecture. Conclusions: These findings support using the Zucker rat strain as a model for vascular changes in DKD. Despite identifying severe hydronephrosis in this population, it had minimal quantifiable impact on renal function or diabetes modelling.