Purpose: Impaired insulin-induced microvascular recruitment in skeletal muscle contributes to insulin resistance in type 2 diabetic disease. Previously, quantification of microvascular recruitment at the capillary level has been performed with either the full image or manually selected region-of-interests. These subjective approaches are imprecise, time-consuming, and unsuitable for automated processes. Here, an automated multiscale image processing approach was performed by defining a vessel diameter threshold for an objective and reproducible analysis at the microvascular level.
Approach: A population of C57BL/6J male mice fed standard chow and studied at age 13 to 16 weeks comprised the lean group and 24- to 31-week-old mice who received a high-fat diet were designated the obese group. A clinical ultrasound scanner (Acuson Sequoia 512) equipped with an 15L8-S linear array transducer was used in a nonlinear imaging mode for sensitive detection of an intravascular microbubble contrast agent.
Results: By eliminating large vessels from the dynamic contrast-enhanced ultrasound (DCE-US) images (above 300 μm in diameter), obesity-related changes in perfusion and morphology parameters were readily detected in the smaller vessels, which are known to have a greater impact on skeletal muscle glucose disposal. The results from the DCE-US images including all of the vessels were compared for three different-sized vessel groups, namely, vessels smaller than 300, 200, and 150 μm in diameter.
Conclusions: Our automated image processing provides objective and reproducible results by focusing on a particular size of vessel, thereby allowing for a selective evaluation of longitudinal changes in microvascular recruitment for a specific-sized vessel group between diseased and healthy microvascular networks.
Diabetes is a major disease and known to impair microvascular recruitment due to insulin resistance. Previous quantifications of the changes in microvascular networks at the capillary level were being performed with either full or manually selected region-of-interests (ROIs) from super-resolution ultrasound (SR-US) images. However, these approaches were imprecise, time-consuming, and unsuitable for automated processes. Here we provided a custom software solution for automated multiscale analysis of SR-US images of tissue microvascularity patterns. An Acuson Sequoia 512 ultrasound (US) scanner equipped with a 15L8-S linear array transducer was used in a nonlinear imaging mode to collect all data. C57BL/6J male mice fed standard chow and studied at age 13-16 wk comprised the lean group (N = 14), and 24-31 wk-old mice who received a high-fat diet provided the obese group (N = 8). After administration of a microbubble (MB) contrast agent, the proximal hindlimb adductor muscle of each animal was imaged (dynamic contrast-enhanced US, DCE-US) for 10 min at baseline and again at 1 h and towards the end of a 2 h hyperinsulinemiceuglycemic clamp. Vascular structures were enhanced with a multiscale vessel enhancement filter and binary vessel segments were delineated using Otsu’s global threshold method. We then computed vessel diameters by employing morphological image processing methods for quantitative analysis. Our custom software enabled automated multiscale image examination by defining a diameter threshold to limit the analysis at the capillary level. Longitudinal changes in AUC, IPK, and MVD were significant for lean group (p < 0.02 using Full-ROI and p < 0.01 using 150 μm-ROI) and for obese group (p < 0.02 using Full-ROI, p < 0.03 using 150 μm-ROI). By eliminating large vessels from the ROI (above 150 μm in diameter), perfusion parameters were more sensitive to changes exhibited by the smaller vessels, that are known to be more impacted by disease and treatment.