Microvascular heterogeneity exploration in core and invasive zones of orthotopic rat glioblastoma via ultrasound localization microscopy.

Background: We studied the microvascular structure and function of in situ glioblastoma using ultrasound localization microscopy (ULM).

Methods: The in vivo study was conducted via craniotomy in six Sprague-Dawley rats. Capillary pattern, capillary hemodynamics, and functional quantitative parameters were compared among tumor core, invasive zone, and normal brain tissue with ex vivo micro-computed tomography (micro-CT) and scanning electron microscopy. Correlations between quantitative parameters and histopathological vascular density (VD-H), proliferation index, and histopathological vascular maturity index (VMI-H) were evaluated. Kruskal-Wallis H, ANOVA, Mann-Whitney U, Pearson, and Spearman correlation statistics were used.

Results: Compared to the tumor core, the invasive zone exhibited higher microvascularity structural disorder and complexity, increased hemodynamic heterogeneity, higher local blood flow perfusion (p ≤ 0.033), and slightly lower average flow velocity (p = 0.873). Significant differences were observed between the invasive zone and normal brain tissue across all parameters (p ≤ 0.001). ULM demonstrated higher microstructural resolution compared to micro-CT and a nonsignificant difference compared to scanning electron microscopy. The invasive zone vascular density correlated with VD-H (r = 0.781, p < 0.001). Vessel diameter (r = 0.960, p < 0.001), curvature (r = 0.438, p = 0.047), blood flow velocity (r = 0.487, p = 0.025), and blood flow volume (r = 0.858, p < 0.001) correlated with proliferation index. Vascular density (r = -0.444, p = 0.044) and fractal dimension (r = -0.933, p < 0.001) correlated with VMI-H.

Conclusion: ULM provided high-resolution, noninvasive imaging of glioblastoma microvascularity, offering insights into structural/functional abnormalities.

Relevance statement: ULM technology based on ultrafast ultrasound can accurately quantify the microvessels of glioblastoma, providing a new method for evaluating the effectiveness of antiangiogenic therapy and visualizing disease progression. This method may facilitate early therapeutic assessment.

Key points: ULM reliably captures the vascular structures and hemodynamic features of glioblastoma in rats. Micro-CT and scanning electron microscopy validated its effectiveness in microvascular non-invasion characterization. ULM is expected to effectively evaluate glioblastoma anti-vascular therapy response.