Correlation of Contrast-Enhanced Ultrasound with Two Distinct Types of Blood Vessels for the Assessment of Angiogenesis in Lewis Lung Carcinoma

Abstract

Objective: The aim of our study was to evaluate tumor angiogenesis in Lewis lung carcinoma (LLC) of mice using a contrast-enhanced ultrasound (CEUS) examination, and to determine the correlation of contrast-enhanced ultrasonographic parameters with different blood vessel markers of microvessel density (MVD).

Materials and Methods: Subcutaneous Lewis lung carcinomas were established in 25 mice, which were evaluated by contrast-enhanced US using SonoVue (a second-generation US contrast agent). The results were recorded as digital video images and the time-intensity curves and hemodynamic parameters were analyzed. Pathological tumor specimens were obtained just after US examination. Tumor specimens were stained with hematoxylin and eosin (H & E) and expression of CD31 and CD34, the different endothelial cell markers, was determined by immunohistochemical straining. Then the relationship between the CEUS parameters and the level of MVD was analyzed.

Results: Two distinct types of microvessels were identified in Lewis lung carcinoma: differentiated (CD34 +) and undifferentiated (CD31 +) vessels. A significant correlation was found between CEUS parameters and undifferentiated MVD (CD31 + vessels) in LLC (P < 0.05). There was a reverse correlation between the different MVDs.

Conclusion: The study showed that among the distinct types of vasculature (CD31 + and CD34 +) in Lewis lung carcinoma, the former correlated with the CEUS parameters. Therefore, CEUS using a second-generation US contrast agent may be useful for the evaluation of tumor angiogenesis of LLC of mice.

Zusammenfassung

Ziel: Studienziel war die Beurteilung der Tumorangiogenese im Lewis-Lungenkarzinom (LLC) von Mäusen unter Verwendung von kontrastverstärktem Ultraschall (CEUS) sowie die Untersuchung, ob CEUS-Parameter mit den verschiedenen Blutgefäßmarkern der Kapillardichte (MVD) korrelieren.

Material und Methoden: Subkutane Lewis-Lungenkarzinome wurden in 25 Mäusen erzeugt und mittels CEUS unter Verwendung von SonoVue (einem US-Kontrastmittel der zweiten Generation) bewertet. Die Ergebnisse wurden als digitale Videobilder aufgenommen und die Zeit-Intensitätskurven sowie hämodynamische Parameter analysiert. Die Tumorpräparate wurden mit Hämatoxylin und Eosin (HE) gefärbt und die Expression der Endothelzellmarker CD31 und CD34 immunhistologisch untersucht. Die Beziehung zwischen den CEUS-Parametern und dem Grad der MVD wurde analysiert.

Ergebnisse: Zwei unterschiedliche Kapillartypen wurden im Lewis-Lungenkarzinom nachgewiesen: Differenzierte (CD34 +) und undifferenzierte (CD31 +)-Gefäße. Ein signifikanter Zusammenhang wurde im LLC zwischen den CEUS-Parametern und den nicht-differenzierten MVDs (CD31 +-Gefäße) gefunden (P < 0,05). Zwischen den verschiedenen MVDs gab es eine umgekehrte Korrelation.

Schlussfolgerung: Die Studie zeigte, dass im Lewis-Lungenkarzinom im Mausmodell beide Typen des Gefäßsystems (CD31 + und CD34 +) der CD31 + Marker mit den CEUS-Parametern korrelierte. Deshalb kann CEUS mit einem Kontrastmittel der zweiten Generation zur Beurteilung der Tumorangiogenese des LLC im Mausmodell genutzt werden.

Key words

tumor - ultrasound - radiobiology

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