Increase in Angiogenesis and Vascularization in Patient-Derived Endometriosis Tissue: Insights from a 3D In Vivo Model

Abstract

Aim

Endometriosis is a gynecological disorder characterized by endometrial-like tissue outside the uterus. This study evaluates the vascularization and proliferation of human endometriosis and endometrium tissues engrafted onto the chorioallantoic membrane of chicken embryos using immunohistochemistry and laser speckle contrast analysis imaging. For the assessment of clinical relevance, a comparison between laboratory and clinical data was performed.

Material and Methods

Tissue samples from 10 patients categorized by #Enzian scores and undergoing endometriosis surgery were investigated in the chorioallantoic membrane model. Hematoxylin-eosin staining and immunohistochemical markers, including CD10, cytokeratin, Ki67, and Caspase-3, assessed cellular structures, proliferation, and apoptosis. Changes in blood perfusion, implemented as a surrogate marker for angiogenesis and vascularization, were analyzed over three days using laser speckle contrast analysis. The fertilized chicken eggs used for the chorioallantoic membrane model were stratified for their gender utilizing an in ovo sexing technique.

Results

Immunohistochemistry confirmed stromal and glandular cells in transplanted tissues. Ki67 indicated variable proliferation, while Caspase-3 identified apoptosis. Perfusion increased significantly in 75% of endometriosis samples. Endometrium from a patient with endometriosis showed increased perfusion, contrasting with stable perfusion in healthy endometrium. Higher #Enzian scores partly correlated with increased vascularization.

Summary

The chorioallantoic membrane model is a viable platform for studying endometriosis vascularization and angiogenesis. Endometriosis tissue showed enhanced vascularization influenced by lesion size and anatomical location, offering insights into disease progression and therapeutic strategies.

Zusammenfassung

Zielsetzung

Endometriose ist eine gynäkologische Erkrankung, die durch endometriumähnliches Gewebe außerhalb der Gebärmutter gekennzeichnet ist. Diese Studie untersucht die Vaskularisation und Proliferation von humanem Endometriose- und Endometriumgewebe, das auf die Chorioallantoismembran (CAM) von Hühnereiern transplantiert wurde, mittels Immunhistochemie und Laser-Speckle-Kontrastanalyse (LASCA). Zur Bewertung der klinischen Relevanz wurde ein Vergleich zwischen experimentellen und klinischen Daten durchgeführt.

Material und Methoden

Gewebeproben von 10 Patientinnen, klassifiziert nach #Enzian-Score und operiert aufgrund von Endometriose, wurden im CAM-Modell untersucht. Zelluläre Strukturen, Proliferation und Apoptose wurden mittels Hämatoxylin-Eosin-Färbung und immunhistochemischer Marker (CD10, Zytokeratin, Ki-67 und Caspase-3) beurteilt. Veränderungen der Durchblutung – als Surrogatmarker für Angiogenese und Vaskularisation – wurden über 3 Tage hinweg mittels LASCA analysiert. Die befruchteten Hühnereier im CAM-Modell wurden mithilfe eines In-ovo-Geschlechtsbestimmungstests hinsichtlich ihres Geschlechts stratifiziert.

Ergebnisse

Die Immunhistochemie bestätigte Stromazellen und Drüsenzellen in den transplantierten Geweben. Ki-67 zeigte eine variable Proliferation, während Caspase-3 Apoptose nachwies. In 75% der Endometrioseproben nahm die Durchblutung signifikant zu. Endometrium einer Patientin mit Endometriose zeigte eine erhöhte Perfusion im Gegensatz zu stabiler Durchblutung in gesundem Endometrium. Höhere #Enzian-Scores korrelierten teilweise mit erhöhter Vaskularisation (p < 0,001).

Schlussfolgerung

Das CAM-Modell stellt eine geeignete Plattform zur Untersuchung der Vaskularisation und Angiogenese bei Endometriose dar. Endometriosegewebe zeigte eine verstärkte Gefäßneubildung, die von der Größe und anatomischen Lokalisation der Läsionen beeinflusst wurde, und liefert somit wertvolle Erkenntnisse über den Krankheitsverlauf und potenzielle Therapieansätze.

Keywords

CAM model - endometriosis - chorioallantoic membrane - 3D in vivo model - human endometriosis tissue - human tissue

Schlüsselwörter

CAM-Modell - Endometriose - Chorioallantoismembran - 3D-in-vivo-Modell - humanes Endometriosegewebe - humanes Gewebe

Publication History

Received: 19 January 2025

Accepted after revision: 02 July 2025

Article published online:
22 July 2025

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