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DOI: 10.1055/s-2006-924118
Georg Thieme Verlag KG Stuttgart · New York
Optimizing the Outcome of Grafted Ovarian Tissue: Exogenous Gonadotropins to Increase Microvessel Density and Follicular Survival
Optimierung der Ergebnisse der Transplantation von Ovarialgewebe: Exogene Gonadotropine zur Erhöhung der mikrovaskulären Dichte und des follikulären ÜberlebensPublication History
Eingang Manuskript: 20.3.2006
Akzeptiert: 27.3.2006
Publication Date:
23 May 2006 (online)
![](https://www.thieme-connect.de/media/gebfra/200605/lookinside/thumbnails/10.1055-s-2006-924118-1.jpg)
Zusammenfassung
Einführung: Die Strategie der Kryokonservierung und Transplantation von ovariellem Gewebe bei malignomerkrankten Frauen zur Erhaltung der Fertilität beinhaltet die Problematik eines ausgeprägten ischämiebedingten Follikelverlustes. Fragestellung: Lässt sich durch die systemische bzw. lokale Applikation angiogeneseinduzierender Substanzen die (Neo-)Vaskularisation nach Transplantation verbessern bzw. der Follikel-Verlust reduzieren? Material und Methoden: A: Drei weiblichen Schafen wurden beide Ovarien explantiert, in 1 mm dicke Scheiben geschnitten und anschließend autolog heterotop in die Bauchwand retransplantiert. Bei einem Schaf wurde das ovarielle Gewebe im Rahmen der Transplantation lokal in eine VEGF (5 µg-)Fibrinkleber-Suspension eingebettet. Einem weiteren Schaf wurden über 2 Wochen nach Transplantation alle 2 Tage 150 IE/hMG systemisch appliziert. Das dritte Schaf blieb ohne Behandlung. Neun Monate später wurde das transplantierte Gewebe zur histologischen Analyse entnommen und das Follikelüberleben miteinander verglichen. B: Zwei weiteren Schafe wurden beide Ovarien explantiert, in entsprechende Scheiben präpariert, und in 20 immun-inkompetente Mäuse xenolog heterotop im Bereich des Rückens transplantiert. Zehn dieser Mäuse erhielten jeden 2. Tag 10 IE hMG systemisch beginnend ab dem Tag der Transplantation bis zur finalen Explantation. Die übrigen 10 Mäuse erhielten keine weitere spezifische Behandlung. Anschließend wurden jeweils 2 Mäuse beider Gruppen an Tag 2, 4, 6, 10 und 14 nach primärer Transplantation euthanasiert, das Transplantat entnommen und die Gefäßdichte in beiden Gruppen zu unterschiedlichen Zeitpunkten miteinander verglichen. Ergebnisse: A: Im Gegensatz zur Applikation von VEGF, welche im Vergleich zur Kontrolle keinen Effekt auf das Follikelüberleben hatte (12 vs. 7,4 %), konnte durch die Applikation von hMG das Follikelüberleben deutlich gesteigert werden (12 % vs. 29 %). B: Weiterhin ergab die Applikation von hMG im Vergleich zur Kontrollgruppe eine signifikant höhere mittlere Gefäßdichte pro mm2 zu sämtlichen untersuchten Zeitpunkten (Tag 2: 22 vs. 13 [p < 0,001], Tag 4: 22 vs. 13 [p < 0,001], Tag 6: 17 vs. 13 [p = 0,006], Tag 10: 28 vs. 12 [p < 0,001], bzw. Tag 14: 35 vs. 16 [p < 0,001]). Schlussfolgerungen: Der ausgeprägte ischämiebedingte Follikelverlust nach Transplantation von ovariellem Gewebe lässt sich durch die systemische Gabe von hMG, welches eine gesteigerte (Neo-)Vaskularisation induziert, deutlich reduzieren.
Abstract
Background: Fertility preservation by ovarian cryopreservation and heterotopic ovarian transplant is hampered by ischemic damage of the transplanted graft and poor oocyte viability. Design: Two longitudinal experiments were done - phase A: to evaluate if the administration of gonadotrophins (hMG) or vascular endothelial growth factor (VEGF) increases follicular survival after heterotopic autologous ovarian transplantation, and phase B: to determine if the administration of hMG enhanced neovascularization by increase in microvessel density in the heterotopic xenologous ovarian transplant. Methods: A: In three sheep, ovaries were removed and immediately transplanted into the abdominal wall. In one sheep, VEGF was administered at the transplantation site during surgery, the second was treated with hMG after transplantation for 2 weeks, and the third sheep served as control. B: The ovaries from two other sheep were removed, prepared in tissue pieces, and immediately grafted into the subcutaneous space of several combined immunodeficient (SCID-)mice (n = 20). Following the transplantation, 10 of these mice (group 1) were treated with hMG until they were sacrificed, the other 10 untreated mice (group 2) served as controls. In each group, two mice were sacrificed at intervals of 2, 4, 6, 10, and 14 days after grafting to permit histologic examination of the grafted tissue. Results: A: Whereas VEGF had no effect on follicular survival after transplantation (12.0 % vs. 7.4 %.), the administration of hMG increased follicular survival (12.0 % vs. 29.0 %). B: The use of hMG resulted in significantly higher numbers of microvessels per mm2 (mean) at all of the time points studied - at 2 days 22 vs. 13 (p < 0.001), 4 days 22 vs. 13 (p < 0.001), 6 days 17 vs. 13 (p = 0.006), 10 days 28 vs. 12 (p < 0.001), and 14 days 35 vs. 16 (p < 0.001), respectively. Conclusion: The improved follicular survival and increase in microvessel density in heterotopic ovarian transplants associated with the use of hMG suggests that neovascularization may be an important mechanism by which hMG improves the survival of ovarian transplants.
Schlüsselwörter
Fertilität - Malignom - Ovar - Transplantation - Angiogenese
Key words
Fertility - cancer - ovary - transplantation - angiogenesis
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Dominik Denschlag
Department of Obstetrics and Gynecology
McGill University Montreal
Royal Victoria Hospital
687 Pine Avenue West
Women's Pavilion, F9.29
Canada, QC
Phone: + 1-514-843-2833
Fax: + 1-514-843-2830
Email: dominik.denschlag@muhc.mcgill.ca