Selective cyclo-oxygenase-2 inhibition induces regression of autologous endometrial grafts by down-regulation of vascular endothelial growth factor-mediated.

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Selective cyclo-oxygenase-2 inhibition induces regression of autologous endometrial grafts by down-regulation of vascular endothelial growth factor-mediated angiogenesis and stimulation of caspase-3-dependent apoptosis Matthias W. Laschke, M.D., Antje Elitzsch, D.V.M., Claudia Scheuer, M.D., Brigitte Vollmar, M.D., Michael D. Menger, M.D. Fertility and Sterility Volume 87, Issue 1, Pages 163-171 (January 2007) DOI: 10.1016/j.fertnstert.2006.05.068 Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

FIGURE 1 Size of endometrial grafts (given in percent of initial size of grafts) on day 14 after autologous transplantation into dorsal skinfold chambers of control (open bars) and NS398-treated (solid bars) Syrian golden hamsters, as assessed by intravital fluorescence microscopy and computer-assisted image analysis. Means ± SEM. *P<.05 vs. control. Laschke. Anti-angiogenic therapy of endometriosis. Fertil Steril 2007. Fertility and Sterility 2007 87, 163-171DOI: (10.1016/j.fertnstert.2006.05.068) Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

Figure 2 (A and B) Intravital fluorescence microscopy of the newly formed microvasculature of endometrial grafts (borders marked by arrows) on day 14 after autologous transplantation into the dorsal skinfold chamber of a control (A) and NS398-treated (B) Syrian golden hamster. In both experimental groups, endometrial grafts are characterized by glomerulum-like microvascular networks. However, the anti-angiogenic effect of NS398 is reflected in decreased microvessel density (B). Blue-light epi-illumination with contrast enhancement by 5% FITC-labeled dextran 150,000 IV. Scale bars: 80μm. (C and D) Vascularized area (%) and microvessel density (cm/cm2) of endometrial grafts after autologous transplantation into dorsal skinfold chambers of control (open circles) and NS398-treated (solid circles) Syrian golden hamsters, as assessed by intravital fluorescence microscopy and computer-assisted image analysis. Means ± SEM. *P<.05 vs. control animals at corresponding time points. aP<.05 vs. day 0 within each individual group. bP<.05 vs. days 0 and 2 within each individual group. cP<.05 vs. days 0, 2, and 4 within each individual group. Laschke. Anti-angiogenic therapy of endometriosis. Fertil Steril 2007. Fertility and Sterility 2007 87, 163-171DOI: (10.1016/j.fertnstert.2006.05.068) Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

FIGURE 3 Western blot analysis of VEGF, PCNA, caspase-3, and activated caspase-3 protein expression (optical density·mm2) of endometrial grafts on day 2 (A) and day 14 (B) after implantation into dorsal skinfold chambers of hormonally synchronized Syrian golden hamsters. Animals were treated with vehicle DMSO (control, open bars) or selective COX-2 inhibitor NS398 (solid bars). Means ± SEM. *P<.05 vs. control. Laschke. Anti-angiogenic therapy of endometriosis. Fertil Steril 2007. Fertility and Sterility 2007 87, 163-171DOI: (10.1016/j.fertnstert.2006.05.068) Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

FIGURE 4 Hematoxylin-eosin-stained cross sections of endometrial grafts on day 14 after transplantation onto striated muscle tissue (arrows) within the dorsal skinfold chamber of a control (A and C) and NS398-treated (B and D) Syrian golden hamster. Grafts are characterized by cyst-like dilated endometrial glands (asterisks) with an intact glandular epithelium surrounded by a vascularized endometrial stroma. Higher magnification of grafts (C and D) reveals red blood cell-perfused blood vessels composed of an intact endothelium and without any signs of fibrin deposition or microthrombus formation. Scale bars: A and B: 120μm; C and D: 30μm. Laschke. Anti-angiogenic therapy of endometriosis. Fertil Steril 2007. Fertility and Sterility 2007 87, 163-171DOI: (10.1016/j.fertnstert.2006.05.068) Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions