1. Volume 130, Issue 7, Pages 2060-2073 (June 2006)
Angiogenesis as a Novel Component of Inflammatory Bowel Disease Pathogenesis 
Silvio Danese, Miquel Sans, Carol de la Motte, Cristina Graziani, Gail West, Manijeh H. Phillips, Roberto Pola, Sergio Rutella, Joe Willis, Antonio Gasbarrini, Claudio Fiocchi 
Gastroenterology 
Volume 130, Issue 7, Pages (June 2006)
DOI: /j.gastro
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

2. Figure 1
Detection of microvasculature in normal and IBD mucosa. The panels show brown immunohistochemical staining for CD31 and von Willebrand/factor VIII in the microvasculature of colonic mucosa and submucosa from histologically normal control, active UC, and active CD tissue. The panels are representative of 8 control, 9 UC, and 8 CD samples.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

3. Figure 2
Computerized morphometric analysis of the microvasculature in control and IBD mucosa. After immunohistochemical staining for CD31 and von Willebrand/factor VIII of the microvasculature of histologically normal control, CD and UC colonic mucosa, sections were analyzed for the total number of vessels/field (microvascular density), percentage section area, mean vessel area, and larger vessel diameter. Data derived from 8 control, 8 CD and 9 UC samples. *P < .05 for UC and CD compared with control; **P < .01 for UC and CD compared with control.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

4. Figure 3
Localization of integrin αVβ3 in IBD mucosa. (A) Confocal micrographs of colonic mucosa microvessels in CD and UC mucosa stained for αVβ3 (red) and nuclei (blue). Figures are representative of 7 CD and 8 UC samples. Eight control samples were also examined but αVβ3 was barely detectable (not shown). (B) Semiquantification of the integrin αVβ3 in microvessels of normal, CD, and UC mucosa. Data derived from 8 control, 7 CD, and 8 UC samples. *P < .01 for CD and UC compared with normal.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

5. Figure 4
Modulation of integrin αVβ3. HIMEC monolayers were left untreated or stimulated with TNF-α, bFGF, and VEGF for 24 hours, after which cells were suspended and αVβ3 expression measured by flow cytometry. The black curve represents the background signal from the isotype control. The figure is representative of 3 separate experiments. Numbers represent the net percentage of positive cells.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

6. Figure 5
Effect of αVβ3 blockade on HIMEC survival. (A) HIMEC monolayers were left untreated or incubated with Vitaxin or RGDV peptide for 1 hour, after which cells were stained with calcein and assessed by fluorescence microscopy. (B) HIMEC monolayers were suspended by gentle pipetting, detached cells stained with propidium iodide (PI) and analyzed by flow cytometry. The figure is representative of 4 separate experiments.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

7. Figure 6
Expression of angiogenic factors in IBD mucosa. Mucosal extracts of control and IBD patients with active and inactive disease were obtained and their content of VEGF assessed by Western blotting (A) and of IL-8 and bFGF measured by ELISA (B and C). The VEGF immunoblot is representative of 6 control, 8 active, and 4 inactive CD, and 8 active and 4 inactive UC samples; IL-8 and bFGF data are derived from 8 control, 8 active, and 6 inactive CD, and 8 active and 6 inactive UC samples. (A) *P < .05 for CD and UC compared with control and for active compared with inactive disease; (B) *P < .05 and **P <.01 for CD and UC compared with control; and (C) *P < .05 and **P < .01 for active compared with inactive disease.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

8. Figure 7
Production of angiogenic factors by LPMCs and HIFs. (A) LPMCs isolated from control mucosa were left untreated or stimulated with LPS or anti-CD3/CD28 monoclonal antibodies; (B) HIF monolayers from control mucosa were left untreated or stimulated with TNF-α. The concentration of VEGF, IL-8, and bFGF was measured by ELISA in the culture supernatants. Data derived from 4 separate LPMC isolates and 4 HIF monolayers. *P < .05 for LPS or anti-CD3/CD28-stimulated compared with unstimulated LPMCs and **P < .01 and ***P <.001 for TNF-α-stimulated compared with unstimulated HIF.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

9. Figure 8
Effect of normal and IBD mucosal extracts on induction of HIMEC migration. HIMECs were seeded on the membrane of a transwell insert and left untreated or exposed to PMA and bFGF as positive controls or different concentrations of extracts from normal CD and UC mucosa. Migrated cells were labeled with calcein and counted by computerized fluorescence microscopy. Data were derived from 5 separate experiments. *P < .05 for CD and UC compared with normal mucosa extracts at both 1:10 and 1:100.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

10. Figure 9
Differential effect of angiogenic factors and αVβ3 blockade on HIMEC migration. (A) HIMECs were seeded on the membrane of a transwell insert and left untreated or exposed to a 1:10 concentration of extracts from normal and IBD mucosa (2 CD and 2 UC). Experiments were performed in the absence and presence of blocking antibodies against IL-8, bFGF, and VEGF. Migrated cells were labeled with calcein and counted by computerized fluorescence microscopy. Data were derived from 4 separate experiments. *P < .05 for anti-VEGF-treated compared with untreated IBD mucosal extracts; **P < .01 for anti-IL-8- or anti-bFGF–treated compared with untreated IBD mucosal extracts. (B) HIMECs were seeded on the membrane of a transwell insert and left untreated or exposed to a 1:10 concentration of extracts from IBD mucosa (2 CD and 2 UC). Experiments were performed in the absence and presence of Vitaxin and RGDV peptide, both of which block the integrin αVβ3. ***P < .001 for Vitaxin- and RGDV peptide–treated compared with untreated IBD mucosal extracts.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

11. Figure 10
Effect of control and IBD mucosal extracts on induction of angiogenesis in vivo. (A) Filter papers containing PBS, VEGF, control, or IBD (2 CD and 1 UC) mucosal extracts were implanted in a corneal pocket, and endothelial staining was performed with fluorescent BS1-lectin. Images are representative of 3 separate experiments. (B) Filter papers containing PBS, VEGF, control, or IBD (1 CD and 1 UC) mucosal extracts were placed on a chick chorioallantoic membrane, and vessel formation was observed with a fiberoptic light source. Images are representative of 2 separate experiments.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions