A comparative study on the effects of tumor necrosis factor‐α (TNF‐α), human angiogenic factor (h‐AF) and basic fibroblast growth factor (bFGF) on the chorioallantoic membrane of the chick embryo

Abstract

The chorioallantoic membrane (CAM) assay is a widely used bioassay for testing angiogenic activities. In the present study we compared the gross and micromorphological effects of three angiogenic factors applied in Elvax carriers on the CAM: Tumor necrosis factor-alpha (TNF-alpha), human angiogenic factor (h-AF), and basic fibroblast growth factor (bFGF). Our question was whether the CAM responds to these factors which have very different actions with a stereotype or with a factor specific reaction. By microangiography and light microscopy, all positive reactions appeared as a spoke-wheel vascular pattern with a bundle of small capillary blood vessels in the center. These vessels were predominantly of a distended type in h-AF and TNF experiments, while narrower capillary vessels followed bFGF application. Chorioallantoic ectoderm and endoderm were thickened by cell accumulation and the mesenchymal stroma of the CAM was edematous and infiltrated with leucocytes in all three reactions. Additionally, bFGF experiments showed areas of densely arranged fibroblasts. Observations in vivo showed chorioallantoic tissue movements as a possible mechanism for the spokewheel vascular pattern. As compared with our results from studies of cytokinetics with bromodeoxyuridine, these current findings indicate that chemotaxis is responsible for the chorioallantoic angiogenic reaction rather than cellular proliferation.

Chorioallantoic membrane capillary bed: A useful target for studying angiogenesis and anti‐angiogenesis in vivo

“…Growth factor Esch et al (1985) Basic fibroblast growth factor (bFGF) or Wilting et al (1991) Fibroblast growth factor-2 (FGF-2) Olivo et al (1992) Ribatti et al (1995) Fett et al (1985 Angiogenin Wilting et al (1991) Olivo et al (1992 Leibovich et al (1987) Tumor necrosis factor alpha (TNF-␣) Olivo et al (1992) Yang andMoses (1990) Transforming growth factor beta (TGF-␤) Wilting et al (1992) Platelet derived growth factor (PDGF) Wilting et al (1992Wilting et al ( , 1993 Vascular endothelial growth factor (VEGF) Ziche et al (1997) Placenta growth factor-1 (PIGF-1) Bouloumie et al (1998) Leptin Ribatti et al (2001 Erythropoietin (EPO) Ramoshebi and Ripamonti (2000) Osteogenic protein-1 (OP-1) Bernardini et al (2000) CC chemokine I-309…”

New Model for the Study of Angiogenesis and Antiangiogenesis in the Chick Embryo Chorioallantoic Membrane: The Gelatin Sponge/ Chorioallantoic Membrane Assay

“…Eight-micrometer serial sections were cut in parallel to the surface of the CAM, stained with a 0.5% aqueous solution of toluidine blue (Merck) and were observed under a Lcitz-Dialux 20 light microscope. On day 12, some CAMs were injected into a large chorioallantoic vein with india ink [22].…”

Pyogenic Granuloma Stimulates Angiogenesis in the Chick Embryo Chorioallantoic Membrane

“…The microscopic and morphometric evidence is more convincing com pared to the macroscopic results because, as suggested by Olivo et al [20], the macroscopic 'spoked wheel' pattern may also be related to cellular accumulation and perhaps CAM contraction, rather than to the formation of new microvessels. Otherwise, it remains unclear whether the reaction of the CAM to a stimulus is a stereotypic reaction or a specific response to certain angiogenic factors.…”

“…At day 12 CAMs were injected intravascularly with india ink according to Olivo et at. [20]. The ink was diluted 1:1 with phosphate-buffered saline, filtered through a filter with 0.8-pm-sized pores, and injected into a large chorioallantoic vein by using a tine glass needle and a micromanipu lator.…”