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  • Title: Generational analysis reveals that TGF-beta1 inhibits the rate of angiogenesis in vivo by selective decrease in the number of new vessels.
    Author: Parsons-Wingerter P, Elliott KE, Farr AG, Radhakrishnan K, Clark JI, Sage EH.
    Journal: Microvasc Res; 2000 Mar; 59(2):221-32. PubMed ID: 10684728.
    Abstract:
    Quantitative analysis of vascular generational branching demonstrated that transforming growth factor-beta1 (TGF-beta1), a multifunctional cytokine and angiogenic regulator, strongly inhibited angiogenesis in the arterial tree of the developing quail chorioallantoic membrane (CAM) by inhibition of the normal increase in the number of new, small vessels. The cytokine was applied uniformly in solution at embryonic day 7 (E7) to the CAMs of quail embryos cultured in petri dishes. After 24 h the rate of arterial growth was inhibited by as much as 105% as a function of increasing TGF-beta1 concentration. Inhibition of the rate of angiogenesis in the arterial tree by TGF-beta1 relative to controls was measured in digital images by three well-correlated, computerized methods. The first computerized method, direct measurement by the computer code VESGEN of vascular morphological parameters according to branching generations G(1) through G(>/=5), revealed that TGF-beta1 selectively inhibited the increase in the number density of small vessels, N(v>/=5) (382 +/- 85 cm(-2) for specimens treated with 1 microg TGF-beta1/CAM for 24 h, compared to 583 +/- 99 cm(-2) for controls), but did not significantly affect other parameters such as average vessel length or vessel diameter. The second and third methods, the fractal dimension (D(f)) and grid intersection (rho(v)), are statistical descriptors of spatial pattern and density. According to D(f) and rho(v), arterial density increased in control specimens from 1.382 +/- 0.007 and 662 +/- 52 cm(-2) at E7 (0 h) to 1.439 +/- 0.013 and 884 +/- 55 cm(-2) at E8 (24 h), compared to 1. 379 +/- 0.039 and 650 +/- 111 cm(-2) for specimens treated with 1 microg TGF-beta1/CAM for 24 h. TGF-beta1 therefore regulates vascular pattern and the rate of angiogenesis in a unique "fingerprint" manner, as do other major angiogenic regulators that include VEGF, FGF-2 (bFGF), and angiostatin. TGF-beta1 did not stimulate angiogenesis significantly at low cytokine concentrations, which suggests that this quail CAM model of angiogenesis is not associated with an inflammatory response.
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