Effects of Thalidomide, Cytochrome P-450 and TNF-.ALPHA. on Angiogenesis in a Three-dimensional Collagen Gel-culture.

Okajimas Folia Anat. Jpn.

Murata

et al. 2003

Self Cite

Summary: Small pieces of mouse aorta were cultured in collagen gels, and the formation of capillary-like tubes from the aortic explant was observed under phase-contrast and transmission electron microscopes. Migration of fibroblastic cells from the aortic explant occurred in 2 days. After about 10 days of culture, capillary-like tubes from the aortic explant were formed in the collagen gels. An immunohistochemical study on the collagen gel culture revealed the expression of fibroblast growth factor 2 (FGF-2) near the aortic explant at the active initial stage, with random migration of fibroblastic cells expressing FGF-9. mRNA was isolated from these cultures, and reverse transcription-polymerase chain reaction (RT-PCR) of the cultures revealed the expressions of both FGF-2 and FGF-9. Based on our results, we propose that FGF-9 is also related to angiogenic events.

Section: Discussionmentioning

confidence: 99%

Angiogenesis and Fibroblast Growth Factors (FGFs) in a Three-Dimensional Collagen Gel Culture

Nagatoro

Okajimas Folia Anat. Jpn.

Murata

et al. 2003

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“…The anti-angiogenic effect of thalidomide depended not on the type of thalidomide or thalidomide concentration, but rather a species-specific difference in the metabolism of the agent, and it was confirmed to have been induced through a suppression of TNF-a (Fujita et al 2002). On the basis of these data, we sought in this study to investigate the potent anti-angiogenic activity of thalidomide using an in vitro angiogenic model with three-dimensional collagen gel cultures of mouse aortae and a human in vitro model with human umbilical vein endothelial cells (HUVECs).…”

Section: Introductionmentioning

confidence: 89%

Anti-angiogenic effects of thalidomide: expression of apoptosis-inducible active-caspase-3 in a three-dimensional collagen gel culture of aorta

Asami

Tanaka

et al. 2004

Histochem Cell Biol

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The anti-angiogenic properties of thalidomide have led to the use of the agent as a remedy for multiple myeloma. Nevertheless, the anti-angiogenic moiety of thalidomide remains unidentified. In this study we examined the anti-angiogenic effects of thalidomide in an in vitro model using a three-dimensional collagen gel culture. Angiogenesis was significantly inhibited when the culture was treated with thalidomide plus cytochrome P-450 (CYP2B4), and the migrating cells and tubules were positive for active-caspase-3 in an accompanying immunohistochemical investigation. Transmission electron microscopic observation also confirmed that active-caspase-3-positive cells demonstrated apoptotic characteristics. This study is the first to morphologically demonstrate the effect of thalidomide in directly inducing the apoptosis of new tubules and migrating cells on a three-dimensional collagen gel culture of aorta. Taken together with earlier findings, our new results indicate that the thalidomide-induced inhibition of angiogenesis involves apoptosis in addition to the suppression of TNF-alpha and inhibition of cell migration from aorta explants, i.e., the factors important for capillarogenesis.

“…Later, angiogenesis was induced in tissues with the use of fibrin and collagen gels (Montesano et al 1985), matrigel, collagen, fibrin and plasma clot (Nicosia and Ottinetti 1990a). Among these various techniques, the culture of blood vessels in collagen gels has been used widely and effectively (Mori et al 1988;Nicosia and Ottinetti 1990b;Akita et al 1997a, b;Fujita et al 2002Fujita et al , 2004Suda et al 2004). In our previous experiment using collagen gels, the morphological and histochemical structures of the tubes formed in the gels were similar to the capillary-like structures of blood capillaries observed in vivo (Akita et al 1997a, b).…”

Section: Introductionmentioning

confidence: 95%

An in vitro model for studying vascular injury after laser microdissection

Komatsu

Tanaka

et al. 2005

Histochem Cell Biol

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We have developed an in vitro model for studying vascular injury. After 7-10 days in a three-dimensional collagen gel culture, capillary-like tubes were formed in the collagen gels. We injured these capillary-like tubes with a laser microdissection system or a scrape method with razors and then examined the collagen gel culture by phase contrast and electron microscopy. After laser injury, profuse necrotic cells were observed around the injured capillary-like tubes and within the tubular lumen compared to the razor injury. We then isolated total RNA from these cultures and prepared cDNA for investigations by quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). Quantitative real time RT-PCR revealed the up-regulation of transcription factor early growth response-1 (Egr-1) after both laser and razor injury, accompanied by the up-regulation of fibroblast growth factor-2 (FGF-2), a proangiogenic factor downstream of Egr-1. The effective laser energy is concentrated on the minute focal spot only. These methods provide a useful in vitro model for studying vascular injury.