Area Vasculosa Of Chick Embryo Research Articles

Morphometric analysis of the branching of the vascular tree in the chick embryo area vasculosa

The chick embryo includes the area vasculosa is subdivided into 2 concentric zones, the inner transparent area pellucida vasculosa and the surrounding less transparent area opaca vasculosa, peripherally limited by the sinus terminalis. In this study, we have analyzed by a modern morphometric approach the total length of the vascular network, the number of vascular branches, of the branching points density, the modality of vessel ramification, and spatial arrangement of the vascular network in four consecutive stages of development of the area vasculosa. The results have shown that there is a significant 15% increase in the total length of the vascular network associated with a progressive increase of the number of vascular branches and of the branching points density. Moreover, the results indicated that vascular spatial disorder significantly decreased during development in area vasculosa, suggesting a more uniform occupancy of the tissue by the vascular pattern. Finally, a more regular pattern of branching was observed, as indicated by the significant decrease of topological disorder of the vascular tree.

Developing a partially perfused liver model using caprine liver explant conditioned to the chicken extra-embryonic perfusion system

The Liver is constantly subjected to mechanical, chemical and pathological insults throughout life, as a result of which there is a common occurrence of various liver diseases. Due to the complex nature of liver architecture, it is not possible to mimic the in-vivo conditions beyond a certain limit. Hence, the development of in-vitro and ex-vivo models to study various liver diseases has gained more importance over the last few decades. The present study aims to develop a semi-perfused liver explant model to give an extended lifetime for studying liver pathology and treatment options. Caprine liver tissue explants were sliced, weighed (25 mg) and placed on the area vasculosa of fertilized chicken eggs. Unfertilized eggs were used as controls. After varying time intervals of incubation on area vasculosa of fertilized chicken eggs, the liver slices were subjected to cell viability assay, lactate dehydrogenase assay and Serum glutamic pyruvic transaminase assays. The results indicate that the viability and functional properties of such semi-perfused liver tissue explants holds good up to 6 h. Finally, the liver tissue explants were pre-treated with FBS, with and without anti-fibrotic drugs, and placed on chick embryo area vasculosa up to 6 h. The anti-fibrotic drug-treated semi perfused liver tissue explant showed a decrease in collagen formation as confirmed by histology and western blot. We deem that the use of extra-embryonic vasculature of chicken bed for extending the life of tissue explants will serve as a cost-effective alternative to animal models to understand disease mechanisms under drug treatments.

Live Imaging and Analysis of Vasoactive Properties of Drugs Using an in-ovo Chicken Embryo Model: Replacing and Reducing Animal Testing

Vasodilation occurs as a result of the relaxation of the smooth muscle cells present in the walls of blood vessels. Various suitable models are available for the analysis of the vasoactive properties of drugs with therapeutic applications. But all these models have limitations, such as ethical issues and high cost. The purpose of this study is to develop an alternative model for studying the vasoactive properties of drugs using an in-ovo chicken embryo model. In the preliminary experiment, we used a well-known vasoconstrictor (adrenaline) and a vasodilator (spermine NoNoate) in the chick embryo area vasculosa and evaluated their concentration-response curve. Adrenaline (10 µM) and spermine NoNoate (10 µM) were administered in different arteries and veins and different positions of the right vitelline artery of the chick embryo. Results showed the middle of the vessel bed of the right vitelline artery having the best vasoactive effect compared to others. Finally, anti-hypertensive drugs, calcium channel blockers, and NOS agonists were administered in the chick embryo area vasculosa to validate the model. Results demonstrate that the chick embryo area vasculosa can be an alternative, robust, and unique in-ovo model for screening of anti-hypertensive drugs in real time.

Investigation of Effects of Essential Oil from Berries of Juniperus excelsa Bieb. Subsp excelsa (Cupressaceae) on Angiogenesis in Shell-less Chick Embryo (CAM) Culture

The aim of this study was to investigate the effect of essential oil of the berries from Juniperus excelsa Bieb. subsp. excelsa (Cupressaceae) to the vascularization systems of the chick embryos in a chorioallantoic membrane model. The berries of J. excelsa were subjected to hydrodistillation. 50 µl of essential oil were applied to the center of the blastodisc using a precision micropipette prior to the transfer of shell-less cultures to the incubator. Antiangiogenic effect of essential oil of the study plant was determined with the use of the chick chorioallantoic membrane model. Numbers of main, branching and capillary blood vessels were counted. In the statistical analysis, when the groups were compared with regard to the main blood vessel (MBV) and branch blood vessel (BBV) development, no significant difference (p>0.05) was observed. However, when the groups were compared with regard to capillary vessel (CV) development, the difference between the data of the groups was found significant (p<0.05). This study examined the effect of exposure J. excelsa essential oil on extraembryonic vascular development in the chick embryo area vasculosa (AV) in shell-less culture for the first time. Essential oil of the berries from J. excelsa has antiangiogenic effect.

Investigation of effects of essential oils of Origanum minutiflorum O Schwarz PH Davis and Cyclotrichium niveum (Labiatae) plants on angiogenesis in shell-less chick embryo culture

The aim of this study was to investigate the effect of essential oils extracted from Origanum minutiflorum and Cyclotrichium niveum (Labiatae) plants to the vascularization systems of the chick embryos in a chorioallantoic membrane model. The aerial parts of O. minutiflorum and C. niveum were subjected to hydrodistillation. Essential oils were applied to the center of the blastodisc of 50 µl using a precision micropipette prior to the transfer of shell-less cultures to the incubator. Antiangiogenic effect of essential oil of the study plants was determined with the use of chorioallantoic membrane model. Numbers of main, branching and capillary blood vessels were counted. In the statistical analysis, when the groups were compared with regard to the main blood vessel (MBV) and branch blood vessel (BBV) development, no significant difference (p > 0.05) was observed. However, when the groups were compared with regard to capillary vessel (CV) development, the difference between the data of the groups was found significant (p < 0.05). One of the tested compounds, C. niveum essential oils, showed antiangiogenic effect; while the other, O. minutiflorum showed no antiangiogenic effect. This study examined the effect of exposure to O. minutiflorum and C. niveum essential oils on extraembryonic vascular development in the chick embryo area vasculosa (AV) in shell-less culture for the first time. Essential oils extracted from C. niveum has antiangiogenic effect; while O. minutiflorum essential oils had no antiangiogenic effect.

αvβ3 Integrin-dependent antiangiogenic activity of resveratrol stereoisomers

Angiogenesis is target for antineoplastic and chemopreventive therapies. The natural phytoalexin resveratrol is found in grapes and red wine as cis and trans stereoisomers. trans-Resveratrol shows antiangiogenic activity, but its mechanism of action is not fully elucidated. Recently, trans-resveratrol has been shown to interact with the beta3 integrin subunit, raising the possibility that inhibition of endothelial alphavbeta3 integrin function may concur to its angiosuppressive activity. To get novel insights about the antiangiogenic activity of resveratrol, we compared cis- and trans-resveratrol stereoisomers for their effect on the angiogenesis process and endothelial alphavbeta3 integrin function. trans-Resveratrol inhibits endothelial cell proliferation and the repair of mechanically wounded endothelial cell monolayers. Also, it prevents endothelial cell sprouting in fibrin gel, collagen gel invasion, and morphogenesis on Matrigel. In vivo, trans-resveratrol inhibits vascularization of the chick embryo area vasculosa and murine melanoma B16 tumor growth and neovascularization. In all the assays, cis-resveratrol exerts a limited, if any, effect. In keeping with these observations, trans-resveratrol, but not cis-resveratrol, inhibits alphavbeta3 integrin-dependent endothelial cell adhesion and the recruitment of enhanced green fluorescent protein-tagged beta3 integrin in focal adhesion contacts. In conclusion, stereoisomery affects the antiangiogenic activity of resveratrol, the trans isomer being significantly more potent than the cis isoform. The different antiangiogenic potential of resveratrol stereoisomers is related, at least in part, to their different capacity to affect alphavbeta3 integrin function. This may have profound implications for the design of synthetic antiangiogenic/angiopreventive phytoalexin derivatives.

The effects of lithium on vascular development in the chick area vasculosa.

The effects of lithium on vascular development were examined using the chick embryo area vasculosa in shell-less culture as an experimental model. Embryos were explanted after 48 h in ovo and LiC1 (50, 100, 150 and 200 microg in 10 microl water) was applied to the centre of the blastodisc. Controls were untreated or given equimolar amounts of NaCl. At 24 h and 48 h after treatment, untreated and NaCl controls were identical, having well developed extraembryonic vessels. At doses of 100 microg and greater, LiCl significantly inhibited normal vascular development and expansion of the area vasculosa in the majority of explants. In many specimens blood islands continued to form but their assembly into primitive vessels was prevented, indicating that lithium affects the mechanism regulating the assembly of vascular endothelium. At the same time the embryos were alive but retarded in development compared with controls. When LiCl (150 microg) was applied to cultures explanted after 72 h in ovo (when the primary vascular network had already formed through vasculogenesis) no adverse effects were seen. This suggests that lithium affects vasculogenesis but not angiogenesis. Treatment with myo-inositol completely reversed the effects of lithium in a time dependent manner indicating that the phosphatidylinositol second messenger cycle may be involved in the cellular events of vasculogenesis. Finally the results of this study show that the yolk sac vasculature is particularly vulnerable to lithium and the consequent effects of this interference on embryonic development are discussed.

217 - Effect of amifostine (Ethyol®) on vascular density of the area vasculosa of chick embryo

217 - Effect of amifostine (Ethyol®) on vascular density of the area vasculosa of chick embryo

Human Recombinant Interferon Alpha-2a Inhibits Angiogenesis of Chick Area Vasculosa in Shell-Less Culture

In vivo suppression of neovascularization by in situ administration of human recombinant interferon alpha 2a (hrIFN-alpha 2a) was tested on the chick embryo area vasculosa (AV). Methylcellulose discs, each containing 5 IU of hrIFN-alpha 2a were implanted onto the AV at Hamburger-Hamilton (HH) stage 13, and inhibition of blood vessel growth was morphometrically evaluated between HH stages 20 and 27. Our results show: (i) a decreased extent of the AV; (ii) a decreased AV total vessel length; (iii) modified percent ratios of different classes of AV vessels having a definite length. The observed effects of treating with hrIFN-alpha 2a are likely attributable to inhibition of endothelial cell proliferation.