An antagonist of integrin alpha v beta 3 prevents maturation of blood vessels during embryonic neovascularization

Experimental data in this study demonstrate that integrin alpha v beta 3 is fundamentally involved in the maturation of blood vessels during embryonic neovascularization (vasculogenesis). Integrin alpha v beta 3 was specifically expressed on the surface of angioblasts during vessel development in quail embryos and vitronectin, a ligand for alpha v beta 3, localized to the basal surface of these cells. More importantly, microinjection of the anti-alpha v beta 3 monoclonal antibody, LM609, disrupted the normal pattern of vascular development. After exposure to LM609 the angioblasts in experimental embryos appeared as clusters of rounded cells lacking normal cellular protrusions. This led to disruption of lumen formation and abnormal vessel patterning. These findings demonstrate that during vasculogenesis ligation of integrin alpha v beta 3 on the surface of primordial endothelial cells is critical for the differentiation and maturation of blood vessels. Similar studies on chicken chorioallantoic membrane showed that LM609 blocks angiogenesis. Together the two studies suggest that integrin alpha v beta 3 plays a role in neovascularization of tissues.

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Angiogenesis in chicken chorioallantoic membrane under hypoxic incubations

Animal Biology ◽

10.1163/15707563-00002540 ◽

2017 ◽

Vol 67 (3-4) ◽

pp. 251-261 ◽

Cited By ~ 1

Author(s):

Hongliang Zhang ◽

Peng Shang ◽

Yawen Zhang ◽

Ying Zhang ◽

Xiaolong Tian ◽

...

Keyword(s):

Embryonic Development ◽

Blood Vessels ◽

Quantitative Pcr ◽

Early Stage ◽

Chorioallantoic Membrane ◽

Developmental Timing ◽

Vascular Density ◽

Hypoxic Conditions ◽

Density Index ◽

Chicken Chorioallantoic Membrane

Under hypoxic conditions, angiogenesis in the chorioallantoic membrane exhibits a sensitive response depending on the developmental timing and intensity and duration of the hypoxia. Furthermore, the effects of hypoxia on vascularization in the chorioallantoic membrane are controversial. In this study, we used microscopy to determine the vascular density index of chicken embryonic chorioallantoic membrane and quantitative PCR to examine the expression of the HIF-1α and VEGFA genes. Two levels of hypoxia (15% and 13% O2) and three durations (days 0–10, 11–18, and 0–18) were applied. The results showed that 13% O2 incubation restrained angiogenesis in the chorioallantoic membrane at an early stage of embryonic development as seen on day 6, but incubation under 13% or 15% O2 efficiently stimulated vascularization of the chorioallantoic membrane as seen on days 16 and 18. Notably, 13% O2 incubation caused visible curling of the blood vessels in the chorioallantoic membrane on day 18. The increased vascular density index under hypoxia was accompanied by an increase in the expression of VEGFA and HIF-1α. Curled blood vessels were observed on day 18 under 13% O2 incubation; however, the curling was not a result of VEGFA overexpression. Thus, the present study helps in elucidating vascularization of the chorioallantoic membrane under hypoxic conditions.

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Transforming growth factor beta 1-induced changes in cell migration, proliferation, and angiogenesis in the chicken chorioallantoic membrane.

The Journal of Cell Biology ◽

10.1083/jcb.111.2.731 ◽

1990 ◽

Vol 111 (2) ◽

pp. 731-741 ◽

Cited By ~ 312

Author(s):

E Y Yang ◽

H L Moses

Keyword(s):

Endothelial Cell ◽

Blood Vessels ◽

Cell Density ◽

Chorioallantoic Membrane ◽

Cell Types ◽

Cellular Responses ◽

Endothelial Cell Proliferation ◽

Tgf Beta ◽

Inhibition Of Proliferation ◽

Chicken Chorioallantoic Membrane

Application of TGF beta 1 (10-100 ng) to the chicken chorioallantoic membrane (CAM) for 72 h resulted in a dose-dependent, gross angiogenic response. The vascular effects induced by TGF beta 1 were qualitatively different than those induced by maximal doses of basic FGF (bFGF) (500 ng). While TGF beta 1 induced the formation of large blood vessels by 72 h, bFGF induced primarily small blood vessels. Histologic analysis revealed that TGF beta 1 stimulated pleiotropic cellular responses in the CAM. Increases in fibroblast and epithelial cell density in the area of TGF beta 1 delivery were observed as early as 4 h after TGF beta 1 treatment. By 8 h, these cell types also demonstrated altered morphology and marked inhibition of proliferation as evidenced by 3H-thymidine labeling. Thus, the TGF beta 1-stimulated accumulation of these cell types was the result of cellular chemotaxis from peripheral areas into the area of TGF beta 1 delivery. Microscopic angiogenesis in the form of capillary sprouts and increased endothelial cell density first became evident at 16 h. By 24 h, capillary cords appeared within the mesenchyme of the CAM, extending towards the point of TGF beta 1 delivery. 3H-thymidine labeling revealed that the growth of these capillary cords was due to endothelial cell proliferation. Finally, perivascular mononuclear inflammation did not become evident until 48 h of treatment, and its presence correlated spatially and temporally with the gross and histological remodelling of newly formed capillary cords into larger blood vessels. In summary, these data suggest that, in the chicken CAM, TGF beta 1 initiates a sequence of cellular responses that results in growth inhibition, cellular accumulation through migration, and microvascular angiogenesis.

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Expression of SPARC during development of the chicken chorioallantoic membrane: evidence for regulated proteolysis in vivo.

Molecular Biology of the Cell ◽

10.1091/mbc.6.3.327 ◽

1995 ◽

Vol 6 (3) ◽

pp. 327-343 ◽

Cited By ~ 69

Author(s):

M L Iruela-Arispe ◽

T F Lane ◽

D Redmond ◽

M Reilly ◽

R P Bolender ◽

...

Keyword(s):

Endothelial Cells ◽

Blood Vessels ◽

Chorioallantoic Membrane ◽

Focal Adhesions ◽

Protein Accumulation ◽

Dependent Manner ◽

Plasmin Activity ◽

Specific Expression ◽

Chicken Chorioallantoic Membrane

SPARC is a secreted glycoprotein that has been shown to disrupt focal adhesions and to regulate the proliferation of endothelial cells in vitro. Moreover, peptides resulting from the proteolysis of SPARC exhibit angiogenic activity. Here we describe the temporal synthesis, turnover, and angiogenic potential of SPARC in the chicken chorioallantoic membrane. Confocal immunofluorescence microscopy revealed specific expression of SPARC protein in endothelial cells, and significantly higher levels of SPARC were observed in smaller newly formed blood vessels in comparison to larger, developmentally older vessels. SPARC mRNA was detected at the earliest stages of chorioallantoic membrane morphogenesis and reached maximal levels at day 13 of embryonic development. Interestingly, steady-state levels of SPARC mRNA did not correlate directly with protein accumulation; moreover, the protein appeared to undergo limited degradation during days 10-15. Incubation of [125I]-SPARC with chorioallantoic membranes of different developmental ages confirmed that extracellular proteolysis occurred during days 9-15, but not at later stages (e.g., days 17-21). Comparison of peptides produced by incubation with chorioallantoic membranes with those generated by plasmin showed an identical pattern of proteolysis. Plasmin activity was present throughout development, and in situ zymography identified sites of plasminogen activator activity that corresponded to areas exhibiting high levels of SPARC expression. Synthetic peptides from a plasmin-sensitive region of SPARC, between amino acids 113-130, stimulated angiogenesis in the chorioallantoic membrane in a dose-dependent manner; in contrast, intact SPARC was inactive in similar assays. We have shown that SPARC is expressed in endothelial cells of newly formed blood vessels in a manner that is both temporally and spatially restricted. Between days 9 and 15 of chorioallantoic membrane development, the protein undergoes proteolytic cleavage that is mediated, in part, by plasmin. SPARC peptides released specifically by plasmin induce angiogenesis in vivo. We therefore propose that SPARC acts as an intrinsic regulator of angiogenesis in vivo.

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An angiogenesis platform using a cubic artificial eggshell with patterned blood vessels on chicken chorioallantoic membrane

PLoS ONE ◽

10.1371/journal.pone.0175595 ◽

2017 ◽

Vol 12 (4) ◽

pp. e0175595 ◽

Cited By ~ 3

Author(s):

Wenjing Huang ◽

Makoto Itayama ◽

Fumihito Arai ◽

Katsuko S. Furukawa ◽

Takashi Ushida ◽

...

Keyword(s):

Blood Vessels ◽

Chorioallantoic Membrane ◽

Chicken Chorioallantoic Membrane

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Antitumor Effects of a Monoclonal Antibody That Binds Anionic Phospholipids on the Surface of Tumor Blood Vessels in Mice

Yearbook of Oncology ◽

10.1016/s1040-1741(08)70262-8 ◽

2006 ◽

Vol 2006 ◽

pp. 378-379

Author(s):

M.S. Gordon

Keyword(s):

Monoclonal Antibody ◽

Blood Vessels ◽

Antitumor Effects ◽

Anionic Phospholipids ◽

Tumor Blood Vessels

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Sustainable Development of Chitosan/Calotropis procera-Based Hydrogels to Stimulate Formation of Granulation Tissue and Angiogenesis in Wound Healing Applications

Molecules ◽

10.3390/molecules26113284 ◽

2021 ◽

Vol 26 (11) ◽

pp. 3284

Author(s):

Muhammad Zahid ◽

Maria Lodhi ◽

Zulfiqar Ahmad Rehan ◽

Hamna Tayyab ◽

Talha Javed ◽

...

Keyword(s):

Wound Healing ◽

Blood Vessels ◽

Granulation Tissue ◽

Chorioallantoic Membrane ◽

Calotropis Procera ◽

Connective Tissues ◽

Blood Capillaries ◽

Chick Chorioallantoic Membrane ◽

Thaw Cycle ◽

Freeze Thaw Cycle

The formation of new scaffolds to enhance healing magnitude is necessarily required in biomedical applications. Granulation tissue formation is a crucial stage of wound healing in which granulation tissue grows on the surface of a wound by the formation of connective tissue and blood vessels. In the present study, porous hydrogels were synthesized using chitosan incorporating latex of the Calotropis procera plant by using a freeze–thaw cycle to stimulate the formation of granulation tissue and angiogenesis in wound healing applications. Structural analysis through Fourier transform infrared (FTIR) spectroscopy confirmed the interaction between chitosan and Calotropis procera. Latex extract containing hydrogel showed slightly higher absorption than the control during water absorption analysis. Thermogravimetric analysis showed high thermal stability of the 60:40 combination of chitosan (CS) and Calotropis procera as compared to all other treatments and controls. A fabricated scaffold application on a chick chorioallantoic membrane (CAM) showed that all hydrogels containing latex extract resulted in a significant formation of blood vessels and regeneration of cells. Overall, the formation of connective tissues and blood capillaries and healing magnitude decreased in ascending order of concentration of extract.

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