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J. L. Dreyfuss, T. F. Gesteira, G. L. A. Cunha, V. J. Coulson-Thomas, C. V. Regatieri, H. B. Nader; Acharan Sulfate: A Heparin-Like Compound With Anti-Angiogenic Effect. A Potential Drug to Treat Eye Neovascular Diseases. Invest. Ophthalmol. Vis. Sci. 2010;51(13):60.
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The heparin-like glycosaminoglycan obtained from the giant African snail Achatina fulica is capable to modulate inflammatory responses without interfering on hemostasis. Thus the objective of this study was to evaluate the anti-angiogenic effects of this new compound on "in vitro" models.
Acharan sulfate was purified and analyzed by Nuclear Magnetic Ressonance and electrospray ionization mass spectrometry (ESI-MS). The ability of endothelial cells form capillary-like structures "in vitro" when plated on top of a reconstituted basement membrane extracellular matrix (Matrigel) was investigated in cells treated with acharan sulfate (0.01, 0.1 and 1 mg/mL). The cytotoxicity of this compound was evaluated by MTT test in retinal pigmented epithelial cells (ARPE-19) and endothelial cell cultures.
Electrospray ionization mass spectrometry (ESI-MS) analysis was used to determine the molecular weight of acharan sulfate-derived oligosaccharide. A doubly charged negative ion at m/z 481.1 was also observed for the tetrasaccharide. 1H NMR spectra spectra were obtained and the signal at 3.27 ppm indicates the presence of glucosamine N-sulfated which is an unusual variation of previously purifed acharan sulfate. Only the 1 mg/mL of acharan sulfate induced a significant decrease (p<0.05) in total length of tubes formed by endothelial cells in Matrigel. No cytotoxic effect was detected in ARPE-19 or endothelial cell culture in all tested doses.
The acharan sulfate was resolved and the in vitro studies showed that the optimal dosage to reduce angiogenesis in vitro was 1 mg/mL and it was no cytotoxic effect in ARPE-19 cells or endothelial cells. These findings suggest that the inhibitory effect of acharan sulphate on angiogenesis and the non-cytotoxic effect on ARPE-19 cells indicates acharan sulfate as a potential compound that can be used in the future as a therapy to control neovascularization
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