Treatment of Diseases and Conditions Caused by Increased Vascular Permeability
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example 1
In Vitro Vascular Permeability Assays
[0095]Objective: TNF-α has been shown to increase endothelial monolayer permeability (Mark, K. S., et al., Life Sciences, 1999, 64(21):1941-1953)In order to test whether ACCS has the ability to affect vascular permeability, initial experiments were performed to determine whether ACCS could reduce the level of permeability of endothelial cells exposed to TNF-α.
[0096]Method: The ability of ACCS to modulate vascular permeability was evaluated using an In Vitro Vascular Permeability Assay (Millipore, Cat. No. ECM640). In this assay, Human vascular endothelial cells (HUVEC) were seeded onto collagen or fibrin-coated semi-permeable membrane inserts and a monolayer of cells was formed which occluded the membrane pores. The inserts were then placed in a receiver well. The cell monolayer can be treated with cytokines, growth factors, or other compounds of interest. In this experiment, the cells were treated with TNF-α and a high molecular weight FITC-labe...
example 2
Evaluate Whether ACCS Can Modulate Increased Vascular Permeability as a Result of Irradiation
[0098]Objective: Radiation is known to increase vascular permeability. Therefore, a second set of experiments was conducted to determine whether ACCS could modulate increased vascular permeability as a result of irradiation from a 5 Gy cesium-137 source.
[0099]Method: HUVECs were exposed to a radiation dose of 5 Gy prior to treatment with ACCS, control media, or endothelial growth media control.
[0100]Results: As shown in Table 3 below, ACCS Lot A-treated cells showed reduced FITC-Dextran fluorescence compared to endothelial growth media control, and control media. These results demonstrate that ACCS is modulating and therefore reducing vascular permeability. Table 4 shows that radiation exposure to the cells induced an increase in vascular permeability which was decreased by ACCS Lot A and Lot B compared to endothelial growth media control, and control media.
TABLE 3Endothelial GrowthMedia Con...
example 3
The Effect of ACCS on Reduction of Vascular Permeability in a Setting Wherein Radiation is Combined with TNF-α
[0101]Objective: Increased vascular permeability due to radiation may result from many stimuli in vivo. Radiation combined with inflammatory molecules may better simulate multiple inflammatory causes of permeability in vivo. To further evaluate the effect of ACCS on reduction of vascular permeability, HUVECs were exposed to both radiation and TNF-α, in various media.
[0102]Method: HUVECs were exposed to 5 Gy radiation and 50 ng / mL TNF-α for 4 hours.
[0103]Results: Both ACCS Lot A and Lot B showed reduced permeability of endothelial cells that were exposed to both 5 Gy radiation and 50 ng / mL TNF-α as compared to endothelial growth media control and control media.
TABLE 5ACCSACCSEndothelialControlLot ALot BGrowthMedia(fluor-(fluor-Media Control(fluor-escence)escence)(fluorescence)escence)5 Gy + 50 ng / mL8.49.325.921.8TNF-α
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