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Antiproliferative activity of fucan nanogel.

Dantas-Santos N, Almeida-Lima J, Vidal AA, Gomes DL, Oliveira RM, Santos Pedrosa S, Pereira P, Gama FM, Oliveira Rocha HA - Mar Drugs (2012)

Bottom Line: The resulting modified material (SNFuc) formed nanosized particles.On the other hand, nanogel improved Chinese hamster ovary (CHO) and monocyte macrophage cell (RAW) non-tumor cell line proliferation in the same concentration range.The antiproliferative effect against tumor cells was also confirmed using the BrdU test.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Biotechnology of Natural Polymers (BIOPOL), Departament of Biochemistry, Federal University of Rio Grande do Norte (UFRN), Natal-RN 59078-970, Brazil. nednaldod@hotmail.com

ABSTRACT
Sulfated fucans comprise families of polydisperse natural polysaccharides based on sulfated L-fucose. Our aim was to investigate whether fucan nanogel induces cell-specific responses. To that end, a non toxic fucan extracted from Spatoglossum schröederi was chemically modified by grafting hexadecylamine to the polymer hydrophilic backbone. The resulting modified material (SNFuc) formed nanosized particles. The degree of substitution with hydrophobic chains was close to 100%, as estimated by elemental analysis. SNFfuc in aqueous media had a mean diameter of 123 nm and zeta potential of -38.3 ± 0.74 mV, as measured by dynamic light scattering. Nanoparticles conserved their size for up to 70 days. SNFuc cytotoxicity was determined using the MTT assay after culturing different cell lines for 24 h. Tumor-cell (HepG2, 786, H-S5) proliferation was inhibited by 2.0%-43.7% at nanogel concentrations of 0.05-0.5 mg/mL and rabbit aorta endothelial cells (RAEC) non-tumor cell line proliferation displayed inhibition of 8.0%-22.0%. On the other hand, nanogel improved Chinese hamster ovary (CHO) and monocyte macrophage cell (RAW) non-tumor cell line proliferation in the same concentration range. The antiproliferative effect against tumor cells was also confirmed using the BrdU test. Flow cytometric analysis revealed that the fucan nanogel inhibited 786 cell proliferation through caspase and caspase-independent mechanisms. In addition, SNFuc blocks 786 cell passages in the S and G2-M phases of the cell cycle.

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Effect of SNFuc nanogel on Chinese hamster ovary (CHO) (A); mouse monocyte macrophage (RAW) (B); human renal cell carcinoma (786) (C); human hepatocellular carcinoma (HepG2) (D); rabbit aorta endothelial cells (RAEC) (E) and human marrow stromal cell line (HS-5) (F) cell proliferation, after 24 h of incubation in different concentrations (50, 100 and 500 µg/mL). Results are expressed as percentage of control cells normalized to 100% in the absence of polysaccharide (mean ± SD of seven determinations). Different letters indicate a significant difference between concentrations of individual sulfated polysaccharides (p < 0.05).
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marinedrugs-10-02002-f005: Effect of SNFuc nanogel on Chinese hamster ovary (CHO) (A); mouse monocyte macrophage (RAW) (B); human renal cell carcinoma (786) (C); human hepatocellular carcinoma (HepG2) (D); rabbit aorta endothelial cells (RAEC) (E) and human marrow stromal cell line (HS-5) (F) cell proliferation, after 24 h of incubation in different concentrations (50, 100 and 500 µg/mL). Results are expressed as percentage of control cells normalized to 100% in the absence of polysaccharide (mean ± SD of seven determinations). Different letters indicate a significant difference between concentrations of individual sulfated polysaccharides (p < 0.05).

Mentions: Antiproliferative activity of SNFuc on CHO, RAW, 786, HepG2, RAEC and HS-5 cells was investigated using the colorimetric MTT assay. Proliferation of non-tumor cells CHO and RAW (Figure 4A,B) was stimulated by the presence of SNFuc, after an incubation period of 24 h. On the other hand, SNFuc showed antiproliferative activity when RAEC cells and tumor cells were used (Figure 5). Additionally, our data suggest that cytotoxicity of fucan nanogel was cell-specific, arising from cell-fucan nanogel interaction. In Figure 6 the SNFuc antiproliferative activity against 786, HepG2 and HS-5 cells was further confirmed using BrdU incorporation as described in the Experimental Section. Proliferation of the 786 tumor cell was the most affected one, showing an inhibition rate of approximately 40% in the presence of SNFuc. This cell line was therefore used in additional tests.


Antiproliferative activity of fucan nanogel.

Dantas-Santos N, Almeida-Lima J, Vidal AA, Gomes DL, Oliveira RM, Santos Pedrosa S, Pereira P, Gama FM, Oliveira Rocha HA - Mar Drugs (2012)

Effect of SNFuc nanogel on Chinese hamster ovary (CHO) (A); mouse monocyte macrophage (RAW) (B); human renal cell carcinoma (786) (C); human hepatocellular carcinoma (HepG2) (D); rabbit aorta endothelial cells (RAEC) (E) and human marrow stromal cell line (HS-5) (F) cell proliferation, after 24 h of incubation in different concentrations (50, 100 and 500 µg/mL). Results are expressed as percentage of control cells normalized to 100% in the absence of polysaccharide (mean ± SD of seven determinations). Different letters indicate a significant difference between concentrations of individual sulfated polysaccharides (p < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3475269&req=5

marinedrugs-10-02002-f005: Effect of SNFuc nanogel on Chinese hamster ovary (CHO) (A); mouse monocyte macrophage (RAW) (B); human renal cell carcinoma (786) (C); human hepatocellular carcinoma (HepG2) (D); rabbit aorta endothelial cells (RAEC) (E) and human marrow stromal cell line (HS-5) (F) cell proliferation, after 24 h of incubation in different concentrations (50, 100 and 500 µg/mL). Results are expressed as percentage of control cells normalized to 100% in the absence of polysaccharide (mean ± SD of seven determinations). Different letters indicate a significant difference between concentrations of individual sulfated polysaccharides (p < 0.05).
Mentions: Antiproliferative activity of SNFuc on CHO, RAW, 786, HepG2, RAEC and HS-5 cells was investigated using the colorimetric MTT assay. Proliferation of non-tumor cells CHO and RAW (Figure 4A,B) was stimulated by the presence of SNFuc, after an incubation period of 24 h. On the other hand, SNFuc showed antiproliferative activity when RAEC cells and tumor cells were used (Figure 5). Additionally, our data suggest that cytotoxicity of fucan nanogel was cell-specific, arising from cell-fucan nanogel interaction. In Figure 6 the SNFuc antiproliferative activity against 786, HepG2 and HS-5 cells was further confirmed using BrdU incorporation as described in the Experimental Section. Proliferation of the 786 tumor cell was the most affected one, showing an inhibition rate of approximately 40% in the presence of SNFuc. This cell line was therefore used in additional tests.

Bottom Line: The resulting modified material (SNFuc) formed nanosized particles.On the other hand, nanogel improved Chinese hamster ovary (CHO) and monocyte macrophage cell (RAW) non-tumor cell line proliferation in the same concentration range.The antiproliferative effect against tumor cells was also confirmed using the BrdU test.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Biotechnology of Natural Polymers (BIOPOL), Departament of Biochemistry, Federal University of Rio Grande do Norte (UFRN), Natal-RN 59078-970, Brazil. nednaldod@hotmail.com

ABSTRACT
Sulfated fucans comprise families of polydisperse natural polysaccharides based on sulfated L-fucose. Our aim was to investigate whether fucan nanogel induces cell-specific responses. To that end, a non toxic fucan extracted from Spatoglossum schröederi was chemically modified by grafting hexadecylamine to the polymer hydrophilic backbone. The resulting modified material (SNFuc) formed nanosized particles. The degree of substitution with hydrophobic chains was close to 100%, as estimated by elemental analysis. SNFfuc in aqueous media had a mean diameter of 123 nm and zeta potential of -38.3 ± 0.74 mV, as measured by dynamic light scattering. Nanoparticles conserved their size for up to 70 days. SNFuc cytotoxicity was determined using the MTT assay after culturing different cell lines for 24 h. Tumor-cell (HepG2, 786, H-S5) proliferation was inhibited by 2.0%-43.7% at nanogel concentrations of 0.05-0.5 mg/mL and rabbit aorta endothelial cells (RAEC) non-tumor cell line proliferation displayed inhibition of 8.0%-22.0%. On the other hand, nanogel improved Chinese hamster ovary (CHO) and monocyte macrophage cell (RAW) non-tumor cell line proliferation in the same concentration range. The antiproliferative effect against tumor cells was also confirmed using the BrdU test. Flow cytometric analysis revealed that the fucan nanogel inhibited 786 cell proliferation through caspase and caspase-independent mechanisms. In addition, SNFuc blocks 786 cell passages in the S and G2-M phases of the cell cycle.

Show MeSH
Related in: MedlinePlus