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Size-dependent cellular toxicity and uptake of commercial colloidal gold nanoparticles in DU-145 cells.

Vedantam P, Huang G, Tzeng TR - Cancer Nanotechnol (2013)

Bottom Line: Plain 20 nm GNPs decrease the percentage of viable cells in 48 and 72 h in log and lag phase of DU-145 cells.It was also observed that the Mn-GNPs were taken up by the DU-145 cells significantly more than the plain GNPs.Protein corona was observed when GNPs were incubated with fetal bovine serum which was confirmed by dynamic light scattering measurements and SDS-PAGE gel.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Clemson University, Clemson, SC 29634 USA.

ABSTRACT

Urinary tract infection (UTI) is a predominant condition in prostate cancer patients. Escherichia coli ORN178 (EC-178) is the uropathogen that causes recurrent infection by binding specifically to adhesins of prostate cancer cells (DU-145 cells). Gold nanoparticles (GNPs) have been used in biodiagnosis of pathogens. In this study, we have investigated the binding time of EC-178 to DU-145 cells, the cytotoxicity and uptake of plain and mannose functionalized and 20 and 200 nm GNPs (d-mannan (Mn)-GNPs). We also investigated the protein corona of GNPs when incubated with fetal bovine serum to study the protein corona which decides the biological fate of the GNPs. It was seen that EC-178 binds and is inside the DU-145 cells by 3 h of incubation period. Plain 20 nm GNPs decrease the percentage of viable cells in 48 and 72 h in log and lag phase of DU-145 cells. It was also observed that the Mn-GNPs were taken up by the DU-145 cells significantly more than the plain GNPs. Protein corona was observed when GNPs were incubated with fetal bovine serum which was confirmed by dynamic light scattering measurements and SDS-PAGE gel.

No MeSH data available.


Related in: MedlinePlus

Coomassie-stained SDS-PAGE gel lanes for protein coated 20 and 200 nm GNPs. FBS is a combination of various proteins. The lanes “20 nm serum and 200 nm serum” show a band for protein bound to the GNPs suggesting the presence of protein corona. The samples incubated with plain DMEM did not show any protein bands (data not shown)
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Fig8: Coomassie-stained SDS-PAGE gel lanes for protein coated 20 and 200 nm GNPs. FBS is a combination of various proteins. The lanes “20 nm serum and 200 nm serum” show a band for protein bound to the GNPs suggesting the presence of protein corona. The samples incubated with plain DMEM did not show any protein bands (data not shown)

Mentions: The hydrodynamic sizes of the plain and protein coated GNPs did increase when incubated with FBS (Fig. 7). The DLS measurements indicate that the protein in FBS did adsorb and hence the increase in diameter. The 20 nm GNPs were found to be of 66.61 ± 2.3 nm size and the 200 nm showed increase in size were of 261.9 ± 5.3 nm. This confirms the presence of protein corona which remained coated even after multiple washes. The zeta values do show good stable protein coated 20 and 200 nm GNPs (Table 1). The SDS-PAGE gel was done to confirm the same (Fig. 8). The 20 and 200 nm GNPs (arrow marks) do confirm the presence of protein corona with a clear thick band. The samples incubated with plain DMEM did not show any bands at all. Hence, it is clear that there was protein bound to the GNPs.Fig. 7


Size-dependent cellular toxicity and uptake of commercial colloidal gold nanoparticles in DU-145 cells.

Vedantam P, Huang G, Tzeng TR - Cancer Nanotechnol (2013)

Coomassie-stained SDS-PAGE gel lanes for protein coated 20 and 200 nm GNPs. FBS is a combination of various proteins. The lanes “20 nm serum and 200 nm serum” show a band for protein bound to the GNPs suggesting the presence of protein corona. The samples incubated with plain DMEM did not show any protein bands (data not shown)
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4544071&req=5

Fig8: Coomassie-stained SDS-PAGE gel lanes for protein coated 20 and 200 nm GNPs. FBS is a combination of various proteins. The lanes “20 nm serum and 200 nm serum” show a band for protein bound to the GNPs suggesting the presence of protein corona. The samples incubated with plain DMEM did not show any protein bands (data not shown)
Mentions: The hydrodynamic sizes of the plain and protein coated GNPs did increase when incubated with FBS (Fig. 7). The DLS measurements indicate that the protein in FBS did adsorb and hence the increase in diameter. The 20 nm GNPs were found to be of 66.61 ± 2.3 nm size and the 200 nm showed increase in size were of 261.9 ± 5.3 nm. This confirms the presence of protein corona which remained coated even after multiple washes. The zeta values do show good stable protein coated 20 and 200 nm GNPs (Table 1). The SDS-PAGE gel was done to confirm the same (Fig. 8). The 20 and 200 nm GNPs (arrow marks) do confirm the presence of protein corona with a clear thick band. The samples incubated with plain DMEM did not show any bands at all. Hence, it is clear that there was protein bound to the GNPs.Fig. 7

Bottom Line: Plain 20 nm GNPs decrease the percentage of viable cells in 48 and 72 h in log and lag phase of DU-145 cells.It was also observed that the Mn-GNPs were taken up by the DU-145 cells significantly more than the plain GNPs.Protein corona was observed when GNPs were incubated with fetal bovine serum which was confirmed by dynamic light scattering measurements and SDS-PAGE gel.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Clemson University, Clemson, SC 29634 USA.

ABSTRACT

Urinary tract infection (UTI) is a predominant condition in prostate cancer patients. Escherichia coli ORN178 (EC-178) is the uropathogen that causes recurrent infection by binding specifically to adhesins of prostate cancer cells (DU-145 cells). Gold nanoparticles (GNPs) have been used in biodiagnosis of pathogens. In this study, we have investigated the binding time of EC-178 to DU-145 cells, the cytotoxicity and uptake of plain and mannose functionalized and 20 and 200 nm GNPs (d-mannan (Mn)-GNPs). We also investigated the protein corona of GNPs when incubated with fetal bovine serum to study the protein corona which decides the biological fate of the GNPs. It was seen that EC-178 binds and is inside the DU-145 cells by 3 h of incubation period. Plain 20 nm GNPs decrease the percentage of viable cells in 48 and 72 h in log and lag phase of DU-145 cells. It was also observed that the Mn-GNPs were taken up by the DU-145 cells significantly more than the plain GNPs. Protein corona was observed when GNPs were incubated with fetal bovine serum which was confirmed by dynamic light scattering measurements and SDS-PAGE gel.

No MeSH data available.


Related in: MedlinePlus