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Peptide-biphenyl hybrid-capped AuNPs: stability and biocompatibility under cell culture conditions.

Connolly M, Pérez Y, Mann E, Herradón B, Fernández-Cruz ML, Navas JM - Nanoscale Res Lett (2013)

Bottom Line: Moreover, these AuNPs have unique biological effects on Hep G2 cells while showing low toxicity.The production of ROS along with supporting optical microscopy images suggests cellular interaction/uptake of these particular AuNPs.Future research efforts should further test this hypothesis, as such interaction/uptake is highly relevant in drug delivery systems.

View Article: PubMed Central - HTML - PubMed

Affiliation: Departamento de Medio Ambiente, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Carretera de la Coruña Km 7,5, Madrid 28040, Spain. yolanda.cortes@urjc.es.

ABSTRACT
In this study, we explored the biocompatibility of Au nanoparticles (NPs) capped with peptide-biphenyl hybrid (PBH) ligands containing glycine (Gly), cysteine (Cys), tyrosine (Tyr), tryptophan (Trp) and methionine (Met) amino acids in the human hepatocellular carcinoma cell line Hep G2. Five AuNPs, Au[(Gly-Tyr-Met)2B], Au[(Gly-Trp-Met)2B], Au[(Met)2B], Au[(Gly-Tyr-TrCys)2B] and Au[(TrCys)2B], were synthesised. Physico-chemical and cytotoxic properties were thoroughly studied. Transmission electron micrographs showed isolated near-spherical nanoparticles with diameters of 1.5, 1.6, 2.3, 1.8 and 2.3 nm, respectively. Dynamic light scattering evidenced the high stability of suspensions in Milli-Q water and culture medium, particularly when supplemented with serum, showing in all cases a tendency to form agglomerates with diameters approximately 200 nm. In the cytotoxicity studies, interference caused by AuNPs with some typical cytotoxicity assays was demonstrated; thus, only data obtained from the resazurin based assay were used. After 48-h incubation, only concentrations ≥50 μg/ml exhibited cytotoxicity. Such doses were also responsible for an increase in reactive oxygen species (ROS). Some differences were observed among the studied NPs. Of particular importance is the AuNPs capped with the PBH ligand (Gly-Tyr-TrCys)2B showing remarkable stability in culture medium, even in the absence of serum. Moreover, these AuNPs have unique biological effects on Hep G2 cells while showing low toxicity. The production of ROS along with supporting optical microscopy images suggests cellular interaction/uptake of these particular AuNPs. Future research efforts should further test this hypothesis, as such interaction/uptake is highly relevant in drug delivery systems.

No MeSH data available.


Related in: MedlinePlus

Peptide-biphenyl hybrid (PBH) ligands used in this study, Tr = Trityl, B = 2, 2’-(bis)carbonylbiphenyl.
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Figure 1: Peptide-biphenyl hybrid (PBH) ligands used in this study, Tr = Trityl, B = 2, 2’-(bis)carbonylbiphenyl.

Mentions: Five AuNPs, (Au[(Gly-Trp-Met)2B], Au[(Gly-Tyr-TrCys)2B], Au[(Gly-Tyr-Met)2B], Au[(Met)2B] and Au[(TrCys)2B]) (Figure 1), were synthesised following the methodology described by Pérez et al. [9] (see Additional file 1). Thus, each PBH (50 μmol) was dissolved in ethanol (20 ml, 2.5 mmol/l) and was added to a solution of HAuCl4 (50 ml, 0.5 mmol/l) in 2-propanol under stirring. After 30 min, a freshly prepared aqueous solution of NaBH4 (4 ml, 50 mmol/l) was added slowly. The mixture was stirred for 2 h at room temperature to afford a red-brown colloidal gold solution. The AuNPs were precipitated by centrifugation for 15 min at 6,000 rpm. The black-brown precipitate was washed with 2-propanol to remove the free ligand and then dried under vacuum. The PBH-capped AuNPs obtained were stable to some cycles of precipitation and re-dispersion and could be easily dispersed in water.


Peptide-biphenyl hybrid-capped AuNPs: stability and biocompatibility under cell culture conditions.

Connolly M, Pérez Y, Mann E, Herradón B, Fernández-Cruz ML, Navas JM - Nanoscale Res Lett (2013)

Peptide-biphenyl hybrid (PBH) ligands used in this study, Tr = Trityl, B = 2, 2’-(bis)carbonylbiphenyl.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3716793&req=5

Figure 1: Peptide-biphenyl hybrid (PBH) ligands used in this study, Tr = Trityl, B = 2, 2’-(bis)carbonylbiphenyl.
Mentions: Five AuNPs, (Au[(Gly-Trp-Met)2B], Au[(Gly-Tyr-TrCys)2B], Au[(Gly-Tyr-Met)2B], Au[(Met)2B] and Au[(TrCys)2B]) (Figure 1), were synthesised following the methodology described by Pérez et al. [9] (see Additional file 1). Thus, each PBH (50 μmol) was dissolved in ethanol (20 ml, 2.5 mmol/l) and was added to a solution of HAuCl4 (50 ml, 0.5 mmol/l) in 2-propanol under stirring. After 30 min, a freshly prepared aqueous solution of NaBH4 (4 ml, 50 mmol/l) was added slowly. The mixture was stirred for 2 h at room temperature to afford a red-brown colloidal gold solution. The AuNPs were precipitated by centrifugation for 15 min at 6,000 rpm. The black-brown precipitate was washed with 2-propanol to remove the free ligand and then dried under vacuum. The PBH-capped AuNPs obtained were stable to some cycles of precipitation and re-dispersion and could be easily dispersed in water.

Bottom Line: Moreover, these AuNPs have unique biological effects on Hep G2 cells while showing low toxicity.The production of ROS along with supporting optical microscopy images suggests cellular interaction/uptake of these particular AuNPs.Future research efforts should further test this hypothesis, as such interaction/uptake is highly relevant in drug delivery systems.

View Article: PubMed Central - HTML - PubMed

Affiliation: Departamento de Medio Ambiente, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Carretera de la Coruña Km 7,5, Madrid 28040, Spain. yolanda.cortes@urjc.es.

ABSTRACT
In this study, we explored the biocompatibility of Au nanoparticles (NPs) capped with peptide-biphenyl hybrid (PBH) ligands containing glycine (Gly), cysteine (Cys), tyrosine (Tyr), tryptophan (Trp) and methionine (Met) amino acids in the human hepatocellular carcinoma cell line Hep G2. Five AuNPs, Au[(Gly-Tyr-Met)2B], Au[(Gly-Trp-Met)2B], Au[(Met)2B], Au[(Gly-Tyr-TrCys)2B] and Au[(TrCys)2B], were synthesised. Physico-chemical and cytotoxic properties were thoroughly studied. Transmission electron micrographs showed isolated near-spherical nanoparticles with diameters of 1.5, 1.6, 2.3, 1.8 and 2.3 nm, respectively. Dynamic light scattering evidenced the high stability of suspensions in Milli-Q water and culture medium, particularly when supplemented with serum, showing in all cases a tendency to form agglomerates with diameters approximately 200 nm. In the cytotoxicity studies, interference caused by AuNPs with some typical cytotoxicity assays was demonstrated; thus, only data obtained from the resazurin based assay were used. After 48-h incubation, only concentrations ≥50 μg/ml exhibited cytotoxicity. Such doses were also responsible for an increase in reactive oxygen species (ROS). Some differences were observed among the studied NPs. Of particular importance is the AuNPs capped with the PBH ligand (Gly-Tyr-TrCys)2B showing remarkable stability in culture medium, even in the absence of serum. Moreover, these AuNPs have unique biological effects on Hep G2 cells while showing low toxicity. The production of ROS along with supporting optical microscopy images suggests cellular interaction/uptake of these particular AuNPs. Future research efforts should further test this hypothesis, as such interaction/uptake is highly relevant in drug delivery systems.

No MeSH data available.


Related in: MedlinePlus