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Investigation of cellular responses upon interaction with silver nanoparticles.

Subbiah R, Jeon SB, Park K, Ahn SJ, Yun K - Int J Nanomedicine (2015)

Bottom Line: When compared with kanamycin, AgNPs exhibited moderate antibacterial activity.The degree to which cellular morphology was altered directly proportional to the level of AgNP-induced cytotoxicity.Together, these data suggest that atomic force microscopy can be used as a potential tool to develop a biomechanics-based biomarker for the evaluation of NP-dependent cytotoxicity and cytopathology.

View Article: PubMed Central - PubMed

Affiliation: Center for Biomaterials, Korea Institute of Science and Technology, Seoul, Republic of Korea ; Department of Biomedical Engineering, Korea University of Science and Technology, Daejon, Republic of Korea.

ABSTRACT
In order for nanoparticles (NPs) to be applied in the biomedical field, a thorough investigation of their interactions with biological systems is required. Although this is a growing area of research, there is a paucity of comprehensive data in cell-based studies. To address this, we analyzed the physicomechanical responses of human alveolar epithelial cells (A549), mouse fibroblasts (NIH3T3), and human bone marrow stromal cells (HS-5), following their interaction with silver nanoparticles (AgNPs). When compared with kanamycin, AgNPs exhibited moderate antibacterial activity. Cell viability ranged from ≤ 80% at a high AgNPs dose (40 µg/mL) to >95% at a low dose (10 µg/mL). We also used atomic force microscopy-coupled force spectroscopy to evaluate the biophysical and biomechanical properties of cells. This revealed that AgNPs treatment increased the surface roughness (P<0.001) and stiffness (P<0.001) of cells. Certain cellular changes are likely due to interaction of the AgNPs with the cell surface. The degree to which cellular morphology was altered directly proportional to the level of AgNP-induced cytotoxicity. Together, these data suggest that atomic force microscopy can be used as a potential tool to develop a biomechanics-based biomarker for the evaluation of NP-dependent cytotoxicity and cytopathology.

No MeSH data available.


Related in: MedlinePlus

AgNPs characterization.Notes: (A) AFM, (B) HR-TEM, and (C) FE-SEM images of AgNPs; both low- and high-magnification images are shown in upper and lower panels, respectively, (D) UV-vis spectra, (E) RMS roughness graph, (F) XRD, (G) size distribution, and (H) elemental analysis of AgNPs. Table summarizes the RMS roughness, ζ potential, adhesion force, and stiffness of the AgNPs. Data are presented as mean ± standard deviation.Abbreviations: AFM, atomic force microscopy; AgNPs, silver nanoparticles; EDXA, energy dispersive X-ray analysis; FE-SEM, field emission scanning electron microscopy; HR-TEM, high-resolution transmission electron microscopy; RMS, root mean square; UV-vis, UV-visible; XRD, X-ray powder diffraction.
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f1-ijn-10-191: AgNPs characterization.Notes: (A) AFM, (B) HR-TEM, and (C) FE-SEM images of AgNPs; both low- and high-magnification images are shown in upper and lower panels, respectively, (D) UV-vis spectra, (E) RMS roughness graph, (F) XRD, (G) size distribution, and (H) elemental analysis of AgNPs. Table summarizes the RMS roughness, ζ potential, adhesion force, and stiffness of the AgNPs. Data are presented as mean ± standard deviation.Abbreviations: AFM, atomic force microscopy; AgNPs, silver nanoparticles; EDXA, energy dispersive X-ray analysis; FE-SEM, field emission scanning electron microscopy; HR-TEM, high-resolution transmission electron microscopy; RMS, root mean square; UV-vis, UV-visible; XRD, X-ray powder diffraction.

Mentions: AgNPs were synthesized via a reduction reaction using a syringe pump-assisted ultrasonication method. Ultrasonics minimized the ionic concentration gradient and enhanced mass transport at the solid sphere–liquid interface, thereby producing homogeneous sphere-shaped NPs. The morphology of the newly synthesized AgNPs was characterized. Figure 1A–C shows the AFM, high-resolution transmission electron microscopy, and FE-SEM images of the sphere-shaped AgNPs, respectively. The corresponding high magnification images are also given in lower panel of Figure 1A–C. All imaging modalities yielded highly similar results. The surface roughness of the AgNPs was moderately smooth, with a root mean square (RMS) roughness of 3.44 nm (Figure 1E). The average size of the AgNPs was 19.5±7.7 nm; the size distribution is shown in Figure 1G and indicates near-homogeneity NP preparations with negligible aggregation. The surface charge (ζ potential) of the AgNPs in the culture media was −18±0.6 mV. Structural confirmation was analyzed using UV spectroscopy, EDX, and XRD. AgNPs were associated with strong surface plasmon resonance absorption curves between 380 nm and 440 nm (Figure 1D). In addition, the XRD (Figure 1F) patterns of AgNPs showed relatively sharp peaks at ~37.68°, ~44.02°, ~64.24°, and ~81.46°, induced by crystalline planes of silver that were assigned as (111), (200), (220), and (222), respectively. Elemental analysis revealed a 99.27% silver content, indicating an impurity-free AgNPs synthesis (Figure 1H). The AFM-FS study yielded an AdF of 2.1±0.3 nN and stiffness of 46.7±5.8 MPa for AgNPs.


Investigation of cellular responses upon interaction with silver nanoparticles.

Subbiah R, Jeon SB, Park K, Ahn SJ, Yun K - Int J Nanomedicine (2015)

AgNPs characterization.Notes: (A) AFM, (B) HR-TEM, and (C) FE-SEM images of AgNPs; both low- and high-magnification images are shown in upper and lower panels, respectively, (D) UV-vis spectra, (E) RMS roughness graph, (F) XRD, (G) size distribution, and (H) elemental analysis of AgNPs. Table summarizes the RMS roughness, ζ potential, adhesion force, and stiffness of the AgNPs. Data are presented as mean ± standard deviation.Abbreviations: AFM, atomic force microscopy; AgNPs, silver nanoparticles; EDXA, energy dispersive X-ray analysis; FE-SEM, field emission scanning electron microscopy; HR-TEM, high-resolution transmission electron microscopy; RMS, root mean square; UV-vis, UV-visible; XRD, X-ray powder diffraction.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4556294&req=5

f1-ijn-10-191: AgNPs characterization.Notes: (A) AFM, (B) HR-TEM, and (C) FE-SEM images of AgNPs; both low- and high-magnification images are shown in upper and lower panels, respectively, (D) UV-vis spectra, (E) RMS roughness graph, (F) XRD, (G) size distribution, and (H) elemental analysis of AgNPs. Table summarizes the RMS roughness, ζ potential, adhesion force, and stiffness of the AgNPs. Data are presented as mean ± standard deviation.Abbreviations: AFM, atomic force microscopy; AgNPs, silver nanoparticles; EDXA, energy dispersive X-ray analysis; FE-SEM, field emission scanning electron microscopy; HR-TEM, high-resolution transmission electron microscopy; RMS, root mean square; UV-vis, UV-visible; XRD, X-ray powder diffraction.
Mentions: AgNPs were synthesized via a reduction reaction using a syringe pump-assisted ultrasonication method. Ultrasonics minimized the ionic concentration gradient and enhanced mass transport at the solid sphere–liquid interface, thereby producing homogeneous sphere-shaped NPs. The morphology of the newly synthesized AgNPs was characterized. Figure 1A–C shows the AFM, high-resolution transmission electron microscopy, and FE-SEM images of the sphere-shaped AgNPs, respectively. The corresponding high magnification images are also given in lower panel of Figure 1A–C. All imaging modalities yielded highly similar results. The surface roughness of the AgNPs was moderately smooth, with a root mean square (RMS) roughness of 3.44 nm (Figure 1E). The average size of the AgNPs was 19.5±7.7 nm; the size distribution is shown in Figure 1G and indicates near-homogeneity NP preparations with negligible aggregation. The surface charge (ζ potential) of the AgNPs in the culture media was −18±0.6 mV. Structural confirmation was analyzed using UV spectroscopy, EDX, and XRD. AgNPs were associated with strong surface plasmon resonance absorption curves between 380 nm and 440 nm (Figure 1D). In addition, the XRD (Figure 1F) patterns of AgNPs showed relatively sharp peaks at ~37.68°, ~44.02°, ~64.24°, and ~81.46°, induced by crystalline planes of silver that were assigned as (111), (200), (220), and (222), respectively. Elemental analysis revealed a 99.27% silver content, indicating an impurity-free AgNPs synthesis (Figure 1H). The AFM-FS study yielded an AdF of 2.1±0.3 nN and stiffness of 46.7±5.8 MPa for AgNPs.

Bottom Line: When compared with kanamycin, AgNPs exhibited moderate antibacterial activity.The degree to which cellular morphology was altered directly proportional to the level of AgNP-induced cytotoxicity.Together, these data suggest that atomic force microscopy can be used as a potential tool to develop a biomechanics-based biomarker for the evaluation of NP-dependent cytotoxicity and cytopathology.

View Article: PubMed Central - PubMed

Affiliation: Center for Biomaterials, Korea Institute of Science and Technology, Seoul, Republic of Korea ; Department of Biomedical Engineering, Korea University of Science and Technology, Daejon, Republic of Korea.

ABSTRACT
In order for nanoparticles (NPs) to be applied in the biomedical field, a thorough investigation of their interactions with biological systems is required. Although this is a growing area of research, there is a paucity of comprehensive data in cell-based studies. To address this, we analyzed the physicomechanical responses of human alveolar epithelial cells (A549), mouse fibroblasts (NIH3T3), and human bone marrow stromal cells (HS-5), following their interaction with silver nanoparticles (AgNPs). When compared with kanamycin, AgNPs exhibited moderate antibacterial activity. Cell viability ranged from ≤ 80% at a high AgNPs dose (40 µg/mL) to >95% at a low dose (10 µg/mL). We also used atomic force microscopy-coupled force spectroscopy to evaluate the biophysical and biomechanical properties of cells. This revealed that AgNPs treatment increased the surface roughness (P<0.001) and stiffness (P<0.001) of cells. Certain cellular changes are likely due to interaction of the AgNPs with the cell surface. The degree to which cellular morphology was altered directly proportional to the level of AgNP-induced cytotoxicity. Together, these data suggest that atomic force microscopy can be used as a potential tool to develop a biomechanics-based biomarker for the evaluation of NP-dependent cytotoxicity and cytopathology.

No MeSH data available.


Related in: MedlinePlus