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. 2018 Oct 4;8(4):87.
doi: 10.3390/bios8040087.

Functionalized Gold Nanoparticles as Biosensors for Monitoring Cellular Uptake and Localization in Normal and Tumor Prostatic Cells

Marianna Pannico  1 , Anna Calarco  2 , Gianfranco Peluso  3 , Pellegrino Musto  4
Affiliations

Functionalized Gold Nanoparticles as Biosensors for Monitoring Cellular Uptake and Localization in Normal and Tumor Prostatic Cells

Marianna Pannico et al. Biosensors (Basel). .

Abstract

In the present contribution the fabrication and characterization of functionalized gold nanospheres of uniform shape and controlled size is reported. These nano-objects are intended to be used as Surface Enhanced Raman Spectroscopy (SERS) sensors for in-vitro cellular uptake and localization. Thiophenol was used as molecular reporter and was bound to the Au surface by a chemisorption process in aqueous solution. The obtained colloidal solution was highly stable and no aggregation of the single nanospheres into larger clusters was observed. The nanoparticles were incubated in human prostatic cells with the aim of developing a robust, SERS-based method to differentiate normal and tumor cell lines. SERS imaging experiments showed that tumor cells uptake considerably larger amounts of nanoparticles in comparison to normal cells (up to 950% more); significant differences were also observed in the uptake kinetics. This largely different behaviour might be exploited in diagnostic and therapeutic applications.

Keywords: SERS; biosensing; nanomaterials; prostate cancer; single cell spectroscopy.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Typical Transmission Electron Microscopy (TEM) micrograph of gold nanoparticles. The insets are the high-magnification TEM of the indicated gold nanoparticles (AuNPs).
Figure 2
Figure 2
Size distribution of AuNPs.
Figure 3
Figure 3
Concentration-normalized Absorbance of AuNPs (blue circles) and AuNPs/TP (red circles) as a function of ageing time.
Figure 4
Figure 4
Blue trace: Raman spectrum of TP (liquid); Red trace: SERS spectrum of the AuNPs/TP colloid (30 μM in TP). The inset represents the SERS spectrum of AuNPs/TP colloid averaged over 20 points (red trace) ± the standard deviation (blue traces). The black trace in the inset represents the SERS spectrum of the native AuNPs.
Figure 5
Figure 5
Cytotoxicity evaluation of AuNPs assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays in PNT2 (A,C) and PC3 (B,D) cells. Cells were incubated with increasing concentrations of Au from 0.1 to 1000 μM (corresponding to 0–2.3 nM in nanoparticles) for 6, 24, 48, and 72 h. In each experiment, four replicates per concentration were tested. The experiment was repeated three times. The data are mean values of three independent experiments (± SEM).
Figure 6
Figure 6
(A) SERS spectrum collected at the point of maximum SERS activity (red spot in the Raman image). (B) SERS intensity map of a PNT2 cell incubated for 2 h with AuNPs/TP. (C) superposition of the visible and SERS images.
Figure 7
Figure 7
(A) SERS intensity map of a PNT2 cell incubated for 6 h with AuNPs/TP. (B) superposition of the visible and SERS images.
Figure 8
Figure 8
(A) SERS intensity map of a PC3 cell incubated for 2 h with AuNPs/TP. (B) superposition of the visible and SERS images.
Figure 9
Figure 9
(A) SERS spectrum collected at the point of maximum SERS activity (bright yellow point in the Raman image). (B) SERS intensity map of a PC3 cell incubated for 6 h with AuNPs/TP. (C) superposition of the visible and SERS images.
Figure 10
Figure 10
I1078 vs. incubation time for PNT2 and PC3 cells.
See this image and copyright information in PMC

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