Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 May 8;46(5):4489-4505.
doi: 10.3390/cimb46050272.

A Novel Approach Using Reduced Graphene Oxide for the Detection of ALP and RUNX2 Osteogenic Biomarkers

Elena Alina Chiticaru  1 , Mariana Ioniță  1   2
Affiliations

A Novel Approach Using Reduced Graphene Oxide for the Detection of ALP and RUNX2 Osteogenic Biomarkers

Elena Alina Chiticaru et al. Curr Issues Mol Biol. .

Abstract

In this work, we propose a new technique involving the modification of commercial screen-printed carbon electrodes with electrochemically reduced graphene oxide to serve as the starting point of a future electrochemical biosensor for the detection of two osteogenic biomarkers: alkaline phosphatase (ALP) and Runt-related transcription factor 2 (RUNX2). The electrodes were characterized after each modification by cyclic voltammetry and electrochemical impedance spectroscopy, showing the appropriate electrochemical characteristics for each modification type. The results obtained from scanning electron microscopy, Raman spectroscopy, X-ray photoelectron spectroscopy, and contact angle measurements are well correlated with each other, demonstrating the successful modification of the electrodes with graphene oxide and its subsequent reduction. The bioreceptors were immobilized on the electrodes by physical adsorption, which was confirmed by electrochemical methods, structural characterization, and contact angle measurements. Finally, the functionalized electrodes were incubated with the specific target analytes and the detection relied on monitoring the electrochemical changes occurring after the hybridization process. Our results indicated that the pilot platform has the ability to detect the two biomarkers up to 1 nM, with increased sensitivity observed for RUNX2, suggesting that after further optimizations, it has a high potential to be employed as a future biosensor.

Keywords: ALP; RUNX2; electrochemistry; electrode modification; osteogenic biomarkers; reduced graphene oxide.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Figures

Figure 1
Figure 1
Schematic representation of the electrochemical detection platform fabrication process.
Figure 2
Figure 2
SEM images of bare (A) SPCE, (B) GO/SPCE, (C) RGO/SCPE, and (D) ssDNA/RGO/SPCE. Images recorded at 20 kX magnification (3 µm scale).
Figure 3
Figure 3
Raman spectra of SPCEs modified with GO and RGO.
Figure 4
Figure 4
High-resolution C1 XPS spectra of SPCE modified with (A) GO, (B) RGO, (C) probe, and (D) target molecules.
Figure 5
Figure 5
High-resolution N1 XPS spectra of SPCE modified with (A) probe molecule and (B) target molecule.
Figure 6
Figure 6
Contact angle variation on (A) unmodified SPCE, (B) GO/SPCE, (C) RGO/SPCE, (D) ssDNA/RGO/SPCE, and (E) dsDNA/RGO/SPCE.
Figure 7
Figure 7
Schematic representation of ssDNA probe binding mechanism to RGO surface.
Figure 8
Figure 8
CV showing the electrochemical reduction in 0.5 M KCl of GO/SPCE.
Figure 9
Figure 9
CV (A) and EIS Nyquist plot (B) recorded in 1 mM [Fe(CN)6]3−/4−, 0.1 M KCl, for bare SPCE, GO/SPCE, RGO/SPCE, and ALP ssDNA/RGO/SPCE.
Figure 10
Figure 10
CV (A) and EIS Nyquist plot (B) recorded in 1 mM [Fe(CN)6]3−/4−, 0.1 M KCl, for ssDNA/RGO/SPCE and after hybridization with 1, 10, 50, and 100 nM ALP target.
Figure 11
Figure 11
CV (A) and EIS Nyquist plot (B) recorded in 1 mM [Fe(CN)6]3−/4−, 0.1 M KCl, for bare SPCE, GO/SPCE, RGO/SPCE, and RUNX2 ssDNA/RGO/SPCE.
Figure 12
Figure 12
CV (A) and EIS Nyquist plot (B) recorded in 1 mM [Fe(CN)6]3−/4−, 0.1 M KCl, for ssDNA/RGO/SPCE and after hybridization with 1, 10, 50, and 100 nM RUNX2 target.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Rani S., Bandyopadhyay-Ghosh S., Ghosh S.B., Liu G. Advances in Sensing Technologies for Monitoring of Bone Health. Biosensors. 2020;10:42. doi: 10.3390/bios10040042. - DOI - PMC - PubMed
    1. Zhang Y., Yu T., Ding J., Li Z. Bone-on-a-chip platforms and integrated biosensors: Towards advanced in vitro bone models with real-time biosensing. Biosens. Bioelectron. 2023;219:114798. doi: 10.1016/j.bios.2022.114798. - DOI - PubMed
    1. Kim M.E., Seon J.K., Kang J.Y., Yoon T.R., Lee J.S., Kim H.K. Bone-Forming Peptide-4 Induces Osteogenic Differentiation and VEGF Expression on Multipotent Bone Marrow Stromal Cells. Front. Bioeng. Biotechnol. 2021;9:734483. doi: 10.3389/fbioe.2021.734483. - DOI - PMC - PubMed
    1. Nasir N.J.N., Arifin N., Noordin K.B.A., Yusop N. Bone repair and key signalling pathways for cell-based bone regenerative therapy: A review. J. Taibah Univ. Med. Sci. 2023;18:1350–1363. doi: 10.1016/j.jtumed.2023.05.015. - DOI - PMC - PubMed
    1. Vimalraj S. Alkaline phosphatase: Structure, expression and its function in bone mineralization. Gene. 2020;754:144855. doi: 10.1016/j.gene.2020.144855. - DOI - PubMed

LinkOut - more resources