Label-free electrochemical immunosensor based on gold nanoparticle/polyethyleneimine/reduced graphene oxide nanocomposites for the ultrasensitive detection of cancer biomarker matrix metalloproteinase-1.

Matrix metalloproteinase-1 (MMP-1) is associated with many types of cancers, including oral, colorectal, and brain cancers. This paper describes the fabrication of an MMP-1 immunosensor based on a gold nanoparticle/polyethyleneimine/reduced graphene oxide (AuNP/PEI/rGO)-modified disposable screen-printed electrode (SPE). A microwave-assisted single-step method was employed for the simultaneous reduction of gold and graphene oxide in a PEI environment to avoid AuNP agglomeration. The crystal structure, chemical composition, optical properties, and interior morphology of the materials were probed by X-ray diffraction, Raman spectroscopy, UV-visible spectrometry, and transmission electron microscopy techniques. To assemble a label-free MMP-1 immunosensor layer-by-layer, 3-mercaptopropionic acid was utilized due to its strong sulfur-gold bonding ability, and its tail end was attached to a carboxyl group, allowing the MMP-1 antibody (anti-MMP-1) to be subsequently cross-linked using the traditional N-(3-dimethylaminopropyl) and N' ethylcarbodiimide hydrochloride method. Differential pulse voltammetry analysis showed a linear relationship with MMP-1 concentration in the range of 1-50 ng ml-1 with an R2 value of ∼0.996 (n = 5, RSD < 5%). This immunosensor was successfully applied for MMP-1 detection in urine, saliva, bovine serum, and cell culture media (HSC-3 & C6) of oral and brain cancers showing results comparable to those of the credible ELISA method.

Oral Cancer Theranostic Application of FeAu Bimetallic Nanoparticles Conjugated with MMP-1 Antibody.

Metastatic oral squamous cell carcinoma (SCC) displays a poor disease prognosis with a 5-year survival rate of 39%. Chemotherapy has emerged as the mainstream treatment against small clusters of cancer cells but poses more risks than benefits for metastatic cells due to the non-specificity and cytotoxicity. To overcome these obstacles, we conjugated antibodies specific for matrix metalloproteinase-1 (MMP-1), a prognostic biomarker of SCC, to iron-gold bimetallic nanoparticles (FeAu NPs) and explored the capability of this complex to target and limit SSC cell growth via magnetic field-induced hyperthermia. Our results showed that 4.32 ± 0.79 nm sized FeAu NPs were superparamagnetic in nature with a saturation magnetization (Ms) of 5.8 emu/g and elevated the media temperature to 45 °C, confirming the prospect to deliver hyperthermia. Furthermore, conjugation with MMP-1 antibodies resulted in a 3.07-fold higher uptake in HSC-3 (human tongue squamous cell carcinoma) cells as compared to L929 (fibroblast) cells, which translated to a 5-fold decrease in cell viability, confirming SCC targeting. Finally, upon magnetic stimulation, MMP-1-FeAu NPs conjugate triggered 89% HSC-3 cellular death, confirming the efficacy of antibody-conjugated nanoparticles in limiting SCC growth. The synergistic effect of biomarker-specific antibodies and magnetic nanoparticle-induced hyperthermia may open new doors towards SCC targeting for improved disease prognosis.