Performance Validation of Fabricated Nanomaterial-Based Biosensor for Matrix Metalloproteinase-8 Protein Detection

Matrix metalloproteinase-8 (MMP-8) is a crucial collagenase enzyme that primarily degrades type I collagen and extracellular glycoproteins, playing a significant role in the pathological processes of periodontal disease. It can serve as a biomarker for early detection and screening of the disease through advanced biosensor technology. The aim of this study was to fabricate and validate the performance of a nanomaterial-based biosensor for detecting MMP-8 protein.

The screen-printed gold electrode was modified with a thin film of 11-mercaptoundecanoic acid using the self-assembled monolayer technique. A biosensor was then created with N-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide to form ester bonds, followed by the immobilization of the antibody MMP-8 and blocking of nonspecific binding. The performance characteristics of a biosensor for detecting MMP-8 concentrations, ranging from 1 to 50 ng/mL, were evaluated using electrochemical techniques with data analysis performed using the NOVA software. Enzyme-linked immunosorbent assay (ELISA) was used as a control.

The results were expressed as mean values ± standard deviation. The coefficient of determination (R ²) was calculated based on the obtained calibration curve.

Electrochemical measurements revealed that the peak current after modifying the thin film on the electrode was lower than the bare electrode. Characterization of biosensors showed an increase in response compared to the previous step. Differential pulse voltammetry measurements indicated that the peak current for MMP-8 concentrations ranging from 1 to 50 ng/mL increased proportionally with concentration. The biosensor demonstrated high sensitivity, with a correlation coefficient of R ² = 0.953 when compared to ELISA (R ² = 1).

A biosensor utilizing nanomaterials has been successfully fabricated for the detection of the MMP-8 protein with high sensitivity. Subsequent research should prioritize the evaluation of its performance in clinical patients alongside an assessment of its specificity and stability. The objective was to advance this biosensor as a reliable diagnostic tool for the screening of periodontitis.