Highly sensitive electrochemical Osteoprotegerin (OPG) immunosensor for assessing fracture healing and evaluating drug efficacy.

Tibial fractures are common long bone injuries requiring effective monitoring for optimal healing. Osteoprotegerin (OPG), as a key marker of bone formation, is closely related to the degree of fracture healing. However, existing detection methods have certain limitations in sensitivity and specificity. This study successfully crafted an exceptionally sensitive electrochemical immunosensor based on COOH-CNFs/Ti₃C₂T_x MXene/PANI-AgNPs nanocomposite material for the quantitative analysis of OPG in serum, providing a methodological basis for auxiliary diagnosis of fracture healing degree and evaluation of drug efficacy. A one-pot hydrothermal method was employed to synthesize and modify the nanocomposite material on gold electrode surfaces, which exhibit high electrochemical activity, low charge transfer resistance, and a large electroactive surface area, thereby enhancing the immunosensor's conductivity and stability, with a wide linear range (10^-17 to 10^-12 g/mL) and a low detection limit (1.94 × 10^-18 g/mL). Methodological validation further confirmed the immunosensor's excellent performance in specificity, reproducibility, and stability. Moreover, the successful application of this immunosensor in detecting OPG in serum samples from actual tibial fracture patients before and after medication demonstrates significant potential for clinical application in assisting the assessment of fracture healing and evaluating the efficacy of orthopedic drugs.