Flexible needle-type Microbiosensor for real-time monitoring traditional acupuncture-mediated adenosine release In vivo

Rapid adenosine (ADO) signaling, on the time frame of seconds, regulates physiological and pathological processes, including the therapeutic efficacy of acupuncture. Nevertheless, standard monitoring strategies are limited by poor temporal resolution. Herein, an implantable needle-type microsensor capable of monitoring ADO release in vivo in response to acupuncture in real time has been developed. Electrocatalytic Prussian Blue nanoparticles, an immobilized multienzyme system, and a permselective poly-o-phenylenediamine-based membrane were used for the sequential modification of the sensing region of the electrode. The resultant sensor can perform amperometric measurements of ADO levels in response to a very low level of applied potential (-0.05 V vs Ag/AgCl). This microsensor also functioned across a broad linear range (0-50 μM) and exhibited good sensitivity (1.1 nA/μM) with a rapid response time of under 5 s. Importantly, the sensor also exhibited good reproducibility and high selectivity. For in vivo animal studies, the microsensor was employed for the continuous assessment of instantaneous ADO release at the ST36 (Zusanli) acupoint when this acupoint was subjected to twirling-rotating acupuncture manipulation. Benefiting from superior sensor in vivo performance and stability, the positive correlation between the variability in acupuncture-induced ADO release and the stimulus intensity levels that affect the clinical benefit can be demonstrated for the first time. Overall, these results highlight a powerful approach to analyzing the in vivo physiological effects of acupuncture, expanding application realm of micro-nano sensor technology on a fast time scale.