Focused ultrasound, as a protocol of cancer therapy,
might induce
extracellular adenosine triphosphate (ATP) release, which could enhance
cancer immunotherapy and be monitored as a therapeutic marker. To
achieve an ATP-detecting probe resistant to ultrasound irradiation,
we constructed a Cu/N-doped carbon nanosphere (CNS), which has two
fluorescence (FL) emissions at 438 and 578 nm to detect ultrasound-regulated
ATP release. The addition of ATP to Cu/N-doped CNS was conducted to
recover the FL intensity at 438 nm, where ATP enhanced the FL intensity
probably via intramolecular charge transfer (ICT) primarily and hydrogen-bond-induced
emission (HBIE) secondarily. The ratiometric probe was sensitive to
detect micro ATP (0.2–0.6 μM) with the limit of detection
(LOD) of 0.068 μM. The detection of ultrasound-regulated ATP
release by Cu,N-CNS/RhB showed that ATP release was enhanced by the
long-pulsed ultrasound irradiation at 1.1 MHz (+37%, p < 0.01) and reduced by the short-pulsed ultrasound irradiation
at 5 MHz (−78%, p < 0.001). Moreover, no
significant difference in ATP release was detected between the control
group and the dual-frequency ultrasound irradiation group (+4%). It
is consistent with the results of ATP detection by the ATP-kit. Besides,
all-ATP detection was developed to prove that the CNS had ultrasound-resistant
properties, which means it could bear the irradiation of focused ultrasound
in different patterns and detect all-ATP in real time. In the study,
the ultrasound-resistant probe has the advantages of simple preparation,
high specificity, low limit of detection, good biocompatibility, and
cell imaging ability. It has great potential to act as a multifunctional
ultrasound theranostic agent for simultaneous ultrasound therapy,
ATP detection, and monitoring.