Accurate and in situ visualization of nickel ions (Ni2+) at inflammatory sites is crucial for early diagnosis, progression monitoring, and targeted intervention of nickel-induced pathological conditions. However, no reports have been reported on the in situ detection of Ni2+ at inflammatory sites. Herein, a programmable allosteric DNA nanomachine (ES-AP-tFNA) with dual-enzyme-mediated signal amplification for in vivo molecular imaging of Ni2+ at inflammatory sites was developed. ES-AP-tFNA can be sequentially cleaved by apurinic/apyrimidinic endonuclease 1 (APE1), which is highly expressed in inflammatory cells, and by the Ni2+-dependent DNAzyme, thereby enabling the in situ detection of Ni2+ within inflammatory microenvironments. First, the locked DNAzyme (S5) is released following the cleavage of the AP site in ES-AP-tFNA by APE1. Subsequently, the DNAzyme hybridizes with S6 through base complementarity and, in the presence of Ni2+, specifically cleaves riboadenosine (rA), resulting in the separation of the fluorophore and quencher. Furthermore, the liberated DNAzyme can repeatedly cleave rA sites on multiple S6 strands, enabling fluorescence signal amplification through a catalytic recycling mechanism. Experimental results demonstrate that ES-AP-tFNA exhibits high sensitivity (LOD: 0.08 U/mL for APE1, 0.25 μM for Ni2+) and specificity. Furthermore, ES-AP-tFNA can rapidly and specifically visualize Ni2+ in inflammatory cells at the implantation sites without causing significant organ toxicity in mouse models implanted with a nickel alloy.
Liu et al. (Tue,) studied this question.