Gold nanoparticles (AuNPs) with simultaneous plasmonic and biocatalytic properties provide a promising approach to developing versatile bioassays. However, the combination of AuNPs' intrinsic enzyme-mimicking properties with their surface-enhanced Raman scattering (SERS) activities has yet to be explored. Here we designed a peroxidase-mimicking nanozyme by in situ growing AuNPs into a highly porous and thermally stable metal-organic framework called MIL-101. The obtained AuNPs@MIL-101 nanozymes acted as peroxidase mimics to oxidize Raman-inactive reporter leucomalachite green into the active malachite green (MG) with hydrogen peroxide and simultaneously as the SERS substrates to enhance the Raman signals of the as-produced MG. We then assembled glucose oxidase (GOx) and lactate oxidase (LOx) onto AuNPs@MIL-101 to form AuNPs@MIL-101@GOx and AuNPs@MIL-101@LOx integrative nanozymes for in vitro detection of glucose and lactate via SERS. Moreover, the integrative nanozymes were further explored for monitoring the change of glucose and lactate in living brains, which are associated with ischemic stroke. The integrative nanozymes were then used to evaluate the therapeutic efficacy of potential drugs (such as astaxanthin for alleviating cerebral ischemic injuries) in living rats. They were also employed to determine glucose and lactate metabolism in tumors. This study not only demonstrated the great promise of combining AuNPs' multiple functionalities for versatile bioassays but also provided an interesting approach to designing nanozymes for biomedical and catalytic applications.
Nanozymes, the nanostructures with enzymatic activities, have attracted considerable attention because, in comparison with natural enzymes, they offer the possibility of lowered cost, improved stability, and excellent recyclability. However, the specificity and catalytic activity of current nanozymes are still far lower than that of their natural counterparts, which in turn has limited their use such as in bioanalysis. To address these challenges, herein we report the design and development of integrated nanozymes (INAzymes) by simultaneously embedding two cascade catalysts (i.e., a molecular catalyst hemin and a natural enzyme glucose oxidase, GOx) inside zeolitic imidazolate framework (ZIF-8) nanostructures. Such integrated design endowed the INAzymes with major advantage in improved catalytic efficiency as the first enzymatic reaction occurred in close (nanoscale) proximity to the second enzyme, so products of the first reaction can be used immediately as substrates for the second reaction, thus overcoming the problems of diffusion-limited kinetics and product instability. The considerable high catalytic activity and stability enabled the INAzymes to efficiently draw a colorimetric detection of glucose with good sensitivity and selectivity. When facilitated with in vivo microdialysis, the INAzyme was successfully used for facile colorimetric visualization of cerebral glucose in the brain of living rats. Moreover, when further combined with microfluidic technology, an integrative INAzyme-based online in vivo analytical platform was constructed. The promising application of the platform was successfully illustrated by continuously monitoring the dynamic changes of striatum glucose in living rats' brain following ischemia/reperfusion. This study developed a useful approach to not only functional nanomaterial design but also advanced platforms developments for diverse targets monitoring.
One of the current challenges in nanozymebased technology is to rationally control the enzyme mimicking activities with suitable modulation strategies to mimic the complexity and functions of natural systems. In this regard, nanoceria has recently emerged as a promising nanozyme because of its unique enzyme mimicking properties. Herein, we demonstrated that the oxidase-like catalytic activity of nanoceria was rationally modulated in situ via protonproducing/consuming enzyme-catalyzed bioreactions, which formed the basis of self-regulated bioassays for determining the corresponding enzyme activity, as well as other important targets, such as nerve agents, drugs, and bioactive ions. More interestingly, the oxidase-like activity of nanoceria was cooperatively modulated with the aid of adenosine triphosphate, thus improving the analytical performance of such selfregulated bioassays. The current study not only demonstrated regulatory strategies to modulate the nanozymes' activities, but also established a facile approach to designing self-regulated bioassays.
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