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. 2022 Jan 28;7(5):4457-4470.
doi: 10.1021/acsomega.1c06322. eCollection 2022 Feb 8.

β-Cyclodextrin-Stabilized Biosynthesis Nanozyme for Dual Enzyme Mimicking and Fenton Reaction with a High Potential Anticancer Agent

Salim Ali  1 , Suranjan Sikdar  2 , Shatarupa Basak  1 , Biplab Rajbanshi  1 , Modhusudan Mondal  1 , Debadrita Roy  1 , Ankita Dutta  3 , Anoop Kumar  3 , Vikas Kumar Dakua  4 , Rinku Chakrabarty  4 , Ashim Roy  4 , Abhinath Barman  4 , Anupam Datta  4 , Pijush K Roy  4 , Bhaskar Chakraborty  5 , Mahendra Nath Roy  1   4
Affiliations

β-Cyclodextrin-Stabilized Biosynthesis Nanozyme for Dual Enzyme Mimicking and Fenton Reaction with a High Potential Anticancer Agent

Salim Ali et al. ACS Omega. .

Abstract

The myth of inactivity of inorganic materials in a biological system breaks down by the discovery of nanozymes. From this time, the nanozyme has attracted huge attention for its high durability, cost-effective production, and easy storage over the natural enzyme. Moreover, the multienzyme-mimicking activity of nanozymes can regulate the level of reactive oxygen species (ROS) in an intercellular system. ROS can be generated by peroxidase (POD), oxidase (OD), and Fenton-like catalytic reaction by a nanozyme which kills the cancer cells by oxidative stress; therefore, it is important in CDT (chemo dynamic therapy). Our current study designed to investigate the enzyme mimicking behavior and anticancer ability of cerium-based nanomaterials because the cerium-based materials offer a high redox ability while maintaining nontoxicity and high stability. Our group synthesized CeZrO4 nanoparticles by a green method using β-cyclodextrin as a stabilizer and neem leaf extract as a reducing agent, exhibiting POD- and OD-like dual enzyme activities. The best enzyme catalytic activity is shown in pH = 4, indicating the high ROS generation in an acidic medium (tumor microenvironment) which is also supported by the Fenton-like behavior of CeZrO4 nanoparticles. Inspired by the high ROS generation in vitro method, we investigated the disruption of human kidney cells by this nanoparticle, successfully verified by the MTT assay. The harmful effect of ROS in a normal cell is also investigated by the in vitro MTT assay. The results suggested that the appreciable anticancer activity with minimal side effects by this synthesized nanomaterial.

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Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1
Figure 1
Schematic representation for the preparation of green synthesized CeZrO4.
Figure 2
Figure 2
(a) XRD spectra of CeZrO4 nanoparticles, (b,c) SEM images of CeZrO4 nanoparticles with different resolutions, (d) SEM image of the CeO2 nanoparticle, (e) SEM image of the ZrO2 nanoparticle, and (f) size distribution of CeZrO4 nanoparticles.
Figure 3
Figure 3
(a) EDAX spectrum of CeZrO4, (b) elemental mappings of cerium, zirconium, and oxygen atoms, (c) FTIR spectra of CeO2, ZrO2, and CeZrO4 nanoparticles, and (d) hydrodynamic particle size distribution of CeZrO4 nanoparticles.
Figure 4
Figure 4
UV–vis spectra of the POD-like activity of CeZrO4 compared to model indicators including (a) TMB and (c) dopamine. (b) POD-like activity of CeZrO4 against different time intervals. (d) pH-dependent POD-like activity of CeZrO4 at a wavelength of 650 nm.
Figure 5
Figure 5
Steady-state kinetic analysis of CeZrO4 as a POD mimetic. (a) Curve of velocity against the TMB concentration in the condition of 10 mM H2O2. (c) Curve of velocity against the H2O2 concentration under conditions of 0.3 mM TMB. (b,d) Double-reciprocal plots of (a,c), respectively. Condition: 250 μg·mL–1 and CeZrO4 in an acetate buffer (pH 5.0) at room temperature. Graphical representation of the POD-mimicking activity of CeZrO4 versus TMB (e).
Figure 6
Figure 6
UV–vis spectral monitoring of the OD-like activity of CeZrO4 compared to model indicators including (a) TMB and (c) dopamine. (b) OD-like activity of CeZrO4 against different time intervals. (d) pH-dependent OD-like activity of CeZrO4 at a wavelength of 650 nm.
Figure 7
Figure 7
Steady-state kinetic analysis of CeZrO4 as an OD mimetic. (a) Curve of velocity against the TMB concentration in Condition: 250 μg·mL–1 and CeZrO4 in an acetate buffer (pH 5.0) at room temperature. (b) Double-reciprocal plots of (a) graphical representation of the OD-mimicking activity of CeZrO4 versus TMB (c).
Figure 8
Figure 8
(a) UV–vis spectral monitoring of degradation of MB by the CeZrO4-mediated Fenton-like reaction. (b) Impact of time on MB degradation by the CeZrO4 nanoparticle-mediated Fenton-like reaction. (c) Degradation % by different synthesized nanomaterials. (d) Influence of scavengers on the MB degradation by the CeZrO4-driven Fenton-like reaction.
Figure 9
Figure 9
Study of the anticancer potential of CeO2, ZrO2, and CeZrO4 nanoparticles through the MTT assay: (a) ACHN cell line (Human Embryonic Kidney cancerous cell line) was treated with different concentrations of CeO2, ZrO2, and CeZrO4, as given in Methods. The bar diagrams show percentage cell toxicity. (b) HEK-293 cell line (Human Embryonic Kidney normal cell line) was treated with different concentrations of CeO2, ZrO2, and CeZrO2, as given in Methods. The bar diagrams show percentage cell viability.
Figure 10
Figure 10
Fluorescence-based ROS production in the presence of IC50 values of CeO2, ZrO2, and CeZrO2 in the ACHN cell line. ACHN cell lines were treated with IC50 (Inhibitory Concentration 50) values of CeO2, ZrO2, and CeZrO4 with (a) blank (control), (b) H2O2, (c) CeO2-0.599 μg/mL (d), ZrO2-1.228 μg/mL (e) CeZrO4-1.24 μg/mL and (f) table shows IC50 values of the ACHN cell line.
Figure 11
Figure 11
Graphical representation of the anticancer activity of synthesized CeZrO4.
See this image and copyright information in PMC

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