Nanocatalytic Tumor Therapy by Biomimetic Dual Inorganic Nanozyme‐Catalyzed Cascade Reaction

Abstract Emerging nanocatalytic tumor therapies based on nontoxic but catalytically active inorganic nanoparticles (NPs) for intratumoral production of high‐toxic reactive oxygen species have inspired great research interest in the scientific community. Nanozymes exhibiting natural enzyme‐mimicking catalytic activities have been extensively explored in biomedicine, mostly in biomolecular detection, yet much fewer researches are available on specific nanocatalytic tumor therapy. This study reports on the construction of an efficient biomimetic dual inorganic nanozyme‐based nanoplatform, which triggers cascade catalytic reactions for tumor microenvironment responsive nanocatalytic tumor therapy based on ultrasmall Au and Fe3O4 NPs coloaded dendritic mesoporous silica NPs. Au NPs as the unique glucose oxidase‐mimic nanozyme specifically catalyze β‐D‐glucose oxidation into gluconic acid and H2O2, while the as produced H2O2 is subsequently catalyzed by the peroxidase‐mimic Fe3O4 NPs to liberate high‐toxic hydroxyl radicals for inducing tumor‐cell death by the typical Fenton‐based catalytic reaction. Extensive in vitro and in vivo evaluations have demonstrated high nanocatalytic‐therapeutic efficacy with a desirable tumor‐suppression rate (69.08%) based on these biocompatible composite nanocatalysts. Therefore, this work paves a way for nanocatalytic tumor therapy by rationally designing inorganic nanozymes with multienzymatic activities for achieving high therapeutic efficacy and excellent biosafety simultaneously.


DOI: 10.1002/advs.201801733
Tumor microenvironment (TME) is generally known to feature abundant unique characteristics such as acidity, [1] hypoxia, [2] inflammation, [3] and overproduced hydrogen peroxide. [4] Based on the unique tumor physiological microenvironment and the rising nanotechnology, a number of promising nanotheranostic strategies of designing TME-specific/responsive nanoplatforms for efficient tumor therapy and precise bioimaging diagnosis have been extensively explored and developed. [5] Tumor cells produce large amounts of hydrogen peroxide (H 2 O 2 ) by the overexpressed superoxide dismutase (SOD) through a catalytic process by superoxide ions as generated from mitochondria. [6] As a typical feature of TME, the overexpressed H 2 O 2 has thus been exploited for triggering responsive drug-releasing in chemotherapy or producing endogenous O 2 for oxygen-favored cancer therapy such as photodynamic therapy, [7] sonodynamic therapy, [8] or radiotherapy. [9] As an emerging efficient cancertherapeutic modality, the proof-of-concept nanocatalytic tumor therapy as first proposed by our group [10] utilizes intratumoral in situ catalytic chemical reactions to produce oxidative stress for inducing cancer-cell death by damaging intracellular biomolecule substances such as proteins, lipids, and DNA. [11] A typical strategy of the proposed nanocatalytic tumor therapy is the representative chemodynamic therapy, [12] which converts less reactive endogenous H 2 O 2 into the most harmful reactive oxygen species (ROS), hydroxyl radicals (·OH) under mildly acidic TME via an intratumoral Fenton or Fenton-like reaction by metal ions. [13] Such an endogenous but direct chemical energy conversion strategy uses no external energy input such as laser, ultrasound, or magnetic field, thus avoiding the limitations of low tissue-penetration depth and nonspecificity of these external triggers on inducing cancer-cell death.
Based on the fact that the intratumoral H 2 O 2 concentration, generally believed to be 50-100 × 10 −6 m, [4b] is too low to produce desirable and sufficient amount of hydroxyl radicals for inducing satisfactory nanocatalytic-therapeutic efficacy via Fenton or Fenton-like reactions, natural glucose oxidase (GOx) was introduced to elevate the intratumoral H 2 O 2 concentration through natural catalytic oxidation of intratumoral glucose to H 2 O 2 and gluconic acid. [14] However, natural GOx possesses several intrinsic drawbacks such as high cost in preparation and purification, and relatively low operational stability, which will undoubtedly hamper its practical biomedical application under complicated and harsh physiological environments. [15] Therefore, exploring and developing natural GOx alternatives with much-enhanced stability and lowered cost is highly desirable and necessary.
Recently, the merging of biology with nanotechnology has motivated extensive research fever for designing functional nanoplatforms that exhibit unique natural enzyme-mimic catalytic activities for a broad range of biomedical applications. [16] As the promising alternatives for natural enzymes, catalytically active nanomaterials, known as the so-called "artificial enzymes" or "nanozymes," have demonstrated numbers of merits over natural enzymes, such as facile fabrication, low cost, and robust stability against severe conditions. [17] So far, a variety of nanomaterials, such as carbon-based nanoparticles (NPs), [18] metal NPs, [19] and metal oxides NPs [16a,20] have been discovered to possess unique enzyme-mimic catalytic activities, which are extensively used in numerous fields, including biomolecular detection, [21] biosensor, [22] antibacterial applications, [23] immunoassays, [24] cancer diagnostics and therapy, [25] and environmental monitoring. [26] Till now, an exceedingly large number of reports on peroxidase (POD)-mimic nanozymes have been reported, while other kinds of enzyme-mimic NPs, such as GOx-mimic nanozymes, glutathione peroxidase-mimic nanozymes, or SOD-mimic nanozymes, have been rarely explored. Fortunately, Rossi and co-workers discovered that Au NPs could catalyze the oxidation of glucose to H 2 O 2 and glucono delta-lactone (GDL) under the presence of dissolved oxygen, which was very similar to the reaction as catalyzed by GOx, suggesting that Au NPs could serve as a mimic for GOx. [27] Since then, owing to the unique GOx-mimic activity, Au NPs have been explored for diverse applications, most of which involve biomolecule (DNA, glucose, etc.) detection, while leaving many hidden applications to be discovered.
Bearing the unique GOx-mimicking catalytic activity of Au NPs and POD-mimicking catalytic performance of Fe 3 O 4 NPs, we herein report, for the first time, on a biomimetic dual inorganic nanozyme-triggered TME-responsive cascade catalytic reaction for efficient nanocatalytic tumor therapy based on an "all-inorganic biocompatible nanosystem", without employing any toxic chemical drug (Scheme 1). Ultrasmall and highly dispersed Au NPs and Fe 3 O 4 NPs were successively integrated into the large pore channels of dendritic mesoporous silica NPs Adv. Sci. 2019, 6, 1801733 Scheme 1. Schematic illustration of "toxic-drug-free" nanocatalytic tumor therapy by biomimetic inorganic nanomedicine-triggered cascade catalytic reaction. The composite nanoplatform with dual inorganic nanozyme activity was fabricated by the sequential loading of Au NPs as GOx-mimic nanozyme and Fe 3 O 4 NPs as POD-mimic nanozyme into the large mesopores of DMSN NPs followed by PEGylation. The therapeutic process by cascade catalytic chemical reaction includes the initial GOx-mimicking Au nanozyme-mediated catalytic oxidation reaction of glucose into H 2 O 2 , which is further utilized as the reactant for the POD-mimicking Fe 3 O 4 nanozyme-based catalytic Fenton reaction to produce highly toxic ·OH and subsequently induce tumor-cell apoptosis.
(DMSN NPs) to construct the composite nanoplatform, i.e., DMSN-Au-Fe 3 O 4 NPs. After further modification with polyethylene glycol (PEG) molecules for improved biocompatibility and physiological stability, DMSN-Au-Fe 3 O 4 NPs could accumulate into tumor tissue via the typical enhanced permeability and retention (EPR) effect, triggering the intratumoral TMEresponsive cascade catalytic reaction.
Acquiring an unusual anaerobic glycolytic behavior to support their metabolism and proliferation, tumor cells demand nutrients, essentially glucose, in a hysterical manner. [28] Therefore, in TME, masses of glucose molecules are transported and accumulated, which provides the possibility of initiating catalytic reaction by utilizing glucose molecules. The intratumoral cascade catalytic reaction is initially triggered by Au NPs, the GOx mimic inorganic nanozyme which catalyzes the intratumoral glucose oxidation, generating large amounts of H 2 O 2 molecules to serve as the substrate for the subsequent catalytic reaction where H 2 O 2 is disproportionated by the coloaded ultrasmall Fe 3 O 4 NPs via the typical Fenton catalytic reaction, liberating high-toxic hydroxyl radicals to effectively induce tumor-cell death. DMSN-Au-Fe 3 O 4 NPs as the potent nanoplatforms are capable of realizing the endogenous cascade reaction for TME-specific and effective nanocatalytic tumor therapy, in a noninvasive and "toxic-drug-free" way, benefiting from the robust biomimetic nanozymes. It is noted that such a cascade reaction with TME acidity-responsiveness would not be triggered under the neutral condition in normal tissue microenvironment, guaranteeing the high tumor-specificity and therapeutic biosafety.
DMSN NPs with unique central-radial pore structures, serving as the robust nanosupports for small NPs, were synthesized by an anion-assisted approach based on the Stöber mechanism and sol-gel chemistry. [29] A high density of highly dispersed and ultrasmall particle-sized Au NPs was confined into the large mesopores of DMSN NPs via an in situ reduction reaction of HAuCl 4 to produce DMSN-Au NPs, followed by the collection and integration of ultrasmall  pore volume of DMSN NPs were measured to be 197.5 m 2 g −1 and 0.9 cm 3 g −1 , respectively, with an average pore diameter of 23.3 nm according to the nitrogen adsorption-desorption isotherm and corresponding pore-size distribution ( Figure S3, Supporting Information). Such a unique branched structure with large pore size and highly accessible surface area of DMSN NPs is highly in favor of the subsequent growth of ultrasmall Au NPs and deposition of Fe 3 O 4 NPs. After functionalized with amine groups by aminopropyltriethoxysilane (APTES) to provide anchoring sites for Au NPs, the mesopores' surface of DMSN NPs was grown with Au NPs via an in situ reduction of auric ions by NaBH 4. TEM and SEM images reveal that ultrasmall Au NPs of ≈1.5 nm in diameter have been well dispersed and immobilized within the mesopores channels of DMSN NPs ( A typical colorimetric method based on 3,3′,5,5′-tetramethylbenzidine (TMB) was introduced as a substrate to test the POD-like catalytic activity of DMSN-Fe 3 O 4 NPs. In the presence of H 2 O 2 , DMSN-Fe 3 O 4 NPs catalyzed the oxidation of TMB to form the oxidized and therefore blue-colored TMB (oxTMB) featuring characteristic absorbances at 370 and 652 nm (Figure 2a). The possible reaction mechanism involves two steps where the OO bond in H 2 O 2 molecule was broken into ·OH followed by TMB oxidation by ·OH to form oxTMB. UV-vis absorption spectroscopy was used to monitor the production of the colorimetric product oxTMB. Negligible absorbance can be observed in the absence of DMSN-Fe 3 O 4 NPs (Figure 2b), suggesting that no oxidation reaction has occurred in the mixture of TMB and H 2 O 2 . However, an apparently blue-colored solution can be obtained after the addition of DMSN-Fe 3 O 4 NPs into TMB-H 2 O 2 mixture solution (pH 6.5) for 10 min with two major absorbance peaks at 370 and 652 nm attributable to oxTMB, which confirms the production of ·OH by DMSN-Fe 3 O 4 NPs and H 2 O 2 .
To further evaluate the catalytic activity of DMSN-Fe 3 O 4 nanomedicine, the steady-state catalytic kinetics was investigated at room temperature in a reaction system containing DMSN-Fe 3 O 4 NPs, TMB, and H 2 O 2 of varied concentrations (5,10,20,30,40, and 50 × 10 −3 m) in Na 2 HPO 4 -citric acid buffer solution (pH 6.5). The time-dependent absorbance variation of the reaction solution was monitored in time-scan mode at 652 nm using a microplate reader (Figure 2c). At each H 2 O 2 concentration, the concentration-changing rate (v) of oxTMB was calculated from the absorbance-changing rate via the Beer-Lambert law, A = εlc (A is the absorbance, ε is the molar absorbance coefficient, l is the path length, and c is the molar concentration) with l = 10 mm and ε of 39 000 m −1 cm −1 for oxTMB. Furthermore, the concentration change rates of TMB were plotted against corresponding H 2 O 2 concentrations, which follows the Michaelis-Menten equation (Figure 2d), known as where v 0 is the initial velocity of the reaction, V max is the maximal velocity of reaction, After confirming the catalytic activity of POD-mimic Fe 3 O 4 nanozyme in generating highly toxic ·OH from H 2 O 2 molecules, the catalytic activity of Au NPs as the GOxmimic nanozyme and the cascade catalytic performance were investigated. As a GOx mimic, Au NPs catalyze the oxidation of glucose in the presence of oxygen, producing gluconic acid and H 2 O 2 (Figure 3a). Therefore, the production of H 2 O 2 and gluconic acid, and the consumption of O 2 were assessed.
First, the reaction product H 2 O 2 generated from glucose catalyzed by Au NPs was qualitatively detected based on the H 2 O 2 -mediated size-enlargement of Au NPs in the presence of HAuCl 4 . [30] DMSN-Au NPs first catalyze the glucose oxidation reaction, and the in situ generated H 2 O 2 reduces HAuCl 4 to Au 0 following the equation The resulting Au species can further deposit on the surface of the initial Au NPs as seeds, causing the enlarged size of initially deposited Au NPs and correspondingly enhanced UV-vis absorbance intensity. According to the UV-vis results, in the presence of glucose and HAuCl 4 , the absorption of Au NPs at 505 nm increased over time (Figure 3b) and the absorption peak slightly red-shifted from 505 to 515 nm in 1 h of reaction (Figure 3c  The other product of glucose oxidation reaction, gluconic acid, was detected using a gluconic acid-specific colorimetric assay based on the reaction between gluconic acid, hydroxylamine, and FeCl 3 , which leads to the formation of a red compound hydroxamate-Fe 3+ with a typical absorbance peak at 505 nm. [31] Specifically, DMSN-Au NPs were incubated with glucose of different concentrations at pH 6.5 for 30 min. After the addition of hydroxylamine and Fe 3+ , the resulting solution turns into red with the major absorbance peak of the reaction solution being at 505 nm and increases with the elevated concentrations of glucose (Figure 3f), which confirms the production of gluconic acid in this Au NPs-catalyzed reaction and implies the catalytic activity of DMSN-Au NPs on glucose oxidation. In order to monitor the level of dissolved oxygen in the reaction system, a dissolved oxygen meter was used. After adding DMSN-Au NPs into the buffer solution of glucose of different  concentrations, the oxygen level in reaction solution declined rapidly owing to the consumption of dissolved oxygen for the glucose oxidation as catalyzed by the loaded Au NPs (Figure 3g).
Based on TMB colorimetric assay (Figure 3a), it has been verified that the self-organized enzymatic cascade reaction can be achieved by DMSN-Au-Fe 3 O 4 NPs without the aid of any natural enzyme. In detail, DMSN-Au-Fe 3 O 4 NPs and glucose solution were mixed and incubated in a TMB solution for 1 h. The reaction solution then turned blue with the absorbance peak at 370 and 652 nm, indicating the production of ·OH originating from the cascade catalysis of Au and Fe 3 O 4 NPs (Figure 3h). To be specific, the Au NPs immobilized within the mesopores of DMSN NPs initially catalyze the oxidation of glucose into H 2 O 2 , which is further utilized as the reactant for Fe 3 O 4 NPsbased catalytic Fenton reaction to produce hydroxyl radicals. Finally, the generated hydroxyl radicals oxidize the colorless TMB into the blue-colored oxTMB. Therefore, DMSN-Au-Fe 3 O 4 NP is indeed a robust composite nanozyme with dual enzymatic functionalities capable of catalyzing the cascade reaction without the assistance of any natural enzyme, i.e., initially catalyzing glucose oxidation to yield gluconic acid and H 2 O 2 , and then catalyzing H 2 O 2 into high-toxic ·OH. Especially, the catalytic performances of DMSN-Au-Fe 3 O 4 NPs in buffer solutions at pH 5.0, 6.5, 7.4, and 8.0 were conducted to investigate the influence of pH. It has been found that the catalytic activity of DMSN-Au-Fe 3 O 4 NPs is pH-dependent, exhibiting better catalytic activity in acid condition than in neutral or alkaline condition ( Figure S10, Supporting Information).
Before assessing the in vitro nanocatalytic therapeutic efficiency, the surface of DMSN-Au-Fe 3 O 4 NPs was modified with methoxy PEG-thiol (mPEG-SH) molecules for improved stability in physiological microenvironment. The Fourier transform infrared spectroscopy (FTIR) was conducted for the characterization of PEGylation. The FTIR spectrum of DMSN-Au-Fe 3 O 4 -PEG NPs shows the stretch of the COC band at 1060, 1110, 1150, 1250, and 1280 cm −1 , as well as the stretch band of CH 2 and CH 3 at 2890 and 2740 cm −1 , respectively, indicating the successful modification of mPEG-SH molecules ( Figure S11, Supporting Information). Attributing to the PEG modification, the DMSN-Au-Fe 3 O 4 -PEG NPs could be well dispersed in water, phosphate buffer solution, simulated body buffer, saline, dulbecco's modified eagle medium (DMEM), and fetal bovine serum, indicating their excellent stability ( Figure  S12, Supporting Information).
Initially, the cytotoxicities of DMSN-Au-Fe 3 O 4 NPs were tested on murine breast cancer 4T1 cells, as well as two normal cell lines, namely brain capillary endothelial cells and human umbilical vein endothelial cells by the standard cell-counting kit 8 assay. The results show that the DMSN-Au-Fe 3 O 4 NPs exhibit negligible effects on the proliferation of the two kinds of normal cells, at the elevated concentration up to 200 µg mL −1 within 12 and 24 h, indicating their high biosafety and biocompatibility for further in vitro and in vivo therapeutic applications ( Figure S13, Supporting Information). In addition, it has been found that DMSN-Au NPs and DMSN-Fe 3 O 4 NPs exhibit no significant inhibition on 4T1 cancer cells proliferation even at the elevated concentration up to 200 µg mL −1 after incubation for 12 and 24 h, at pH 6.5 (Figure 4a,b) and 7.4 ( Figure S14, Supporting Information). Importantly, when incubated with DMSN-Au-Fe 3 O 4 NPs, the cell viabilities exhibit a marked decline in a dose-dependent manner with an inhibition rate on 4T1 tumor cell viability being higher than 75% at 200 µg mL −1 (Figure 4c). This result indicates that the cytotoxicity of DMSN-Au-Fe 3 O 4 NPs should be attributed to the cascade catalytic reactions by Au and Fe 3 O 4 NPs, both of which are indispensable to trigger the generation of sufficiently high amount of toxic ·OH to induce the apoptosis of 4T1 tumor cells. In detail on the related therapeutic mechanism, H 2 O 2 , as produced by the reaction between glucose and oxygen under the catalysis by GOx mimic-Au NPs in cell culture medium, is further catalyzed by Fe 3 O 4 -based Fenton nanocatalysts to disproportionate, producing ·OH for killing the cancer cells. To evaluate the intracellular ·OH generation, a ROS fluorescence probe 2′,7′-dichlorofluorescin diacetate (DCFH-DA), which turns into 2′,7′-dichlorofluorescein (DCF) with strong green fluorescence in the presence of ROS, was introduced to stain cancer cells in order to reveal the intracellular ROS production. At pH 6.5, 4T1 cancer cells exhibit negligible fluorescence when coincubated only with high doses of glucose in DMEM or with DMSN-Au-Fe 3 O 4 NPs in glucose-free DMEM for 1 h. Comparatively, strong green fluorescence of 4T1 cancer cells can be observed after coincubation with DMSN-Au-Fe 3 O 4 NPs in the presence of glucose (Figure 4d), implying the massive intracellular ROS production by DMSN-Au-Fe 3 O 4 NPs and glucose under weakly acidic condition. Furthermore, it has been found that the DCF fluorescence intensity is dependent on the pH value of cell culture medium and incubation duration. As the incubation duration is prolonged from 15 min to 1 h, the DCF fluorescence intensity of cells increases accordingly when incubated with DMSN-Au-Fe 3 O 4 NPs and glucose under the weakly acidic condition (pH 6.5). However, in the neutral condition (pH 7.4), much weaker DCF fluorescence intensity is observed even after prolonged incubation (Figure 4e). The DCF fluorescence can be further monitored by flow cytometry (Figure 4f), which demonstrates that the DCF fluorescence increases by prolonging the incubation duration of DMSN-Au-Fe 3 O 4 NPs. The CLSM and flow cytometry results demonstrate that the intracellular ROS production as catalyzed by DMSN-Au-Fe 3 O 4 NPs has been substantially promoted in the mildly acidic environment as compared to that in the neutral condition.
Especially, the intracellular nanocatalytic therapeutic mechanism of DMSN-Au-Fe 3 O 4 nanoplatform was investigated by the typical flow cytometric apoptosis assay after stained with FITC-labeled Annexin V and propidium iodide (PI). According   (Figure 5a). [31] Importantly, after i.v. administration, DMSN-Au-Fe 3 O 4 NPs efficiently accumulated into the tumor tissue with the passive accumulation efficacy of 3.49% ID g −1 in 2 h and 1.80% ID g −1 in 12 h (Figure 5b) based on the typical EPR effect for tumor-passive targeting and accumulation. [32] Encouraged by the high efficiency of in vitro nanocatalytic therapy for killing cancer cells, the prolonged blood circulation, and potent tumor accumulation effect, the in vivo nanocatalytic therapeutic efficiency of DMSN-Au-Fe 3 O 4 NPs was evaluated against the 4T1 breast tumor xenograft on nude mice. Eighteen 4T1 tumor-bearing mice (tumor volume ≈ 50 mm 3 ) were randomly separated into three groups (n = 6 per group). Saline (control) and DMSN-Au-Fe 3 O 4 NPs at different doses (10 and 20 mg kg −1 ) were i.v. administrated to investigate the therapeutic performance and related in vivo mechanism. During a 15 day therapeutic period, the body weights of mice in two therapeutic groups show no significant difference from those of mice in the control group (Figure 5c). The tumor volumes of mice in each group were recorded using a digital caliper, and the digital photos of mice were taken every 2 days after the i.v. injection( Figure S19, Supporting Information). It has been found that the i.v. administration of DMSN-Au-Fe 3 O 4 NPs shows a dose-dependent tumor-growth inhibition effect during the therapeutic period (Figure 5d,e), with the inhibition rates of 45.96% and 69.08% at the doses of 10 and 20 mg kg −1 , respectively. This substantial tumor inhibition effect is attributed to the high-toxic hydroxyl radicals as produced by the endogenous cascade reaction triggered by the biomimetic Au and Fe 3 O 4 coloaded nanoplatforms under the mildly acidic TME, effectively inducing tumor cell death. It should be noted that the constructed nanocomposites are highly biocompatible without any toxic substance used, and the toxic effect can only be triggered under the mildly acidic TME, which means that this DMSN-Au-Fe 3 O 4 -based nanocatalytic tumor therapy features high tumor specificity and excellent therapeutic biosafety. The neutral condition of normal tissue will not trigger such a cascade catalytic reaction, therefore no toxic effect will be induced to cause noticeable damages to normal cells/tissues.
In order to reveal the detailed therapeutic mechanism by tumor-pathological analysis, hematoxylin and eosin (H&E), terminal deoxynucleotidyl transferase mediated dUTP nickend labeling (TUNEL) and Ki-67 antibody staining of dissected tumor tissues from each group were conducted. H&E and TUNEL staining results exhibit severe damage and necrosis of tumor cells in two therapeutic groups in comparison to the control group (Figure 5f). Ki-67 antibody staining results reveal the suppressed proliferative activities of tumors cells in two therapeutic groups while there is almost no significant adverse effect on cell proliferation in the control group. Furthermore, H&E staining of the major organs (heart, liver, spleen, lung, and kidney) conducted after the therapeutic period shows no noticeable pathological side effects on major organs of mice in therapeutic groups ( Figure S20, Supporting Information), indicating the high biocompatibility and therapeutic biosafety of DMSN-Au-Fe 3 O 4 NPs as nanocatalytic agents.
Additionally, the rapid clearance of therapeutic nanomaterials is highly favorable for the clinical translation, which can avoid In summary, this work reports on the effective TME-responsive catalytic cascade reactions for nanocatalytic tumor-specific therapy with marked therapeutic efficacy and biosafety based on an inorganic biomimetic DMSN-Au-Fe 3 O 4 composite nanoplatforms. The in situ grown Au NPs within the large mesopores of DMSN NPs as a GOx-mimic nanozyme specifically catalyze β-D-glucose oxidation into gluconic acid and H 2 O 2 under aerobic conditions, while the produced H 2 O 2 can subsequently be catalyzed by the coloaded Fe 3 O 4 -based Fenton nanocatalysts to produce high-toxic hydroxyl radicals which substantially induce tumor-cell death afterward. Both the comprehensive in vitro cell-level and in vivo tumor-bearing mice xenograft evaluations have demonstrated the efficient nanocatalytic tumor

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Adv. Sci. 2019, 6, 1801733 therapy on killing the cancer cells and suppressing the tumor growth by as high as 69.08% of inhibition rate without any toxic substance being used, in highly TME-responsive and tumorspecific manners. The high biocompatibility, high therapeutic biosafety, and easy excretion of these DMSN-Au-Fe 3 O 4 composite nanoplatforms have also been verified which guarantees their further clinical translation. Therefore, the present biomimetic dual inorganic nanocomposite-triggered cascade reaction strategy for TME-responsive and effective nanocatalytic tumor therapy not only establishes a paradigm of "toxic-drug-free" endogenous and noninvasive nanocatalytic biomedicine, but also takes an important step forward in developing biomimetic nanoplatforms with multienzyme mimicking catalytic activities for tumor-specific therapies.

Supporting Information
Supporting Information is available from the Wiley Online Library or from the author.