Fe-doped biodegradable dendritic mesoporous silica nanoparticles for starvation therapy and photothermal-enhanced cascade catalysis in tumor therapy.

Combination therapies have attracted significant attention because they address the limitations of monotherapy while improving overall efficacy. In this study, we designed a novel nanoplatform, named GOx@Fe-DMSN@PDA (GFDP), by integrating Fe2+ into dendritic mesoporous silica nanoparticles (DMSN) and selecting glucose oxidase (GOx) as the model drug loaded into the DMSN pores. Additionally, we coated the surface of the DMSN with polydopamine (PDA) to confer pH/near infrared (NIR) light-responsive controlled-release behavior and photothermal therapy (PTT). The introduction of Fe2+ into the DMSN framework greatly improved biodegradability and enhanced the peroxidase (POD)-like activity of GFDP. In addition, GOx could consume glucose and generate hydrogen peroxide (H2O2) within tumor cells to facilitate starvation therapy and enhance cascade catalysis. The PDA coating provided the DMSN with an intelligent response release ability, promoting efficient photothermal conversion and achieving the PTT effect. Cellular tests showed that under NIR light irradiation, GFDP exhibited a synergistic effect of PTT-enhanced starvation therapy and cascade catalysis, with a half-maximal inhibitory concentration (IC50) of 2.89 µg/mL, which was significantly lower than that of GFDP without NIR light irradiation (18.29 µg/mL). The in vivo anti-tumor effect indicated that GFDP could effectively accumulate at the tumor site for thermal imaging and showed remarkable synergistic therapeutic effects. In summary, GFDP is a promising nanoplatform for multi-modal combination therapy that integrates starvation therapy, PTT, and cascade catalysis.

CoNiCoNC tumor therapy by two-ways producing H2O2 to aggravate energy metabolism, chemokinetics, and ferroptosis.

The effectiveness of chemokinetic therapy nanozymes is severely constrained because of the low H2O2 levels in the tumor microenvironment. Unlike other self-produced H2O2 nanozymes, the N-CNTs-encapsulated CoNi alloy (CoNiCoNC) with glucose oxidase and lactate oxidase activities has two ways to produce H2O2. It can facilitate the transformation of glucose and lactic acid into H2O2 simultaneously. First, the H2O2 generation pathway is favorable for aggravating energy metabolism. Second, some produced H2O2 can be decomposed by CoNiCoNC to H2O and O2 with the 4e- pathway to alleviate the TME hypoxia. Third, H2O2 can be catalyzed to form OH to enhance reactive oxygen species (ROS) content. Through proteomic analysis, nanozymes substantially impact the metabolic pathways of cancer cells because of their aggravating energy metabolism. The high levels of ROS can cause mitochondrial lipid peroxidation and cellular ferroptosis. Consequently, the two-way H2O2-selective nanoenzymatic platform realizes the synergistic effect of starvation therapy and chemokinetics.

Conductive single enzyme nanocomposites prepared by in-situ growth of nanoscale polyaniline for high performance enzymatic bioelectrode.

Enzyme-based electrochemical biosensors hold great promise for applications in health/disease monitoring, drug discovery, and environmental monitoring. However, inherently non-conductive nature of proteinaceous enzymes often hampers effective electron transfer at enzyme-electrode interface, limiting biosensor performance of enzyme bioelectrodes. To address this problem, we present an approach to synthesize polyaniline (PAN)-based conductive single enzyme nanocomposites of glucose oxidase (GOx) (denoted as PAN-GOx). To prevent multimerization of enzymes during nanocomposite synthesis and enable single enzyme wrapping, we activate GOx surface with phenylamine groups based on the programmed diffusion of reactants in the reaction solution. Subsequent in-situ polymerization enables the synthesis of nanoscale conductive PAN layer (∼2.7 nm thickness) grafted from individual GOx molecule. PAN-GOx retains 83% and 74% of its specific activity and catalytic efficiency, respectively, compared to free GOx, while demonstrating a ∼500% improved conductivity. Furthermore, PAN-GOx-based glucose biosensors show an approximately 16- and 3-fold higher sensitivity compared to biosensors prepared by using free GOx and a mixture of PAN and GOx, respectively. This study provides a facile method to fabricate conductive single enzyme nanocomposites with enhanced electron transfer, which can potentially be further modified and/or compounded with conductive materials for demonstrating high performance enzymatic bioelectrodes.

Screening Germplasms and Detecting Quantitative Trait Loci for High Sucrose Content in Soybean.

Sucrose is a desirable component of processed soybean foods and animal feed, and thus, its content is used as an important characteristic for assessing the quality of soybean seeds. However, few studies have focused on the quantitative trait loci (QTLs) associated with sucrose regulation in soybean seeds. This study aims to measure the sucrose content of 1014 soybean accessions and identify genes related to high sucrose levels using QTL analysis. Colorimetric analysis based on the enzymatic reaction of invertase (INV) and glucose oxidase (GOD) was employed to test the germplasms. A total of six high-sucrose genetic resources (IT186230, IT195321, IT263138, IT263276, IT263286, and IT276521) and two low-sucrose genetic resources (IT025668 and IT274054) were identified. Two F2:3 populations, IT186230 × IT025668 and Ilmi × IT186230, were then established from these germplasms. QTL analysis identified four QTLs (qSUC6.1, qSUC11.1, qSUC15.1, and qSUC17.1), explaining 7.3-27.6% of the phenotypic variation in the sugar content. Twenty candidate genes were found at the four QTLs. Notably, Glyma.17G152300, located in the qSUC17.1 QTL region, exhibited a 17-fold higher gene expression in the high-sucrose germplasm IT186230 compared to the control germplasm Ilmi, confirming its role as a major gene regulating the sucrose content in soybean. These results may assist in marker-assisted selection for breeding programs that aim to develop soybean lines with a higher sucrose content.

A pH-Sensitive Glucose Oxidase and Hemin Coordination Micelle for Multi-Enzyme Cascade and Amplified Cancer Chemodynamic Therapy.

Chemodynamic therapy (CDT) is an emerging therapeutic paradigm for cancer treatment that utilizes reactive oxygen species (ROS) to induce apoptosis of cancer cells but few biomaterials have been developed to differentiate the cancer cells and normal cells to achieve precise and targeted CDT. Herein, a simple cascade enzyme system is developed, termed hemin-micelles-GOx, based on hemin and glucose oxidase (GOx)-encapsulated Pluronic F127 (F127) micelles with pH-sensitive enzymatic activities. Histidine-tagged GOx can be easily chelated to hemin-F127 micelles via the coordination of histidine and ferrous ions in the center of hemin by simple admixture in an aqueous solution. In tumor microenvironment (TME), hemin-micelles-GOx exhibits enhanced peroxidase (POD)-like activities to generate toxic hydroxyl radicals due to the acidic condition, whereas in normal cells the catalase (CAT)-like, but not POD-like activity is amplified, resulting in the elimination of hydrogen peroxide to generate oxygen. In a murine melanoma model, hemin-micelles-GOx significantly suppresses tumor growth, demonstrating its great potential as a pH-mediated enzymatic switch for tumor management by CDT.

Enzymatic Synthesis of Isotopically Labeled Hydrogen Peroxide for Mass Spectrometry-Based Applications.

Methionine oxidation is involved in multiple biological processes including protein misfolding and enzyme regulation. However, it is often challenging to measure levels of methionine oxidation by mass spectrometry, in part due to the prevalence of artifactual oxidation that occurs during the sample preparation and ionization steps of typical proteomic workflows. Isotopically labeled hydrogen peroxide (H218O2) can be used to block unoxidized methionines and enables accurate measurement of in vivo levels of methionine oxidation. However, H218O2 is an expensive reagent that can be difficult to obtain from commercial sources. Here, we report a method for synthesizing H218O2 in-house. Glucose oxidase catalyzes the oxidation of ß-d-glucose and produces hydrogen peroxide in the process. We took advantage of this reaction to enzymatically synthesize H218O2 from 18O2 and assessed its concentration, purity, and utility in measuring methionine oxidation levels by mass spectrometry.

Paper immobilized BSA-decorated gold nanoclusters for single-step optical sensing of glucose and cholesterol without cross-reactivity.

Minimally invasive methods for detecting glucose, cholesterol and hydrogen peroxide are crucial for monitoring the nutritional and health status of humans and animals. The peroxidase mimetic activity by nanozymes is one of the versatile methods for detecting glucose, cholesterol, hydrogen peroxide, and other biomolecules. However, the strict requirement of acidic pH limits their sensing and interfacing ability with natural enzymes. The present study developed bovine serum albumin (BSA) coated gold nanoclusters (AuNC) immobilized on paper fabric to enable single-step visual detection of glucose, cholesterol and hydrogen peroxide in complex biological fluids like serum and milk. The BSA-AuNC suspension and immobilized paper fabric synergistically interface with the natural oxidative enzymes, glucose oxidase or cholesterol oxidase, at physiological pH. The concomitant loss in the fluorescent intensity of BSA-AuNC-loaded paper fabric exposed to the generated hydrogen peroxide (glucose/glucose oxidase or cholesterol/cholesterol oxidase) was directly proportional to the concentration of glucose or cholesterol. These reactions enabled simple visual detection as well as quantification of hydrogen peroxide, glucose and cholesterol using Image-J software and common smartphone-based mobile applications. The detection ability of BSA-AuNC-embedded paper fabric is specific and remains unaltered in the presence of similar oxidase enzymes or similar substrate analogues. With these unique features, the BSA-AuNC embedded paper fabric stands out as a prominent analytical device with enormous potential as a simple, user-friendly detection tool for monitoring biomolecules that are important to health, nutrition, and environmental safeguarding.

Focused starvation of tumor cells using glucose oxidase: A comprehensive review.

Starvation therapy targets the high metabolic demand of tumor cells. It primarily leans over the consumption of intracellular glucose and simultaneous blockade of alternative metabolic pathways. The strategy involves the use of glucose oxidase (GOx) for catalyzing the conversion of glucose into gluconic acid and hydrogen peroxide. Under these conditions, metabolic re-programming of tumor cells enables the utilization of substrates such as amino acids, fatty acids and lipids. This can be overcome by co-administration of chemo-, photo- and immuno-therapeutics together with glucose oxidase. Targeted delivery of glucose oxidase at tumor site can be enabled with the use of nanoformulations. In this review, we highlight that the outcomes of starvation therapy can be improved using rationally developed nano-formulations. It is possible to load synergistically acting bioactives in these formulations and deliver in site-specific manner and hence achieve the elimination of tumors cells with greater efficacy.

The importance of factorial design on the optimization of biosensor performance: immobilization of glucose oxidase as a case study.

Conventionally, the optimization of glucose biosensors is achieved by varying the concentrations of the individual reagents used to immobilize the enzyme. In this work, the effect and interaction between glucose oxidase enzyme (GOx), ferrocene methanol (Fc), and multi-walled carbon nanotubes (MWCNTs) at different concentrations were investigated by a design of experiments (DoE). For this analysis, a factorial design with three factors and two levels each was used with the software RStudio for statistical analysis. The data were obtained by electrochemical experiments on the immobilization of GOx-Fc/MWCNT at different concentrations. The results showed that the factorial DoE method was confirmed by the non-normality of the residuals and the outliers of the experiment. When examining the effects of the variables, analyzing the half-normal distribution and the effects and contrasts for GOx-Fc/MWCNT, the factors that showed the greatest influence on the electrochemical response were GOx, MWCNT, Fc, and MWCNT:Fc, and there is a high correlation between the factors GOx, MWCNT, Fc, and MWCNT:Fc, as shown by the analysis of homoscedasticity and multicollinearity. With these statistical analyses and experimental designs, it was possible to find the optimal conditions for different factors: 10 mM mL-1 GOx, 2 mg mL-1 Fc, and 15 mg mL-1 MWCNT show a greater amperometric response in the glucose oxidation. This work contributes to advancing enzyme immobilization strategies for glucose biosensor applications. Systematic investigation of DoE leads to optimized immobilization for GOx, enables better performance as a glucose biosensor, and allows the prediction of some outcomes.

Molecular Photoswitching Unlocks Glucose Oxidase for Synergistically Reinforcing Fenton Reactions for Antitumor Chemodynamic Therapy.

We have developed a new type of nanoparticles with potent antitumor activity photoactivatable via the combination of molecular photoswitching of spiropyran (SP) and enzymatic reaction of glucose oxidase (GOx). As two key processes involved therein, Fe(III)-to-Fe(II) photoreduction in Fe(III) metal-organic frameworks (MOFs) brings about the release of free Fe2+/Fe3+ while the photoswitching of SP to merocyanine (MC) unlocks the enzymatic activity of GOx that was pre-passivated by SP. The release of free Fe3+ boosts its hydrolysis and therefore enables the acidification of microenvironment, which is further reinforced by one of the products of the GOx-mediated glucose oxidation reaction, gluconic acid (GlcA). Based on the generation of Fe2+ and acidic milieu together with another product of the oxidation reaction, hydrogen peroxide (H2O2), these two processes jointly present triple enabling factors for generating lethal hydroxyl radicals (•OH) species via Fenton reactions and therefore oxidative stress capable of inhibiting tumor. The antitumor potency of such nanoparticle is verified in tumor-bearing model mice in vivo, proclaiming its potential as a potent and safe agent based on the unique mechanism of optically manipulating enzyme activity for synergistic antitumor therapeutics with high spatial precision, enhanced efficacy and minimized side effects.

Development of a sequential release nanomaterial for co-delivery of hypoxia-induced tirapazamine and HIF-1α siRNA in cancer therapy.

Unlike traditional drug carriers, sequential drug delivery systems can release different drugs in order, with the first released drug providing a prerequisite for the later released drug to maximize its function, thereby achieving stronger anti-tumor effects. Herein we constructed a temporal sequential system designated TPZ@MSN/HIF-1α siRNA@PDA@GOx (MTRPG) in which mesoporous silica nanoparticles were used as cores to load hypoxia induced chemotherapy drug tirapazamine (TPZ) and gene targeted nucleic acid drug HIF-1α siRNA, polydopamine (PDA) as acid -responsive coating as well as to realize photothermal therapy, and glucose oxidase (GOx) as the outermost layer to achieve starvation therapy and construct a deepened hypoxia to activate TPZ. Through in vitro and in vivo experiments, we demonstrated that the first released glucose oxidase catalyzed the oxidation of glucose, achieving starvation treatment while reducing the acidic environment and further exacerbating hypoxia in tumor cells. The reduced acidic conditions enabled the degradation of PDA, resulting in the release of loaded HIF-1α siRNA and TPZ. At the same time, PDA could also exert photothermal therapy under 808 nm near-infrared (808 nm NIR) laser irradiation. The later released hypoxia induced chemotherapy drug TPZ amplifies its anti-tumor activity under intensified hypoxia conditions. Meanwhile, the released HIF-1α siRNA interfered with the up-regulated HIF-1α induced by the deepened hypoxia condition, which caused hypoxia tolerance in tumors, reduced its expression activity, and achieved synergistic killing of tumor cells with chemotherapy. This work provides an effective multimodal synergistic therapy strategy to promote tumor therapeutic index, which may possess a promising future in clinical application.

A Self-Powered Enzymatic Glucose Sensor Utilizing Bimetallic Nanoparticle Composites Modified Pencil Graphite Electrodes as Cathode.

Enzymatic biofuel cells (EBFC) are promising sources of green energy owing to the benefits of using renewable biofuels, eco-friendly biocatalysts, and moderate operating conditions. In this study, a simple and effective EBFC was presented using an enzymatic composite material-based anode and a nonenzymatic bimetallic nanoparticle-based cathode respectively. The anode was constructed from a glassy carbon electrode (GCE) modified with a multi-walled carbon nanotube (MWCNT) and ferrocene (Fc) as a conductive layer coupled with the enzyme glucose oxidase (GOx) as a sensitive detection layer for glucose. A chitosan layer was also applied to the electrode as a protective layer to complete the composite anode. Chronoamperometry (CA) results show that the MWCNT-Fc-GOx/GCE electrode has a linear relationship between current and glucose concentration, which varied from 1 to 10 mM. The LOD and LOQ were calculated for anode as 0.26 mM and 0.87 mM glucose, respectively. Also the sensitivity of the proposed sensor was calculated as 25.71 µ A/mM. Moreover, the studies of some potential interferants show that there is no significant interference for anode in the determination of glucose except ascorbic acid (AA), uric acid (UA), and dopamine (DA). On the other hand, the cathode consisted of a disposable pencil graphite electrode (PGE) modified with platinum-palladium bimetallic nanoparticles (Nps) which exhibit excellent conductivity and electron transfer rate for the oxygen reduction reaction (ORR). The constructed EBFC was optimized and characterized using various electroanalytical techniques. The EBFC consisting of MWCNT-Fc-GOx/GCE anode and Pt-PdNps/PGE cathode exhibits an open circuit potential of 285.0 mV and a maximum power density of 32.25 µW cm-2 under optimized conditions. The results show that the proposed EBFC consisting of an enzymatic composite-based anode and bimetallic nanozyme-based cathode is a unique design and a promising candidate for detecting glucose while harvesting power from glucose-containing natural or artificial fluids.

Glucose Oxidase Coupling with Pistol-Like DNAzyme Based Colorimetric Assay for Sensitive Glucose Detection in Tears and Saliva.

Non-invasive monitoring of glucose levels in tears and saliva is crucial for diagnosing and predicting various illnesses, such as diabetic nephropathy. However, the capability of the current glucose detection methods to identify small amounts of glucose with a high sensitivity remains a significant obstacle. This study proposes a simple, visual technique for sensitively detecting glucose levels from tears and saliva using glucose oxidase (GOx) to catalyze glucose and pistol-like DNAzyme (PLDz) to enhance the signal. In particular, the ß-D-glucose present in the samples serves as the initial molecule that GOx identifies and catalyzes to generate gluconic acid and hydrogen peroxide (H2O2). The H2O2 induces the self-cleavage of PLDz, activating the "part b" sequence. This activation initiates catalytic hairpin assembly (CHA) and releases the DNAzyme section in the H1 probe. The DNAzyme acts as a peroxidase analog, facilitating the catalysis of the 3,3',5,5'-tetramethylbenzidine (TMB)-hydrogen peroxide (H2O2) system and resulting in color changes. The proposed method exhibits a broad detection range of six orders of magnitude and a low limit of 0.32 µM for glucose detection. Furthermore, the proposed method was highly effective in detecting glucose in saliva and tears, suggesting that it could potentially diagnose hyperglycemia-related disorders in clinical environments.

Glucose Oxidase-Coated Calcium Peroxide Nanoparticles as an Innovative Catalyst for In Situ H2O2-Releasing Hydrogels.

In situ forming and hydrogen peroxide (H2O2)-releasing hydrogels have been considered as attractive matrices for various biomedical applications. Particularly, horseradish peroxidase (HRP)-catalyzed crosslinking reaction serves efficient method to create in situ forming hydrogels due to its advantageous features, such as mild reaction conditions, rapid gelation rate, tunable mechanical strength, and excellent biocompatibility. Herein, a novel HRP-crosslinked hydrogel system is reported that can produce H2O2 in situ for long-term applications, using glucose oxidase-coated calcium peroxide nanoparticles (CaO2@GOx NPs). In this system, CaO2 gradually produced H2O2 to support the HRP-mediated hydrogelation, while GOx further catalyzed the oxidation of glucose for in situ H2O2 generation. As the hydrogel is formed rapidly is expected and the H2O2 release behavior is prolonged up to 10 days. Interestingly, hydrogels formed by HRP/CaO2@GOx-mediated crosslinking reaction provided a favorable 3D microenvironment to support the viability and proliferation of fibroblasts, compared to that of hydrogels formed by either HRP/H2O2 or HRP/CaO2/GOx-mediated crosslinking reaction. Furthermore, HRP/CaO2@GOx-crosslinked hydrogel enhanced the angiogenic activities of endothelial cells, which is demonstrated by the in vitro tube formation test and in ovo chicken chorioallantoic membrane model. Therefore, HRP/CaO2@GOx-catalyzed hydrogels is suggested as potential in situ H2O2-releasing materials for a wide range of biomedical applications.

Synthesis and comparison of the performance of two different water-soluble phthalocyanine based electrochemical biosensor.

Herein, a comparative study between novel water-soluble phthalocyanine-based biosensors was performed for the application of glucose sensing. For this purpose, two different copper (II) and manganese (III) phthalocyanines and their water-soluble derivatives were synthesized, and then their role as a supporting material for enzyme immobilization was evaluated by comparing their sensor performances. Two different phthalocyanine (AP-OH2-MnQ (MnPc) and AP-OH2-CuQ (CuPc)) were tested using electrochemical biosensor with immobilized glucose oxidase (GOx). To the best of our knowledge, the related water-soluble phthalocyanine-based glucose biosensors were attempted for the first time, and the developed approach resulted in improved biosensor characteristics. The constructed biosensors GE/MnPc/GOx and GE/CuPc/GOx showed good linearity between 0.003-1.0 mM and 0.05-0.4 mM, respectively. The limit of detection was estimated at 0.0026 mM for the GE/MnPc/GOx and 0.019 mM for the GE/CuPc/GOx. KMapp and sensitivity values were also calculated as 0.026 mM and 175.043 µAmM-1 cm-2 for the GE/MnPc/GOx biosensor and 0.178 mM and 117.478 µAmM-1 cm-2 for the GE/CuPc/GOx biosensor. Moreover, the fabricated biosensors were successfully tested to detect glucose levels in beverages with high recovery results. The present study shows that the proposed water-soluble phthalocyanines could be a good alternative for quick and cheap glucose sensing with improved analytical characteristics.

Multi-walled carbon nanotubes functionalized with a new Schiff base containing phenylboronic acid residues: application to the development of a bienzymatic glucose biosensor using a response surface methodology approach.

An innovative supramolecular architecture is reported for bienzymatic glucose biosensing based on the use of a nanohybrid made of multi-walled carbon nanotubes (MWCNTs) non-covalently functionalized with a Schiff base modified with two phenylboronic acid residues (SB-dBA) as platform for the site-specific immobilization of the glycoproteins glucose oxidase (GOx) and horseradish peroxidase (HRP). The analytical signal was obtained from amperometric experiments at - 0.050 V in the presence of 5.0 × 10-4 M hydroquinone as redox mediator. The concentration of GOx and HRP and the interaction time between the enzymes and the nanohybrid MWCNT-SB-dBA deposited at glassy carbon electrodes (GCEs) were optimized through a central composite design (CCD)/response surface methodology (RSM). The optimal concentrations of GOx and HRP were 3.0 mg mL-1 and 1.50 mg mL-1, respectively, while the optimum interaction time was 3.0 min. The bienzymatic biosensor presented a sensitivity of (24 ± 2) × 102 µA dL mg-1 ((44 ± 4) × 102 µA M-1), a linear range between 0.06 mg dL-1 and 21.6 mg dL-1 (3.1 µM-1.2 mM) (R2 = 0.9991), and detection and quantification limits of 0.02 mg dL-1 (1.0 µM) and 0.06 mg dL-1 (3.1 µM), respectively. The reproducibility for five sensors prepared with the same MWCNT-SB-dBA nanohybrid was 6.3%, while the reproducibility for sensors prepared with five different nanohybrids and five electrodes each was 7.9%. The GCE/MWCNT-SB-dBA/GOx-HRP was successfully used for the quantification of glucose in artificial human urine and commercial human serum samples.

Dual-mode detection of glucose based on pistol-like DNAzyme-mediated exonuclease-assisted signal cycle.

Detecting glucose accurately and sensitively from clinical samples like tears and saliva is still difficult. We have created a sensor that can detect glucose with high sensitivity and accuracy by combining the use of glucose oxidase (GOx) to catalyze glucose, a pistol-like DNAzyme (PLDz) to transform the signal, gold nanoparticles (AuNPs) to enhance the optical properties and the exonuclease-III (Exo-III) to amplify the signal. As a result, the proposed method exhibits a low detection limit of 7.5 pM and a wide detection range covering seven orders of magnitude. The suggested dual-mode strategy provides a sensitive, precise and specific detection method for glucose. Another advantage is that the dual-mode technique significantly improves the precision and consistency of the measurements, demonstrating its immense potential for use in biomedical research and clinical diagnostics.

In brief, glucose oxidation, facilitated by GOx, results in the creation of H₂O₂. The self-cleavage function of PLDz is triggered upon detection of the generated H₂O₂. The two procedures initiated by GOx and PLDz are crucial in precisely detecting glucose and turning glucose signals into nucleic acid signals. The PLDz fragments obtained can facilitate Exo-III-assisted signal amplification, resulting in the release of fluorophores into the solution and the aggregation of AuNPs. The probes were intricately constructed to prevent hydrolysis by Exo-III under ideal conditions, resulting in a low background and high sensitivity.

Designing Mesoporous Prussian Blue@zinc Phosphate Nanoparticles with Hierarchical Pores for Varisized Guest Delivery and Photothermally-Augmented Chemo-Starvation Therapy.

Background: With the rapid development of nanotechnology, constructing a multifunctional nanoplatform that can deliver various therapeutic agents in different departments and respond to endogenous/exogenous stimuli for multimodal synergistic cancer therapy remains a major challenge to address the inherent limitations of chemotherapy. Methods: Herein, we synthesized hollow mesoporous Prussian Blue@zinc phosphate nanoparticles to load glucose oxidase (GOx) and DOX (designed as HMPB-GOx@ZnP-DOX NPs) in the non-identical pore structures of their HMPB core and ZnP shell, respectively, for photothermally augmented chemo-starvation therapy. Results: The ZnP shell coated on the HMPB core, in addition to providing space to load DOX for chemotherapy, could also serve as a gatekeeper to protect GOx from premature leakage and inactivation before reaching the tumor site because of its degradation characteristics under mild acidic conditions. Moreover, the loaded GOx can initiate starvation therapy by catalyzing glucose oxidation while causing an upgradation of acidity and H2O2 levels, which can also be used as forceful endogenous stimuli to trigger smart delivery systems for therapeutic applications. The decrease in pH can improve the pH-sensitivity of drug release, and O2 can be supplied by decomposing H2O2 through the catalase-like activity of HMPBs, which is beneficial for relieving the adverse conditions of anti-tumor activity. In addition, the inner HMPB also acts as a photothermal agent for photothermal therapy and the generated hyperthermia upon laser irradiation can serve as an external stimulus to further promote drug release and enzymatic activities of GOx, thereby enabling a synergetic photothermally enhanced chemo-starvation therapy effect. Importantly, these results indicate that HMPB-GOx@ZnP-DOX NPs can effectively inhibit tumor growth by 80.31% and exhibit no obvious systemic toxicity in mice. Conclusion: HMPB-GOx@ZnP-DOX NPs can be employed as potential theranostic agents that incorporate multiple therapeutic modes to efficiently inhibit tumors.

Investigation of Direct Electron Transfer of Glucose Oxidase on a Graphene-CNT Composite Surface: A Molecular Dynamics Study Based on Electrochemical Experiments.

Graphene and its variants exhibit excellent electrical properties for the construction of enzymatic interfaces. In particular, the direct electron transfer of glucose oxidase on the electrode surface is a very important issue in the development of enzyme-based bioelectrodes. However, the number of studies conducted to assess how pristine graphene forms different interfaces with other carbon materials is insufficient. Enzyme-based electrodes (formed using carbon materials) have been extensively applied because of their low manufacturing costs and easy production techniques. In this study, the characteristics of a single-walled carbon nanotube/graphene-combined enzyme interface are analyzed at the atomic level using molecular dynamics simulations. The morphology of the enzyme was visualized using an elastic network model by performing normal-mode analysis based on electrochemical and microscopic experiments. Single-carbon electrodes exhibited poorer electrical characteristics than those prepared as composites with enzymes. Furthermore, the composite interface exhibited 4.61- and 2.45-fold higher direct electron efficiencies than GOx synthesized with single-carbon nanotubes and graphene, respectively. Based on this study, we propose that pristine graphene has the potential to develop glucose oxidase interfaces and carbon-nanotube-graphene composites for easy fabrication, low cost, and efficient electrode structures for enzyme-based biofuel cells.

Gas Cluster Ion Beam-Assisted Deposition for Fabricating Dry Multilayers Containing Enzyme and Substrate with On-Demand Release.

Gas cluster ion beam (GCIB)-assisted deposition is used to build multilayered protein-based structures. In this process, Ar3000-5000+ clusters bombard and sputter molecules from a reservoir (target) to a collector, an operation that can be sequentially repeated with multiple targets. The process occurs under a vacuum, making it adequate for further sample conservation in the dry state, since many proteins do not have long-term storage stability in the aqueous state. First of all, the stability in time and versatility in terms of molecule selection are demonstrated with the fabrication of peptide multilayers featuring a clear separation. Then, lysozyme and trypsin are used as protein models to show that the activity remaining on the collector after deposition is linearly proportional to the argon ion dose. The energy per atom (E/n) of the Ar clusters is a parameter that was also changed for lysozyme deposition, and its increase negatively affects activity. The intact detection of larger protein molecules by SDS-PAGE gel electrophoresis and a bioassay (trypsin at ≈25 kDa and glucose oxidase (GOx) at ≈80 kDa) is demonstrated. Finally, GOx and horseradish peroxidase, two proteins involved in the same enzymatic cascade, are successively deposited on ß-d-glucose to build an on-demand release material in which the enzymes and the substrate (ß-d-glucose) are combined in a dry trilayer, and the reaction occurs only upon reintroduction in aqueous medium.