Endogenous GSH Research Articles

An ultrasensitive fluorescent platform for monitoring GSH variation during ischemic stroke

The growing evidence suggests that the disturbance of redox homeostasis in the brain tissue contributes to the process of tissue damage following ischemic stroke (IS). Tracking redox process in situ is critical for diagnosis, treatment and drug design of IS. Monitoring of substances in the reduction system is more suitable than detection of molecules of oxidation system, such as reactive oxygen species (ROS), due to their difficult capture of fleeting existence. Glutathione (GSH), a dominated component of reduction system, has higher concentration than other reduced substances and is the major cellular antioxidant for maintaining the redox balance but dramatically decreases during cerebral ischemia. Efficient tools can realize the real-time tracing of endogenous GSH at ultra low concentration during ischemic damage are significant and indispensable for dynamic monitoring the process of IS. Till now, only a few fluorescent probes have been reported for imaging GSH. The direct visualization of reductive substances in the cerebral vasculature of live mice is still a significant challenge. More importantly, due to the presence of the blood–brain barrier (BBB), monitoring GSH at ultralow concentration in brain tissue seems more difficult, which tremendously restricts the development of new strategies for targeting, detecting, and imaging GSH in vivo. In present study, we successfully synthesized a series of BODIPY-based fluorescent probes, WD-1-SH, WD-2-SH, and WD-3-SH, and detected many properties, including the capacity to penetrate the BBB, sensitivity to GSH and tissue penetration depth of excitation, and emission wavelengths. In vitro experiments revealed that among the probes, WD-1-SH exhibited the highest sensitivity to reducing substances; however, WD-2-SH exhibited higher sensitivity in vivo. These desired photophysical and pharmacokinetic properties endow our fluorescent probes with the capability to monitor brain tissue low concentration GSH after injury with excellent temporal and spatial resolution. Due to these favourable properties, our fluorescent probes hold great promise for visualizing endogenous reducing substances in a variety of pathophysiological processs in vitro and in vivo, such as during stroke induced ferroptosis.

Self-cascade nanohybrids boost cell ferroptosis stress for tumor radiosensitization therapy

Although ferroptosis plays a potential role in sensitizing tumor radiotherapy, its radiosensitization efficacy is still limited by insufficient endogenous H2O2 and excess glutathione (GSH) in the tumor microenvironment (TME). Herein, we engineered a “four-in-one” self-cascade nanohybrid composed of MnO2 nanodots-encompassed and bovine serum albumin (BSA)-coated gold nanoclusters (BAM) for radiosensitization therapy of tumors by reprogramming tumor cell fate to ferroptosis and reshaping TME. The MnO2 nanodots possessed fascinating glucose oxidase (GOx)-mimicking activity to effectively catalyze tumor endogenous glucose into H2O2, could further decompose H2O2 into hydroxyl radicals (HO·) and O2 in response to acidic TME, and simultaneously depleted the endogenous GSH, breaking the redox homeostasis in TME. As a result, lipid peroxides (LPO) in cell membranes and ferroptosis stress of tumor cells were significantly up-regulated. Synergy with the hypoxia relief and enhancement by the gold nanocores, radiotherapy completely inhibited the tumor progression. Moreover, the gold nanocores were successfully radiolabeled with 64Cu, and PET imaging revealed that BAM had high tumor-targeting efficiency and was renally clearable. Our study provides a unique perspective on radiosensitization therapy by regulating tumor cell fate to ferroptosis through rational design of a self-cascade nanozyme.

Crosstalk Between Iron and Sulfur Homeostasis Networks in Arabidopsis.

The widespread deficiency of iron (Fe) and sulfur (S) is becoming a global concern. The underlying mechanisms regulating Fe and S sensing and signaling have not been well understood. We investigated the crosstalk between Fe and S using mutants impaired in Fe homeostasis, sulfate assimilation, and glutathione (GSH) biosynthesis. We showed that chlorosis symptoms induced by Fe deficiency were not directly related to the endogenous GSH levels. We found dynamic crosstalk between Fe and S networks and more interestingly observed that the upregulated expression of IRT1 and FRO2 under S deficiency in Col-0 was missing in the cad2-1 mutant background, which suggests that under S deficiency, the expression of IRT1 and FRO2 was directly or indirectly dependent on GSH. Interestingly, the bottleneck in sulfite reduction led to a constitutively higher IRT1 expression in the sir1-1 mutant. While the high-affinity sulfate transporter (Sultr1;2) was upregulated under Fe deficiency in the roots, the low-affinity sulfate transporters (Sultr2;1, and Sultr2;2) were down-regulated in the shoots of Col-0 seedlings. Moreover, the expression analysis of some of the key players in the Fe–S cluster assembly revealed that the expression of the so-called Fe donor in mitochondria (AtFH) and S mobilizer of group II cysteine desulfurase in plastids (AtNFS2) were upregulated under Fe deficiency in Col-0. Our qPCR data and ChIP-qPCR experiments suggested that the expression of AtFH is likely under the transcriptional regulation of the central transcription factor FIT.

A Study on the Safety and Effects of Amorpha fruticosa Fruit Extract on Spontaneously Hypertensive Rats with Induced Type 2 Diabetes.

Metabolic syndrome is characterized by a variety of diagnostic criteria: obesity, dyslipidemia, type 2 diabetes, and arterial hypertension. They contribute to the elevated risk of cardiovascular morbidity and mortality. The potential for Amorpha fruticosa L. (Fabaceae) to improve diabetes and metabolic disease is promising, based on in vitro tests. This is why a further investigation of the species is needed. Additionally, a toxicity review in relation to safety revealed that to date, there are no published data regarding the toxicity of A. fruticosa towards humans. This species could provide abundant and cheap resources because it is an aggressive invasive plant that grows almost unrestrictedly. The objective of this study was to evaluate the acute toxicity of a purified extract of A. fruticosa (EAF), and to assess its antioxidant, antihypertensive, and antihyperglycemic activity in streptozotocin-induced diabetic spontaneously hypertensive rats (SHRs). The EAF was slightly toxic (LD50 = 2121 mg/kg, b.w.) when administered orally, and moderately toxic (LD50 = 316 mg/kg, b.w.) at intraperitoneal administration, both in mice. The oral administration of EAF (100 mg/kg) for 35 days to SHRs caused significant decreases in the systolic pressure, blood glucose levels, and MDA quantity. It also increased the hepatic level of the endogenous antioxidant GSH, not only in diabetic SHRs, but also in the control group. An additional potential benefit to human health might be conferred through the environmental management of A. fruticosa based on its large-scale use for medicinal purposes, as this aggressive invasive species brings problems to natural habitats in many European countries.

A Biodegradable High-Efficiency Magnetic Nanoliposome Promotes Tumor Microenvironment-Responsive Multimodal Tumor Therapy Along with Switchable T2 Magnetic Resonance Imaging.

We explored the catalytic activity and magnetic resonance imaging (MRI) capacity of Cu-doped ultrasmall iron oxides with different doping ratios. Then, we screened a highly efficient ultrasmall active catalyst (UAC). Subsequently, a biodegradable magnetic nanoliposome was developed for multimodal cancer theranostics through pH-sensitive liposome coating of these UACs. Upon entering the body, the magnetic nanoliposomes significantly prolonged the metabolic time of UACs and promoted their accumulation in tumors. Then, the strong photothermal (PT) effect of the magnetic nanoliposome quickly ablated the tumor, showing promising PT therapy. Upon entering tumor cells, the magnetic nanoliposome rapidly degraded into many UACs and released chemotherapeutic drugs, contributing to chemotherapy. In addition, UACs not only catalyzed Fenton-type reaction to produce excessive reactive oxygen species (ROS) but also inhibited the synthesis of endogenous GSH by inactivating glutamyl cysteine ligase, contributing to cancer ferroptosis. Furthermore, the assembly-dissociation process of UACs showed the function of magnetic relaxation switches, significantly enhancing tumor MRI signal change, achieving a more accurate diagnosis of the tumor. Therefore, this magnetic nanoliposome splits into many UACs upon drug release and regulates the tumor microenvironment to overproduce ROS for enhanced synergistic tumor theranostics, which provides a strategy for developing next-generation magnetic catalysts with biodegradability and multimodal antitumor theranostics.

Gold nanostructured membranes to concentrate low molecular weight thiols, a proof of concept study

Thiols are very important molecules in the biomedical field involved for example in redox homeostasis. Their detection and quantification remain difficult due to their poor stability (oxidation) linked to their strong reactivity towards other thiols (by the formation of S-S bonds) or other interfering molecules in the medium. Cellulose membranes with immobilized gold nanoparticles (AuNP) were developed to capture and quantify thiols in simple and complex matrices. This device was first optimized and characterized in terms of nanostructuration and thiol adsorption. N-Acetylcysteine (NAC) and reduced glutathione (GSH), chosen as model molecules, were filtered through the device demonstrating a maximal adsorption capacity of 270 and 60 nmol respectively. In a second step, the adsorbed species were subjected to ligand exchange using a more reactive thiol, dithiothreitol. The results showed release rates of approximately 90% for NAC and GSH. Finally, the amount of endogenous GSH in rat plasma was determined without any pretreatment. For the first time to our knowledge, a nanostructured device for the capture, selective and sensitive quantification of thiols is proposed. This device is easy to handle and overcomes matrix effects. Moreover, the very large concentration factor induced by this technology will be a valuable asset to decrease the quantification limits of analytical methods.

Taify Pomegranate Juice (TPJ) Abrogates Acrylamide-Induced Oxidative Stress Through the Regulation of Antioxidant Activity, Inflammation, and Apoptosis-Associated Genes.

Acrylamide (ACR) has various effects on biological systems, including oxidative stress and its associated metabolic disorders. Previous research reports that plants growing at high altitude have a different profile of antioxidants. In the current report, the Taify pomegranate juice (TPJ) of the Taify pomegranate growing at the Taif region (high altitude), Saudi Arabia, was investigated for its protective activity from ACR-induced oxidative stress. Rats were treated with ACR, TPJ, or TPJ+ACR, and various assays, including blood chemistry, liver function biomarkers, gene expression of endogenous antioxidant enzymes, oxidative stress regulatory genes, inflammation biomarkers, and apoptosis, were estimated using biochemical, real-time PCR, histopathological, and immunohistochemical analysis. TPJ showed a protective function of ACR-induced alteration of AST, ALT, GGT, urea, total proteins, albumin, MDA, and NO. It also increased the level of the endogenous antioxidative enzymes, including SOD, catalase, and GSH. It showed anti-inflammatory activity by reduction the TNF-α, IL-6 secretion and the enhancing of IL-10 levels. At the gene expression level, TPJ upregulated the expression of endogenous antioxidant genes (SOD and catalase) and of antioxidant-regulating genes Nrf2 and HO-1; downregulated the expression of inflammatory genes TGF-β1, COX2, and the apoptotic gene caspase-3; and upregulated the expression of antiapoptotic gene Bcl2. At the histological level, TPJ showed a protective effect from the ACR-induced hepatic histological damage. Results of this study conclude that TPJ has a protective effect from ACR-induced oxidative stress and its associated metabolic alterations through its antioxidant and anti-inflammatory activities.

Highly selective fluorescent detection platform based on isoquinoline Schiff base ligand monitors glutathione in biological systems

Aberrant levels of intracellular glutathione (GSH) have been held responsible for various disease and pathological processes, and existing evidence underscores the significance of GSH detection in biological systems. The recognition, discrimination, and precise detection of GSH in the cellular matrix and living organisms can revolutionize our understanding of the physiological and pathological processes of GSH. Herein, we reported a new isoquinoline Schiff base fluorescence probe, N'-((7-(diethylamino)-2-oxo-2H-chromene)methylene) isoquinoline-3-carbohydrazide (NCMC), which can selectively recognize GSH by its ensemble complex NCMC-Cu2+ with a low nanomolar level detection limit (4.16 × 10−8 M) in the aqueous environment. Moreover, the binding behavior and “ON-OFF-ON” fluorescence probing pattern were illustrated in detail by UV–vis titration, ESI-MS spectroscopy, and density functional theory calculations. Notably, the nontoxic NCMC-Cu2+ ensemble platform showed great potential for detecting GSH distribution in cells and zebrafish larvae, and can also be applied in differentiating endogenous GSH in both cancer cells and normal cells, which may represent a promising GSH fluorescence probe in bioimaging and disease diagnosis.

In-vivo two-photon visualization and quantitative detection of redox state of cancer.

Glutathione (GSH), the most common and abundant antioxidant in the body, is particularly concentrated in cancer cells (2-10 mM). This concentration is approximately 1000 times that of normal cells, making GSH a specific tumor marker. Overexpression of GSH is critical for mapping the redox state of cancer cells. However, there are few probes and detection methods responsive to GSH that can quantitatively visualize GSH in vivo in two-photon excitation fluorescence (TPEF) imaging mode. The experimental results show that TPEF-GSH could not only target GSH in tumors, but also establish the quantitative relationship between TPEF signal and GSH concentration. We explored the optimal two-photon excitation wavelength of TPEF-GSH, the optimal cell incubation duration with TPEF-GSH, the best imaging time point for GSH in cells, and the quantitative relationship between the TPEF signal and the changes in GSH concentrations. In zebrafish embryo and zebrafish experiments, the ratiometric value of TPEF-GSH increased with the decrease of GSH concentration. Microinjection and co-incubation were used to verify whether the ratiometric value could quantify endogenous GSH in tumor-bearing zebrafish, and the obtained GSH levels were 4.66 mM and 5.16 mM, respectively. The ratio TPEF probe could accurately visualize and quantify GSH in vivo, reflecting the redox status of the tumor. The design of the ratiometric molecular probe provides a reliable strategy for the development of TPEF nanoprobe in vivo. In this article, a new GSH sensitive molecular probe, TPEF-GSH, has been developed with good specificity and sensitivity. TPEF-GSH was successfully used to image cancer cells in vitro and tumor-bearing zebrafish in vivo, and to further detect GSH levels.

Assessment of metabolic activation of felbamate in chimeric mice with humanized liver in combination with in vitro metabolic assays.

Felbamate (FBM) is an antiepileptic drug that has minimal toxicity in preclinical toxicological species but has a serious idiosyncratic drug toxicity (IDT) in humans. The formation of reactive metabolites is common among most drugs associated with IDT, and 2-phenylpropenal (2-PP) is believed to be the cause of IDT by FBM. It is important to consider the species difference in susceptibility to IDT between experimental animals and humans. In the present study, we used an in vitro and in vivo model system to reveal species difference in IDT of FBM. Human cytochrome P450 (CYP) and carboxylesterase (CES) expressing microsomes were used to clarify the isozymes involved in the metabolism of FBM. The remaining amount of FBM was significantly reduced in incubation with microsomes expressing human CYP2C8, 2C9, 2E1, and CES1c isozymes. Chimeric mice with humanized liver are expected to predict IDT in humans. Therefore, metabolite profiles in chimeric mice with humanized liver were investigated after administration of FBM. Metabolites after glutathione (GSH) conjugation of 2-phenylpropenal (2-PP), which is the reactive metabolite responsible for FBM-induced IDT, were detected in chimeric mice plasma and liver homogenate. Mass spectrometry imaging (MSI) visualizes distribution of FBM and endogenous GSH, and GSH levels in human hepatocyte were decreased after administration of FBM. In this study, we identified CYP and CES isozymes involved in the metabolism of FBM and confirmed reactive metabolite formation and subsequent decrease in GSH using humanized animal model. These results would provide useful information for the susceptibility to IDT between experimental animals and humans.

In situ monitoring of functional activity of extracellular matrix stiffness-dependent multidrug resistance protein 1 using scanning electrochemical microscopy.

Extracellular matrix (ECM) stiffness affects the drug resistance behavior of cancer cells, while multidrug resistance protein 1 (MRP1) on the cell membrane confers treatment resistance via actively transporting drugs out of cancer cells. However, the relationship between ECM stiffness and MRP1 functional activity in cancer cells remains elusive, mainly due to the technical challenge of in situ monitoring. Herein, we engineered in vitro cancer cell models using breast cancer cells (MCF-7 and MDA-MB-231 cells) as the reprehensive cells on polyacrylamide (PA) gels with three stiffness, mimicking different developmental stages of cancer. We in situ characterized the functional activity of MRP1 and investigated the effect of ECM stiffness on MRP1 of cancer cells before and after vincristine treatment using scanning electrochemical microscopy (SECM) with ferrocenecarboxylic acid (FcCOOH) as the redox mediator and endogenous glutathione (GSH) as the indicator. The SECM results show that the functional activity of MRP1 is enhanced with increasing ECM stiffness, and the MRP1-mediated vincristine efflux activity of MCF-7 cells is more affected by ECM stiffness than that of MDA-MB-231 cells. This work, for the first time, applied SECM to in situ and quantitatively monitor the functional activity of MRP1 in cancer cells in different tumor mechanical microenvironments, which could help to elucidate the mechanism of matrix stiffness-dependent drug resistance behavior in cancer cells.

A water-soluble fluorescent probe for monitoring mitochondrial GSH fluctuations during oxidative stress.

In this research, we constructed a styrylpyridine derivative-based fluorescent probe MITO-PQDNs to monitor mitochondrial glutathione (GSH). The probe MITO-PQDNs could react rapidly (20 min) with GSH in PBS buffer and exhibited a strong fluorescence signal (586 nm) as well as a significant Stokes shift (200 nm). Moreover, MITO-PQDNs could quantitatively detect GSH with high sensitivity (LOD = 253 nM). Meanwhile, MITO-PQDNs possessed favorable biocompatibility and could detect both endogenous and exogenous GSH in MCF-7 cells. Above all, MITO-PQDNs enabled the detection of fluctuations in mitochondrial GSH concentrations during oxidative stress.

White tea modulates antioxidant defense of endurance-trained rats.

The interest in nutritional strategies that may counteract the deleterious oxidative effects induced by strenuous exercises is remarkable. Herein, the impact of white tea (Camellia sinensis) (WT), a polyphenol-rich beverage, on antioxidant status in endurance-trained rats after one session of exhaustive exercise were evaluated. Male Wistar rats were divided into groups, which received: control groups - water, and testing groups - WT1 (0.25%; w/v) or WT2 (0.5%; w/v). Drinks were consumed, ad libitum, for 5 or 10 weeks, concomitantly with the running training. Exhaustive running tests were applied before and after the experimental periods. WT intake increased the serum antioxidant capacity of rats in a dose-dependent manner (P < 0.001), which was unaccompanied by the activity of endogenous antioxidant enzymes SOD, GPx, and GR, and GSH content. Inflammatory markers in serum [IL-1β (P = 0.004) and IL-6 (P = 0.001)] could be downregulated by tea intake. In liver tissue, lower levels of lipid oxidation (P < 0.05) and improved antioxidant defenses (SOD, GPx, GR, and GSH, P < 0.05) were related to the consumption of 10.13039/100010269WT in both doses, supporting protective effects in this responsible metabolic organ. In conclusion, long-term consumption of WT could be a promising adjuvant to exercise-stress management, emphasizing its ability to regulate antioxidant responses and prevent oxidative tissue damage.

Initiation and Prevention of Biological Damage by Radiation-Generated Protein Radicals.

Ionizing radiations cause chemical damage to proteins. In aerobic aqueous solutions, the damage is commonly mediated by the hydroxyl free radicals generated from water, resulting in formation of protein radicals. Protein damage is especially significant in biological systems, because proteins are the most abundant targets of the radiation-generated radicals, the hydroxyl radical-protein reaction is fast, and the damage usually results in loss of their biological function. Under physiological conditions, proteins are initially oxidized to carbon-centered radicals, which can propagate the damage to other molecules. The most effective endogenous antioxidants, ascorbate, GSH, and urate, are unable to prevent all of the damage under the common condition of oxidative stress. In a promising development, recent work demonstrates the potential of polyphenols, their metabolites, and other aromatic compounds to repair protein radicals by the fast formation of less damaging radical adducts, thus potentially preventing the formation of a cascade of new reactive species.

Harnessing in situ glutathione for effective ROS generation and tumor suppression via nanohybrid-mediated catabolism dynamic therapy

The overexpression of glutathione (GSH) in cancer cells has long been regarded as the primary obstacle for reactive oxygen species (ROS)-involved anti-tumor therapies. To solve this issue, a ferric ion and selenite-codoped calcium phosphate (Fe/Se–CaP) nanohybrid here is fabricated to catabolize endogenous GSH, instead of directly deleting it, to trigger a ROS storm for tumor suppression. The selenite component in Fe/Se–CaP can catabolize GSH to superoxide anion (O2•−) and hydroxyl radicals (•OH) via cascade catalytic reactions, elevating oxidative stress while destroying antioxidant system. The doped Fe can further catalyze the soaring hydrogen peroxide (H2O2) originated from O2•− to •OH via Fenton reactions. Collectively, Fe/Se–CaP mediated self-augmented catabolism dynamic therapy finally induces apoptosis of cancer cells owing to the significant rise of ROS and, combined with CaP adjuvant, evokes adaptive immune responses to suppress tumor progression, providing an innovative train of thought for ROS-involved anti-tumor therapies.

Combination of Ocimum sanctum extract and Levetiracetam ameliorates cognitive dysfunction and hippocampal architecture in rat model of Alzheimer’s disease

Alzheimer’s disease (AD) is the most common neurodegenerative disease which affects more than 40 million people worldwide with progressive loss of memory and cognitive functions. It is reported, persistent AD is also one of the main causes of epilepsy in elders and comorbidity of both these will contribute to worsening the health status of AD patients. Recently, herbal plants with potent neuroprotective and antioxidant properties were used for increasing the quality of life in neurodegenerative disease patients. The present study was conceptualized to access the protective effect of Ocimum sanctum extract (OSE) and Levetiracetam (LEV) and their combination (OSE+LEV) against AD and epilepsy associated with AD in the rat AD model. AD was induced in aged male Wistar albino rats with Amyloid-β (Aβ) by intracerebroventricular administration. The results reveal, treatment with OSE, LEV and OSE+LEV significantly reversed the memory impairment, increases the BDNF expressions and decreases AChE activity in Aβ induced AD animals. Expression of A-β and p-tau in the hippocampus was significantly reduced in treatment group when compared to the control animals. Treatment with OSE and OSE+LEV also restored the hippocampal architecture by ameliorating the neuronal count in CA1, CA3 and DG regions. It also observed that treatment has decreased the excitoneurotoxicity evidenced by decreased glutamate and increased GABA levels and thus provided protection against epilepsy. Treatment groups also exhibited a potent antioxidant activity when tested endogenous antioxidant enzymes SOD, GSH and LPO in the brain hippocampus. Our findings provide evidence for use of OSE, LEV and OSE+LEV against AD and epilepsy associated with AD in Aβ induced AD animal model. However, further clinical studies are required to prove the use of OSE, LEV and OSE+LEV in the management of AD and AD-associated epilepsy in human volunteers.

An Ultrasmall Fe3 O4 -Decorated Polydopamine Hybrid Nanozyme Enables Continuous Conversion of Oxygen into Toxic Hydroxyl Radical via GSH-Depleted Cascade Redox Reactions for Intensive Wound Disinfection.

Nanozyme-based chemodynamic therapy (CDT) for fighting bacterial infections faces several major obstacles including low hydrogen peroxide (H2 O2 ) level, over-expressed glutathione (GSH) in infected sites, and inevitable damage to healthy tissue with abundant nonlocalized nanozymes. Herein, a smart ultrasmall Fe3 O4 -decorated polydopamine (PDA/Fe3 O4 ) hybrid nanozyme is demonstrated that continuously converts oxygen into highly toxic hydroxyl radical (•OH) via GSH-depleted cascade redox reactions for CDT-mediated bacterial elimination and intensive wound disinfection. In this system, photonic hyperthermia of PDA/Fe3 O4 nanozymes can not only directly damage bacteria, but also improve the horseradish peroxidase-like activity of Fe3 O4 decorated for CDT. Surprisingly, through photothermal-enhanced cascade catalytic reactions, PDA/Fe3 O4 nanozymes can consume endogenous GSH for disrupting cellular redox homeostasis and simultaneously provide abundant H2 O2 for improving •OH generation, ultimately enhancing the antibacterial performance of CDT. Such PDA/Fe3 O4 can bind with bacterial cells, and reveals excellent antibacterial property against both Staphylococcus aureus and Escherichia coli. Most interestingly, PDA/Fe3 O4 nanozymes can be strongly retained in infected sites by an external magnet for localized long-term in vivo CDT and show minimal toxicity to healthy tissues and organs. This work presents an effective strategy to magnetically retain the therapeutic nanozymes in infected sites for highly efficient CDT with good biosafety.

Glutathione-Responsive Chemodynamic Therapy of Manganese(III/IV) Cluster Nanoparticles Enhanced by Electrochemical Stimulation via Oxidative Stress Pathway.

Escalating the level of reactive oxygen species (ROS) in a tumor microenvironment is one of the effective strategies to improve the efficacy of anticancer therapy. In this work, manganese cluster nanoparticles (Mn12) encapsulated with heparin (Mn12-heparin) were developed as a chemodynamic therapeutic agent for cancer treatment by raising ROS levels in tumor cells via cascade reactions. The manganese cluster is a cluster of mixed valence (III/IV) with acetate as the ligand. The cluster is readily subject to reduction by glutathione (GSH) to release Mn(II), which reacts with H2O2 to generate hydroxyl radicals via a Fenton-like pathway. The generation of hydroxyl radicals could be enhanced by the stimulation of an external alternative electric field during which GSH acts as an electron mediator to enhance the release of Mn(II) from the cluster. The relatively high levels of both H2O2 and GSH and the acidic environment in tumor cells strengthen its specificity when the manganese cluster system is employed to suppress or eliminate tumors. Both in vitro and in vivo results suggest that, in addition to the cytotoxicity imposed by the raised ROS level due to the presence of Mn(II) species, the depletion of endogenous GSH leads indirectly to the inhibition of glutathione peroxidase 4 (GPX4), consequently raising the lipid peroxidation (LPO) level to cause ferroptosis. The apoptosis and ferroptosis jointly render the manganese-based agent potent efficacy with tumor-targeting specificity in antitumor treatment under electric stimulation.

2D Piezoelectric Bi2 MoO6 Nanoribbons for GSH-Enhanced Sonodynamic Therapy.

Reducing the scavenging capacity of reactive oxygen species (ROS) and elevating ROS production are two primary goals of developing novel sonosensitizers for sonodynamic therapy (SDT). Hence, ultrathin 2D Bi2 MoO6 -poly(ethylene glycol) nanoribbons (BMO NRs) are designed as piezoelectric sonosensitizers for glutathione (GSH)-enhanced SDT. In cancer cells, BMO NRs can consume endogenous GSH to disrupt redox homeostasis, and the GSH-activated BMO NRs (GBMO) exhibit an oxygen-deficient structure, which can promote the separation of electron-hole pairs, thereby enhancing the efficiency of ROS production in SDT. The ultrathin GBMO NRs are piezoelectric, in which ultrasonic waves introduce mechanical strain to the nanoribbons, resulting in piezoelectric polarization and band tilting, thus accelerating toxic ROS production. The as-synthesized BMO NRs enable excellent computed tomography imaging of tumors and significant tumor suppression in vitro and in vivo. A piezoelectric Bi2 MoO6 sonosensitizer-mediated two-step enhancement SDT process, which is activated by endogenous GSH and amplified by exogenous ultrasound, is proposed. This process not only provides new options for improving SDT but also broadens the application of 2D piezoelectric materials as sonosensitizers in SDT.

W-Doped TiO2 Nanorods for Multimode Tumor Eradication in Osteosarcoma Models under Single Ultrasound Irradiation.

Sonosensitizers play crucial roles in the controlled production of reactive oxygen species (ROS) under ultrasound (US) irradiation with high tissue-penetration depth for noninvasive solid tumor therapy. It is desirable to fabricate structurally simple yet multifunctional sonosensitizers from ultrafine nanoparticles for ROS-based multimode therapy to overcome monomode limitations such as low ROS production yields and endogenous reductive glutathione (GSH) to ROS-based treatment resistance. We report the facile high-temperature solution synthesis of ultrafine W-doped TiO2 (W-TiO2) nanorods for exploration of their sonodynamic, chemodynamic, and GSH-depleting activities in sonodynamic-chemodynamic combination tumor therapy. We found that W5+ and W6+ ions doped in W-TiO2 nanorods play multiple roles in enhancing their ROS production. First, W doping narrows the band gap from 3.2 to 2.3 eV and introduces oxygen and Ti vacancies for enhancing their sonodynamic performance. Second, W5+ doping endows W-TiO2 nanorods with Fenton-like reaction activity to produce •OH from endogenous H2O2 in the tumor. Third, W6+ ions reduce endogenous GSH to glutathione disulfide (GSSG) and, in turn, form W5+ ions that further enhance their chemodynamic activity, which greatly modifies thae oxidation-reduction tumor microenvironment in the tumor. In vivo experiments display the excellent ability of W-TiO2 nanorods for enhanced tumor eradication in human osteosarcoma models under single US irradiation. Importantly, the ultrafine nanorod morphology facilitates rapid excretion from the body, displaying no significant systemic toxicity. Our work suggests that multivalent metal doping in ultrafine nanomaterials is an effective and simple strategy for the introduction of new functions for ROS-based multimode therapy.