Deep Tissue Penetration Research Articles

Near-Infrared Optogenetic Module for Conditional Protein Splicing

Optogenetics has emerged as a powerful tool for spatiotemporal control of biological processes. Near-infrared (NIR) light, with its low phototoxicity and deep tissue penetration, holds particular promise. However, the optogenetic control of polypeptide bond formation has not yet been developed. In this study, we introduce a NIR optogenetic module for conditional protein splicing (CPS) based on the gp41-1 intein. We optimized the module to minimize background signals in the darkness and to maximize the contrast between light and dark conditions. Next, we engineered a NIR CPS gene expression system based on the protein ligation of a transcription factor. We applied the NIR CPS for light-triggered protein cleavage to activate gasdermin D, a pore-forming protein that induces pyroptotic cell death. Our NIR CPS optogenetic module represents a promising tool for controlling molecular processes through covalent protein linkage and cleavage.

Abstract 17025: Investigating Retinal Neurovascular Interactions via Multi-View Light-Sheet Microscopy

Introduction: Vascular retinopathy is a leading cause of preventable blindness in diabetes and retinopathy of prematurity. While most studies focus on individual neural or vascular structures, there remains a knowledge gap in understanding the retinal neurovascular interactions after injury and during repair (Fig. 1A). We seek to demonstrate 3-D neurovascular interactions with a multi-view light-sheet imaging system. Methods and Results: Intact 3-D retinas from 2-month-old mice were fixed and permeabilized followed by the optical clearance (CytoVista). Retinal vessels were stained with biotinylated isolectin B4 for microvasculature and Alexa Fluor 488-conjugated streptavidin (Fig. 1B-C) for labeling, while bipolar cells were stained with anti-PKCα (rabbit host) for neurons and Alexa Fluor Plus 555-conjugated goat anti-rabbit IgG for labeling. Refractive index matching was performed to 1.53. Multi-view light-sheet imaging was performed at 5 different angles followed by 3-D registration and reconstruction. The point spread function (PSF) demonstrates a spatial resolution of less than 5 μm (Fig. 1B, inlet). This enabled 3-D co-registration of the vasculature (Fig. 1D, in green) in relation to the bipolar cells (Fig. 1D, in red) for visualization. Conclusion: We revealed the 3-D neuro-microvascular network providing both deep tissue penetration and high spatial resolution for investigating hyperoxia and diabetes-associated vascular retinopathy.

Amplification of oxidative damage using near-infrared II-mediated photothermal/thermocatalytic effects for periodontitis treatment.

Periodontitis is a biofilm-related disease characterized by damage to the periodontal tissue and the development of systemic diseases. However, treatment of periodontitis remains unsatisfactory, especially with deep-tissue infections. This study describes rationally designed multifunctional photothermocatalytic agents for near-infrared-II light-mediated synergistic antibiofilm treatment, through modification of Lu-Bi2Te3 with Fe3O4 and poly(ethylene glycol)-b-poly(l-arginine) (PEG-b-PArg). Notably, 1064-nm laser irradiation led to photothermal/thermocatalytic effects, resulting in the synergistic generation of reactive oxygen species (ROS) and reactive nitrogen species (RNS) and consequent damage to the biofilm. This treatment was based on the thermoelectric and photothermal conversion properties of Lu-Bi2Te3, the peroxidase-like catalytic capacity of Fe3O4, and the guanidinium polymer, PEG-b-PArg. Oxidative damage to biofilm was further enhanced by H2O2, resulting in the effective elimination of biofilm both in vitro and in vivo. These findings suggest that this synergistic therapeutic strategy is effective for the clinical treatment of periodontitis. STATEMENT OF SIGNIFICANCE: The current treatment for periodontitis involves time-consuming and labor-intensive clinical scaling of the teeth. The present study is the first to assess the efficacy of a photothermal catalyst for periodontitis treatment. This used near-infrared-II light at 1064 nm to induce oxidative damage in the biofilm, resulting in its degradation. The synergistic photothermal/thermoelectric effect produced deep tissue penetration and was well tolerated, and can kill the biofilm formed by periodontitis pathogens up to 5 orders of magnitude, effectively treating the biofilm-induced periodontitis.

An Activatable Prodrug Nanosystem for Ultrasound-Driven Multimodal Tumor Therapy and Metastasis Inhibition.

Ultrasound, featuring deep tissue penetration and noninvasiveness, offers a new opportunity to activate functional materials in a tumor-selective manner. However, very few direct ultrasound-responsive redox systems are applicable under therapeutic ultrasound (1MHz). Herein, the investigations on nanoprodrug of DHE@PEG-SS-DSPE are reported, which exhibit glutathione-activated release of dihydroethidium (DHE) in tumor cells. DHE is stable with good biosafety and is transformed into cytotoxic ethidium to induce DNA damage under medical ultrasound irradiation, accompanied by the generation of reactive oxygen species. Further, DHE@PEG-SS-DSPE could effectively induce ferroptosis through glutathione depletion, lipid peroxide accumulation, and downregulation of glutathione peroxidase 4. In vivo studies confirmed that DHE@PEG-SS-DSPE nanoparticles effectively inhibit both the growth of solid tumors and the expression of metastasis-related proteins in mice, thus effectively inhibiting lung metastasis. This DHE-based prodrug nanosystem could lay a foundation for the design of ultrasound-driven therapeutic agents.

FRET-based two-photon benzo[a] phenothiazinium photosensitizer for fluorescence imaging-guided photodynamic therapy

To overcome the conflict between the long-wavelength excitation and high singlet oxygen quantum yield of photosensitizers, we conjugated a two-photon fluorophore, tetrahydroquinoxaline coumarin (TQ), and an efficient photodynamic therapeutic agent, benzo[a]phenothiazinium (NBS-NH2), through a hexamethylene linker to build a two-photon photosensitizer, TQ-NBS. In TQ-NBS, TQ served as an energy donor and NBS-NH2 acted as an energy acceptor; and TQ-NBS was a Förster resonance energy transfer (FRET) cassette with a 92.8% efficiency. The large two-photon absorption cross-section of TQ allowed photosensitizer TQ-NBS to work in a 900 nm two-photon excitation (TPE) mode, which greatly benefited the deep tissue penetration in PDT treatment. Meanwhile, the excellent phototoxicity and near-infrared fluorescence of NBS-NH2 was kept in TQ-NBS under a TPE mode via a FRET process. Photosensitizer TQ-NBS exhibited a high phototoxic efficacy in living cells and tumor-bearing mice.

Indocyanine Green‐Glycogen Conjugates for Near‐Infrared‐Triggered Photothermal Cancer Therapy

AbstractPhotothermal reagents sensitive to near‐infrared (NIR) light are promising imaging agents and therapeutics for anticancer applications because of the deep tissue penetration of NIR light, allowing for spatiotemporal control over the therapeutic activity, with minimal damage to normal tissues. Herein, a new class of NIR‐sensitive biopolymer‐based nanoparticles is presented by covalently conjugating indocyanine green (ICG) onto the surface of naturally occurring glycogen nanoparticles. The resulting ICG‐glycogen conjugates exhibit a markedly enhanced aqueous stability in comparison to free ICG molecules. Furthermore, an efficient light‐to‐heat conversion is enabled by ICG‐glycogen conjugates, as evidenced by the elevated temperatures of their aqueous solutions upon exposure to NIR light. Critically, ICG‐glycogen conjugates are capable of cell internalization, and under NIR irradiation the effective eradication of breast cancer cells, demonstrating their potential in photothermal therapy for cancer.

Developing a novel benzothiazole-based red-emitting probe for intravital imaging of superoxide anion

Superoxide anion (O2•−), the first generated reactive oxygen species (ROS), is a critical player in cellular signaling network and redox homeostasis. Imaging of O2•−, particularly in vivo, is of concern for further understanding its roles in pathophysiological and pharmacological events. Herein, we designed a novel probe, (E)-4-(5-(2-(benzo[d]thiazol-2-yl)-2-cyanovinyl)furan-2-yl)phenyl trifluoromethane-sulfonate (BFTF), by modifying hydroxyphenyl benzothiazole (a widely used dye scaffold) which includes insertion of both an acrylonitrile unit and a furan ring to extend the total π-conjugation system and to enhance push-pull intramolecular charge transfer process, and utilization of trifluoromethanesulfonate as the response unit. Toward O2•−, the probe features near-infrared fluorescent emission (685 nm), large Stokes shift (135 nm), and deep tissue penetration (300 μm). With its help, we successfully mapped preferential generation of O2•− in HepG2 cells over L02 cells, as well as in A549 over BEAS-2B cells by β-lapachone (an anticancer agent that generates O2•−), and more importantly, visualized overproduction of O2•− in living mice with liver injury induced by acetaminophen (a well-known analgesic and antipyretic drug).

Rational Design of Tunable Near-Infrared Oxazine Probe with Large Stokes Shift for Leucine Aminopeptidase Detection and Imaging.

Near-Infrared (NIR) fluorescence imaging with the advantages of deep tissue penetration and minimum background, has been widely employed and developed in the study of biological applications. However, small Stokes shifts, difficulty in optical tuning, and pH sensitivity are still the major limitations faced by current NIR dyes. To solve these problems, we rationally designed a pH insensitive amino-tunable NIR oxazine fluorophore DQF-NH2 , which exhibited large Stokes shift (125 nm) accompanied with NIR excitation/emission due to the introduction an asymmetrical alternating vibronic feature. By benefiting from the excellent photophysical properties of DQF-NH2 , we have successfully constructed the probe DQF-NH2 -LAP with the ability to detect endogenous LAP. Bioimaging assays demonstrated that DQF-NH2 -LAP can not only effectively detect LAP in living cells, but also was successfully applied to image tumor tissue in vivo. We anticipate that the functionalizable dye DQF-NH2 may be a potential new NIR dye platform with an optically tunable group for the development of future desirable probes for bioimaging.

Near-Infrared-II In Vivo Visualization and Quantitative Tracking of Micro/Nanoplastics in Fish.

As emerging contaminants, micro/nanoplastics (MNPs) are widely present in aquatic environments and are often ingested by aquatic organisms. However, the in vivo trafficking and fate of MNPs remain largely unknown. Here, we developed near-infrared (NIR) aggregated-induced emission (AIE) fluorophore-labeled microplastics (2 μm) and nanoplastics (100 nm) as models of MNPs. This model was based on the NIR-AIE technique with strong emission at the second near-infrared (NIR-IIII) window, which overcomes the interference of autofluorescence and observation artifacts in the detection of commercial fluorescent-labeled particles. Due to its deep tissue penetration and noninvasiveness, the dynamic process of accumulation and transport of MNPs in individuals can be tracked with NIR imaging. We then directly visualized and quantified the uptake and depuration processes of MPs and NPs in zebrafish. The results showed that the MPs and NPs were mainly accumulated in the fish gut, and the distribution was heterogeneous. MPs tended to accumulate more in the fore and mid areas of the gut compared with NPs. Besides, both MPs and NPs could accumulate in large quantities locally in the gut and might cause intestinal obstruction. MNPs accumulated slowly during the initial exposure followed by rapid and sustained accumulation in gut. Based on these kinetic accumulation and depuration, we developed a refined toxicokinetic (TK) model to describe the dynamic changes in the uptake and depuration of MNPs. Overall, this study proposed a MNP model based on the NIR-AIE technique, which provided a reliable tracer technology for the visualization, tracking and quantification of MNPs in vivo.

Nanoparticles – A Booming Drug Delivery System in Chemotherapy

The current study proposed the “Nanoparticles -A Booming Drug Delivery System in Chemotherapy” is a Novel targeted approach which enhances the efficacy of chemotherapeutic agents by reducing the dose-related side effect as well as mortality rate a in patients due to its non-immunogenic, nontoxic nature. Drug bioavailability, drug solubility, drug biodistribution, drug resistance brought on by treatment, and nonspecific toxicity can all be improved with the development of nanoparticle chemotherapeutic drug delivery applications based on nanotechnology. It possesses active as well as passive targeting of tumour cells. Due to this reason, a wide range of chemotherapeutic agents like cisplatin, taxol, doxorubicin, and carboplatin are extensively utilized for treating cancer. Deep tissue penetration of nanoparticles is found to increase the enhanced permeability and retention (EPR) effect. There are some limitations with conventional drug delivery system which is minimized by utilizing nanoparticles as a drug delivery system. The current review has focused on targeted strategies and novel approaches in cancer treatment with nanoparticles.

Near-Infrared Afterglow ONOO--Triggered Nanoparticles for Real-Time Monitoring and Treatment of Early Ischemic Stroke.

Early detection and drug intervention with the appropriate timing and dosage are the main clinical challenges for ischemic stroke (IS) treatment. The conventional therapeutic agents relay fluorescent signals, which require real-time external light excitation, thereby leading to inevitable autofluorescence and poor tissue penetration. Herein, we report endogenous peroxynitrite (ONOO-)-activated BDP-4/Cur-CL NPs that release NIR afterglow signals (λmax 697 nm) for real-time monitoring of the progression of ischemia reperfusion (I/R) brain injury while releasing curcumin for the safe treatment of IS. The BDP-4/Cur-CL NPs exhibited bright NIR afterglow luminescence (maximum 732-fold increase), superb sensitivity (LOD = 82.67 nM), high energy-transfer efficiency (94.6%), deep tissue penetration (20 mm), outstanding antiapoptosis, and anti-inflammatory effects. The activated NIR afterglow signal obtained in mice with middle cerebral artery occlusion (MCAO) showed three functions: (i) the BDP-4/Cur-CL NPs are rapidly activated by endogenous ONOO-, instantly illuminating the lesion area, distinguishing I/R damage from normal areas, which can be successfully used for endogenous ONOO- detection in the early stage of IS; (ii) real-time reporting of in situ generation and dynamic fluctuations of endogenous ONOO- levels in the lesion area, which is of great value in monitoring the evolutionary mechanisms of IS; and (iii) dynamic monitoring of the release of curcumin drug for safe treatment. Indeed, the released curcumin effectively decreased apoptosis, enhanced survival, alleviated neuroinflammation, reduced brain tissue loss, and improved the cognition of MCAO stroke mice. This work is the first example of afterglow luminescence for early diagnosis, real-time reporting, drug tracing, and treatment for IS.

A tumor microenvironment responsive titanium-based nanosonosensitizer for combination sonodynamic-immunotherapy with calcium ion overload

Ultrasound (US)-triggered sonodynamic therapy (SDT) has become a promising method and has attracted considerable attention for cancer therapy due to its high accuracy, deep tissue penetration, and few side effects. However, the therapeutic efficacy of SDT is limited by the hypoxic tumor microenvironment (TME) and the low quantum yield of sonosensitizers. Immunogenic cell death (ICD) induced by calcium ion overload has attracted widespread attention because it can trigger antitumor immune responses and reconstitute tumor immune microenvironment (TIM).In this study, a TME-responsive CaCO3@Pt-TiO2 nanocomposite (CaPT) was engineered, which combined platinum (Pt) nanoparticles and CaCO3 nanoparticles to amplify oxidative stress in cancer cells for cancer sonodynamic immunotherapy. The Pt nanoparticles not only enhanced the reactive oxygen species (ROS) quantum yield of TiO2 during SDT but also catalyzed the decomposition of H2O2 to overcome tumor hypoxia. The CaCO3@Pt-TiO2 nanocomposite responded to the TME and released a large amount of Ca2+, which amplified oxidative stress and triggered robust ICD activation. This process greatly contributed to inflammatory cell infiltration and shifted the TIM from cold to hot. The experimental results confirmed and demonstrated the synergistic therapeutic effects of sonodynamic-immunotherapy in vitro and in vivo. The antitumor mechanisms of sonodynamic immunotherapy were demonstrated by RNA sequencing. This study paves the way for Ti-based nanosensitizers, providing a novel strategy for sonodynamic-cooperated immunotherapy.

Synthesis and sonodynamic performance of spinel ferrites

Sonodynamic therapy has emerged as one of the most valuable treatment methods with low side effects and deep tissue penetration. As a representative inorganic sonosensitizer, spinel-type ferrites have many advantages such as high reactive oxygen species (ROS) production efficiency and stability. However, current studies have not explored how metal doping impacts the sonodynamic effect of spinel ferrites. Herein, we synthesized three kinds of spinel ferrite nanoparticles (ZnFe2O4, CoFe2O4, and MgFe2O4) and systematically investigated their sonodynamic performance. It is shown that three spinel ferrites are capable of producing singlet oxygen under ultrasound irradiation. Hydrogen peroxide further enhances the production of singlet oxygen, increasing the production rate by 1.05–2.31 times. The simultaneous existence of Co2+/Co3+ and Fe2+/Fe3+ redox couples in CoFe2O4 could convert more efficiently hydrogen peroxide into oxygen through catalytic reaction, providing more oxygen to promote singlet oxygen production. Thus, CoFe2O4 exhibits higher production efficiency than ZnFe2O4 and MgFe2O4. In vitro experiments demonstrate the good biocompatibility of CoFe2O4 nanoparticles and high ability to effectively kill cancer cells under ultrasound irradiation. In addition, CoFe2O4 shows good therapeutic efficacy in animal models of colon cancer. These findings highlight the great potential of ferrite sonosensitizer for cancer treatment.

Targeted Near-Infrared Fluorescence Imaging of Liver Cancer using Dual-Peptide-Functionalized Albumin Particles.

Fluorescence imaging is an emerging strategy for preoperative diagnosis and intraoperative resection. In particular, owing to their outstanding spatial resolution and deep-tissue penetration, imaging agents in the near-infrared (NIR)-II window (1000-1700 nm) have received intensive interest for biomedical applications. However, NIR II-based imaging agents for targeted visualization of hepatocellular carcinoma (HCC) have barely been barely developed. Here, we report the construction of structurally uniform, biocompatible human serum albumin (HSA)-based particles orthogonally modified with two functional peptides as a carrier for the delivery of NIR-II imaging agents to HCC cell-derived solid tumor in vivo. Cysteine conjugation combined with host-guest chemistry enables the orthogonal introduction of two functionally independent peptides to HSA-based nanoparticles. One of these peptides targets glypican-3 (GPC-3), a specific biomarker of HCC, and the other facilitates the escape of the nanoparticles from macrophagic phagocytosis. Series of cellular and in vivo assays were carried out to demonstrate the efficacy of the dual-peptide-functionalized HSA nanoparticles for targeted NIR-II fluorescence imaging of HCC.

Near-Infrared Photothermal Manipulates Cellular Excitability and Animal Behavior in Caenorhabditis elegans.

Near-infrared (NIR) photothermal manipulation has emerged as a promising and noninvasive technology for neuroscience research and disease therapy for its deep tissue penetration. NIR stimulated techniques have been used to modulate neural activity. However, due to the lack of suitable in vivo control systems, most studies are limited to the cellular level. Here, a NIR photothermal technique is developed to modulate cellular excitability and animal behaviors in Caenorhabditis elegans in vivo via the thermosensitive transient receptor potential vanilloid 1 (TRPV1) channel with an FDA-approved photothermal agent indocyanine green (ICG). Upon NIR stimuli, exogenous expression of TRPV1 in AFD sensory neurons causes Ca2+ influx, leading to increased neural excitability and reversal behaviors, in the presence of ICG. The GABAergic D-class motor neurons can also be activated by NIR irradiation, resulting in slower thrashing behaviors. Moreover, the photothermal manipulation is successfully applied in different types of muscle cells (striated muscles and nonstriated muscles), enhancing muscular excitability, causing muscle contractions and behavior changes in vivo. Altogether, this study demonstrates a noninvasive method to precisely regulate the excitability of different types of cells and related behaviors in vivo by NIR photothermal manipulation, which may be applied in mammals and clinical therapy.

Polarization-sensitive Two-photon Microscopy for a Label-free Amyloid Structural Characterization.

Compared to its one-photon counterpart, two-photon excitation is beneficial for bioimaging experiments because of its lower phototoxicity, deeper tissue penetration, efficient operation in densely packed systems, and reduced angular photoselection of fluorophores. Thus, the introduction of polarization analysis in two-photon fluorescence microscopy (2PFM) provides a more precise determination of molecular organization in a sample compared to standard imaging methods based on linear optical processes. In this work, we focus on polarization-sensitive 2PFM (ps-2PFM) and its application in the determination of molecular ordering within complex bio-structures-amyloid spherulites. Neurodegenerative diseases such as Alzheimer's or Parkinson's are often diagnosed through the detection of amyloids-protein aggregates formed due to an impaired protein misfolding process. Exploring their structure leads to a better understanding of their creation pathway and consequently, to developing more sensitive diagnostic methods. This paper presents the ps-2PFM adapted for the determination of local fibril ordering inside the bovine insulin spherulites and spherical amyloidogenic protein aggregates. Moreover, we prove that the proposed technique can resolve the three-dimensional organization of fibrils inside the spherulite.

Biomimetic liposomes as a pH/ROS cascade-responsive nanoagent with high selectivity for breast carcinoma in photothermal therapy

With the recent advances in nanomaterials, photothermal therapies (PTT) enabled by deep-tissue penetration with novel nanoagents have offered a new hope for treating cancers. Nevertheless, PTT applications in conventional approaches remain translationally restricted due to their non-specificity and collateral pro-inflammatory effects during implementation. To address these drawbacks, we here report Lipo-PTO as the prototype of a new class of liposome-functionalized photothermal theragnostic agents, which can eradicate cancer cells in vivo with high selectivity. Lipo-PTO is activated by low pH and superoxide (O2•–) to specifically deliver and convert preloaded cyanine molecules to release Cy-NH in the tumour microenvironments, eventually enabling the activation of photothermal properties to induce tumour ablation. In further studies on Lipo-PTO for treating breast cancer (BC) in mouse models, Lipo-PTO usage was characterized by a high degree of biocompatibility, optimal retention time pharmacokinetics and tumour regression with minimal collateral damages. This study here thus illustrates an innovative strategy to target intrinsic attributes of tumour microenvironments for improved therapeutic precision and efficacy in PTT applications.

NIR light-facilitated bone tissue engineering.

In the last decades, near-infrared (NIR) light has attracted considerable attention due to its unique properties and numerous potential applications in bioimaging and disease treatment. Bone tissue engineering for bone regeneration with the help of biomaterials is currently an effective means of treating bone defects. As a controlled light source with deeper tissue penetration, NIR light can provide real-time feedback of key information on bone regeneration in vivo utilizing fluorescence imaging and be used for bone disease treatment. This review provides a comprehensive overview of NIR light-facilitated bone tissue engineering, from the introduction of NIR probes as well as NIR light-responsive materials, and the visualization of bone regeneration to the treatment of bone-related diseases. Furthermore, the existing challenges and future development directions of NIR light-based bone tissue engineering are also discussed. This article is categorized under: Diagnostic Tools > In Vivo Nanodiagnostics and Imaging Implantable Materials and Surgical Technologies > Nanotechnology in Tissue Repair and Replacement.

Medical Applications and Advancement of Near Infrared Photosensitive Indocyanine Green Molecules.

Indocyanine green (ICG) is an important kind of near infrared (NIR) photosensitive molecules for PTT/PDT therapy as well as imaging. When exposed to NIR light, ICG can produce reactive oxygen species (ROS), which can kill cancer cells and pathogenic bacteria. Moreover, the absorbed light can also be converted into heat by ICG molecules to eliminate cancer cells. In addition, it performs exceptionally well in optical imaging-guided tumor therapy and antimicrobial therapy due to its deeper tissue penetration and low photobleaching properties in the near-infrared region compared to other dyes. In order to solve the problems of water and optical stability and multi-function problem of ICG molecules, composite nanomaterials based on ICG have been designed and widely used, especially in the fields of tumors and sterilization. So far, ICG molecules and their composite materials have become one of the most famous infrared sensitive materials. However, there have been no corresponding review articles focused on ICG molecules. In this review, the molecular structure and properties of ICG, composite material design, and near-infrared light- triggered anti-tumor, and antibacterial, and clinical applications are reviewed in detail, which of great significance for related research.

Doping Engineering to Modulate Lattice and Electronic Structure for Enhanced Piezocatalytic Therapy and Ferroptosis.

Piezocatalytic therapy, which generates reactive oxygen species (ROS) under mechanical force, has garnered extensive attention for its use in cancer therapy owing to its deep tissue penetration depth and less O2 -dependence. However, the piezocatalytic therapeutic efficiency is limited owing to the poor piezoresponse, low separation of electron-hole pairs, and complicated tumor microenvironment (TME). Herein, a biodegradable, porous Mn-doped ZnO (Mn-ZnO)nanocluster with enhanced piezoelectric effect is constructed via doping engineering. Mn-doping not only induces lattice distortion to increase polarization but also creates rich oxygen vacancies (OV ) for suppressing the recombination of electron-hole pairs, leading to high-efficiency generation of ROS under ultrasound irradiation. Moreover, Mn-doped ZnO shows TME-responsive multienzyme-mimicking activityandglutathione (GSH) depletion ability owing to the mixed valence of Mn (II/III), further aggravating oxidative stress. Density functional theory calculations show that Mn-doping can improve the piezocatalytic performance and enzyme activity of Mn-ZnO due to the presence of OV . Benefiting from the boosting of ROS generation and GSH depletion ability, Mn-ZnO can significantly accelerate the accumulation of lipid peroxide and inactivate glutathione peroxidase 4 (GPX4) to induce ferroptosis. The work may provide new guidance for exploring novel piezoelectric sonosensitizers for tumor therapy.