INTERNATIONAL JOURNAL OF LATEST TECHNOLOGY IN ENGINEERING,

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ISSN 2278-2540 | DOI: 10.51583/IJLTEMAS | Volume XIV, Issue XI, November 2025

Enzyme-Mimic Nanomaterials in Biomedicine: Catalytic Mechanisms,

Functional Platforms, and Translational Potential.

Ashlesh D Alva^1*, Praveen BM², Riya Nair³, Shraddha Rani M⁴, Vikhyath Bangera S⁵, Pavana

Krishnamoorthy⁶

¹Department of Medical Physics, Srinivas Institute of Allied Health Sciences, Srinivas University,

Mangalore, India

²Department of Chemistry, Srinivas Institute of Engineering and Technology, Srinivas University,

Mangalore, India.

³⁴⁵Srinivas Institute of Allied Health Science, Srinivas University, Mukka, India.

⁶Department of Physiology, Srinivas Institute of Allied Health Sciences, Srinivas University, Mangalore,

India.

DOI: https://doi.org/10.51583/IJ L T EMAS.2025.1411000038

Received: 18 November 2025; Accepted: 27 November 2025; Published: 05 December 2025

ABSTRACT

The emergence of enzyme-mimicking nanomaterials (nanozymes) represents a transformative development in

biomedical research, offering catalytic versatility, enhanced stability, and cost-effective scalability compared to

natural enzymes. This review consolidates current advances in the design, synthesis, and biomedical deployment

of nanozymes, particularly those based on noble metals, metal oxides, and functional nanocomposites. A

systematic comparative analysis was conducted across diverse studies to elucidate the catalytic mechanisms,

structural-functional relationships, and environmental adaptability of these nanomaterials.

The literature was critically evaluated with emphasis on the physicochemical factors governing nanozyme

activity such as particle size, surface ligands, and crystal facets and their modulation strategies. Trends indicate

that noble metal nanoparticles (e.g., Au, Pt, Pd) and metal oxides (e.g., Fe₃O₄, CeO₂, V₂O₅) exhibit peroxidase,

oxidase, and catalase-like functions, with performance often surpassing their biological counterparts under

physiological conditions. Moreover, 2D nanomaterials and Prussian blue analogues demonstrate significant

promise as tunable catalytic platforms.

Functionally, nanozymes are proving integral in areas such as tumor theranostics, antibacterial therapy,

antioxidation, and bioorthogonal catalysis. Applications range from in situ ROS modulation for cancer treatment

to programmable catalysis in cellular imaging and drug activation. Despite these advances, challenges remain in

enhancing substrate specificity, minimizing cytotoxicity, and fully elucidating mechanistic pathways.

In conclusion, nanozymes hold substantial potential to reshape therapeutic and diagnostic modalities. Future

research must focus on integrating simulation-driven design, expanding the scope of enzyme mimicry, and

ensuring biosafety in complex biological environments. Addressing these gaps could accelerate the clinical

translation of nanozyme-based technologies, establishing them as cornerstone tools in next-generation

biomedical applications.

Keywords: Nanozymes, Catalysis, Theranostics, Antioxidants, Nanomedicine

INTRODUCTION

In recent years, the field of nanobiotechnology has witnessed a paradigm shift with the advent of enzyme-

mimicking nanomaterials, commonly referred to as nanozymes (Pant et al., 2024). These synthetic

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nanostructures, capable of replicating the catalytic functions of natural enzymes, have garnered considerable

attention due to their exceptional stability, tunable activity, and cost-efficient production. Unlike their biological

counterparts, which often suffer from structural fragility, limited reusability, and environmental sensitivity,

nanozymes offer robust catalytic performance under a broad range of physiological and environmental

conditions. This has positioned them at the forefront of biomedical research, where there is a growing demand

for smart, adaptive, and multifunctional materials capable of addressing complex diagnostic and therapeutic

challenges (Nagendran et al., 2024).

The significance of nanozymes extends far beyond their chemical novelty. Their integration into biomedical

applications ranging from tumor imaging and targeted therapy to antimicrobial treatment, antioxidant

intervention (Tao et al., 2020; Zhong et al., 2023), and in vivo bioorthogonal catalysis marks a critical

advancement in translational nanomedicine. These materials are not only enhancing the precision and efficiency

of existing clinical protocols but also enabling entirely new therapeutic paradigms, such as hypoxia-tolerant

photodynamic therapy and stimuli-responsive drug activation within the tumor microenvironment. Moreover,

their potential in combating antibiotic resistant pathogens and mitigating oxidative stress opens avenues for

addressing pressing public health concerns (Li et al., 2025).

Despite these promising developments, a number of critical knowledge gaps persist. Current literature often

presents fragmented insights into the structure activity relationships, catalytic mechanisms, and regulatory

factors that govern nanozyme behaviour. In particular, the heterogeneity in material composition, particle

morphology, and surface chemistry across studies complicates the establishment of universal design principles.

Furthermore, while several proof-of-concept studies have demonstrated the feasibility of nanozyme based

systems, their long-term biocompatibility, in vivo kinetics, and molecular-level interaction pathways remain

incompletely understood (Aldrich et al., 2023).

This review is therefore both timely and necessary. It seeks to provide a consolidated and critical account of the

diverse classes of nanozymes, with particular emphasis on their synthetic strategies, catalytic performance

modulation, and emerging roles in biomedicine. By integrating findings from recent investigations including

noble metal nanoparticles, metal oxides, 2D materials, and Prussian blue analogues this article aims to delineate

the fundamental principles guiding nanozyme functionality and to highlight translational opportunities and

unresolved challenges.

The objective of this review is to synthesize current advances in enzyme-mimicking nanomaterials and assess

their biomedical applications through a comprehensive, mechanistically informed lens, thereby offering

direction for future research and development in this rapidly evolving domain.

Objectives

1. To critically synthesize the recent advancements in the design, synthesis, and catalytic behaviour of

enzyme-mimic nanomaterials, with emphasis on their structure function relationships and physicochemical

regulation.

2. To evaluate the biomedical relevance of nanozymes across domains such as tumor theranostics,

antibacterial therapy, antioxidation, and bioorthogonal catalysis, highlighting their multifunctionality and

advantages over natural enzymes.

3. To identify existing challenges such as toxicity, specificity, and in vivo biostability and propose future

directions that integrate interdisciplinary strategies for enhancing the clinical translatability of nanozyme-

based platforms.

Enzyme Mimic Nanomaterials

Enzyme-mimic nanomaterials, or nanozymes, are a class of synthetic nanostructures engineered to replicate the

catalytic functions of natural enzymes. These materials exhibit activities such as oxidase, peroxidase, catalase,

and superoxide dismutase, yet they surpass biological enzymes in terms of stability, cost-efficiency, and

environmental tolerance. Their unique physicochemical properties derived from nanoscale dimensions, surface

tunability, and multifunctional architectures enable applications across diagnostics, therapeutics, and biosensing.

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This section critically examines two major categories of nanozymes: noble metal nanoparticles and metal oxide-

based systems, emphasizing their mechanisms, biomedical relevance, and translational challenges (Lai et al.,

2025; Xu et al., 2025; S. Zhang et al., 2025).

Noble Metal Nanoparticles with Catalytic Behaviours

Noble metal nanoparticles (NMNPs), particularly those composed of gold (Au), silver (Ag), platinum (Pt), and

palladium (Pd), have emerged as a pivotal class of nanozymes owing to their superior catalytic versatility, surface

modifiability, and electron transfer dynamics. Their intrinsic enzyme-like activities have been harnessed for a

range of biomedical applications including tumor diagnostics, pathogen detection, and therapeutic modulation

of reactive oxygen species (ROS) (Kashtiaray et al., 2025a; Malekzad et al., 2016).

Catalytic Functionality and Mechanisms

Among NMNPs, Au nanoparticles (AuNPs) were among the first reported to exhibit oxidase-mimetic behaviour,

notably in glucose oxidation. The mechanistic basis lies in their ability to adsorb electron-rich substrates and

mediate oxygen activation via surface complexes, facilitating redox transformations similar to those of glucose

oxidase (Sen et al., 2024a). This process is highly pH-dependent and regulated by the nanoparticle’s surface

chemistry. Notably, DNA-functionalized AuNPs demonstrated stereoselective catalysis, exhibiting

enantioselective oxidation of glucose isomers a functionality unattainable by most natural enzymes.

Ag nanoparticles similarly demonstrate peroxidase and oxidase like activity. Electron spin resonance

spectroscopy has revealed their ROS generating capabilities during hydrogen peroxide decomposition,

implicating AgNPs in potential antimicrobial and biosensing functions (He et al., 2011). Their catalytic output

is highly responsive to environmental parameters such as pH and ionic species. For instance, the presence of

Hg²⁺ ions dramatically enhance their catalytic response in chromogenic assays, a feature exploited in metal ion

detection platforms.

Pt nanoparticles, known for their catalytic robustness, mimic peroxidase activity effectively. Their reactivity can

be fine-tuned through control over particle morphology, ligand chemistry, and size. Studies have shown that

ultrasmall Pt nanocubes exhibit strong thermal and pH tolerance, making them suitable for physiological

applications. Moreover, DNA templating strategies allow size and activity modulation, providing an added layer

of control over catalytic kinetics and substrate affinity (Abdelhamid et al., 2020; Sen et al., 2024b).

Pd nanoparticles extend this paradigm by offering tunable oxidase and catalase-like activities. The catalytic

behaviour is facet-dependent; Pd (111) surfaces exhibit superior redox capabilities compared to higher-energy

surfaces. Ligand modification further alters their solubility and interaction profiles, expanding their usability in

aqueous biological media. Pd-based nanozymes have been deployed in bioassays for cancer biomarkers and even

demonstrated dual functionalities acting both as sensors and therapeutic agents (R. Zhang et al., 2025a).

Biomedical Relevance and Translational Insights

The utility of noble metal nanozymes in medicine hinges on their ability to function under biologically relevant

conditions. For instance, Au@Pt core-shell nanoparticles have been integrated into ELISA platforms with

performance metrics rivaling traditional horseradish peroxidase (HRP). Furthermore, programmable Au

assemblies, directed by DNA nanotechnology, have facilitated single-particle catalytic imaging a significant step

toward real-time, intracellular biochemical monitoring (Shamsabadi et al., 2024).

In antimicrobial strategies, AgNPs exhibit selective bactericidal action when modulated with external ions or

surface ligands, showing promise in infection control and biosafety applications. Pt nanozymes, on the other

hand, contribute to glucose monitoring in diabetic diagnostics and have been adapted for blood-based clinical

assays (Sen et al., 2024c).

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Synthesis Strategies and Challenges

Despite these advancements, the synthesis of NMNPs with consistent size, shape, and surface chemistry remains

a bottleneck. Wet-chemical routes often lead to heterogeneous populations, which can result in variable catalytic

activity. Advances in biomolecule-assisted synthesis particularly using DNA or peptides have improved control

over morphology and biocompatibility. However, scalability and batch reproducibility remain significant

challenges (Desai et al., 2025).

Another major consideration is the biocompatibility and potential toxicity of noble metals upon long-term

exposure. While surface modifications (e.g., PEGylation) mitigate immunogenicity and improve circulation

time, systematic in vivo studies are still required to understand biodistribution, clearance, and chronic effects

(Santhanakrishnan et al., 2024).

Current Trends and Future Directions

The field is now pivoting toward hybrid nanozyme systems where NMNPs are embedded within or conjugated

to responsive platforms, such as hydrogels, photosensitive polymers, or smart vesicles. Such systems allow

spatiotemporal control of enzymatic activity opening up avenues for stimuli-triggered therapeutic interventions.

Moreover, efforts to combine catalytic and plasmonic properties are pushing the boundaries of integrated

diagnostics and therapy, particularly in the realm of cancer theranostics (Cao et al., 2023).

In summary, noble metal nanoparticles offer a unique platform for enzyme mimicry, distinguished by their

robustness, tunability, and multifunctionality. While their biomedical potential is profound, further refinement

in design strategies and safety evaluation is essential to transition these materials from laboratory to clinic (Bayda

et al., 2020; Truong et al., 2024).

Metal Oxide Nanoparticles with Catalytic Behaviours

Metal oxide nanoparticles constitute another major class of nanozymes with remarkable enzyme-like functions,

offering catalytic activities that span peroxidase, catalase, superoxide dismutase (SOD), and glutathione

peroxidase (GPx) mimetics. Their versatile redox properties, chemical stability, and scalable synthesis have

positioned them as compelling alternatives to natural enzymes, particularly in biomedical contexts requiring

sustained functionality under physiological stress or complex biofluid environments (X. Wang, 2022; R. Zhang

et al., 2022).

Catalytic Mechanisms and Functional Diversity

The prototypical metal oxide nanozyme is magnetite (Fe₃O₄), first reported to exhibit intrinsic peroxidase-like

activity under mild conditions. Its catalytic behaviour stems from the Fenton-like reaction between Fe²⁺/Fe³⁺

redox pairs and hydrogen peroxide (H₂O₂), generating reactive hydroxyl radicals. Notably, the catalytic output

is particle size-dependent, with smaller nanoparticles displaying higher activity due to increased surface-to-

volume ratio and surface defect density. Beyond spherical forms, morphology-specific variants such as truncated

octahedrons have shown superior catalytic performance due to the enhanced exposure of active crystal facets (L.

Gao et al., 2007a).

Fe₃O₄-based nanozymes have been applied in biosensing platforms, immunoassays, and choline detection

systems. Their ferromagnetic properties further facilitate recovery and reuse, enhancing their appeal for

diagnostic and therapeutic devices. However, their activity is often nonspecific and can be influenced by surface

fouling in biological media, highlighting the need for surface functionalization or protective coatings (L. Gao et

al., 2007a).

Cerium oxide nanoparticles (CeO₂), or nanoceria, represent a distinctive category of SOD- and catalase-

mimicking nanozymes. Their unique redox-switching capability between Ce³⁺ and Ce⁴⁺ oxidation states allow

them to scavenge superoxide radicals and decompose H₂O₂, maintaining redox homeostasis in cells. This

regenerative redox cycling enables continuous catalytic activity, mimicking the function of antioxidant enzymes.

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However, Ce³⁺ content on the surface is highly sensitive to synthetic conditions, and improper surface oxidation

can severely impair catalytic performance.

PEGylation of nanoceria has emerged as a successful strategy to enhance their dispersion in aqueous media,

improve biocompatibility, and preserve Ce³⁺ surface content. Moreover, nanoceria’s catalytic activity can be

regulated through interactions with biomolecules such as DNA, which when adsorbed, can inhibit surface redox

reactions introducing possibilities for smart regulation in biosensing or targeted therapy (Abokyi et al., 2020).

Manganese dioxide (MnO₂) and its derivatives (e.g., Mn₃O₄) offer another line of redox-active nanozymes. MnO₂

nanoparticles have demonstrated dual oxidase and peroxidase activities and have been deployed in colorimetric

immunoassays, wound disinfection, and antioxidant therapies. Their GPx-like activity has been used to modulate

oxidative stress, with applications in neuroprotection and anti-inflammatory treatments. Like Fe₃O₄, MnO₂

nanozyme activity is structure-dependent, and biological performance improves with protein templating or

surface engineering, as seen in bovine serum albumin-stabilized formulations.

Vanadium pentoxide (V₂O₅) nanowires also stand out for their GPx-like activity. Their catalytic efficiency

depends not only on their size and aspect ratio but also on the dominant exposed crystal facet, with the {010}

surface showing the highest reactivity. Density functional theory (DFT) simulations have elucidated the facet-

dependent electron transfer processes, aligning experimental findings with theoretical predictions. V₂O₅

nanozymes have shown effectiveness in intracellular ROS regulation, offering therapeutic benefits in

degenerative and inflammatory diseases (Alrobaian, 2023).

Biomedical Applications and Functional Integration

Metal oxide nanozymes are finding expanding roles in tumor theranostics, antimicrobial treatments, and

oxidative stress management. Their ability to catalyze ROS production or scavenging in situ enables dual

functionality: pro-oxidant strategies for cancer therapy and antioxidant strategies for cellular protection.

In tumor applications, Fe₃O₄ and MnO₂ nanozymes have been employed for enhanced imaging and targeted

therapy. When conjugated with tumor-targeting ligands or embedded in stimuli-responsive platforms, they can

amplify oxidative stress within the tumor microenvironment, selectively inducing apoptosis. Simultaneously,

their inherent MRI contrast-enhancing properties or colorimetric responses facilitate real-time monitoring

(Mohapatra & Park, 2023).

For anti-bacterial applications, nanozymes such as MnO₂ and CeO₂ demonstrate high catalytic activity in

generating bactericidal species (e.g., hydroxyl radicals) or disrupting bacterial redox balance. Importantly, these

systems maintain activity in biofilm-associated infections, where conventional antibiotics often fail. Recent

designs incorporate charge-switchable or light-responsive elements to modulate selectivity toward Gram-

positive or Gram-negative bacteria.

Antioxidant therapies also benefit from metal oxide nanozymes. Mn₃O₄ and V₂O₅ nanoparticles have been shown

to attenuate oxidative damage to DNA, proteins, and lipids, supporting their role in combating oxidative-stress-

related pathologies such as neurodegeneration, cardiovascular disease, and aging-related dysfunction (Jiang et

al., 2022).

Limitations and Design Challenges

Despite significant progress, several issues temper the clinical translation of metal oxide nanozymes. First, their

catalytic specificity remains lower than that of natural enzymes, limiting their utility in highly selective

biochemical processes. Second, concerns persist regarding their long-term accumulation and potential toxicity,

particularly for materials like V₂O₅ and Mn-based systems. Although surface modifications (e.g., PEG,

biomolecule coatings) improve compatibility, comprehensive toxicological assessments remain limited (Chen et

al., 2013; Qu et al., 2014).

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Another challenge lies in standardizing activity assays. Disparities in experimental conditions such as pH, buffer

composition, and substrate concentrations complicate direct comparisons across studies. Moreover, the impact

of protein corona formation in vivo on catalytic activity is underexplored and could significantly alter

functionality (Liu et al., 2021).

Emerging Directions

Emerging trends in metal oxide nanozyme research include the development of hybrid platforms that integrate

multiple enzymatic activities (e.g., SOD + GPx + catalase), enabling synergistic effects in complex oxidative

environments. Molecular imprinting techniques and bio-inspired coatings are being investigated to enhance

substrate specificity and reduce off-target effects. Simultaneously, computational modelling and machine

learning are aiding in predictive design, allowing researchers to tailor nanoparticle composition and morphology

for optimized catalytic profiles.

In conclusion, metal oxide-based nanozymes offer a rich landscape of catalytic functionalities with substantial

potential in biomedical science. Their adaptability, tunability, and multifunctionality continue to drive

innovation, though further refinement in biocompatibility, specificity, and regulatory understanding will be

essential for their successful integration into clinical practice (Z. Wang et al., 2025).

Other Nanomaterials with Catalytic Behaviours

Beyond noble metals and metal oxides, a diverse range of nanomaterials including carbon-based nanostructures,

two-dimensional (2D) materials, Prussian blue analogues, and metal–organic frameworks (MOFs) have been

identified as potent enzyme mimics. These materials expand the functional landscape of nanozymes by offering

novel surface chemistries, structural tunability, and unique physicochemical interactions with biological

substrates.

Graphene oxide (GO), a widely studied carbon-based nanomaterial, demonstrates intrinsic peroxidase-like

activity largely attributed to its surface carboxyl and carbonyl groups. These functionalities create catalytic sites

capable of facilitating hydrogen peroxide decomposition and substrate oxidation in colorimetric assays. Unlike

transition metal-based nanozymes, the catalytic mechanisms in carbon materials are often governed by electron

transfer processes localized at defect sites or functional groups, enabling reaction pathways distinct from metal-

centered catalysis. This difference allows for broader environmental adaptability and pH tolerance, which are

valuable for biosensing and diagnostic platforms (Yusuf et al., 2022).

Similarly, other 2D nanomaterials like molybdenum disulfide (MoS₂) have gained attention due to their layered

structure and ability to catalyze ROS generation. MoS₂ exhibits peroxidase-like behaviour that can be exploited

for photothermal antimicrobial therapies and oxidative stress modulation. Its surface can also be functionalized

with polymers or biomolecules to enhance specificity and biocompatibility.

Prussian blue nanoparticles (PBNPs), known for their multienzyme mimicry particularly peroxidase, catalase,

and SOD-like activities exhibit strong redox activity across a broad pH range. Their iron-cyanide framework

facilitates electron relay mechanisms, enabling efficient ROS scavenging and oxidative stress management in

biomedical applications. Unlike many other nanozymes, PBNPs demonstrate exceptional catalytic efficiency

and low toxicity, positioning them as promising agents for inflammation therapy and biosensing (Gunatilake et

al., 2021).

Despite their advantages, challenges persist. Carbon-based and MOF nanozymes often suffer from limited

catalytic turnover rates compared to metal-based counterparts. Additionally, their catalytic activity may be highly

sensitive to structural defects, surface oxidation, or biomolecule adsorption, complicating reproducibility.

Nevertheless, innovations such as ligand-directed assembly, hybrid nanostructures, and computational design

are rapidly improving their functional performance (Gunatilake et al., 2023).

In sum, these alternative nanomaterials introduce new catalytic mechanisms and application modalities into the

enzyme-mimetic domain. Their structural diversity and functional versatility present powerful opportunities for

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developing next-generation nanozymes tailored for complex biomedical and environmental systems (Gunatilake

et al., 2025).

Biomedical Applications of Enzyme Mimic Nanomaterials

The emergence of nanozymes as synthetic enzyme mimics has ushered in a transformative era in biomedical

science. Their catalytic adaptability, physicochemical stability, and biofunctionality under physiological

conditions make them attractive candidates for therapeutic and diagnostic applications. This section explores the

major biomedical domains where nanozymes have shown pronounced utility.

Tumor Theranostics

Enzyme-mimicking nanomaterials have become pivotal agents in cancer theranostics, offering a unique

convergence of diagnostic imaging and therapeutic efficacy. Central to their functionality is the ability to

modulate the tumor microenvironment (TME) through catalytic generation of reactive oxygen species (ROS),

particularly via peroxidase or oxidase-like reactions that exploit elevated endogenous hydrogen peroxide levels

within tumours. This catalytic ROS amplification induces oxidative stress selectively in malignant cells, leading

to apoptosis without harming adjacent healthy tissue.

Beyond cytotoxicity, nanozymes are employed to reverse hypoxia, a common feature of solid tumours that

impairs conventional treatments. Catalase-mimicking nanomaterials decompose H₂O₂ into oxygen, thereby

enhancing photodynamic therapy (PDT) and radiotherapy outcomes. Some nanozymes are further functionalized

with photosensitizers or chemotherapeutic agents to enable synergistic therapy leveraging catalytic oxygen

production to potentiate drug efficacy (R. Wang et al., 2025).

Nanozymes have also shown remarkable potential in tumor imaging. Magnetic metal oxide nanozymes (e.g.,

Fe₃O₄) provide contrast in magnetic resonance imaging (MRI), while plasmonic metal-based nanozymes (e.g.,

Au@Pt) enable high-resolution optical imaging. Their integration into stimuli-responsive platforms allows

targeted release and catalytic activation only within the TME, enhancing specificity and minimizing systemic

toxicity.

Collectively, nanozymes serve as multifunctional agents capable of targeted delivery, catalytic activation, and

image-guided therapy offering a modular and intelligent alternative to traditional single-function cancer

treatments (Kashtiaray et al., 2025b).

Anti-bacteria

Nanozymes have emerged as innovative antimicrobial agents capable of addressing bacterial resistance and

biofilm-associated infections through catalytic generation of bactericidal species. Their primary mechanism

involves peroxidase-like activity, wherein nanozymes catalyze the decomposition of H₂O₂ into highly reactive

hydroxyl radicals (•OH), effectively disrupting bacterial membranes, proteins, and genetic material.

Unlike conventional antibiotics, nanozymes do not rely on specific metabolic pathways, thereby minimizing the

risk of resistance development. Their activity can be modulated by environmental pH, metal ions, or external

stimuli (e.g., light), enabling context-specific bacterial killing. This is particularly advantageous in

heterogeneous infections where Gram-positive and Gram-negative bacteria coexist.

Recent designs incorporate charge-switchable nanozymes that preferentially interact with specific bacterial

surfaces based on cell wall composition. Light-activated nanozymes further enhance selectivity, enabling

programmable targeting through differential surface charge modulation and controlled ROS generation (Javed

et al., 2025).

In addition to planktonic bacteria, nanozymes effectively target biofilms complex microbial communities

notoriously resistant to antibiotics. Their catalytic activity facilitates biofilm penetration and oxidative

degradation of the extracellular matrix, exposing embedded bacteria to immune clearance or secondary

therapeutics.

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The biocompatibility and durability of nanozymes make them suitable for wound healing applications, surgical

coatings, and antimicrobial textiles. As resistance to conventional agents rises, nanozymes provide a robust,

tunable, and mechanistically distinct platform for antibacterial intervention (Ge et al., 2024).

Antioxidation

In biological systems, maintaining redox homeostasis is vital for cellular integrity. Nanozymes capable of

mimicking antioxidant enzymes such as superoxide dismutase (SOD), catalase, and peroxidase have

demonstrated promise in mitigating oxidative stress associated with numerous pathologies, including

neurodegeneration, cardiovascular disorders, and chronic inflammation.

SOD-mimicking nanozymes catalyze the dismutation of superoxide anions (O₂⁻•) into hydrogen peroxide, which

is subsequently decomposed by catalase-like nanozymes into oxygen and water thus neutralizing two major

classes of ROS in a cascade fashion. Cerium oxide (CeO₂) nanoparticles are notable in this regard, with their

reversible Ce³⁺/Ce⁴⁺ redox cycling enabling continuous ROS scavenging under physiological conditions (X. Gao

et al., 2024).

These antioxidant nanozymes offer enhanced stability compared to natural enzymes, remaining active across

broader pH and temperature ranges. Moreover, their surface can be engineered to enable intracellular delivery

and organelle-specific localization, allowing targeted protection of mitochondria or nuclei from oxidative insults.

In preclinical models, nanozymes have been shown to prevent DNA damage, lipid peroxidation, and protein

oxidation hallmarks of oxidative pathology. Their integration into nanocarriers or hydrogels has further improved

tissue retention and therapeutic efficacy.

Nonetheless, modulation of catalytic activity remains essential to avoid over-suppression of physiological ROS,

which are involved in signalling processes. Therefore, rational design of nanozymes with feedback-regulated or

stimuli-responsive behaviour is key to their safe and effective use in antioxidation therapies (L. Gao et al.,

2007b).

Bioorthogonal Catalysis

Bioorthogonal catalysis involves chemical reactions that proceed within living systems without perturbing native

biochemical processes. In this context, nanozymes serve as artificial catalysts capable of mediating selective

transformations such as prodrug activation or fluorophore release within targeted biological niches.

Encapsulated metal catalysts within functionalized nanoparticles (e.g., Au or Pd nanozymes) can be engineered

for bioorthogonal transformations, leveraging their robustness and structural confinement to maintain catalytic

activity in the crowded intracellular environment. These systems mimic natural allosteric regulation by using

surface ligands or host–guest interactions to gate substrate access, thus preventing off-target effects (Huang et

al., 2019).

One significant application is the in-situ activation of therapeutics such as cleavage of protective groups from

prodrugs triggered by the nanozyme at the disease site. This localized catalysis enhances drug efficacy while

reducing systemic exposure. Similarly, non-fluorescent precursors can be converted into imaging probes in vivo,

allowing real-time visualization of biological processes or disease progression.

The stability of nanozymes under oxidative, proteolytic, and variable pH conditions ensures functionality in

complex biological environments, such as tumours, inflamed tissues, or infection sites. Light-activated or pH-

responsive nanozymes offer additional layers of control, enabling temporal precision in catalysis (Bird et al.,

2021).

The field is rapidly evolving, with emerging designs incorporating supramolecular assemblies, photo responsive

elements, and biodegradable scaffolds. These advancements underscore the growing utility of nanozymes in

non-invasive, site-specific catalysis laying the groundwork for dynamic control over therapeutic and diagnostic

functions in vivo.

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CONCLUSION

The advancement of enzyme-mimicking nanomaterials, or nanozymes, represents a compelling shift in

biomedical innovation, offering a synthetic route to catalytic functionality that rivals and, in many cases, exceeds

that of natural enzymes. Over the past decade, significant progress has been made in engineering nanozymes

with finely tuned activity profiles, environmental responsiveness, and biocompatibility. These materials,

spanning noble metals, metal oxides, 2D nanostructures, and hybrid composites, have demonstrated remarkable

versatility across key biomedical domains such as cancer theranostics, antibacterial therapy, oxidative stress

regulation, and in vivo catalytic transformations.

Nanozymes overcome many intrinsic limitations of natural enzymes, including poor thermal stability,

susceptibility to proteolysis, and narrow pH activity ranges. Their physicochemical robustness, coupled with

surface modifiability and scalable synthesis, makes them ideal candidates for clinical translation and integration

into multifunctional therapeutic platforms. A consistent trend across the reviewed literature is the use of

nanozymes not only as catalytic mimics but also as intelligent agents capable of dual or even triple functionality

diagnosis, therapy, and real-time monitoring within a single system.

The biomedical potential of nanozymes is further reinforced by their adaptability. Advances in nanostructure

design, stimuli-responsive activation, and bioorthogonal catalysis are pushing the boundaries of precision

medicine. The integration of nanozymes into smart delivery vehicles, biosensors, and microenvironment-

sensitive platforms underscores their interdisciplinary relevance, particularly at the interface of nanotechnology,

materials science, and molecular medicine.

Looking forward, the next phase of nanozyme research must focus on enhancing biological specificity,

minimizing off-target interactions, and addressing long-term biosafety. Collaboration across fields including

computational modelling, immunology, and regulatory science will be essential to accelerate clinical adoption.

With rigorous design principles and translational foresight, nanozymes are poised to become foundational tools

in next-generation biomedicine, enabling catalysis-driven solutions to some of the most complex challenges in

diagnostics and therapy.

LIMITATIONS

Despite the notable progress, several limitations continue to constrain the full translational potential of enzyme-

mimic nanomaterials. Chief among these is the issue of biocompatibility and potential long-term toxicity,

especially for noble metal and transition metal-based nanozymes. While surface modifications can improve

biostability and reduce immunogenicity, comprehensive in vivo toxicity profiling remains insufficient across

much of the existing literature (R. Zhang et al., 2025b).

Another persistent challenge is the limited reproducibility and standardization in nanozyme synthesis. Variability

in particle size, morphology, and surface chemistry can significantly affect catalytic behaviour, hindering batch-

to-batch consistency and comparative evaluation. Moreover, while numerous studies have demonstrated efficacy

in vitro, in vivo validations particularly in clinically relevant models are still sparse. The lack of controlled, large-

scale clinical trials further delays regulatory approval and widespread clinical adoption (Sen et al., 2024d).

Additionally, the catalytic efficiency of many nanozymes, though high in isolated systems, can be suppressed in

complex biological environments due to biomolecule adsorption, ion interference, or rapid clearance. Addressing

these issues through rational design and mechanistic understanding is critical (Feng et al., 2024).

Overcoming these challenges will not only enhance nanozyme reliability but also unlock their broader

application in clinical diagnostics, targeted therapy, and regenerative medicine. Acoordinated effort toward safer,

smarter, and more controllable nanozymes will ultimately define their role in future healthcare systems (Cordani

et al., 2025).

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MANAGEMENT & APPLIED SCIENCE (IJLTEMAS)

ISSN 2278-2540 | DOI: 10.51583/IJLTEMAS | Volume XIV, Issue XI, November 2025

Conflict of Interest

All authors confirm that they have no financial, personal, or professional conflicts of interest to declare in relation

to the content of this manuscript.

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