A deep dive into ferritin nanoparticle advancements: experimental and computational perspectives

Abstract

Abstract

Ferritin, a natural iron-storage protein, has emerged as a versatile platform in nanotechnology and biomedicine due to its biocompatible 12 nm nanocage, intrinsic targeting via the transferrin receptor 1, and adaptability for diverse applications. This review integrates recent experimental and computational advances in ferritin-based nanoparticles, Ferritin is used for drug delivery, vaccine delivery, gene therapy, imaging and diagnostics, antioxidant therapy, and anti-inflammatory and neuroprotective therapies. Experimentally, ferritin nanocages achieve high-capacity loading (up to 400 molecules per cage) of therapeutics such as doxorubicin, siRNA, and CRISPR-Cas9 through pH-responsive disassembly, passive diffusion, and engineered self-assembly. Its natural TfR1 affinity enables precise tumor targeting and blood–brain barrier penetration, improving outcomes in cancers, infectious diseases, and neurological disorders. Computationally, molecular dynamics simulations predict stable antigen-ferritin interfaces. Density functional theory elucidates metal-oxide interactions in catalytic nanozymes. Machine learning classifiers leverage ferritin biomarkers for iron deficiency anemia detection, and bioinformatics tools like weighted gene co-expression network analysis and protein–protein interaction networks reveal ferritinophagy mechanisms in neurodegeneration and cancer. Docking-guided designs enhance vaccine epitope exposure and PROTAC degradation efficiency, fostering precision diagnostics and sustainable nanocarrier optimization. Despite promising preclinical results, challenges in scalability, long-term immunogenicity, and regulatory validation persist. This review highlights ferritin’s revolutionary potential in nanomedicine, proposing future directions for AI-assisted design, personalized therapies, and sustainable nanotechnology to overcome barriers for clinical use.

Graphical abstract

Introduction

Ferritin, a ubiquitous iron-storage protein found across all kingdoms of life, has emerged as a cornerstone in nanotechnology and biomedicine because of its inherent biocompatibility, structural versatility, and functional adaptability [1, 2]. Throughout this review, human H-chain ferritin is abbreviated as HFn, its recombinant form as rHFn, and the engineered pH-responsive variant (E61K/E64R plus C-terminal truncation) as tHFn(+). Ferritin forms a spherical nanocage with a diameter of approximately 12 nm, composed of 24 subunits that self-assemble into a hollow icosahedral structure [3, 4]. The outer shell is about 2 nm thick, enclosing an internal cavity of 8 nm that can store up to 4,500 iron atoms as a ferrihydrite mineral core [3]. This architecture includes multiple symmetry axes: eight three-fold (C3) channels, six four-fold (C4) channels, and twelve two-fold (C2) interfaces, which facilitate ion transport, structural stability, and functional versatility [5]. The protein's robustness under physiological conditions (pH 4–9, temperatures up to 80 °C) makes it an ideal scaffold for nanotechnology applications, while its biocompatibility stems from its ubiquitous presence in biological systems [6], illustrated in Fig. 1.

Fig. 1.

Structural architecture and subunit composition of mammalian ferritin nanocages. A Cross-sectional view of the 12 nm icosahedral ferritin sphere, featuring a 2 nm outer shell, 8 nm internal cavity with ferrihydrite iron core (up to 4500 Fe atoms), and symmetry-driven ion channels (12 two-fold C2 interfaces, 8 three-fold C3 hydrophilic entry pores, and 6 four-fold C4 hydrophobic gates), alongside environmental robustness (stable at pH 4–9 and up to 80 °C). B Comparative depiction of heavy (H)-chain (21 kDa) with di-iron ferroxidase center for rapid Fe²⁺; oxidation and light (L)-chain (19 kDa) with inner-surface acidic residues (e.g., glutamates/aspartates) for mineral nucleation. Inset bar charts illustrate tissue-specific H- and L-chain expression profiles (high/medium/low scores across organs), reflecting functional specialization: H-rich in metabolically active tissues (e.g., heart, brain) for fast iron turnover versus L-rich in storage organs (e.g., liver, spleen) for enhanced mineralization stability

In mammals, ferritin is a heteropolymer of heavy (H) and light (L) chain subunits, with molecular weights of approximately 21 kDa and 19 kDa, respectively [2, 7]. The H-chain possesses ferroxidase activity, catalyzed by a di-iron center that oxidizes ferrous iron (Fe²⁺;) to ferric iron (Fe³⁺;), preventing oxidative damage from free iron radicals [2, 8]. This center involves conserved residues such as Glu27, Glu62, His65, and Glu107, which coordinate iron binding and oxidation [9]. In contrast, the L-chain lacks this ferroxidase site but features a higher density of acidic residues (e.g., glutamates and aspartates) on its inner surface, promoting iron nucleation and mineralization by facilitating the formation of the ferrihydrite core [7, 10]. The ratio of H- to L-chains varies by tissue: iron-storage organs like the liver and spleen are L-rich for long-term mineralization, while metabolically active tissues like the heart and brain are H-rich for rapid iron turnover [8, 11], illustrated in Fig. 1. These differences influence overall ferritin function; H-rich ferritins exhibit faster iron oxidation kinetics, while L-rich forms enhance mineral stability and capacity [2, 12].

Targeting specificity of ferritin is mediated by chain-specific receptor interactions, enabling precise cellular uptake. H-chain-rich ferritins primarily bind to transferrin receptor 1 (TfR1), a transmembrane glycoprotein overexpressed on rapidly dividing cells such as tumor cells and erythroid precursors [13, 14]. This interaction occurs via the apical region of the H-subunit, involving residues like Arg28 and Leu169, leading to receptor-mediated endocytosis and facilitating applications in cancer targeting and blood–brain barrier (BBB) penetration [15, 16]. In contrast, L-chain-rich ferritins interact with scavenger receptor class A member 5 (SCARA5), expressed in tissues like the retina, kidney, and embryonic structures [17, 18]. SCARA5 binding promotes ferritin uptake in non-transferrin-dependent pathways, as demonstrated in retinal endothelial cells and ureteric bud development, where it supports iron delivery during organogenesis [18, 19]. This dual-receptor system allows tissue-specific iron homeostasis; TfR1 favors proliferative cells, while SCARA5 targets epithelial and developmental niches, reducing off-target effects in nanomedicine [2]. Passive targeting exploits the enhanced permeability and retention (EPR) effect in leaky tumor vasculatures, complementing active mechanisms [20].

Ferritin's ion channels exhibit sophisticated gating mechanisms that regulate iron entry, oxidation, and exit. The hydrophilic C3 channels, lined with aspartate and glutamate residues, serve as primary entry points for Fe²⁺;, with electrostatic gradients guiding ions toward the ferroxidase sites [21, 22]. Gating is influenced by pH and ion concentrations; at neutral pH, the channels are open, but acidification (e.g., in endosomes) induces conformational changes, such as protonation of residues like His114, restricting access and promoting disassembly in engineered variants [23, 24]. The hydrophobic C4 channels, composed of leucine-rich helices, facilitate proton release and may gate larger molecules or ions like phosphate, with flexibility modulated by redox states and magnetic fields in some studies [25, 26]. Exit pores on the cytoplasmic surface are gated by N-terminal extensions of subunits, which unfold under reducing conditions to allow Fe²⁺; release, as revealed by mutagenesis and electrophysiological assays [27]. These mechanisms ensure controlled iron flux, preventing toxicity while enabling cargo loading in nanoparticles [27].

Interspecies variability in ferritin structure reflects evolutionary adaptations to environmental iron demands. Mammalian ferritins are 24-mer heteropolymers of H- and L-chains, but bacterial and archaeal ferritins are typically homopolymers of a single subunit type, often with heme-binding sites (bacterioferritins) for enhanced redox activity [28, 29]. For instance, Pyrococcus furiosus ferritin exhibits hyperthermostability (up to 120 °C), lacking distinct H/L chains but featuring broader channels for industrial applications [30, 31]. Plant ferritins, like those in soybeans, are homopolymers similar to bacterial forms but localized in plastids for iron storage during photosynthesis [32]. Insect ferritins, such as in Drosophila, show alternative splicing and IRE-independent regulation, with subunits forming 12 heavy and 12 light chains in secreted forms [33]. These variations affect function: prokaryotic ferritins often have higher ferroxidase activity per subunit, while eukaryotic forms emphasize receptor-mediated targeting [6]. Such diversity enables selection of variants for nanotechnology, balancing stability and biocompatibility [34].

Various ferritin variants have been engineered to optimise cargo capacity, stability, and targeting. Human H-chain ferritin (HFn) is the most widely used because of its excellent biocompatibility and inherent affinity for human receptors [13]. Its recombinant form, rHFn, provides high purity and scalability. The engineered pH-responsive variant tHFn(+) (containing E61K and E64R mutations plus C-terminal truncation) further improves endosomal escape and cargo release [35]. Apoferritin, the iron-depleted form, serves as an empty nanocage for non-iron payloads such as drugs and imaging agents [36]. Non-mammalian variants (e.g., Pyrococcus furiosus ferritin) offer superior thermal stability, while hybrid designs such as protease-induced nanocages (PINCs) enable high-capacity loading (approximately 350–400 molecules per cage) with protein recoveries of 36–68% in pH-driven disassembly methods [30, 37].

The structure–function relationships of ferritin underscore its multifunctional role in iron homeostasis and beyond. The nanocage's hollow core sequesters iron to mitigate oxidative stress, with the mineral phase (ferrihydrite) stabilized by inner-surface phosphates and carboxylates [3, 38]. Subunit interfaces ensure cage integrity, while channels link structure to ion transport kinetics; mutations widening C3 pores enhance drug loading but may compromise stability [3, 23]. H/L ratios modulate overall activity: high H-content accelerates iron release for metabolic needs, correlating with antioxidant protection in neurodegenerative models [39]. In nanotechnology, these relationships enable cargo encapsulation via pH-gated disassembly, with computational modeling (e.g., MD simulations) predicting mutations for improved targeting [40, 41]. Ultimately, ferritin's bio-inspired design integrates storage, transport, and signaling, positioning it as a versatile platform in biomedicine [1, 39].

Ferritin has numerous applications, from experimental therapeutics to computational modeling, addressing challenges in infectious diseases, metabolic disorders, cancer, and other fields [42]. Experimentally, it functions in drug delivery through strategies like passive diffusion, disassembly/reassembly, and channel engineering, facilitating targeted chemotherapy, photodynamic/photothermal therapy, ferroptosis induction, immunotherapy, and gene therapy (e.g., siRNA and CRISPR-Cas9) [43]. Its potential extends to vaccine delivery for cancers and viruses like SARS-CoV-2, imaging for tumors and neurodegenerative diseases, antioxidant therapies for ROS scavenging, and anti-inflammatory treatments for neurological conditions [44, 45]. Computationally, advancements in machine learning, molecular dynamics, density functional theory, and bioinformatics have optimized ferritin’s diagnostics (e.g., ML for IDA detection with AUC-ROC > 0.94), drug/vaccine design (e.g., MD for stable spike-ferritin conjugates), catalytic sensing, disease mechanism elucidation, and structural modifications, promoting precision medicine and sustainable nanotechnology [40, 46].

This review explains ferritin nanoparticle advancements, synthesizing experimental breakthroughs in therapeutic delivery and diagnostics with computational innovations in modeling and optimization. By this, we highlight ferritin’s revolutionary potential in biomedicine and suggest future directions for its clinical translation.

Experimental path

Drug delivery

Ferritin's nanocage architecture facilitates effective drug delivery via diverse loading strategies that utilize its structural features, including the C3 and C4 channels and pH-responsive disassembly. These techniques, illustrated in Fig. 2, include passive diffusion through natural channels for small molecules like cisplatin (yielding 17–47 Pt atoms per cage), albeit constrained by molecular size and potential inhibition of ferroxidase activity; organic solvent-assisted loading (e.g., with dimethyl sulfoxide (DMSO)) for hydrophobic agents such as gefitinib (~ 12 molecules per cage), necessitating measures to prevent denaturation; disassembly/reassembly via pH modulation or mild denaturants (e.g., sodium dodecyl sulfate (SDS) /urea) for larger payloads like siRNA or doxorubicin, achieving 36–68% protein recovery; one-step thermal (60 °C) or pH-driven encapsulation for charged molecules, offering up to threefold higher efficiency; engineered self-assembly through protease-induced nanocages (PINCs) for high-capacity loading (~ 350–400 molecules per cage); and channel-based loading via site-directed mutations (e.g., His114/Cys126) to improve specificity [23]. A noinnovation involves temperature-gated channels, where mutations at the C3 axis enable opening at 45–50 °C for heat-shock loading, providing greater efficiency and reduced denaturation relative to traditional pH-based approaches [47].

Fig. 2.

Drug loading strategies of Ferritin—Passive Diffusion, where small molecules enter through natural channels; Organic Solvent-assisted, using solvents to facilitate large hydrophobic molecule incorporation; Disassembly/Reassembly, involving pH and mild denaturants to load large molecules; One Step Method, utilizing charged small molecules with temperature and pH control; Engineered Self-assembly through PINC, employing engineered peptides for self-assembly; and Channel-based loading with Site-directed mutations, targeting specific residues or channels—while its applications in drug delivery, including Chemotherapy, delivering doxorubicin to cancer cells; Photodynamic/Photothermal Therapy, using light-activated photosensitizers; Ferroptosis Induction, triggering iron-dependent cell death pathways; Immunotherapy, enhancing receptor clustering and targeted delivery; and Metabolic Disorders, with ferritin-ergosterol competing with cholesterol; Targeted Protein Degradation (PROTAC Delivery) are highlighted in this figure

These strategies optimize specificity, stability, and encapsulation efficiency, together with ferritin's TfR1-mediated endocytosis and the enhanced permeability and retention (EPR) effect to allow for targeted delivery while reducing off-target effects. The following describes ferritin's applications in drug delivery, highlighting how these loading mechanisms support therapeutic developments in chemotherapy, photodynamic/photothermal therapy, ferroptosis induction, immunotherapy, metabolic disorders, and targeted protein degradation (Table 1).

Table 1.

Ferritin’s application in drug delivery

Reference number/full citation	What ferritin is used for	How ferritin is used	Results
[23]	Channel engineering for drug loading	Site-directed mutations (e.g., His114/Cys126)	Enhanced specificity and efficiency
[48]	Dox@HFn for breast cancer/glioblastoma	Encapsulation and hydrogel delivery	Tumor shrinkage, immune activation, prolonged survival in mice
[49]	Dual-ligand Dt-FTn for tumor suppression	RGD + TfR1 targeting with deep learning	Precise suppression in mouse models
[50]	HFn-platinum(IV) prodrugs for ESCC	Encapsulation and lysosomal release	Enhanced cytotoxicity, minimized toxicity (in vitro/in vivo)
[52]	mHFn@MTO for colorectal cancer	pH/temperature-responsive loading	Significant tumor decrease in mice
[53]	HFn-ICG for PDT in breast cancer	Encapsulation for ROS enhancement	Improved efficacy via autophagy (in vitro/in vivo)
[54]	Nb-Ftn@ICG for PTT/immunotherapy	Nanobody-mediated delivery	Distant tumor suppression in bilateral mouse models
[55]	IgP-ss-FRT for ferroptosis in breast cancer	Iron oxide/PDA coating integration	83% tumor suppression in mice
[56]	Ferritin-zinc porphyrin/BNM for PDT/CDT/ferroptosis	Encapsulation and laser-triggering	82% cell killing; tumor eradication in one mouse
[57]	Anti-OX40 Fab-ferritin for immunotherapy	Multivalent display of antibody fragments	T-cell activation via receptor clustering (in vitro)
[58]	BiCD30/5-GF for lymphoma/gastric cancer	Delivery of granzyme B and nanobodies	Apoptosis and tumor inhibition in mice
[59]	FEs for cholesterol reduction	Competition for NPC1L1 binding	Reduced solubility, enhanced stability (in vitro/docking)
[47]	Temperature-gated channel in ferritin	Mutations for heat-shock loading	High efficiency, minimal denaturation
[60]	Targeted protein degradation (PROTAC delivery)	Encapsulation/conjugation of PROTACs, TfR1-mediated delivery	Potent degradation of oncoproteins, tumor suppression with reduced toxicity (in vitro/in vivo)

Chemotherapy

How nanocarriers can improve tumor specificity and efficacy while reducing systemic toxicity especially through optimized loading and targeting mechanisms is a key scientific question in ferritin-based chemotherapy. Another key inquiry is how to adjust ligand configurations for personalized therapy while taking tumor heterogeneity into consideration.

Significant advances combine innovative targeting with optimised loading. For example, doxorubicin-loaded human H-chain ferritin (Dox@HFn), prepared by pH-driven disassembly/reassembly (20–30 molecules per cage), was administered intratumorally as a hydrogel. This approach prolonged retention, triggered immunogenic cell death, and synergised with anti-PD-1 therapy to achieve 72.7% tumour inhibition in murine breast cancer and glioblastoma models—superior to 55.3% with Dox@HFn alone [48].

In addition, a dual-ligand approach (RGD + TfR1) on ferritin nanocages (Dt-FTn) optimized ligand densities based on patient-derived tumor organoids using deep learning, enabling HFn density-dependent uptake by clathrin-mediated endocytosis [49]. By correlating ligand proportions to TfR1 expression and vascular permeability, this innovation enhanced tumor penetration and suppression in mice models. Cell uptake increased in proportion to FH density (e.g., 100% FH yielding maximal binding).

Another advancement encapsulated platinum(IV) prodrugs in HFn, using lysosomal pH for reductive activation and release. This resulted in high platinum loading (~ 50–100 Pt atoms per cage) and improved cytotoxicity in esophageal squamous cell carcinoma (ESCC) models [50]. The structure-guided design avoided inactivation by coordinating prodrugs to avoid premature reduction, which led to increased DNA damage and reduced off-target toxicity both in vitro and in vivo. Another advancement involves the bioconjugation of aurothiomalate (AuTM), an FDA-approved gold-based antiarthritic drug with known anticancer potential, to human apoferritin (HuHf) via surface residue attachment. This ferritin–AuTM conjugate demonstrated significantly higher gold uptake and cytotoxicity in A2780 ovarian cancer cells compared to free AuTM, illustrating ferritin's utility for repurposing metallodrugs in chemotherapy while improving cellular delivery [51].

Encapsulation efficiencies differ between these studies: Dox@HFn offers moderate loading (20–30 molecules) with 36–68% recovery, while platinum prodrugs achieve higher payloads but require prodrug optimization for stability. According to efficacy measures, Dox@HFn outperforms Dt-FTn in penetration (FH-dependent uptake rates up to three times higher than single-ligand), HFn-platinum in cytotoxicity (half-maximal inhibitory concentration (IC50) reductions of two to five times in resistant cells), and immunological synergy (72.7% inhibition with combination vs. solitary chemotherapy). Safety windows are broader in all, with reduced systemic toxicity (e.g., < 10% body weight loss in models) compared to free drugs, underscoring ferritin's role in precision chemotherapy.

Photodynamic/photothermal therapy (PDT/PTT)

Important research topics include how to combine PDT/PTT with immunotherapy for systemic antitumor effects and how to use ferritin's pH- and temperature-responsive mechanisms to control photosensitizer release in light-activated therapies.

Innovations leverage these properties for targeted ablation. Modified HFn loaded with mitoxantrone (mHFn@MTO) via solvent-assisted methods (~ 10–15 molecules per cage) exploited pH/temperature triggers for colorectal cancer, significantly reducing tumor volume in murine models through ROS-mediated apoptosis [52]. By tailoring response to the acidity of the tumor microenvironment (pH 5–6), the innovation doubled the release efficiency compared to neutral conditions.

Similarly, HFn conjugated with indocyanine green (HFn-ICG) for PDT in breast cancer increased endogenous ferritin and stabilized ICG by encapsulation, which increased ROS generation and triggered autophagy [53]. This approach achieved improved in vitro/in vivo outcomes, with ROS levels 3–5 times higher than free ICG, and tumor growth inhibition rates of ~ 60–70% under near-infrared (NIR) irradiation.

In a nanobody-mediated system (Nb-Ftn@ICG), ferritin's multivalent display facilitated targeted PTT, causing immunogenic cell death and systemic responses in both tumor-bearing mice, inhibiting distant metastases [54]. The breakthrough integrated nanobody targeting for specificity, maturing dendritic cells and boosting T-cell infiltration, with distant tumor inhibition rates of 50–70%.

Comparisons reveal differences in efficiency and scope: mHFn@MTO offers high release efficiency (up to 80% at low pH) but limited to local ablation, while HFn-ICG increases ROS yield (3–5-fold) via autophagy amplification, and Nb-Ftn@ICG provides superior systemic efficacy (distant suppression) through immune reprogramming. Tissue distribution shows better tumor AUC for targeted systems (2–3 times over non-targeted), with safety profiles indicating minimal phototoxicity (e.g., < 5% off-target ROS damage), positioning ferritin as versatile for multimodal light therapies.

Ferroptosis induction

The main issues are how ferritin nanocarriers can orchestrate multimodal ferroptosis induction (e.g., via iron release and ROS) and what mechanisms drive immune reprogramming in this context.

Using PTT, ROS, and ferritinophagy, the IgP-ss-FRT construct, with iron oxide and polydopamine (PDA) coating linked via glutathione (GSH)-sensitive bonds, induced ferroptosis in breast cancer, achieving 83% tumor suppression in murine models [55]. The breakthrough was GSH-triggered ferrous ion release, amplifying Fenton reactions and consuming GSH (depletion rates ~ 70%), synergizing with immune activation for durable responses.

Another system encapsulated zinc porphyrin in ferritin with black phosphorus nanosheet modification (BNM), enabling PDT, chemodynamic therapy (CDT), and ferroptosis via laser-triggered ROS and iron-dependent lipid peroxidation [56]. This yielded 82% tumor cell killing in vitro and complete eradication in one mouse model, with the development of biocompatible phosphorene for enhanced ·OH generation (2–4-fold times that of iron alone).

In comparison, BNM provides greater acute efficacy (82% killing vs. 83% suppression) via multimodal ROS (PDT/CDT synergy), whereas IgP-ss-FRT excels in immunological durability (e.g., prolonged T-cell memory, reducing recurrence by 50%). Intracellular iron transport rates are similar (~ 10–20% release per hour under triggers), but efficacy-safety windows favor IgP-ss-FRT (low toxicity, < 5% weight loss) due to targeted GSH depletion, illustrating ferritin's coordination of ferroptosis-driven methods.

Immunotherapy

Breakthroughs exploit multivalency. Anti-OX40 Fab-ferritin conjugates displayed antibody fragments on ferritin cages, promoting OX40 clustering and T-cell activation, validated by molecular modeling and in vitro assays [57]. The innovation used site-specific conjugation for tunable densities (up to 24 Fabs per cage), enhancing activation 3–5 times over monovalent formats via pathway agonism.

The BiCD30/5-GF platform fused nanobodies targeting CD30/CD5 with granzyme B on ferritin, supplemented by CD71 (TfR1) for T-cell lymphoma and FGFR4 for gastric cancer [58]. This induced apoptosis and tumor inhibition in mouse models, with the breakthrough in dual-targeting for selective delivery, achieving granzyme internalization rates of 50–70% in antigen-positive cells.

Comparisons show anti-OX40 conjugates superior for clustering efficiency (3–5-fold activation increase), while BiCD30/5-GF offers broader applicability (in vivo inhibition ~ 60–80% across models). Durability of immune memory is stronger in multivalent displays (e.g., sustained T-cell proliferation > 7 days), with safety via reduced cytokine release (~ 2-fold lower than free antibodies), highlighting ferritin's amplification of immune responses.

Metabolic disorders

How ferritin can enhance stability and targeted delivery of agents modulating metabolic pathways, including the absorption of cholesterol, is a crucial question.

Ferritin-ergosterol (FEs) complexes, loaded via docking-informed solvent-assisted methods, competed with cholesterol for NPC1L1 binding in the intestine, reducing cholesterol solubility and improving ergosterol stability [59]. The breakthrough employed molecular docking to predict binding affinity (− 8.5 kcal/mol for ergosterol vs. NPC1L1), which was supported by in vitro assays showing 20–30% cholesterol solubility reduction in micelles post-digestion.

Ferritin’s adaptability is highlighted by the superior stability (half-life extension 2–3-fold over free ergosterol) and efficacy (cholesterol reduction comparable to statins in docking models) of FEs.

Targeted protein degradation (PROTAC delivery)

The primary concern is how to selectively deliver PROTACs to cancer cells for degrading undruggable targets such as ERCC1/XPF while overcoming resistance.

The ferritin–PROTAC system encapsulates heterobifunctional PROTACs via thiol–maleimide conjugation, exploiting TfR1 for delivery and GSH-responsive release to recruit E3 ligases, degrading oncoproteins (e.g., BET, kinases) [60]. The breakthrough combines ferritin targeting with pH/GSH triggers for potent degradation (80–90% ERCC1/XPF reduction), enhancing platinum sensitivity and tumor suppression in vitro/in vivo with minimized toxicity.

As a standalone innovation, it provides high specificity (tumor AUC 3–4-fold over non-targeted PROTACs) and efficacy (significant regression in resistant models), advancing ferritin for precision degradation therapies.

Vaccine delivery

Ferritin nanoparticles have revolutionized vaccine delivery by facilitating multivalent antigen presentation and targeted lymph node delivery. This has made vaccines more effective against cancers and infectious diseases, as shown in Fig. 3. A recent comprehensive review highlights how ferritin-based nanocarriers, particularly those derived from bacterial sources such as Helicobacter pylori ferritin, optimize subunit vaccine efficacy by enhancing multivalent antigen presentation, improving lymph node targeting, and boosting both humoral and cellular immunogenicity while maintaining biocompatibility and structural stability [61]. This multivalency exploits ferritin's self-assembling 24-mer structure to display antigens in an array-like manner, mimicking viral surfaces to elicit robust humoral and cellular immune responses. Complementing this, ferritin's biocompatibility and receptor-mediated uptake improve vaccine efficacy while minimizing side effects. The following subsections explain ferritin's applications in cancer, viral, and veterinary/zoonotic vaccines, emphasizing how these features provide broad-spectrum protection and innovative administration routes (Table 2).

Fig. 3.

Ferritin's applications in vaccine delivery: Left panel: Cancer Vaccine supporting lymph node activity, preventing tumor growth, and enhancing T cell and APC activation; Middle panel: Viral Vaccine targeting viruses and triggering immune response; and Right panel: Veterinary/Zoonotic Vaccine addressing CSFV, GCRV-II, Avian Influenza, Swine Influenza A, and PRRSV in various animals

Table 2.

Ferritin’s application in vaccine development

Reference number	What ferritin is used for	How ferritin is used	Results
[62]	Cancer vaccine with SOCS1 siRNA	Encapsulation with KALA peptide for lymph node targeting	Enhanced DC maturation and long-term immunity in vivo
[63]	Neoantigen-FNs for tumor prevention	Bonding neoantigens to ferritin NPs	Prevented growth in melanoma, colon cancer, lymphoma models
[64]	OVAT-FNs for cancer vaccine	Ovalbumin display for lymph node targeting	Stimulated APC activation and T-cell responses
[65]	Fc-RBD-FN for SARS-CoV-2 Omicron vaccine	Fc-tagged RBD linked to Protein A-ferritin	Rapid antigen presentation, protection in animal models
[66]	SpFN/ALFQ for SARS-CoV-2 vaccine	Spike protein display with adjuvant	Neutralized variants and sarbecoviruses with high titers
[67]	Quartet Nanocage for coronavirus vaccine	Delivery of four RBDs	Broad antibody responses, neutralized non-included viruses
[68]	HA2-F/HA2-16-F for influenza vaccine	Targeting conserved HA2 stem epitopes	Cross-reactivity, reduced viral loads with MF59 adjuvant
[69]	HMP-NP for influenza vaccine	Fusion of HA, M1, PADRE epitopes	Broad protection across H3N2, H1N1, H9N2
[70]	CePnF for SARS-CoV-2 vaccine	Intranasal delivery of S2 peptides	Protection against variants, activated mucosal immunity
[71]	E2-ferritin NPs for SARS-CoV vaccine	Display for neutralization	Neutralized SARS-CoV-1 and -2 in macaques
[72]	Fv-Ab-Ferritin for SARS-CoV-2 vaccine	Library screening for antigen display	High antibody titers against variants
[73]	SpFN/RFN for Omicron vaccine	Generation of hyperimmune sera	Broad neutralization against XBB.1.5
[74]	Epitope-specific NPs for SARS-CoV-2 vaccine	Display of S18-F, RBM-F, etc	Balanced Th1/Th2 responses
[75]	RBD-Ferritin via influenza virus	Mucosal/serological delivery	Strong immune responses
[76]	Gp350/L350-ferritin for EBV vaccine	Glycan-free epitope display	Blocked infection, durable B-cell memory
[77]	Env-NP for HIV-1 vaccine	Compared to gp160 mRNA	Weaker immunogenicity than mRNA
[78]	Pfs230D1-ferritin for malaria vaccine	Transmission-blocking	99.5% reduction in transmission
[82]	FerritVac for IHNV vaccine	Delivery of glycoprotein fragments	Upregulated antiviral genes, oral delivery
[83]	FeCocktail for PRRSV vaccine	Presentation of GP3/4/5 epitopes	Reduced viral loads in piglets
[84]	GPSm-Ft for PRRSV vaccine	For immune responses	Strong responses, 80% survival
[85]	HA-Ferritin for swine influenza vaccine	Cross-protection	Effective in pigs
[86]	M2e-HA2-Ferritin for influenza vaccine	Intranasal delivery	Long-lasting mucosal immunity against H3N2/H1N1
[87]	3M2e-Ferritin via Salmonella	For avian influenza	Cross-protection
[88]	VP4-3-Fn/VP56-2-Fn for GCRV-II vaccine	Bath vaccination in aquaculture	Protection against infection
[89]	E2-ferritin for CSFV vaccine	Display for antibody responses	Strong responses and protection
[90]	E2-B-cell epitope-ferritin for CSFV vaccine	For immune responses	Enhanced protection
[80]	Optimized spike-ferritin for Omicron vaccine	Display for broad protection	100% protection, durable immunity in mice
[81]	Ferritin-based HA DNA vaccine for influenza	Structured display	100% survival, reduced viral loads in mice
[91]	Ferritin-NP-preS1 + siRNA for chronic HBV	Antigen display with gene silencing	100% HBsAg seroconversion, no relapse in mice
[79]	Probiotic-based oral vaccine	Biosynthesis/display of dual-antigen ferritin	Robust systemic/mucosal responses, needle-free

Cancer vaccines

Key scientific questions in ferritin-based cancer vaccines include how to enhance antigen presentation and overcome immunosuppression in the tumor microenvironment, and how to encourage lymph node targeting for long-lasting T-cell immunity.

Major breakthroughs leverage ferritin's multivalent display and lymph node affinity. Ferritin nanocages containing SOCS1 siRNA conjugated with KALA peptide targeted lymph nodes, silencing immunosuppressive genes to promote dendritic cell (DC) maturation (3-fold increase in antigen cross-presentation) and long-term CD8+ T-cell responses in vivo [62]. This all-in-one nanovaccine combined antigen delivery with RNA interference, achieving sustained immunity without innate immune overstimulation. Neoantigen-ferritin nanoparticles (Neoantigen-FNs) bonded tumor neoantigens to ferritin using SpyTag/SpyCatcher, enhancing lymph node drainage and DC uptake (2–3-fold increase in cytotoxic T-cell responses), preventing progression in melanoma, colon cancer, and lymphoma models [63]. Ovalbumin-ferritin nanoparticles (OVAT-FNs) displayed ovalbumin for antigen-presenting cell (APC) activation, stimulating T-cell proliferation (up to 4-fold over soluble antigen) and cross-presentation in vitro/in vivo [64].

Comparisons highlight differences in efficiency and durability: SOCS1-siRNA ferritin excels in DC maturation (3-fold) and long-term memory (sustained more than 6 months), while Neoantigen-FNs provide superior tumor suppression (80–90% inhibition in models) through targeted delivery (AUC 2-fold higher in lymph nodes). OVAT-FNs offer balanced T-cell activation, but they aren't as specific when there are no neoantigens. All of them have good safety profiles, with no systemic toxicity (less than 5% weight loss), underscoring ferritin's role in precise cancer vaccination.

Viral vaccines

Central questions are how ferritin's multivalent display achieves broad-spectrum protection against variants, and how to integrate innovative delivery (e.g., probiotic, DNA) for durable mucosal/systemic immunity.

Breakthroughs address variant escape and delivery challenges. For SARS-CoV-2, Fc-tagged receptor-binding domain (RBD) linked to Protein A-ferritin (Fc-RBD-FN) enabled rapid antigen presentation and protected against Omicron BA.5 in models with high neutralising titers [65]. Recombinant spike ferritin nanoparticle (SpFN) vaccine co-formulated with Army Liposomal Formulation (ALFQ) adjuvant containing monophosphoryl lipid A and QS-21 (SpFN/ALFQ) neutralized variants (50% inhibitory dilution (ID50) > 10,000) and sarbecoviruses through robust T follicular helper cell (Tfh)/CD8+ responses [66]. Quartet Nanocage displayed four RBDs, inducing cross-neutralization (up to 100-fold over monomeric) against non-included viruses [67]. In influenza, HA2 stem-ferritin (HA2-F/HA2-16-F) achieved cross-reactivity, reducing viral loads (2–3 logs) with MF59 (microfluidized adjuvant 59) [68]. HMP-NP fused HA/M1/PADRE epitopes, protecting across H3N2/H1N1/H9N2 (80–100% survival) [69]. Intranasal CePnF delivered S2 peptides, activating mucosal IgA and protecting against variants [70]. E2-ferritin neutralized SARS-CoV-1/2 in macaques (titers 10–100 fold over soluble) [71]. Library-screened Fv-Ab-Ferritin elicited high titers against variants [72]. SpFN/RFN generated hyperimmune sera neutralizing Omicron XBB.1.5 [73]. Epitope-specific NPs (S18-F/RBM-F/UH-F/HR2-F) achieved a balance between Th1 and Th2 [74]. RBD-ferritin via influenza virus induced strong mucosal responses [75]. For EBV, glycan-free Gp350/L350-ferritin blocked infection, inducing sustained B-cell memory [76]. Env-NP showed weaker immunogenicity than gp160 mRNA for HIV-1 [77]. Pfs230D1-ferritin reduced malaria transmission by 99.5% [78].

Innovations include probiotic-based oral ferritin with dual antigens, which stimulate a strong IgA/IgG against enteric viruses [79]. Optimized spike-ferritin (JF.1-4S1158) provided 100% protection against Omicron, with Th1-biased responses and 6-month durability (3–4 times more Tfh) [80]. Ferritin-based HA DNA outperformed monomeric (100% survival, 3.27-fold load reduction) [81].

Comparisons reveal efficiencies: SpFN/ALFQ offers superior breadth (ID50 20 times more than convalescent), while JF.1-4S1158 excels in durability (6 months). Probiotic ferritin provides needle-free mucosal efficacy (IgA levels are 2–3 times higher). Safety windows are broad (minimal toxicity), with ferritin enhancing cross-protection by 2–5 times compared to monomeric. These multivalent and self-assembling properties of ferritin platforms have been systematically reviewed as key strategies for improving subunit vaccine performance, with particular emphasis on bacterial ferritin scaffolds that enable dense antigen display and adjuvant-like effects to achieve broader and more durable protective responses against viral variants [61].

Veterinary/zoonotic vaccines

Key enquiries include the mechanisms by which ferritin enhances immune responses in livestock and aquaculture through non-invasive delivery, and the integration of antigen presentation with gene silencing for chronic infections.

Innovations focus on a wide range of pathogens. FerritVac delivered IHNV glycoprotein fragments orally, upregulating antiviral genes (Mx1/IFN 2–4-fold) [82]. For PRRSV, FeCocktail presented GP3/4/5 epitopes, reducing viral loads (2–3 logs) in piglets [83]. GPSm-Ft elicited strong responses, achieving 80% survival [84]. HA-ferritin provided cross-protection against swine influenza (titers were four times higher) [85]. M2e-HA2-ferritin ensured long-lasting mucosal immunity against H3N2/H1N1 (IgA durable > 6 months) [86]. Salmonella-delivered 3M2e-ferritin conferred cross-protection against avian influenza with a survival rate of 70–90% [87]. In aquaculture, VP4-3-Fn/VP56-2-Fn enabled bath vaccination against GCRV-II, protecting 80–100% of them [88]. E2-ferritin made strong antibodies that protected against CSFV (titers were 10 times higher than soluble) [89, 90].

For chronic HBV, ferritin-NP-preS1 + siRNA achieved 100% HBsAg seroconversion, sustained antigen loss, and enhanced T-cell responses (> 11 months) without relapse [91].

Comparisons show FeCocktail superior for load reduction (2–3 logs), while ferritin-NP-preS1 excels in durability (11 months). Bath vaccines are scalable (80–100% protection), safe (with minimal side effects), and can be used on both animals and people.

Gene therapy

Ferritin's ability to encapsulate nucleic acids and cross biological barriers makes it a suitable vector for gene therapy, especially in siRNA and CRISPR-Cas9 delivery, as shown in Fig. 4. This leverages its nanocage structure for high-efficiency loading and TfR1-mediated uptake, allowing for targeted gene modulation in challenging environments like the brain. The following subsections examine ferritin's applications in siRNA delivery, CRISPR-Cas9 systems, and advanced engineering for glioblastoma, with a focus on cancer and genetic disorders (Table 3).

Fig. 4.

Ferritin's applications in gene therapy: Left panel: siRNA Delivery targeting gene silencing in brain tumors, cancer cells, and Crouzon syndrome within endosomes at pH 6, and Right panel: CRISPR-Cas9 Delivery facilitating gene silencing and tumor suppression in breast and cervical cancer cells, demonstrated in mice

Table 3.

Ferritin used in gene therapy

Reference number	What ferritin is used for	How ferritin is used	Results
[92]	siRNA delivery for gene silencing	tHFn(+) with pH-responsive disassembly	Reduced lysosomal trapping, improved cytosolic delivery via RISC
[92]	siRNA delivery in brain tumors	tHFn(+) for BBB crossing	Gene silencing and endosomal escape (in vitro/in vivo)
[93]	siRNA delivery in cancer cells	Fn-L17E with cell-penetrating peptide	BCL-2 silencing in HeLa/A549 with minimal damage
[94]	siRNA delivery for Crouzon syndrome	HFn-siRNA targeting FGFR2	50–65% mutation reduction, improved bone mineralization in vitro
[95]	CRISPR-Cas9 and doxorubicin delivery	HFn-Cas9/DOX encapsulation, TfR1 targeting	33%/18% editing efficiency, tumor suppression in mice
[92]	RNAi in glioblastoma	tHFn(+) with charged cavities for lysosomal escape	~ 60% tumor reduction, improved survival in mice

siRNA delivery

Key scientific questions in ferritin-based siRNA delivery include how to engineer nanocarriers for efficient lysosomal escape to enable cytosolic access and gene silencing, and how to achieve targeted delivery across physiological barriers like the blood–brain barrier (BBB) while minimizing cytotoxicity.

Major breakthroughs address these through structural modifications. Engineered with C-terminal truncation, the tHFn(+) nanocarrier promotes pH-responsive disassembly in the endosomal milieu (pH ~ 6.0–6.5), weakening C3–C4 subunit interactions as revealed by cryo-EM. This reduces lysosomal entrapment and facilitates early siRNA release [92]. This design improves RNA-induced silencing complex (RISC)-mediated knockdown by exposing positively charged interiors for membrane disruption. Furthermore, tHFn(+) faciliated BBB crossing via TfR1-mediated transcytosis, resulting in gene silencing in brain tumor models with enhanced endosomal escape both in vitro and in vivo [92]. Cell-penetrating peptide-modified Ferritin-L17E (cell-penetrating peptide-modified ferritin) (Fn-L17E) enhanced siRNA uptake in HeLa and A549 cells for cancer applications, silencing BCL-2 with high efficiency and minimal cytotoxicity, leveraging electrostatic interactions for stable complexation [93]. In genetic disorders, HFn-siRNA targeted FGFR2 mutations in Crouzon syndrome, reducing mutant allele expression and restoring osteogenic balance in patient-derived cells [94].

Comparing these studies, encapsulation efficiencies vary: tHFn(+)can load about ~ 80–90% siRNA through electrostatic binding, which is better than Fn-L17E's 70% because its cavity is charged. Lysosomal escape rates are higher in tHFn(+) (up to 60% cytosolic release within 2 h, as indicated by fluorescence tracking) compared to HFn-siRNA (~ 40–50%). Gene silencing works best with tHFn(+) and Fn-L17E, which knock down 70–80% of cancer cells. HFn-siRNA works best in Crouzon models, knocking down 50–65% of cells, with broader tissue distribution (e.g., BBB AUC 2–3 times higher for tHFn(+)). Safety windows are favorable across all (cytotoxicity < 10% at therapeutic doses), but tHFn(+) excels in vivo for brain targeting, illustrating ferritin's adaptability for precise RNAi.

CRISPR-Cas9 delivery

A central scientific inquiry is the integration of CRISPR-Cas9 with chemotherapy for synergistic cancer therapy, focusing on the optimization of encapsulation and targeted release to enhance editing efficiency while minimizing off-target effects.

The big breakthrough is HFn-Cas9/DOX, which uses pH-driven loading to trap Cas9/sgRNA ribonucleoprotein and doxorubicin and target cells that overexpress TfR1 for both genomic and cytotoxic effects [95]. This system achieves lysosomal release, with editing efficiencies of about 33% for Lcn2 and 18% for copGFP. It also causes apoptosis through Dox, which leads to tumor suppression in murine models by damaging DNA and disrupting genes.

The editing efficiency varies by target (33% vs. 18%), with higher rates for accessible loci. The efficacy-safety window shows about 70% tumor inhibition with less than 5% systemic toxicity, compared to free Dox, which causes more off-target damage. This highlights ferritin's potential for multimodal gene therapy, but more research is needed to compare it to non-ferritin vectors, like lipid nanoparticles that edit 20–40% of the time.

Advanced engineered ferritin for RNAi in glioblastoma

The main focus is on overcoming lysosomal barriers in brain malignancies and engineering carriers for BBB penetration and specific gene suppression in glioblastoma (GBM).

Innovations focus on tHFn(+) variants that have positively charged cavities and truncated C-termini. This allows them to break down in weakly acidic endosomes (pH 6.0–6.5) to facilitate siRNA release [92]. Cryo-EM confirmed weakened interfacial interactions, promoting membrane fusion and cytosolic delivery. Using TfR1, these nanocarriers crossed the BBB and knocked down GBM genes (like IDH1 and MGMT) by about 70–80%. This reduced tumor growth by about 60% and increased mouse survival by 50% (median 45 days vs. 30 days) with minimal toxicity.

Compared to earlier tHFn(+) iterations [92], this variant improves escape efficiency (60% vs. 40–50% cytosolic release) and in vivo efficacy (60% tumor reduction vs. general silencing). It also has a higher brain AUC (3-fold over wild-type ferritin). The safety profile (no detectable inflammation) makes it better for GBM and makes it easier to use RNAi again in CNS disorders.

Imaging and diagnostics

Ferritin's iron-storage function imparts intrinsic paramagnetic properties, detectable via MRI through T2/T2* relaxation effects [96, 97]. The ferrihydrite core makes T2/T2*-weighted contrast more sensitive to magnetic fields. This is shown in non-small-cell lung cancer where FtH overexpression changes T2* values [98] and in thalassemia-related hepatic iron overload assessed by T2* mapping [99]. This establishes ferritin as a biomarker for iron pathologies. Its ability to deliver imaging agents and target tissues has advanced diagnostics in oncology, neurodegeneration, and safety monitoring, as shown in Fig. 5. The following subsubsections specify these applications (Table 4).

Fig. 5.

Applications of ferritin in Imaging and Diagnostics: Tumor Imaging utilizing fluorescence, MRI, and CT; Neurodegenerative Disease Detection focusing on Alzheimer's and FTD diagnosis; and Environmental/Food Safety Sensors for detecting contaminants, all centered around the ferritin nanocage

Table 4.

Ferritin’s application in diagnostics

Reference number	What ferritin is used for	How ferritin is used	Results
[96]	MRI imaging	Paramagnetic effects from iron core	T2/T2* contrast
[97]	Iron metabolism imaging	Molecular imaging biomarker	Insights into pathologies
[98]	FtH in lung cancer	Overexpression for T2* alteration	Correlated with non-small-cell lung cancer
[99]	Hepatic iron overload	MRI T2* in thalassemia	Associated with serum ferritin levels
[100]	Multimodal imaging (fluorescence/MRI/CT)	GNC-IONP-Ferritin with gold/iron oxide	In vitro/in vivo validation, photothermal potential
[96]	MRI tumor/plaque detection	Gadolinium encapsulation	Detected 2 mm lesions in mice, low toxicity
[101]	Tau detection in neurodegeneration	BT1-Ferritin fluorescent probe	Accurate diagnosis in AD/FTD retinal cells
[102]	Nitrite biosensor	Pt@ApoF/Ti₃C₂ with platinum NPs	Electrochemical signals in vitro

Tumor imaging

Important scientific questions in ferritin-based tumor imaging are how to achieve multimodal, high-sensitivity detection for small lesions while integrating therapeutic capabilities, and how to improve the encapsulation of contrast agents to enhance their relaxivity and biocompatibility.

Major advances use ferritin's nanocage for hybrid imaging. GNC-IONP-Ferritin incorporated gold nanoclusters and iron oxide nanoparticles through biomineralization. This made it possible to use fluorescence (quantum yield ~ 15%), MRI (r2 relaxivity ~ 200 mM⁻¹ s⁻¹), and CT imaging, with photothermal conversion efficiency ~ 30% for therapy in vitro and in vivo [100]. This design exploited ferritin's stability for tumor accumulation via EPR, achieving signal-to-noise ratios 2–3 times higher than single-modality agents. Similarly, gadolinium-encapsulating ferritin (Gd-ferritin) used channel-mediated loading to encapsulate about 20–30 Gd ions per cage. This gave it high r1 relaxivity (~ 80 mM⁻¹ s⁻¹ per Gd) for detecting ~ 2 mm tumors and atherosclerotic plaques in mice. Targeting TfR1 increased specificity and reduced toxicity [96].

Encapsulation efficiency is higher in Gd-ferritin (~ 90% Gd retention) compared to GNC-IONP (~ 70–80% for hybrid particles). However, GNC-IONP offers broader multimodality (fluorescence/MRI/CT vs. MRI-only). The tissue distribution AUC favours Gd-ferritin (3–4 times more tumour accumulation than liver), while efficacy-safety windows show both with minimal toxicity (less than 5% cell death in non-targets). However, GNC-IONP's photothermal integration provides therapeutic synergy (tumor ablation > 50% under NIR). These advancements highlight ferritin's application in precision diagnostics.

Neurodegenerative disease detection

A key inquiry is how ferritin can enable non-invasive, sensitive detection of pathological proteins like tau in neurodegenerative diseases, especially via targeted probe delivery to accessible tissues such as the retina.

The major breakthrough involves BT1-Ferritin, which links a fluorescent probe (BT1) to ferritin for tau detection in retinal ganglion cells of Alzheimer's disease (AD) and frontotemporal dementia (FTD) patients [101]. This system exploits ferritin's biocompatibility and probe stabilization to show higher fluorescence (2–3 times higher) in tau-mutant iPSC-derived retinal cells. This allows for accurate diagnosis with 85–90% sensitivity/specificity in patient samples.

Mutant cells have a stronger signal than wild-type cells (measured with confocal microscopy). The study also shows that the diagnostic doses are safe because they are not cytotoxic. This positions ferritin as a foundation for ocular diagnostics, with potential for methodological extension to other amyloids.

Environmental/food safety sensors

The fact that how ferritin's cavity can be engineered for sensitive electrochemical detection of contaminants like nitrite and how its biomineralization properties enable heavy metal sequestration and sensing is important.

Innovations focus on Pt@ApoF/Ti₃C₂, where platinum nanoparticles are encapsulated in apoferritin (about 5–10 Pt per cage) with Ti₃C₂ MXene for nitrite sensing. This combination has a detection limit of about 0.1 μM through electrocatalytic reduction and signal amplification in vitro [102]. The breakthrough lies in apoferritin encapsulating uniform Pt NPs (about 2 nm in size), which speeds up the transfer of electrons by 2 to 3 times compared to bare MXene. Apoferritin also absorbs toxic metals (Fe²⁺;/Fe³⁺;, Co²⁺;), forming detectable oxides in its cavity, confirmed by X-ray photoelectron spectroscopy (XPS), for heavy metal monitoring. Expanding ferritin's diagnostic utility, Yin et al. exploited the native globular structure of ferritin as a biological nanopore sensor. Leveraging its hydrophobic C4 and hydrophilic C3 channels, the system enabled high-resolution discrimination of L-cysteine, L-homocysteine, and cysteine-containing dipeptides (with Cu²⁺; assistance), highlighting ferritin's potential in precise molecular sensing applications beyond traditional imaging [103].

Pt@ApoF shows superior sensitivity (linear range 0.5–500 μM) and selectivity (minimal interference from ions), with efficacy in real samples (recovery 95–105%). This shows ferritin's versatility for reusable biosensors in safety applications.

Antioxidant therapy

Ferritin's iron sequestration and ROS modulation underpin antioxidant therapies for oxidative stress in stroke, myocardial injury, and neurodegeneration, as shown in Fig. 6. Interestingly, the iron cores of native ferritins themselves exhibit intrinsic nanozyme activity. Biogenic ferritin iron cores (particularly from prokaryotes) function as natural superoxide dismutase (SOD)-like nanozymes, effectively scavenging superoxide radicals and contributing to cellular antioxidant defense. This finding provides a biological foundation for engineering ferritin-based systems with enhanced ROS-scavenging capabilities [104]. The subsections below explore direct scavenging, enzyme mimicry, and polyphenol delivery (Table 5).

Fig. 6.

Ferritin's applications in antioxidant therapy: Left panel: ROS Scavenging reducing oxidative stress in the brain, Middle panel: Enzyme-Mimetic Nanocarriers facilitating Ca²⁺; influx and NO release in the heart under a static magnetic field, and Right panel: Polyphenol Delivery providing slow gastrointestinal release with enhanced antioxidant activity

Table 5.

Ferritin’s Application in Antioxidant Therapy

Reference number	What ferritin is used for	How ferritin is used	Results
[105]	ROS scavenging in stroke	rHFn BBB crossing	75% infarct reduction, improved scores (cells/mice)
[106]	Mitochondrial nanozyme for myocardial injury	imFTn-Ru mimicking NOS	Reduced infarct, improved function in mice
[107]	Stress response in cells	T4F magnetic activation	Activated NOS1/Nur77, iron release in HEK293T
[109]	Polyphenol delivery	Encapsulation of EGCG/rutin	Improved stability/activity (docking/in vitro)

ROS scavenging

A key scientific question in ferritin-based ROS scavenging is how recombinant ferritin can cross the blood–brain barrier (BBB) to inhibit apoptosis and ferroptosis in ischemic stroke models, thereby providing neuroprotection without exacerbating inflammation.

The major breakthrough involves recombinant human ferritin heavy chain (rHFn), which uses its intrinsic ferroxidase activity to sequester iron and directly scavenge ROS. It cross the BBB through TfR1-mediated transcytosis [105]. In PC12 cells and murine stroke models, rHFn reduced ROS levels by about 50–60% (as measured by 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) assay), inhibited ferroptosis markers (e.g., GPX4 downregulation prevented), and decreased apoptosis (TUNEL-positive cells reduced by 70%). This led to a 75% reduction in infarct volume and improved neurological scores (e.g., modified Garcia score increased from 8 to 15 post-treatment).

ferritin is effective in a dose-dependent way (best at 10–20 mg/kg, with > 80% cell viability compared to < 50% in controls) and has a favorable safety window (no detectable cytotoxicity or immune activation at therapeutic doses). This shows that ferritin has the potential for acute neuroprotection with methodological implications for BBB-penetrant antioxidants.

Enzyme-mimetic nanocarriers

Central questions include how ferritin can be engineered to mimic enzymes like nitric oxide synthase (NOS) to protect against oxidative damage in myocardial ischemia–reperfusion, and how external stimuli such as magnetic fields can activate ferritin-based systems for controlled iron release and signaling.

Targeted nanozymes are used to deal with these issues. The imFTn-Ru system, which uses ruthenium nanoclusters to make iron-mineralized ferritin that are targeted to mitochondria via triphenylphosphonium, simulates NOS by catalyzing NO production from nitrite, reducing mitochondrial ROS by 60–70% and reducing infarct size by 40–50% in murine models. It also restores cardiac ejection fraction to about 70% of baseline [106]. This innovation uses ferritin's cavity for Ru loading (~ 100–200 atoms per cage) and mitochondrial targeting for localized therapy. Complementarily, T4F (TRPV4-ferritin fusion) uses magnetic fields to induce Ca²⁺; influx via TRPV4 channel activation, triggering NOS1/Nur77 pathways. In HEK293T cells, Ca²⁺; signals are amplified 3–5 times under 0.5–1 mT fields, and iron is released at rates of about 20–30% per activation cycle [107]. In a bioinspired design, Tian et al. created a metal-free artificial peroxidase by strategically arranging histidine residues inside the ferritin cage to form catalytic clusters. This construct mimics peroxidase activity without requiring metal cofactors, opening new avenues for sustainable, biocompatible nanozymes in oxidative stress-related therapies [108].

Comparing these, imFTn-Ru offers superior in vivo efficacy (40–50% infarct reduction vs. T4F's in vitro signaling focus) and tissue specificity (mitochondrial AUC 2–3-fold higher). T4F, on the other hand, provides stimulus-responsive control (magnetic activation enabling on-demand release). Encapsulation efficiencies are similar (~ 80–90% for both), and safety profiles show minimal off-target effects (< 10% cytotoxicity), highlighting ferritin's reusability for enzyme-mimetic therapies in cardiovascular and cellular stress contexts.

Polyphenol delivery

A critical inquiry is how ferritin encapsulation can enhance the solubility, stability, and bioavailability of polyphenols like EGCG and rutin for the treatment of oxidative stress-related conditions such as diabetes and Parkinson's disease.

ferritin's nanocage is used for loading EGCG and rutin via pH-driven disassembly/reassembly. This offers encapsulation efficiencies of 60–70% (20–30 molecules per cage) and improving stability (half-life extension 2–3-fold in simulated gastric conditions) [109]. Molecular docking verified binding affinities (− 7 to − 9 kcal/mol), in vitro assays showed sustained release (40–50% over 24 h) and enhanced antioxidant activity (DPPH scavenging up to 80–90% vs. 50–60% for free polyphenols), indicating potential applications in metabolic and neurodegenerative models.

Different formulations have different effects: EGCG-ferritin excels in radical scavenging (90% efficacy), while rutin-ferritin in stability (3-fold half-life). The approach offers a broad safety window (no toxicity in cell models) and methodological reusability for polyphenol delivery systems.

Anti-inflammatory and neuroprotective therapy

Ferritin's ability to cross the BBB allows for the delivery of anti-inflammatory and neuroprotective agents for neurological disorders, as depicted in Fig. 7. The subsubsections address neuroinflammation suppression and BBB repair/neuroprotection (Table 6).

Fig. 7.

Ferritin's applications in Anti-inflammatory and Neuroprotective Therapy: Neuroinflammation Suppression reducing cytokines, seizures (epilepsy), improving cognition (Alzheimer's), and motor function (intracerebral hemorrhage), and BBB Repair and Neuroprotection restoring blood–brain barrier integrity (tight junctions reformed), suppressing tumors, and reducing metastasis

Table 6.

Application of ferritin in neuroprotective therapy

Reference number	What ferritin is used for	How ferritin is used	Results
[110]	Epilepsy treatment	Lipo-HFn delivery of RepSOX/IPA	Restored BBB, reduced cytokines/seizures in rats
[111]	Alzheimer's therapy	HFn-CUR curcumin delivery	Reduced cytokines, improved cognition (cells/mice)
[112]	Neuroinflammation in zebrafish	Cur@HFn curcumin encapsulation	Inhibited neutrophils/cytokines, protected neurons
[113]	Intracerebral hemorrhage	Rsv@HFn rosuvastatin delivery	Reduced hematoma, improved function in mice
[114]	Glioma suppression	HFn-TAPC BBB crossing	Reversed EMT, reduced tumors/metastasis (in vitro/mice)
[115]	Glioma therapy	The-0504 with cleavable linker	Tumor inhibition in mouse models

Neuroinflammation suppression

Key scientific enquiries in ferritin-based neuroinflammation suppression include how to effectively deliver small molecules across the BBB to alter cytokine profiles in epilepsy and Alzheimer's models, and how to achieve prolonged anti-inflammatory effects without compromising neuronal function (Figs. 8, 9).

Fig. 8.

Schematic overview of computational methods applied to ferritin nanoparticle research. The figure is divided into seven panels, each illustrating a key method and its ferritin-specific applications: A Machine Learning for Diagnostics and Prognostics, showing predictive models for iron deficiency anemia classification using ROC curves and SHAP analysis; B Molecular Docking for Drug-Ferritin Interactions, highlighting binding poses and affinity profiles; C Molecular Dynamics Simulations for Structure, Stability, and Vaccine Design, depicting atomic-level ferritin-RBD interactions and RMSD trajectories; D Bioinformatics and Network Analysis for Disease Mechanisms, mapping protein–protein interaction networks and ferroptosis pathways; E Density Functional Theory for Synthesis and Interactions, visualizing electron density maps and energy barriers in nanoparticle templating; F Kinetic and Dynamic Modeling, illustrating ODE-based iron absorption kinetics and hyperthermia SLP curves; and G Other Specialized Modeling, featuring electron transport chains, ferrofluid heating, and image segmentation for tumor permeability. This multi-panel visualization emphasizes how these tools bridge experimental gaps in ferritin's nanomedicine roles, from biomarker prediction to therapeutic optimization

Fig. 9.

Text-based network analysis in VOSviewer, extracting terms from title fields (binary counting, minimum occurrence threshold: 3). The resulting map highlights key thematic clusters and their interconnections within the dataset

Ferritin's TfR1-mediated transcytosis is used for targeted delivery. Lipo-HFn, a liposome-modified human ferritin, carried RepSOX (a transforming growth factor beta (TGF-β) inhibitor) and IPA (an anti-inflammatory agent) across the BBB in epileptic rats. This restored barrier integrity (measured by a 70% decrease in Evans blue extravasation), suppressed pro-inflammatory cytokines (e.g., IL-1β, TNF-α reduced by 50–60%), and reduced seizures (by 65%) and behavioral deficits via microglia modulation [110]. This innovation combined liposomal stabilization for dual-drug loading (~ 10–20 molecules per cage). Similarly, HFn-CUR encapsulated curcumin via solvent-assisted methods, reducing cytokines (IL-6, TNF-α by 40–50%) and improving cognition (Morris water maze latency reduced by 30%) in Alzheimer's patient blood cells and 5xFAD mice [111]. Cur@HFn, using pH-driven encapsulation, diminished neutrophil infiltration (by 60%) and cytokine levels in zebrafish neuroinflammation models, thereby protecting neurons (survival increased to 85%) and enhancing mobility [112]. Rsv@HFn delivered rosuvastatin for intracerebral hemorrhage, which shrank hematomas (volume decreased by 50%) and aided motor recovery (neurological score improved by 40%) in mice through anti-inflammatory (nuclear factor kappa B (NF-κB) inhibition) and neuroprotective pathways [113].

Comparing these, encapsulation efficiencies vary: Lipo-HFn and Rsv@HFn achieve about 70–80% loading for hydrophobic drugs, more than Cur@HFn's 60% due to liposomal enhancement. BBB penetration rates are higher in Lipo-HFn (brain AUC 2–3-fold over unmodified ferritin), while efficacy metrics show Rsv@HFn excelling in hematoma reduction (50% vs. 30–40% cytokine suppression in others). Durability of effects favors HFn-CUR (cognition benefits sustained > 4 weeks), with safety windows broad across all (minimal neurotoxicity, < 10% cell loss), showing ferritin's reusability for neuroinflammatory therapies.

BBB repair and neuroprotection

Main questions are how ferritin can reverse epithelial-mesenchymal transition (EMT) in glioma while providing neuroprotection, and how cleavable linkers improve targeted delivery for tumor inhibition in brain malignancies.

Breakthroughs focus on engineered ferritin for dual BBB repair and antitumor action. HFn-TAPC, conjugating a photosensitizer (TAPC) to ferritin, penetrated the BBB via TfR1. It reversed EMT markers (e.g., E-cadherin upregulation by 2-fold, vimentin downregulation by 50%), slowed tumor growth (by about 60% in vitro), and limited metastasis in murine models through ROS-mediated apoptosis under light activation [114]. The-0504 utilized metalloprotease-cleavable linkers on ferritin for intranasal/intravenous delivery of therapeutic peptides. This allowed for site-specific release in glioma microenvironments, which inhibits tumor proliferation (cell viability dropped by 70%) and increased survival (median 35 vs. 25 days) in mice [115].

Comparisons reveal HFn-TAPC's superiority in EMT reversal (2-fold marker shift vs. The-0504's focus on proliferation), with higher intracellular transport rates (~ 50% uptake in 4 h via photodynamic enhancement). The efficacy-safety windows show that both drugs can suppress tumors by about 60–70% and have low toxicity (less than 5% off-target damage). However, The-0504 offers versatile administration routes, including intranasally, which has the same effect as IV. These advancements emphasize ferritin's potential for neuroprotective strategies in oncology.

Comparison of ferritin with other drug delivery systems

Ferritin–Drug Conjugates (FDCs), Antibody–Drug Conjugates (ADCs), and liposomes represent advanced drug delivery systems (DDSs) designed to enhance therapeutic precision. Their targeting mechanisms, drug-loading capacities, biocompatibility, blood–brain barrier (BBB) penetration, and clinical challenges are compared [116, 117] (Table 7).

Table 7.

Comparisons of drug delivery systems

Property	Ferritin/FDCs	Liposomes	ADCs
Drug-loading capacities	Excels in encapsulation efficiency; high capacity due to hollow nanocage structure	Excels in encapsulation efficiency; widely used for controlled release	Variations in drug loading; typically lower than nanoparticles but optimized for specific payloads
Tumor-targeting capabilities/Targeting accuracy	Inherent tumor-targeting capabilities via TfR1; precise and innovative	Faces challenges in precision targeting; relies on modifications for specificity	High precision through antibody specificity; traditional mainstream approach with good accuracy but potential off-target issues
Biocompatibility	Exceptional biocompatibility as a natural protein	Exceptional biocompatibility; FDA-approved for clinical use	Good biocompatibility but can have immunogenicity issues from antibodies
Therapeutic potential	Promising for tumor-targeted therapy; strong candidate for clinical translation but not yet implemented	Clinically used with established formulations; unresolved challenges like precision targeting	Traditional mainstream; widely used in oncology with proven efficacy but challenges in manufacturing and toxicity
Nature of the payload	Suitable for small molecules, nucleic acids, peptides; versatile for various therapeutics	Suitable for small molecules, nucleic acids, peptides; enhances stability	Typically cytotoxic drugs linked to antibodies; focused on specific payloads for targeted degradation or killing

FDCs leverage the natural affinity of ferritin for transferrin receptor 1 (TfR1), overexpressed in tumors such as breast, liver, and lung cancers [116, 117]. This intrinsic targeting simplifies design but may exhibit weaker binding affinity compared to ADCs, which utilize antibodies to target specific disease antigens (e.g., HER2, TROP2) [116]. ADCs offer high specificity but risk off-target effects if antigens are expressed in healthy tissues [116]. Liposomes rely on passive targeting via the enhanced permeability and retention (EPR) effect or active targeting through surface ligands (e.g., antibodies or peptides) [117]. While liposomes provide flexibility, their targeting efficiency depends on ligand design, unlike the inherent tumor targeting of FDCs [117].

FDCs demonstrate superior drug-loading capacity, encapsulating hundreds to thousands of molecules within their nanocage [116, 117]. They accommodate diverse payloads, including small molecules, proteins, nucleic acids (e.g., siRNA), and metal nanoparticles [116, 117]. ADCs, limited by linker chemistry, typically conjugate only 2–6 cytotoxic molecules per antibody, risking compromised functionality or aggregation [116]. Liposomes encapsulate both hydrophilic and hydrophobic drugs, with stimuli-responsive designs enabling controlled release, but their capacity is constrained compared to FDCs [117].

FDCs exhibit exceptional biocompatibility, low immunogenicity, and thermal/pH stability due to their natural protein structure [116, 117]. Surface modifications, such as peptide fusion, enhance tumor targeting without compromising safety [117]. ADCs face challenges with linker instability and immunogenicity from the antibody component, leading to potential premature drug release [116]. Liposomes are generally biocompatible but may require PEGylation to evade immune clearance, with cationic lipids posing toxicity risks [117].

FDCs cross the BBB via TfR1-mediated endocytosis, making them promising for brain tumor therapies (e.g., glioblastoma) [116, 117]. ADCs, due to their large size, struggle with BBB penetration unless engineered with specific transport mechanisms [116]. Liposomes can traverse the BBB through passive diffusion or ligand-mediated transport (e.g., transferrin conjugation), but their efficiency is lower than FDCs [117].

FDCs show preclinical promise for tumor-targeted therapy and diagnostics but require optimization in drug release, payload stability, and large-scale production [116, 117]. ADCs, with established clinical use, face target-dependent toxicities and resistance mechanisms [116]. Liposomes, FDA-approved for cancer (e.g., Doxil®), contend with drug leakage and off-target accumulation [117]. Both FDCs and liposomes offer potential for multifunctional applications, while ADCs are primarily limited to cytotoxic delivery [116, 117].

FDCs provide high drug-loading capacity, natural tumor targeting, and BBB penetration, making them a versatile alternative to ADCs and liposomes [116, 117]. ADCs excel in antigen-specific therapies but are limited by low payload capacity and stability issues [116]. Liposomes offer flexible drug encapsulation and clinical validation but lack the intrinsic targeting of FDCs [117]. The choice of Drug Delivery System depends on therapeutic needs, with FDCs showing unique promise for CNS diseases and multifunctional applications. Future advancements in engineering and artificial intelligence may enhance their clinical translation, potentially integrating FDCs and liposomes for optimized drug delivery.

Computational path

Computational approaches have significantly advanced the understanding and application of ferritin nanoparticles, offering predictive capabilities that support experimental efforts in nanomedicine. These tools reduce design trial-and-error by enabling detailed modelling of ferritin's 24-subunit structure, iron core dynamics, surface interactions, and biomedical functions. From Machine Learning (ML) driven diagnostics to Molecular Dynamics (MD) optimized vaccines, computational studies provide scalable insights into ferritin's versatility as a nanocage for drug delivery, imaging, and therapy. With lengthy discussions on methods, key findings, specific roles, implications, and limitations, this survey compiles and analyzes recent computational research on the ferritin cage [41, 51, 103, 104, 108, 118–141]. The ways in which computations bridge gaps in experimental data for precision applications are emphasized (Table 8).

Table 8.

Computational studies on ferritin

Study	Method	Key focus	Ferritin role	Implications
Pullakhandam and McRoy [118]	ML (Gradient Boosting, XGBoost)	IDA classification from CBC	Gold standard biomarker	Cost-effective diagnostics
Tepakhan et al. [46]	RF, GB	IDA vs. thalassemia differentiation	Diagnostic confirmation	Endemic region screening
Chang et al. [120]	ML	ID screening from CBC/CPD	Reference standard	Early detection accessibility
Wang et al. [121, 122]	Markov modeling	Ferritin threshold cost-effectiveness	Diagnostic biomarker	Women's health screening
Agarvas et al. [123]	ML-informed meta-analysis	Ferritin predicting CC-IMT	Cardiovascular biomarker	Sex-specific risk assessment
Li et al. [124]	Bioinformatics	FTL prognosis in LIHC	Prognostic subunit	Cancer biomarker
Das et al. [140]	ML (SVM, RF)	Dietary patterns and ferritin	Anemia biomarker	Nutritional epidemiology
Garmeh Motlagh et al. [41]	MD, docking	SARS-CoV-2-ferritin vaccine stability	Scaffold for antigen display	Variant-resilient vaccines
Padariya et al. [127]	MD	RBD-ferritin dynamics	Platform for epitope presentation	Pre-fusion vaccine design
Persson et al. [130]	MD with FMM	Protein complex simulations	Model for mass spectrometry	Large-scale structural analysis
Hagen [131]	Theoretical modeling	Iron core magnetism	Magnetic system	Quantum bioelectronics
Fu et al. [132]	Network pharmacology, MD	Salidroside mechanism via ferritinophagy	Autophagy target	Gastric cancer therapy
Jiang et al. [133]	Bioinformatics	Ferroptosis in IDD	Hub gene in pathogenesis	Disc degeneration biomarkers
Lin et al. [134]	RNA analysis	ATF3/ferroptosis in OA	Related RNA expression	Osteoarthritis progression
Al-Garawi et al. [135]	DFT, docking	Metal oxides on ferritin	Target biomarker	Anemia treatment NPs
Strom et al. [136]	Simulations	Ferritin in BRD4 condensates	Platform for regulation	Gene therapy modeling
Masison and Mendes [137]	ODE modeling	Iron transport block	Driver of absorption	Metabolic disease mechanisms
Wang et al. [42, 153]	Review with modeling	Structural modifications	Nanoparticle scaffold	Biomedicine applications
Song et al. [139]	Modeling	Bovine ferritin extraction	Sustainable resource	Nutrient delivery
Zhu et al. [141]	Image-segmentation ML	NP permeability in tumors	Nanocage for delivery	Nanomedicine optimization
Masoomi Nomandan et al. [144]	MD, docking	Glyco-RBD-ferritin design	Stability enhancer	Microbial vaccine refinement
Zain Ul Abidin et al. [159]	DFT review	Ferrite synthesis	Nanocluster template	Biomedical electronics
Bera et al. [147]	Theoretical	Electron transport on silicon	Semiconducting core	Bioelectronics hybrids
Raouf et al. [148]	Numerical FEA	Ferrofluid for hyperthermia	MNP model	Cancer therapy optimization
Weidenbacher et al. [149]	Modeling	His-tag insertion for purification	Vaccine scaffold	Scalable production
Song et al. [154]	Review with modeling	Ferritin as nanoplatform	Multifunctional carrier	Detection and delivery
Rodrigues et al. [155]	Review	Functionalization for vaccines	Antigen display platform	Pathogen protection
Lee et al. [156]	Review	Ferritin scaffold for biotherapeutics	Protein cage	Drug/imaging/vaccines
Ravishankar et al. [157]	Modeling	NP fate in macrophages	Cellular carrier	Biomedical responses
Zhang and Fan [138]	Review	Ferritin nanomedicine for cancer	Targeted carrier	Therapy modalities
Zhang et al. [129]	Review	Ferritin as nanocarrier	Encapsulation platform	Food/nutrition/medicine
Cao et al. [125]	Review	Self-assembled ferritin for vaccines	Antigen delivery scaffold	Immune enhancement
Wang et al. [2]	Review	Functional ferritin for applications	Nanocarrier	Delivery/imaging/theranostics
Ricci et al. [146]	Modeling	Iron loading in ferritin	Structural template	Nanotech applications
Zhang et al. [119]	Review	Ferritin applications in biology	Nanoparticle carrier	Vaccine development
Zhao et al. [143]	CADD, docking	SP94-ferritin-doxorubicin	Targeted conjugate	Cancer drug delivery
Moglia et al. [151]	Computational reduction	Silver NP in ferritin	Template for synthesis	Biomedical stability
Xia et al. [158]	Computational SAR	Ferritin design and SAR	Nanocage scaffold	Nutrition/drug delivery
Lucignano and Ferraro [152]	Review with modeling	Bioactive delivery methods	Loading platform	Therapeutic encapsulation

Several computational methods have been employed to advance ferritin research. ML models, such as random forest, gradient boosting, and XGBoost, are used for predictive analytics in diagnostics and nutritional epidemiology, enabling classification, regression, and biomarker identification with high accuracy (e.g., AUC-ROC values). MD simulations provide atomic-level insights into structural dynamics, interactions, and stability, often combined with cryo-EM for validation. Density functional theory (DFT) calculates electronic structures and energy barriers, particularly for catalytic properties and nanoparticle reactivity. Weighted gene co-expression network analysis (WGCNA) and protein–protein interaction (PPI) networks, derived from bioinformatics, identify gene modules and hub proteins in disease contexts. Molecular docking predicts binding affinities and conformations for drug-ferritin interactions. RNA-seq analysis processes transcriptomic data to link ferritin to pathways like ferroptosis. Kinetic modeling, using ordinary differential equations (ODEs), quantifies dynamic processes such as iron absorption. These methods collectively enable optimization, prediction, and mechanistic understanding across ferritin applications as illustrated in Fig. 8.

Machine learning for diagnostics and prognostics

Machine learning algorithms have been essential in utilizing ferritin as a biomarker for accurate, non-invasive diagnostics of iron disorders and related diseases, frequently incorporating large datasets for high predictive performance. For example, Gradient Boosting and XGBoost models trained on NHANES (19,000+ records) and Kenyan datasets classified IDA based on CBC features like hemoglobin and RDW. They achieved 0.98 recall and 0.87 precision-recall area under the curve (PR-AUC), and SHapley Additive exPlanations (SHAP) analysis revealed that low hemoglobin was the most important predictor. Ferritin was only used for labeling (< 15 µg/L), allowing CBC-only predictions that enhance accessibility in low-resource settings, though external validation highlighted precision variability (0.62), suggesting need for diverse cohorts to mitigate bias [118]. Random forest and gradient boosting also identified IDA from thalassaemia in 1143 Thai anaemic patients with 90.7% accuracy and 0.953 AUC-ROC, surpassing RBC formulas. Ferritin (< 30 ng/mL) confirmed labels but was excluded from features, emphasizing ML's efficiency in endemic areas where traditional tests are costly, yet class imbalance required synthetic minority over-sampling technique (SMOTE), indicating potential overfitting risks [46]. Using ferritin < 30 ng/mL as ground truth, ML models screened for ID by combining CBC and CPD, yielding high AUROC in hospital data. This method facilitates routine testing, but lacks specified model types, limiting reproducibility. It also underscores ferritin's role as a reliable reference for training robust classifiers [120].

Markov simulations evaluated ferritin thresholds for ID screening in women of reproductive age. They found that < 25 µg/L was cost-effective (incremental cost-effectiveness ratio (ICER) $940 per quality-adjusted life year (QALY) oral, $1700/QALY IV) using NHANES data. Ferritin defined ID, capturing 38.6% prevalence, but assumptions about utilities may underestimate real-world variability, highlighting modeling's value for policy [121]. An expanded lifetime Markov model confirmed < 25 µg/L optimality (ICER $680/QALY). It incorporated EQ-5D and sensitivity analysis. Ferritin thresholds improved QALYs, but NHANES bias toward US populations suggests need for global adaptations [122]. ML-informed meta-analysis pooled cohorts to predict CC-IMT from ferritin/transferrin in females (R² = 0.447), revealing non-linear associations. Ferritin is a good inflammatory biomarker for cardiovascular risk stratification, though sex-specificity requires further studies [123]. TCGA/GEO bioinformatics correlated high FTL with poor LIHC prognosis (AUC 0.77), linking it to immune infiltration and ferroptosis. FTL's ferritin subunit role helps with targeted therapies, but dataset limitations call for multi-omics validation [124]. ML (SVM (Support Vector Machine), RF (Random Forest)) associated diets with ferritin in pregnant women (70–76% accuracy), identifying protective patterns; ferritin as an anemia marker supports nutritional interventions, yet small sample sizes (n = 100) limit generalizability [140]. The NANO.PTML IFPTML-DT model predicted NP delivery for neurodegenerative diseases (AUROC 0.97) by combining 4403 NDD assays with 260 NP cytotoxicity data. Ferritin variants emerged as candidates, accelerating screening but requiring toxicity checks [142].

Molecular docking, molecular dynamics and modeling for structure, stability, and vaccine design

Molecular dynamics and structural modeling provide atomic-resolution views of ferritin's assembly, cargo interactions, and vaccine efficacy. This guides optimizations for stability and immunogenicity. Molecular docking followed by MD simulations were used to design a monomeric SARS-CoV-2 spike-ferritin vaccine, predicting stable complexes with exposed upward RBD conformations for optimal ACE2 binding. Ferritin's scaffold allowed for epitope exposure, predicting broad immunity across variants, though solvent effects in simulations may make it seem more flexible than it really is [41]. Similarly, molecular docking confirmed stable bonds between ZnO nanoparticles and ferritin, positioning ferritin as a key target for anemia therapeutics by modulating iron levels and enhancing NP stability [135]. However, in vivo toxicity assessments remain essential to validate these predictions. In another application, computer-aided drug design (CADD) docking optimized SP94-ferritin-doxorubicin linker conformations, improving targeted antitumor delivery and ferritin-mediated efficacy. This structural prediction guides conjugate design but highlights the need for integrated pharmacokinetic simulations to capture in vivo dynamics [143]. RBD-ferritin MD demonstrated enhanced B38 affinity (6 H-bonds) with asymmetric chain fluctuations. Ferritin stabilized pre-fusion conformations, promising for protective vaccines, but short simulation times limit long-term dynamics capture [127]. The fast multipole method MD simulated ferritin for native mass spectrometry, accurately matching CCS (159 nm²). Ferritin served as a model that validated large-scale gas-phase modeling and improved structural proteomics, yet computational cost restricts routine use [130]. Theoretical EPR-based re-evaluation of ferritin's iron core magnetism challenged the giant-spin model, promoting higher-order terms; ferritin's core as a quantum system informs bioelectronics, but experimental discrepancies highlight model limitations [131].

Glycan addition at ferritin-glyco-RBD interface via MD improved stability, with glycans acting as "glue". Ferritin enhanced microbial vaccine designs, reducing aggregation risks, though the variability of glycans needs experimental confirmation [144]. Combined MD and experiments on engineered ferritin nanovaccines tested insertion sites, and found that the C-terminus was optimal for hydrophobicity and tolerance. Ferritin scaffold made the vaccines more stable and guided multivalent displays, but antigen-specific effects require tailored simulations [145]. Ferritin displaying surface proteins showed dynamic RBD subdomains stabilized by ACE2/B38. Ferritin as a scaffold for epitope presentation supports RBD vaccines, which could have effects on nanoreactors. However, interface mutations could alter assembly [127]. Modeling ferritin at varying iron loads revealed structural changes for nanotech. Ferritin as a template aids cargo design, but pH-dependent shifts need integrated MD-DFT [146]. Theoretical modeling of ferritin electron transport on silicon showed semiconductor gaps (0.8–2.6 eV), which made bioelectronics hybrids possible. Ferritin's core facilitated conductance, promising for sensors, though surface effects in vivo remain unmodelled [147]. Numerical finite element analysis modeled ferritin-like ferrofluids for hyperthermia, correlating SLP with frequency; ferritin optimized thermal response for cancer therapy, but tissue variability suggests patient-specific adaptations [148]. His-tag modeling in ferritin simplified vaccine purification, maintaining multivalency; ferritin's amenability supports scalable production, reducing costs, yet tag placement may affect immunogenicity [149].

Bioinformatics and network analysis for disease mechanisms

Bioinformatics analyses have mapped ferritin's involvement in pathways like ferroptosis and autophagy, identifying therapeutic targets. Network pharmacology and MD showed Salidroside inducing ferritinophagy via NCOA4 in gastric cancer, promoting ferroptosis. Targeting ferritin degradation makes anticancer drugs work better, but in vitro validation is crucial for clinical translation [132]. Ferroptosis hub genes in IDD included ferritin-related genes (AUC 0.792–0.900), linking iron to degeneration. Ferritin informs biomarkers, aiding regenerative strategies, though single-dataset reliance limits robustness [133]. RNA analysis tied ATF3/ferritin RNAs to OA progression and infiltration. Ferritin expressions can be used as biomarkers to guide interventions, but immune correlations need more studies [134]. Simulations of ferritin in BRD4 condensates revealed chromatin nucleation; ferritin serves as a platform for modelling gene regulation, presenting possibilities for epigenetic therapies, although condensate dynamics require longer MD simulations [136]. ODE modeling positioned ferritin as the primary iron absorption block driver in enterocytes; its regulatory role elucidates metabolic disorders and provides simulation-derived forecasts for iron therapies [137]. Ultrasound extraction modeling confirmed bovine ferritin's stability and iron-binding for nutrients. Ferritin as a resource supports sustainable delivery and uses bioinformatics for sequence optimization [139].

DFT and theoretical studies for synthesis and interactions

Density functional theory models ferritin's electronic structure and interactions, optimizing synthesis and binding. DFT on metal oxides showed ZnO's high reactivity (dipole 5.80 D) modulating ferritin levels [135]. DFT for cobalt ferrite showed magnetic and electronic properties for biomedicine. Ferritin-like structures inform catalysis, expanding applications, though dopant effects require hybrid DFT-MD [150]. Photochemical reduction DFT modeled silver NP formation in ferritin, boosting stability; ferritin as a template for biomedical NPs, with green synthesis advantages, yet yield optimization is key [151]. Bioactive delivery review incorporated DFT for loading efficiencies. Ferritin's cage models encapsulation, supporting therapeutics, integrating with MD for dynamic predictions [152].

Other computational modeling for applications

Diverse modeling supports ferritin's multifunctional uses. Image-segmentation ML analyzed ferritin NP permeability in 67,000 tumor vessels and improved transendothelial transport. Ferritin nanocages enhanced low-permeability delivery, advancing targeted nanomedicine, with genetic engineering implications [141]. Antimalarial docking in ferritin confirmed selective release; ferritin cage for smart therapeutics, minimizing side effects, but in vivo efficacy requires pharmacokinetic modeling [126]. MD-guided charge engineering controlled ferritin-GFP loading. Ferritin acted as a host for supramolecular delivery, enabling controlled release. However, electrostatics in biological media need refinement [128]. Structural modifications review used modeling for biomedicine; utilizing ferritin scaffolds for drug delivery and imaging, guiding multifunctional NPs [42, 153]. Ferritin nanoplatform modeling for detection and delivery highlighted cage versatility. Ferritin can be used for bioassay and theranostics, and it can tumor-targeting via TfR1 [154]. Functionalization review improved ferritin vaccines by using ferritin for antigen display, enhancing immunogenicity [155]. Ferritin scaffold modeling for biotherapeutics supported drug, imaging, and vaccine development; ferritin's biocompatibility enhances this process [156]. NP fate modeling in macrophages revealed ferritin responses; ferritin for biomedical targeting, informing safety [157]. Ferritin nanomedicine review for cancer used modeling for therapy modalities; ferritin for targeted delivery, reducing toxicity [138]. Ferritin as nanocarrier review with loading models for food/medicine; ferritin encapsulation for nutrients/drugs [129]. Self-assembled ferritin modeling for vaccines focused on boosting the immune system; ferritin scaffold for antigen delivery [125]. Functional ferritin review theoretically designed for applications; ferritin nanocarrier for delivery and imaging [2]. Molecular engineering docking for bacterioferritin encapsulation; a ferritin cage for metals and drugs, expanding bionanotech [146]. Ferritin SAR advancements computationally guided design; ferritin nanocage for nutrition/drug SAR [158].

Future directions in computational ferritin research

Integrating AI with multi-scale modeling promises personalized ferritin nanomedicines, predicting patient-specific responses. Challenges include computational resource demands and validation gaps, necessitating hybrid experimental-computational workflows. Diplomatic interdisciplinary collaborations will find a balance between predictive power and real-world applicability, fostering empathetic advancements that address diverse global health needs.

Literature search strategy

This review discusses recent advancements in ferritin nanoparticle applications, combining experimental breakthroughs with computational innovations. To ensure a complete and unbiased overview, we followed a structured literature review protocol guided by the PICO (Population, Intervention, Comparison, Outcome) framework and PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines, as shown in Fig. 10. The search focused on ferritin's roles in biomedicine, emphasizing post-2024 publications while incorporating foundational pre-2024 studies for context. No original experimental or computational data were generated; all insights are derived from published literature.

Fig. 10.

Flowchart of methodology for ferritin nanoparticle literature review

Literature search strategy

A through search was conducted across multiple databases, including PubMed, Scopus, Web of Science, and Google Scholar, to find related peer-reviewed articles. The search combined key terms related to ferritin nanoparticles and their applications, using Boolean operators: ("ferritin nanoparticle" OR "ferritin nanocage" OR "apoferritin") AND ("cancer therapy" OR "chemotherapy" OR "photodynamic therapy" OR "photothermal therapy" OR "immunotherapy" OR "ferroptosis" OR "vaccine delivery" OR "SARS-CoV-2 vaccine" OR "influenza vaccine" OR "gene therapy" OR "siRNA delivery" OR "CRISPR-Cas9" OR "bioimaging" OR "diagnostics" OR "antioxidant therapy" OR "anti-inflammatory" OR "drug delivery" OR "computational modeling" OR "molecular dynamics" OR "machine learning" OR "density functional theory" OR "bioinformatics"). Additional filters were applied for relevance, such as "experimental" OR "computational" to capture both domains (see Fig. 9).

The primary search was limited to publications from 2024 onwards to prioritize emerging advancements, yielding over 200 initial results. To provide foundational context (e.g., ferritin's structural properties and early applications), we extended backward searches to include pre-2024 papers cited in recent works. The search was done on August 10, 2025, with a total of 350 articles screened.

Inclusion/exclusion criteria

Articles were included if they met the following criteria: (1) focused on ferritin nanoparticles as the primary platform for biomedical applications; (2) reported experimental outcomes (e.g., in vitro/in vivo drug delivery, vaccine efficacy, imaging, or therapy) or computational methods (e.g., machine learning for diagnostics, molecular dynamics for structural optimization, or density functional theory for catalytic modeling); (3) published in English; (4) original research, reviews, or meta-analyses providing empirical data or novel insights; and (5) relevant to human health, animal models, or translational nanotechnology.

Exclusion criteria encompassed: (1) studies not centered on ferritin (e.g., general nanoparticle reviews without ferritin-specific data); (2) non-English publications; (3) editorials, letters, or opinion pieces lacking substantive data; and (4) purely theoretical works without experimental or computational validation. For computational studies, we excluded those without direct application to ferritin's biomedical roles (e.g., unrelated protein modeling). This resulted in 128 articles selected for full-text review and synthesis.

Data extraction and synthesis

Data extraction was performed by reviewer (E.R.) using a standardized template to minimize bias. Extracted elements included: study design (experimental vs. computational), ferritin variant (e.g., human HFn, recombinant apoferritin), application area (e.g., drug delivery, vaccine design, diagnostics), key methodologies (e.g., disassembly/reassembly loading, molecular dynamics simulations), outcomes (e.g., encapsulation efficiency, AUC-ROC for ML models, tumor inhibition rates), and limitations. Discrepancies were resolved through discussion with the senior author (M.A.I.).

Synthesis was thematic, organized into experimental (e.g., chemotherapy, vaccine delivery) and computational (e.g., diagnostics, structural modifications) sections. Where applicable, quantitative data (e.g., loading capacities, survival rates in animal models) were collected for comparison. Narrative construction integrated findings across domains, highlighting synergies (e.g., MD-guided experimental designs) and gaps (e.g., clinical translation barriers).

Limitations in methodology

This review's focus on recent (post-2024) literature may overlook earlier foundational works, though key pre-2024 studies were included for completeness. Database biases (e.g., English-language preference) and publication bias could underrepresent ongoing challenges. The interdisciplinary scope (experimental and computational) required broad search terms, potentially introducing noise; however, through screening prevented this. Studies on ferritin's non-biomedical applications (e.g., pure materials science) were excluded, narrowing the synthesis. Future reviews could add quantitative meta-analyses if more homogeneous data emerge.

Discussion

Ferritin nanoparticles have become one of the most promising and versatile platforms in modern nanomedicine. Their natural 24-mer nanocage structure makes them stand out among protein-based, lipid, and synthetic carriers because it has excellent biocompatibility, targets tumors and the blood–brain barrier through transferrin receptor 1 (TfR1), has high cargo capacity (routinely 20–400 molecules per cage depending on strategy), and can be assembled and disassembled in response to changes in pH or temperature. This review synthesizes experimental breakthroughs in therapeutic delivery, imaging, and immunomodulation with computational advances in structural optimization, diagnostics, and predictive modeling. These domains collectively demonstrate ferritin’s potential to address longstanding challenges in precision medicine while revealing critical translational gaps.

Synergy between experimental and computational perspectives

A defining strength of recent ferritin research is the productive integration of wet-lab innovation and in silico guidance. Experimental loading strategies—passive diffusion, organic solvent assistance, pH-driven disassembly/reassembly, protease-induced nanocages (PINCs), and site-directed channel mutagenesis—have attained high encapsulation efficiencies and controlled release, as evidenced by doxorubicin@HFn hydrogels (72.7% tumor inhibition when combined with anti-PD-1), platinum(IV) prodrugs, and siRNA/CRISPR payloads in tHFn(+) variants that promote endosomal escape.

Computational tools have accelerated and rationalized these developments. Molecular dynamics (MD) simulations have predicted that antigen-ferritin interfaces are stable (e.g., spike-RBD or glyco-RBD conjugates with low RMSD and favorable epitope exposure), that insertion sites for multivalent display are guided, and that charge-engineering for GFP or PROTAC loading is possible. Machine learning models, including XGBoost and random forest classifiers, have used ferritin as a ground-truth biomarker to get high AUC-ROC (> 0.94) for iron deficiency anemia detection from routine CBC data. Network pharmacology and DFT studies have also shown how NCOA4-mediated ferritinophagy and metal-oxide interactions work.

This hybrid workflow reduces empirical trial-and-error, shortens design cycles, and enhances predictability. For example, docking-informed ergosterol loading for cholesterol competition or MD-validated temperature-gated channels. This creates a feedback loop in which computational predictions inform experimental constructs, and empirical outcomes refine models, accelerating development of “smart” ferritin variants.

Therapeutic versatility and preclinical efficacy

Ferritin nanocages are great for multiple therapeutic modalities. In chemotherapy and phototherapy, TfR1-mediated active targeting combined, in conjunction with the EPR effect, results in superior tumor accumulation and reduced systemic toxicity relative to free drugs. Dual-ligand (RGD + TfR1) designs optimized via deep learning, ferroptosis-inducing constructs (e.g., IgP-ss-FRT achieving 83% suppression), and PROTAC-ferritin conjugates that degrade ERCC1/XPF demonstrate multimodal synergy. Photodynamic/photothermal systems (HFn-ICG, Nb-Ftn@ICG) further integrate ablation with immunogenic cell death and abscopal effects.

Vaccine applications are especially advanced. Ferritin’s multivalent display mimics viral geometry, eliciting robust humoral and cellular responses. SARS-CoV-2 spike- or RBD-ferritin nanoparticles (SpFN/ALFQ, Quartet Nanocage, optimized JF.1-4S1158) have been shown to neutralize a wide range of variants and sarbecoviruses. Several candidates are now in Phase 1 trials. Similar successes in influenza, EBV, malaria (Pfs230D1-ferritin), and veterinary platforms (e.g., PRRSV, CSFV, GCRV) underscore platform adaptability. Probiotic- or DNA-based ferritin vaccines further enable needle-free mucosal delivery.

Gene therapy benefits from ferritin’s ability to encapsulate nucleic acids and cross the BBB. Engineered tHFn(+) variants achieve efficient lysosomal escape and ~ 70–80% knockdown of GBM targets (IDH1, MGMT), while HFn-Cas9/DOX systems combine editing with chemotherapy. Antioxidant and anti-inflammatory applications take advantage of native ferroxidase activity and ROS scavenging. For example, rHFn reduces stroke infarct volume by 75%, and curcumin- or rosuvastatin-loaded variants reduce neuroinflammation and hemorrhage sequelae.

Compared with liposomes and antibody–drug conjugates (ADCs), Ferritin-drug conjugates (FDCs) have a higher payload capacity, superior BBB penetration, and inherent TfR1 targeting. However, they may not be as specific to antigens as engineered ADCs. These attributes make ferritin particularly attractive for CNS malignancies and multifunctional theranostics.

Diagnostic and imaging advantages

Ferritin's iron core is paramagnetic, which makes intrinsic T2/T2* MRI contrast possible. Its cage serves as a template for multimodal agents like Gd, gold nanoclusters, fluorescent probes. Hybrid constructs attain high relaxivity and signal-to-noise ratios for small lesions (approximately 2 mm) and atherosclerotic plaques. BT1-ferritin probes detect pathological tau in retinal cells with 85–90% sensitivity in neurodegeneration. Electrochemical biosensors (Pt@ApoF/Ti₃C₂) reach sub-micromolar detection limits for contaminants, illustrating non-medical utility.

Machine learning further amplifies diagnostic power, using ferritin levels as reliable labels to train accessible CBC-based classifiers for iron disorders and cardiovascular risk.

Clinical translation and ongoing trials

Despite robust preclinical evidence supporting ferritin nanoparticles as versatile platforms for drug delivery, vaccine design, and targeted therapies, clinical translation remains in its early stages, with the majority of human data centered on vaccine applications rather than therapeutic nanocages.

Several ferritin-based nanoparticle vaccines have advanced to Phase 1 clinical evaluation, demonstrating favorable safety profiles and immunogenicity. For Epstein-Barr virus (EBV), the gp350-Ferritin nanoparticle vaccine (NCT04645147) completed primary enrollment and dosing by mid-2025, with preliminary results indicating mild to moderate reactogenicity and robust gp350-specific antibody responses in both EBV-seronegative and seropositive adults [160]. Building on this, newer Phase 1 studies are evaluating multivalent approaches, including gH/gL/gp42-Ferritin with or without gp350-Ferritin (NCT06908096, recruiting as of late 2025) and adjuvanted gp350-Ferritin in seronegative individuals (NCT05683834, active but not recruiting) [161, 162]. These trials aim to block multiple EBV entry pathways, potentially preventing infectious mononucleosis and associated malignancies.

In the SARS-CoV-2 space, the spike ferritin nanoparticle (SpFN) vaccine adjuvanted with ALFQ elicited strong neutralizing responses in nonhuman primates and progressed to a Phase 1 first-in-human trial (NCT04784767), which reported acceptable safety, reactogenicity, and immunogenicity in 2023–2024 publications [163, 164]. Broader platform safety is being assessed in dedicated ferritin nanoparticle studies (NCT05903339) [161].

For influenza, ferritin-based candidates such as H1ssF (stabilized stem ferritin nanoparticle) entered Phase 1 evaluation in 2026 (NCT07340047), comparing safety and responses against licensed seasonal vaccines [165]. Preclinical extensions to other coronaviruses, including a stabilized MERS-CoV spike ferritin nanoparticle that conferred robust protection in animal models, position the platform for pandemic preparedness [166].

Manufacturing advancements, including patented optimized production methods [167], support scalability toward GMP-compliant batches. However, challenges persist: most trials remain Phase 1, with limited long-term immunogenicity data, adjuvant optimization needs, and no Phase 2/3 efficacy readouts for ferritin-specific candidates as of early 2026. Immunogenicity assessments must align with regulatory guidance on therapeutic proteins [168], while ethical frameworks emphasize equitable access and risk–benefit monitoring in nanomedicine [169, 170].

Overall, these early clinical signals validate ferritin's biocompatibility and adjuvant-sparing potential, paving the way for broader therapeutic applications (e.g., targeted PROTAC or gene delivery) once vaccine platforms mature.

Challenges and barriers to clinical translation

Despite compelling preclinical data, several hurdles remain. Scalability is limited by variable protein recovery (36–68% in disassembly methods) and the necessity for standardized GMP processes. Heterogeneity in tumor EPR effects is evident in single-vessel analyses which shows low permeability in ~ 70% of models. This makes targeting less reliable. Biodistribution studies reveal natural accumulation in liver, spleen, and kidneys, which raises concerns about off-target effects and clearance kinetics. Long-term immunogenicity, though generally low due to human-sequence homology, requires further evaluation, especially for repeated dosing or non-human variants. Regulatory pathways demand comprehensive toxicology, pharmacokinetics, and manufacturing consistency data that many studies have not yet fully addressed.

Most evidence derives from murine or zebrafish models. Human receptor expression (TfR1, SCARA5) and tumor microenvironments are different. This shows the need for more representative organoids, patient-derived xenografts, and non-human primate studies. Clinical activity remains concentrated in vaccine platforms, with ongoing Phase 1 trials for EBV gp350/gH/gL/gp42-ferritin, SARS-CoV-2, and related candidates. Therapeutic delivery is still behind.

Future directions

To realize ferritin’s full potential, several priorities emerge. AI-assisted design, which combines multi-scale MD, DFT, and ML with high-throughput screening, can predict patient-specific responses, optimize mutations for stability and release, and accelerate personalized nanocages. Hybrid systems, like ferritin-liposome, ferritin-MXene, or stimuli-responsive fusions, may combine advantages of multiple platforms. Focus should expand beyond oncology to metabolic disorders, chronic infections (e.g., HBV ferritin-NP + siRNA), and global health applications (affordable anemia diagnostics, veterinary vaccines).

Sustainable production from recombinant or by-product sources (e.g., bovine liver ultrasound extraction) and green synthesis routes will enhance scalability and environmental compatibility. Strict head-to-head comparisons with approved nanomedicines (Doxil®, ADCs) and standardized reporting of loading efficiency, release kinetics, and biodistribution metrics will facilitate regulatory progress. Finally, interdisciplinary efforts involving clinicians, regulatory experts, and ethicists are essential to navigate safety, equity, and access in diverse populations.

Broader implications

Ferritin nanoparticles exemplify bio-inspired nanotechnology: they are a naturally evolved protein that has been changed through smart engineering and computer modeling to meet the medical needs of the twenty-first century. Their ability to integrate diagnosis, therapy, and prevention within a single, biocompatible scaffold offers a pathway toward truly multifunctional precision nanomedicine. While challenges in scalability, standardization, and clinical validation persist, the accelerating convergence of experimental versatility and computational predictive power positions ferritin as a cornerstone platform for addressing cancer, infectious diseases, neurological disorders, and beyond. Continued investment in translational research will determine whether this ancient iron-storage protein becomes a transformative tool in modern healthcare.

Conclusions

Ferritin nanoparticles is utilized as a platform in biomedicine and nanotechnology, due to their biocompatibility, targeting specificity via transferrin receptor 1 (TfR1), and multiple loading strategies. Experimental advancements have shown ferritin’s efficacy in drug delivery, vaccine development, gene therapy, imaging, diagnostics, and antioxidant/anti-inflammatory treatments, addressing challenges in cancer, infectious diseases, metabolic disorders, and neurological conditions. Strategies such as pH-responsive disassembly, engineered self-assembly, and channel modifications enable high-capacity loading (up to 400 molecules/cage) and precise delivery, while TfR1-mediated uptake and the enhanced permeability and retention (EPR) effect ensure effective targeting with minimal off-target effects. Computational approaches, including machine learning, molecular dynamics, density functional theory, and bioinformatics, have further improved ferritin’s capabilities, achieving high diagnostic accuracy (e.g., AUC-ROC > 0.94 for iron deficiency anemia), stable vaccine designs, and enhanced catalytic properties. Despite these breakthroughs, challenges in scalability, long-term immunogenicity, and regulatory validation remain, improved production yields, comprehensive toxicology studies, and standardized GMP protocols should be done. The combination of experimental and computational methods, coupled with emerging AI-assisted design, promises to accelerate personalized medicine and sustainable nanotechnology applications. Ferritin’s potential to transform precision therapeutics, diagnostics, and global health demonstrates the need for continued research to overcome translational barriers and realize its full clinical impact.

Abbreviations

ADC

Antibody–drug conjugate

AI

Artificial intelligence

ALFQ

Army liposome formulation with QS-21

APC

Antigen-presenting cell

AUC-ROC

Area under the curve-receiver operating characteristic

BBB

Blood–brain barrier

BLFer

Bovine liver ferritin

BNM

Black phosphorus nanosheet modified (context-specific compound)

BRD4

Bromodomain-containing protein 4

C3

Three-fold (channel)

C4

Four-fold (channel)

Cas9

CRISPR-associated protein 9

CADD

Computer-aided drug design

CCS

Collision cross-section

CC-IMT

Common carotid intima-media thickness

CDT

Chemodynamic therapy

CePnF

Conserved epitope peptide nanocage ferritin

CRISPR

Clustered regularly interspaced short palindromic repeats

CSFV

Classical swine fever virus

CPD

Clinical patient data

DC

Dendritic cell

DCFH-DA

2′,7′-Dichlorodihydrofluorescein diacetate

DFT

Density functional theory

DMSO

Dimethyl sulfoxide

Dox@HFn

Doxorubicin-loaded human ferritin

DPPH

2,2-Diphenyl-1-picrylhydrazyl

Dt-FTn

Dual-ligand targeted ferritin nanoparticle

EBV

Epstein-barr virus

EMT

Epithelial–mesenchymal transition

EPR

Enhanced permeability and retention

ESCC

Esophageal squamous cell carcinoma

Fab

Fragment antigen-binding

Fc-RBD-FN

Fc-Tagged receptor-binding domain ferritin nanoparticle

FDCs

Ferritin-drug conjugates

FEs

Ferritin-ergosterol

FGFR

Fibroblast growth factor receptor

FMM

Fast multipole method

Fv-Ab-Ferritin

Fragment variable-antibody ferritin

GCRV-II

Grass carp reovirus type II

GMP

Good manufacturing practices

Gp350

Glycoprotein 350

GPSm-Ft

GP5 mosaic ferritin

HA

Hemagglutinin

HA2-F

Hemagglutinin 2 stem ferritin

HER2

Human epidermal growth factor receptor 2

HF

Human ferritin (recombinant form)

HFn

Human ferritin (H-chain rich)

HIV-1

Human immunodeficiency virus type 1

HMP-NP

Hemagglutinin, matrix protein 1 nanoparticle

HR2-F

Heptad repeat 2 ferritin

IC50

Half maximal inhibitory concentration

ICG

Indocyanine green

ICER

Incremental cost-effectiveness ratio

ICH

International Council for Harmonisation

IDA

Iron deficiency anemia

IgP-ss-FRT

Iron oxide-polydopamine-coated ferritin

IHNV

Infectious hematopoietic necrosis virus

IPA

Indolepropionic acid

iPSC

Induced pluripotent stem cell

LIHC

Liver hepatocellular carcinoma

M2e

Matrix protein 2 ectodomain

MD

Molecular dynamics

MF59

Microfluidized adjuvant 59

mHFn@MTO

Modified human ferritin loaded with mitoxantrone

ML

Machine learning

Mx1

Myxovirus resistance protein 1

NCOA4

Nuclear receptor coactivator 4

Nb-Ftn@ICG

Nanobody-mediated ferritin with indocyanine green

NDD

Neurodegenerative disease

Neoantigen-FNs

Neoantigen-ferritin nanoparticles

NPC1L1

Niemann-pick C1-like 1

NPs

Nanoparticles

ODEs

Ordinary differential equations

OVAT-FNs

Ovalbumin-targeted ferritin nanoparticles

PADRE

Pan-DR epitope

PDA

Polydopamine

PDT

Photodynamic therapy

Pfs230D1

Plasmodium falciparum surface protein 230 domain 1

PICO

Population, intervention, comparison, outcome

PINCs

Protease-induced nanocages

PPI

Protein–protein interaction

PRISMA

Preferred reporting items for systematic reviews and meta-analyses

PRRSV

Porcine reproductive and respiratory syndrome virus

PR-AUC

Precision-recall area under the curve

PTT

Photothermal therapy

QALY

Quality-adjusted life year

RBD

Receptor-binding domain

RBM-F

Receptor-binding motif ferritin

RF

Random forest

rHFn

Recombinant human heavy ferritin

RMSD

Root mean square deviation

ROS

Reactive oxygen species

S18-F

Spike protein 18 ferritin

SARS-CoV-2

Severe acute respiratory syndrome coronavirus 2

SDS

Sodium dodecyl sulfate

SHAP

SHapley Additive exPlanations

siRNA

Small interfering RNA

SLP

Specific loss power

SMOTE

Synthetic minority over-sampling technique

SOCS1

Suppressor of cytokine signaling 1

SpFN

Spike ferritin nanoparticle

SVM

Support vector machine

TfR1

Transferrin receptor 1

Tfh

T follicular helper (cell)

tHFn(+)

Truncated human ferritin (mutated, pH-responsive)

UH-F

Universal helix ferritin

VP4-3-Fn

Viral protein 4–3 ferritin

VP56-2-Fn

Viral protein 56–2 ferritin

WGCNA

Weighted gene co-expression network analysis

XGBoost

Extreme gradient boosting

XPS

X-ray photoelectron spectroscopy

Author contributions

M.A.I. designed the study, supervised the project and edited the manuscript. E.R. collected the data, wrote the manuscript, and provided figures. Both authors have read the manuscript thoroughly.

Funding

No Funding was received.

Data availability

No datasets were generated or analysed during the current study.

Declarations

Ethics approval and consent to participate

Not applicable.

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.