Biomaterial‐Integrated Electroporation for Therapeutic Delivery: From Gene Editing to Tumor Ablation and Immune Modulation

ABSTRACT

Electroporation has evolved from a membrane‐permeabilization method into a versatile therapeutic platform for intracellular delivery, locoregional tumor intervention, and bioelectrically regulated treatment. Depending on pulse intensity and duration, electroporation operates in two distinct modes: reversible electroporation (RE), which transiently permeabilizes the plasma membrane to enable delivery of nucleic acids, proteins, and small molecules while preserving cell viability, and irreversible electroporation (IRE), which causes permanent membrane damage for non‐thermal tissue ablation. Increasingly, the therapeutic scope of electroporation is being expanded through integration with biomaterials, including nanocarriers, hydrogels, soft conductors, and micro/nanoengineered bioelectronic interfaces. These material‐assisted strategies improve cargo protection, field confinement, local retention, tissue conformity, and spatiotemporal control, thereby extending electroporation beyond conventional transfection toward gene editing, engineered cell manufacturing, electrochemotherapy, tumor ablation, immune modulation, and transdermal or localized delivery. In this Review, we summarize the biophysical principles of RE and IRE, discuss how biomaterials reshape electroporation performance across therapeutic settings, compare the design logic of major biomaterial‐assisted electroporation platforms, and highlight key translational challenges, including pulse‐material compatibility, manufacturing scalability, in vivo dosimetry, and regulatory complexity.

Biomaterial‐integrated electroporation establishes a programmable therapeutic interface that coordinates electric‐field modulation, transient or irreversible membrane permeabilization, cargo stabilization, and immune remodeling. This Review highlights how nanocarriers, hydrogels, and bioelectronic interfaces expand electroporation from conventional delivery and ablation toward precision gene engineering, spatiotemporally controlled drug penetration, tumor intervention, and antitumor immunity.

1. Introduction

Electroporation has progressed from a membrane‐permeabilization technique into a versatile therapeutic platform for intracellular delivery, non‐viral cell engineering, locoregional tumor ablation, and bioelectrically regulated treatment. Its broad applicability arises from the ability of pulsed electric fields to transiently or permanently disrupt membrane integrity, thereby enabling two mechanistically distinct therapeutic modes: reversible electroporation (RE), which permits intracellular transport while preserving cell viability, and irreversible electroporation (IRE), which induces permanent membrane damage for non‐thermal tissue ablation. On this basis, electroporation has been increasingly explored for gene editing, engineered immune cell manufacturing, electrochemotherapy, tumor intervention, and immune modulation, positioning it at the intersection of bioelectronics, delivery science, and precision medicine.

Despite these advantages, standalone electroporation remains limited by several intrinsic constraints. In bulk systems, the electric‐field distribution is often heterogeneous, which can lead to simultaneous under‐treatment and over‐treatment within the same biological target. In delicate primary cells or stem cells, aggressive pulse conditions may compromise viability and long‐term function. In addition, electroporation can open the membrane, but it cannot by itself protect fragile therapeutic cargoes, control their release behavior, maintain their local retention, or ensure compatibility with irregular, deformable, and dynamic in vivo tissues. These recurring limitations have slowed the broader translation of electroporation from a powerful physical technique into a fully programmable therapeutic modality.

It is precisely at this point that biomaterials become transformative. Nanocarriers, hydrogels, soft conductors, conductive depots, and micro/nanoengineered bioelectronic interfaces do not simply accompany electroporation as passive carriers or coatings; rather, they define how electric fields, cargoes, and tissues interact. By modulating local field distribution, stabilizing and organizing therapeutic payloads, enabling spatial targeting and local retention, and improving tissue conformity and procedural adaptability, biomaterials can compensate for key weaknesses of standalone electroporation and substantially broaden its therapeutic reach. In this view, electroporation is better understood not as a single transfection tool, but as a biomaterial‐enabled therapeutic interface whose performance emerges from the co‐design of pulse input, material properties, and biological context.

This perspective also helps clarify the necessity of the present Review. Recent high‐quality reviews have separately advanced the field by summarizing electroporation technologies in general, focusing specifically on micro/nano‐electroporation platforms for gene therapy, or emphasizing the immunological and oncologic implications of IRE. However, these themes are often treated in parallel rather than integrated through a common design framework. Broad electroporation reviews tend to emphasize biophysics, device evolution, and clinical translation at large; micro/nano‐electroporation reviews focus primarily on device‐level innovation for gene delivery; and IRE‐centered reviews concentrate on tumor ablation, microenvironment remodeling, and immunological consequences. What remains less explicitly synthesized is how biomaterials themselves reshape electroporation performance across these otherwise separated therapeutic domains, and what comparative design principles emerge when nanoparticle‐assisted systems, hydrogel‐based platforms, and micro/nanoelectrode interfaces are considered side by side.

The present Review is therefore intended to fill this gap in a timely manner. Over the past several years, the field has entered a convergence phase in which micro/nano‐electroporation, non‐viral genome engineering, biomaterial‐gated drug delivery, IRE‐driven immune modulation, and translational standardization have all advanced rapidly and are beginning to inform one another. Against this background, a review organized around how biomaterials reshape electroporation performance is both necessary and timely. Here, we first summarize the biophysical principles that distinguish RE from IRE, and then discuss the major therapeutic applications of electroporation in gene delivery, tumor ablation, drug penetration, and immune activation. We next examine how biomaterial integration changes electroporation through electric‐field modulation, cargo stabilization, spatial targeting, local retention, and improved in vivo compatibility. Building on this framework, we compare the distinct design logic of micro/nanoelectrode interfaces, nanoparticle‐assisted systems, and hydrogel‐based platforms, and finally discuss key translational challenges, including pulse–material compatibility, manufacturability, in vivo dosimetry, and regulatory complexity. In doing so, the present Review seeks to reconceptualize electroporation, transitioning it from an application‐driven modality toward a programmable, biomaterial‐integrated interface for spatially controlled precision medicine.

2. Fundamental Principles of Electroporation

2.1. Reversible Electroporation (RE)

The molecular mechanism of RE is governed by the electric field‐mediated dynamic modulation of the cell membrane. When an external electric field is applied, the transmembrane potential (TMP) rises rapidly, leading to polarization of the phospholipid bilayer (Figure 1A). Once the TMP reaches a critical threshold of approximately 0.2–0.5 V, the resting membrane potential is neutralized, causing lipid rearrangement and the transient formation of hydrophilic nanopores [1]. These pores enable the passive diffusion or electrophoretic transport of ions, drugs, and genetic materials across the membrane (Figure 1B). Once the electric field is withdrawn, the TMP decays, and the lipid bilayer self‐reorganizes to reseal the transient pores, restoring membrane integrity and maintaining cell viability.

FIGURE 1.

(A) The application of an exogenous electric field modulates the cellular TMP. (B) Low transmembrane potential induces reversible perforation for delivery. (C) High‐voltage pulses induce apoptosis via IRE while sparing vasculature and ECM. The illustration was created with BioRender.com.

The dynamics of pore formation and recovery are highly sensitive to the pulse parameters, such as field strength, duration, and pulse repetition frequency. When the TMP exceeds a critical upper limit (>1 V), irreversible membrane damage occurs, rendering the cell incapable of recovery and ultimately leading to death [2]. The principal advantage of RE is its parameter‐dependent controllability. By modulating pulse strength, duration, and frequency, researchers can precisely balance membrane permeability and cell viability, thereby creating a highly tunable platform for electrochemotherapy and gene delivery applications such as plasmid DNA transfection.

2.2. Irreversible Electroporation (IRE)

In contrast, IRE employs high‐voltage electric pulses (typically 1500–3000 V) delivered in microsecond‐to‐millisecond durations, which generate permanent nanoscale defects within the phospholipid bilayer of the cell membrane [3]. These irreversible pores disrupt ionic and osmotic homeostasis, leading to programmed cell death [4].

Distinct from traditional thermal ablation methods, IRE induces non‐thermal, physically mediated cell destruction, thereby avoiding protein denaturation, coagulative necrosis, and fibrotic scarring. Importantly, it preserves the structural integrity of the extracellular matrix (ECM), including collagen and elastin fibers, which maintain the mechanical stability and conductivity of the treated tissue [5, 6] (Figure 1C). This unique tissue‐selective preservation provides IRE with remarkable clinical advantages in the treatment of solid tumors adjacent to vital anatomical structures such as blood vessels, nerves, and biliary or pancreatic ducts. By enabling precise ablation of malignant cells while safeguarding the vascular and neural architecture, IRE effectively overcomes the major limitations of thermal ablation techniques [7]. Consequently, IRE has emerged as a next‐generation non‐thermal ablation modality that enables selective tumor eradication with minimal collateral damage and excellent compatibility for integration with systemic immuno‐ and chemotherapeutic regimens.

Clinically, the distinction between RE and IRE is best understood through their intended therapeutic endpoints. RE is primarily used when transient membrane permeabilization is desired without loss of cell viability, such as in gene electrotransfer, DNA vaccination, and ex vivo engineering of therapeutic immune cells, including CAR‐T and TCR‐T products. In these settings, pulse parameters are tuned to maximize intracellular delivery of plasmid DNA, mRNA, or ribonucleoprotein cargos while preserving functional recovery of the target cells. By contrast, IRE is employed when permanent membrane damage is therapeutically beneficial, most notably for non‐thermal tumor ablation. Its ability to induce apoptosis while sparing major vessels, ducts, and nerves has made it particularly attractive for tumors located near critical structures, including pancreatic, hepatic, and prostate cancers. Taken together, the distinction between reversible and IRE is not merely operational, but defines two fundamentally different therapeutic logics: one centered on transient membrane access and cargo delivery, and the other on permanent membrane disruption and selective tissue ablation. This mechanistic dichotomy explains why electroporation has proven adaptable across such diverse biomedical applications. At the same time, it also underscores a central lesson for the rest of this review: the biological outcome of electroporation cannot be separated from pulse design, tissue context, and the interface through which the electric field is delivered.

3. Therapeutic Applications of Electroporation

Electroporation has emerged as a versatile and controllable tool for the manipulation of cellular behavior, enabling targeted delivery of therapeutic molecules, genetic materials, and regulatory factors. By adjusting the strength and duration of the applied electric pulses, electroporation can either transiently increase membrane permeability for intracellular delivery or induce irreversible damage for selective ablation. These properties have expanded electroporation from a conventional transfection method into a broad therapeutic platform spanning gene editing, tumor ablation, drug‐delivery enhancement, and immune activation.

3.1. Electroporation in Gene Therapy

Gene therapy represents a next‐generation precision medicine approach that delivers therapeutic genetic material into target cells to correct defective or dysregulated genes [8]. Traditional gene delivery methods, including viral transduction and liposome‐mediated transport, are often limited by restricted cargo capacity, unpredictable genomic integration, and the risk of undesired immune activation. In contrast, electroporation offers a non‐viral, physical, and highly controllable strategy for gene introduction into mammalian cells. By applying a short series of electric pulses, negatively charged biomacromolecules such as plasmid DNA, mRNA, or CRISPR/Cas9 ribonucleoprotein (RNP) complexes can be driven directly across the cell membrane and into the cytoplasm or nucleus [9] (Figure 2A). Since its first use by Neumann et al. [10] in the 1980s for plasmid transfection of murine lymphoma cells, continuous optimization of parameters such as field strength, pulse width, and pulse number has markedly improved transfection efficiency while minimizing cytotoxicity.

FIGURE 2.

(A) Application mechanism of gene electrotransfer. Reproduced with permission [9]. Copyright 2024, Nanoscale. (B) Comparison of localized and bulk electroporation on cellular gene expression analyzed by scRNA‐seq. Reproduced with permission [30]. Copyright 2023, Materials Today Bio. (C) A genome editing delivery system that achieves gene editing by delivering exosomes carrying Cas9RNP into mice through electroporation. Reproduced with permission [12]. Copyright 2022, Science Advances.

Electroporation has become a cornerstone technique for genome editing, particularly in the delivery of CRISPR/Cas9 systems into primary cells, stem cells, and immune cells that are resistant to chemical transfection. For example, Maresch et al. [11] developed an electroporation‐based multiplex editing method enabling simultaneous modification of up to 15 oncogenes within individual pancreatic cells, thereby establishing new in vivo tumorigenesis models. Wan et al. [12] engineered an exosome–RNP system in which Cas9 ribonucleoproteins were encapsulated into exosomes via electroporation, achieving liver‐specific genome editing both in vitro and in vivo (Figure 2C). This approach addressed a long‐standing challenge of CRISPR delivery in systemic applications and provided a clinically translatable genome‐editing strategy.

In DNA vaccination, electroporation‐mediated gene transfer (GET) markedly enhances intracellular delivery and antigen expression, resulting in robust immune activation. Roos et al. [13] demonstrated that intradermal electroporation of a prostate cancer DNA vaccine encoding prostate‐specific antigen(PSA) significantly increased gene expression (by 100–1000‐fold) and elicited higher levels of PSA‐specific CD8⁺ T‐cell responses compared to conventional DNA injection. Notably, only specific electroporation parameters that induced strong gene expression also maximized immunogenicity, underscoring the necessity of parameter optimization to balance transfection efficiency and immune potency. In vivo electroporation can dramatically enhance plasmid DNA delivery, substantially amplifying both humoral and cellular immune responses across animal models [14]. Electroporation‐based DNA vaccination has since demonstrated substantial efficacy across multiple disease models, including melanoma [15], cervical cancer [16], hepatitis [17, 18], and HIV/AIDS [19, 20], effectively stimulating both B‐cell and T‐cell immunity.

Moreover, electroporation has been successfully applied in CAR‐T cell engineering, facilitating the insertion of chimeric antigen receptor (CAR) genes through either plasmid or transposon vectors without viral integration. Optimized electroporation conditions can markedly increase CAR‐T cell production efficiency while enhancing their antitumor activity compared with conventional viral methods [21]. Cai et al. [22, 23, 24] used electroporation to deliver anti‐DLL3 CAR circRNA, generating CAR‐T cells capable of nearly 100% cytotoxicity against small‐cell lung cancer (SCLC), with improved RNA stability and expression relative to mRNA. Similarly, electroporation has been pivotal in TCR‐T cell engineering, where Marson et al. [25, 26, 27] utilized CRISPR–Cas9 electroporation to replace endogenous T cell receptor (TCR) genes, thereby enabling tumor‐specific recognition and potent cytotoxicity both in vitro and in vivo. This non‐viral manufacturing route not only improves biosafety but also enables flexible reprogramming and rapid clinical scalability.

Stem cells such as induced pluripotent stem cells (iPSCs), human embryonic stem cells (hESCs), and hematopoietic stem cells (HSCs) present significant challenges for genetic engineering due to their low transfection efficiency and high sensitivity to stress. Electroporation has emerged as a highly efficient, integration‐free physical delivery method with immense potential for stem cell gene editing. For instance, Xu et al. [28] employed a tubular electroporation system to deliver Cas9 ribonucleoproteins (RNPs) together with single‐stranded DNA templates into iPSCs, achieving a homology‐directed repair (HDR) efficiency of 42% and successful gene knock‐in. Furthermore, established protocols have demonstrated that coupling electroporation with chemically modified sgRNAs can enhance CRISPR editing efficiency while circumventing the risk of exogenous DNA integration [29]. However, electroporation still poses challenges related to cellular stress and poor post‐transfer viability. For instance, iPSCs exhibit only ∼3% survival after electroporation, although overexpression of the anti‐apoptotic protein BCL‐XL can increase survival to approximately 90% and elevate HDR efficiencies to 15%–25%. To address this fundamental bottleneck, Mukherjee et al. [30] utilized single‐cell RNA sequencing (scRNA‐seq) to demonstrate that localized electroporation (LEP) enables intracellular delivery of biomolecules without inducing widespread cell death or activating stress‐response pathways that could compromise long‐term cellular physiology (Figure 2B). Compared with bulk electroporation, LEP better preserves cell viability and structural integrity.

Collectively, electroporation offers a versatile, scalable, and high‐efficiency platform for gene delivery across diverse therapeutic contexts. However, field strength and pulse parameters must be carefully tailored to each cell type to balance transfection efficiency with viability. Integration with nanocarriers such as exosomes or biomaterial scaffolds may further enhance tissue specificity and promote the advancement of personalized precision therapeutics.

3.2. Electroporation for Precise Tumor Ablation

IRE has redefined tumor ablation by providing a non‐thermal, ECM‐sparing method to induce apoptosis through controlled electric‐field exposure. Preclinical studies across various solid tumor models have validated IRE's efficacy in achieving precise tumor ablation while preserving critical adjacent structures, offering promising strategies for anatomically complex malignancies.

In pancreatic cancer models, IRE demonstrates potent tumoricidal effects with a favorable safety profile. José et al. [31] reported that IRE induced tumor cell apoptosis, microvascular disruption, and lymphocyte infiltration in an orthotopic murine pancreatic cancer model, significantly prolonging survival. Zhang et al. [32] observed apoptotic body formation and membrane damage within 30 min post‐IRE, as evidenced by TUNEL and Caspase‐3 staining. Lee et al. [33] validated IRE safety in a porcine model, showing intact peripancreatic vessels and nerves with no major complications. Similarly, the application of IRE in porcine liver preserved adjacent portal veins, hepatic arteries, and gallbladder structures, with only transient hepatic function changes [34]. Collectively, these studies indicate that IRE achieves precise ablation through apoptosis, rendering it particularly suitable for highly vascularized tumors near critical structures (Figure 3A).

FIGURE 3.

Multimodal therapeutic applications of electroporation in oncology. (A) IRE induces precise, non‐thermal apoptotic ablation while preserving neurovascular integrity. (B) Electrochemotherapy potentiates drug efficacy via permeabilization‐enhanced uptake and vascular lock‐mediated retention. (C) Electroporation triggers immunogenic cell death and remodels the microenvironment to elicit systemic antitumor immunity. The illustration was created with BioRender.com.

In hepatic tumors, Miller et al. [35] demonstrated that high‐intensity electric fields can effectively inactivate liver cancer cells. Lee et al. [34] observed clear ablation margins and preserved bile ducts and vasculature in a porcine liver model, with TUNEL and BCL‐2 staining confirming apoptosis as the primary mechanism of cell death. To enhance ablation efficacy, Isfort et al. [36] combined IRE with drug‐eluting bead transarterial chemoembolization (DEB‐TACE), significantly expanding the ablation volume, while Fang et al. [37] preheated tissues using radiofrequency ablation (RFA) to lower the electric field threshold, suggesting synergistic approaches for large or anatomically challenging tumors.

IRE also exhibits tissue‐selective ablation in renal and urological tumors. Wendler et al. [38] observed that 28 days post‐IRE in the porcine kidney model, necrotic regions were replaced by scar tissue, while the collecting system and ureters remained intact, exhibiting urothelial regeneration. Sorokin et al. [39] further confirmed these findings, demonstrating that IRE spares the ureters and intestines, accompanied by self‐resolving postoperative inflammation and intact mucosa and muscular layers, highlighting IRE's advantages in maintaining urinary function.

In prostate cancer, IRE achieves tumor ablation while preserving organ function. Onik et al. [40] reported collagen replacement within ablation zones in canine models, with retention of erectile and urinary functions. Kim et al. [41] found peak apoptosis at 24 h post‐ablation in rat models, consistent with MRI and histopathology findings. Tsivian et al. [42] further confirmed vascular and neurovascular bundle integrity in canine models post‐IRE, preserving erectile function and underscoring its safety.

Clinical studies have demonstrated the efficacy of IRE in pancreatic, hepatic, renal, and prostate tumors, with local control rates above 80% reported in selected cohorts, although treatment outcomes are strongly influenced by tumor type, lesion size, electrode placement, and follow‐up duration. IRE's advantage lies not only in its precision but also in its immunogenic potential. The process induces immunogenic cell death (ICD), releasing tumor‐associated antigens (TAAs) and danger‐associated molecular patterns (DAMPs) that stimulate antigen‐presenting cells (APCs) and promote cytotoxic T cell recruitment. This immune activation effect positions IRE as a promising partner for immune checkpoint inhibitors (ICIs) and CAR‐T cell therapy, potentially transforming ablation zones into in situ vaccination sites. The next phase of research should focus on dose‐field modeling, synergistic immunotherapy combinations, and the use of real‐time bioelectrical feedback to ensure consistent tumor eradication with immune activation.

3.3. Electroporation‐Enhanced Drug Penetration

In the context of small‐molecule drug delivery, electroporation can significantly enhance the intracellular uptake of chemotherapeutic agents. Electrochemotherapy (ECT) exemplifies this application as a local therapeutic approach that combines electroporation with chemotherapy to transiently increase the permeability of tumor cell membranes, thereby facilitating the entry of otherwise membrane‐impermeable cytotoxic drugs and inducing cell death [43] (Figure 3B). Okino et al. [44] demonstrated that high‐voltage pulse‐mediated delivery of bleomycin into tumor cells led to significant tumor shrinkage, whereas either high‐voltage pulses or bleomycin alone exhibited minimal anticancer effects, highlighting the synergistic potential of this approach. Drugs commonly used in ECT, such as bleomycin and cisplatin, exert their cytotoxic effects intracellularly following membrane resealing. In addition to enhancing cell permeability, electric pulses induce a transient vascular lock effect, retaining chemotherapeutic agents within the treatment area and minimizing systemic washout [45, 46, 47]. Consequently, ECT can increase the local cytotoxicity of bleomycin by up to 1000‐fold and cisplatin by approximately 80‐fold, enabling the use of lower drug concentrations while minimizing systemic or local adverse effects [48, 49, 50].

Building on these principles, IRE similarly modulates tumor cell membranes and the tumor microenvironment to enhance chemotherapeutic delivery. The combination of IRE and chemotherapeutic agents yields profound synergistic effects. By disrupting the dense tumor stroma, IRE enhances drug penetration and promotes intracellular uptake while preserving the integrity of surrounding normal tissue, ultimately improving therapeutic outcomes and prolonging survival. In contrast, standalone IRE is limited by heterogeneous electric field distribution, with peripheral tumor regions receiving suboptimal field strengths, which may leave residual cells and increase the risk of local recurrence and metastasis. Similarly, conventional systemic chemotherapy suffers from poor tumor selectivity and can induce significant off‐target toxicity. These delivery barriers are particularly pronounced in pancreatic cancer, where the dense stromal microenvironment impedes drug delivery. Addressing this, Bhutiani et al. [51] demonstrated in animal models that IRE ablation increased uptake of gemcitabine in the treated region. Clinical studies by Martin et al. [52] further confirmed that IRE following induction chemotherapy in patients with locally advanced pancreatic cancer (LAPC) enhanced tumor control, as postoperative adjuvant chemotherapy synergistically eradicated microscopic residual disease, reducing local recurrence and prolonging survival.

Overall, electroporation‐enhanced chemotherapy offers a precise strategy to improve intratumoral drug delivery. Future research should focus on optimizing pulse parameters to achieve selective and deeper tissue targeting, integrating electroporation with nanoparticle‐ or hydrogel‐based carriers for sustained drug release, and employing image‐guided systems for spatiotemporal control of drug dosing. Despite the demonstrated synergistic benefits of IRE combined with chemotherapeutics, key mechanistic questions remain unresolved, including how IRE reshapes the tumor microenvironment to facilitate drug penetration and how different chemotherapeutic agents interact with electroporation to maximize efficacy. Moreover, as a localized intervention, strategies for combining IRE with systemic chemotherapy to control metastatic lesions remain underexplored. Addressing these gaps will be essential to translating electroporation‐based chemo‐enhancement into safe, precise, and clinically effective therapies.

3.4. Electroporation‐Activated Anti‐Tumor Immunity

Beyond its direct cytotoxic effects on tumor tissues, IRE has garnered attention for its potent immunomodulatory capacity. By inducing nanoscale membrane pores, IRE triggers immunogenic cell death (ICD), driving the release of previously sequestered tumor‐associated antigens and specific damage‐associated molecular patterns (DAMPs) [5]. Among the most widely used ICD‐associated markers are surface exposure of calreticulin, extracellular release of ATP, and liberation of high mobility group box‐1 protein (HMGB1). Calreticulin functions as an “eat‐me” signal that promotes phagocytic uptake of dying tumor cells, ATP serves as a chemoattractant and maturation signal for dendritic cells, and HMGB1 can activate pattern‐recognition pathways that enhance antigen processing and presentation. Together, these signals facilitate the recruitment and maturation of antigen‐presenting cells, promote cross‐priming of tumor‐specific CD8⁺ T‐cells, and help translate local tumor destruction into a systemic immune response [53] (Figure 3C). Crucially, because IRE preserves the local microvascular architecture, it facilitates efficient immune cell infiltration. This process ultimately transforms the ablation zone into an in situ tumor vaccine capable of eliciting systemic antitumor immunity [54, 55].

Mechanistically, IRE promotes both innate and adaptive immune activation. In the context of innate immunity, IRE enhances dendritic cell (DC) maturation and antigen presentation, increases the infiltration of DCs, NK cells, and M1‐like macrophages within tumors, and reduces immunosuppressive cells such as myeloid‐derived suppressor cells (MDSCs) and regulatory T cells (Tregs), thereby reversing the immunosuppressive tumor microenvironment [56]. For instance, Dai et al. [57] demonstrated in murine hepatocellular carcinoma models that IRE markedly elevated splenic IFN‐γ⁺CD8⁺ T‐cell levels and enhanced the density of CD8⁺ T‐cells and DCs in peri‐ablation zones, accompanied by a concomitant decrease in PD‐1⁺ T cells and Tregs. Similarly, He et al. [58] observed that increasing electric‐pulse intensity in locally advanced pancreatic cancer models significantly elevated effector CD8⁺ T‐cell frequencies in the spleen and lymph nodes, indicating that IRE parameters can tune immune activation.

Concurrently, to bolster adaptive immunity, IRE increases CD8⁺ and CD4⁺ T‐cell infiltration within the ablation zone and secondary lymphoid organs, while reducing circulating Tregs and PD‐1⁺ T cells. In immune‐suppressive tumors such as pancreatic ductal adenocarcinoma (PDAC), Zhao et al. [59] reported that combining IRE with PD‐1 blockade promoted CD8⁺ T‐cell proliferation, reversed stroma‐mediated immunosuppression through DAMP release, and induced durable antitumor memory responses. Wu et al. [60] further demonstrated that IRE synergized with PD‐L1 blockade to activate CD11b⁺CD103⁻ type 2 conventional dendritic cells (cDC2s), enhancing antigen presentation to CD4⁺ T cells and promoting their differentiation into Th1 cells, which in turn recruited NK cells to strengthen antitumor immunity. Collectively, these findings underscore that IRE not only reshapes the local tumor milieu but also orchestrates a systemic and sustained antitumor immune response, positioning it as a promising bridge between local ablation and immunotherapy.

Despite the potent “in situ vaccination” effect of IRE that markedly enhances tumor immunogenicity, the immune responses it induces remain subject to dynamic suppression. For example, IFN‐γ signaling—while critical for CD8⁺ T‐cell activation—can simultaneously upregulate PD‐L1 expression on tumor cells [61]; furthermore, clinical studies indicate that post‐IRE Treg depletion is transient, recovering to baseline within 2–3 weeks [62]. Therefore, future investigations should focus on decoding the spatiotemporal interplay between cancer‐associated fibroblasts (CAFs), immune cells, and extracellular matrix components following IRE, as well as optimizing the timing and sequencing of IRE combined with immune adjuvants or checkpoint inhibitors to overcome rebound immunosuppression. Electroporation thus represents not only a precise ablation technique but also a powerful immune‐priming strategy for solid tumors. Advancing this understanding is essential to fully realizing the dual functional potential of IRE.

Overall, the studies summarized in this section establish electroporation as a multifunctional therapeutic modality capable of enabling gene transfer, non‐thermal tumor ablation, intratumoral drug uptake, and immune activation within a single biophysical framework. A key lesson emerging across these application areas is that the principal strength of electroporation, namely its direct physical control over membrane permeability, also gives rise to several important limitations, including cell stress and loss of viability under bulk pulsing, heterogeneous electric‐field distribution in complex tissues, poor spatial selectivity, insufficient cargo protection during delivery, and limited adaptability to challenging in vivo microenvironments [2]. These recurring bottlenecks provide the central rationale for integrating electroporation with biomaterials, which can elevate electroporation from a powerful physical technique into a more selective, protective, and context‐adapted therapeutic platform. Consequently, these inherent limitations also provide the organizational logic for the following sections, in which biomaterials are discussed according to the specific performance dimensions they reshape in electroporation‐enabled therapy (Tables 1 and 2).

TABLE 1.

Major electroporation modes and their therapeutic positioning.

Electroporation mode	Principle	Typical uses	Key advantages	Main limitations	Representative refs.
Reversible electroporation (RE)	Transient membrane permeabilization below the irreversible damage threshold; pores reseal after pulse withdrawal	Gene electrotransfer, DNA/RNA vaccination, CRISPR/RNP delivery, ex vivo immune‐ or stem‐cell engineering, electrochemotherapy.	Non‐viral, tunable, broadly cargo‐compatible; enables intracellular delivery while preserving cell recovery.	Requires careful balance between delivery efficiency and long‐term cell fitness; bulk fields may cause heterogeneous poration; limited cargo protection and retention.	[1, 2, 9, 13, 14, 21, 28, 30, 43, 44, 45, 46, 47, 48, 49, 50]
Irreversible electroporation (IRE)	High‐voltage pulses induce permanent membrane disruption and non‐thermal cell death.	Locoregional tumor ablation, especially pancreatic, hepatic, renal, prostate, and tumors near vessels, ducts, or nerves.	Non‐thermal ablation; relative preservation of ECM and critical anatomical structures; can promote immunogenic tumor remodeling.	Requires accurate field planning and electrode placement; risks include incomplete marginal ablation, edema, inflammation, muscle contraction, arrhythmia, and electrode‐associated injury.	[3, 4, 5, 6, 7, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 85]
Micro/nano‐electroporation	Micro/nanostructured interfaces confine the electric field to local membrane regions, reducing bulk‐field exposure.	Single‐cell delivery, adherent‐cell transfection, sensitive primary‐cell editing, protein/RNP delivery, and high‐throughput ex vivo cell engineering.	Higher spatial precision, lower voltage demand, improved viability, and better dose control than conventional bulk electroporation.	Limited treatment volume; complex fabrication, sterilization, and device integration; scalability to large tissues remains challenging.	[73, 74, 75, 76, 78, 84, 91, 92, 93, 94, 151, 152]
High‐frequency irreversible electroporation (H‐FIRE)	Bipolar high‐frequency pulse bursts induce IRE‐like ablation while reducing neuromuscular stimulation.	Non‐thermal ablation of solid tumors; investigated for prostate, liver, brain, lung, and other anatomically sensitive lesions.	Reduces neuromuscular stimulation compared with conventional monopolar IRE and may improve procedural tolerability.	Ablation outcome is highly waveform‐dependent; treatment planning and clinical evidence are less mature; thermal accumulation and incomplete ablation still require monitoring.	[63, 64, 65, 66, 67, 68, 69, 70, 71, 72]

Abbreviations: EP, electroporation; RE, reversible electroporation; IRE, irreversible electroporation; H‐FIRE, high‐frequency irreversible electroporation; RNP, ribonucleoprotein.

TABLE 2.

Biomaterial‐assisted electroporation strategies and their translational positioning.

Strategy	Representative materials/platforms	Core design logic	Main application focus	Translational strengths	Key limitations/safety concerns	Representative refs.
Nanocarrier‐assisted electroporation	Liposomes, LNPs, exosomes, polymeric nanoparticles, metal/oxide nanomaterials	Couples membrane permeabilization with cargo protection, targeting, and field‐responsive release	Gene/RNP delivery, DNA vaccination, chemo‐enhancement, immunomodulatory payload delivery	Improves cargo stability and intracellular availability; enables co‐delivery and local pharmacologic amplification	Variable biodistribution and clearance; pulse‐induced carrier destabilization; possible oxidative stress or immune activation	[12, 77, 79, 80, 86, 87, 89, 90, 97, 98, 99]
Hydrogel/depot‐assisted electroporation	Injectable hydrogels, conductive hydrogels, soft depots, hydrogel electronic interfaces	Converts transient electroporation into localized and sustained therapeutic exposure	Locoregional drug/gene delivery, post‐ablation immunotherapy, transdermal delivery, regenerative therapy	Prolongs local retention; reduces washout; improves tissue conformity; supports staged release	Invasive placement for deep tissues; degradation products, swelling, conductivity drift, and chronic inflammation require evaluation	[81, 82, 83, 118, 119, 126, 150, 154, 155]
Micro/nanoelectrode and microfluidic interfaces	Nanopores, nanostraws, nanochannels, hollow nanoelectrodes, CMOS microelectrode arrays, continuous‐flow chips	Miniaturizes the poration zone to achieve field confinement and low‐voltage precision delivery	Single‐cell manipulation, ex vivo immune/stem‐cell engineering, protein/RNP delivery, high‐throughput editing	Enhances spatial precision and cell viability; reduces bulk‐field stress; compatible with sensitive cells	Fabrication complexity; limited treatment volume; scaling and sterilization challenges; electrode‐induced pH shift or local damage	[73, 74, 75, 76, 78, 84, 91, 92, 93, 94, 151, 152]
Piezoelectric or externally triggered electroactive materials	Piezoelectric films, ferroelectric particles, ultrasound‐responsive nanoplatforms, bioresorbable electroactive devices, wearable microneedle patches	Transduces ultrasound/mechanical input into local electrical cues for membrane sensitization and gated release	Wireless electro‐sensitization, on‐demand drug release, sonodynamic therapy, local immunotherapy	Enables spatiotemporal control without conventional wired electrodes; integrates release, stimulation, and multimodal therapy	Electrical output at the tissue–material interface is often insufficiently quantified; ultrasound attenuation and coupling loss limit reproducibility; long‐term implant stability remains uncertain	[104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 161, 162, 163]

Abbreviations: LNPs, lipid nanoparticles; RNP, ribonucleoprotein; CMOS, complementary metal–oxide–semiconductor.

4. How Biomaterials Reshape Electroporation Performance

As outlined above, electroporation has matured into a highly versatile therapeutic platform encompassing intracellular delivery, non‐viral cell engineering, electrochemotherapy, and non‐thermal tumor ablation. Nevertheless, the efficacy of standalone electroporation remains inherently constrained by several recurrent limitations. In conventional, device‐centric electroporation paradigms, cells frequently endure severe biophysical stress and viability loss; electric field distributions can become highly heterogeneous within complex tissues; therapeutic payloads lack adequate protection during transit; and overall performance frequently deteriorates upon translation into anatomically intricate or mechanically dynamic in vivo environments. Collectively, these challenges underscore a fundamental insight: electroporation performance is dictated not solely by electrical pulse parameters, but critically by the physical and biological interfaces where the pulse, cargo, and host tissue converge. It is precisely at this nexus that biomaterials become indispensable. Rather than serving merely as passive carriers or inert device coatings, functional biomaterials fundamentally reshape the electroporation process by actively modulating local electric fields, stabilizing and regulating cargo, dictating the spatiotemporal accumulation of therapeutics, and enhancing biomechanical compatibility with the in vivo milieu. This conceptual framework provides a logical bridge to the subsequent sections, wherein the diverse applications of biomaterial‐assisted electroporation are explored as distinct strategies for leveraging these shared, performance‐enhancing capabilities.

4.1. Electric‐Field Modulation and Interface Engineering

The most fundamental way biomaterials reshape electroporation is by changing how the electric field is distributed and coupled to target membranes. In conventional bulk electroporation, relatively large electrode spacing and nonuniform tissue conductivity can create regions of undertreatment and overtreatment, contributing to poor selectivity and viability loss. Micro/nanoengineered interfaces effectively mitigate these limitations by miniaturizing the effective poration region. Nanostraws, nanopores, hollow nanoelectrodes, and insulated capacitive microelectrodes improve membrane contact, confine the field to smaller areas, and facilitate lower‐voltage operation with better spatial precision and cellular survival [73, 74, 75]. These features make microscale electroporation especially attractive for single‐cell manipulation, adherent‐cell delivery, and ex vivo engineering of sensitive cell populations, although current systems are still limited by restricted processing throughput and manufacturing complexity [76]. At the materials level, conductive nanomaterials can further modulate local electrophysical conditions. Gold nanoparticles, for example, have been shown to act as local field amplifiers or “virtual microelectrodes,” improving transfection efficiency while reducing the need for harsher macroscopic field parameters [77]. Biomaterials can also mitigate electrochemical side effects that become more prominent at small scales, including local pH shifts and electrode‐induced toxicity; non‐faradaic insulated interfaces are particularly relevant here [78]. From a translational standpoint, this category is best viewed as transitioning electroporation from a macroscopic pulse‐delivery method toward a membrane‐matched interface technology. It is therefore the dominant design logic behind many of the micro/nano‐electroporation systems discussed in subsequent sections for precise gene delivery and cell engineering.

4.2. Cargo Stabilization and Formulation Control

Although electroporation effectively permeabilizes the plasma membrane, it lacks the intrinsic capacity to protect or optimize therapeutic cargo during the critical transport window. This limitation becomes particularly important when the payload is fragile, diffusion‐limited, or multicomponent, as is often the case for mRNA, circRNA, proteins, CRISPR/Cas9 ribonucleoproteins, immune adjuvants, or combination nanomedicines [12]. Biomaterials reshape electroporation performance in this context by conditioning the cargo prior to pulse exposure and preserving its functional state during and after delivery. Nanoparticles, liposomes, exosomes, polymeric micelles, and polyelectrolyte nanocomplexes can condense nucleic acids, shield them from degradation, tune surface charge and size for optimal cellular interaction, and support the co‐loading of inhibitors, adjuvants, or imaging agents [79, 80]. In bulk electroporation, such formulation control helps compensate for the otherwise non‐selective nature of membrane permeabilization; in micro/nano‐electroporation, it improves the match between small‐area poration and intracellular trafficking; and in IRE‐associated delivery settings, it allows pharmacologic or immunologic payloads to be spatially and temporally coupled to the ablation event rather than administered as disconnected therapies.

Importantly, this role of biomaterials transcends generic enhancements in delivery efficiency. It specifically addresses a core weakness of stand‐alone electroporation: achieving electrical access to the cytosol without equivalent control over cargo integrity, release behavior, or synchronized co‐delivery. Consequently, cargo stabilization and formulation control form the conceptual basis for many of the subsequent sections on gene modification, drug penetration, and immune modulation. Their main translational challenge is that carrier performance must remain predictable under the intended pulse regime, since strong electric fields can destabilize lipid membranes, alter polymer structure, or modify release kinetics.

4.3. Spatial Targeting and Local Retention

A third major limitation of stand‐alone electroporation is that membrane permeabilization is transient, whereas therapeutic benefit often depends on sustained local availability. Electroporation alone cannot guarantee where a payload accumulates, how long it remains near the target, or whether it is still present when the biological consequences of electroporation—such as stromal disruption, vascular lock, or post‐ablation immune activation—become most relevant. Biomaterials address this challenge by enabling spatial targeting, depot formation, and time‐coordinated release. At the nanoscale, ligand‐functionalized or tissue‐tropic carriers improve regional accumulation and cell selectivity, which is especially valuable when electroporation must be combined with targeted intracellular delivery. At the tissue scale, hydrogels and depot‐forming matrices provide a different but complementary solution: they can retain nucleic acids, drugs, cytokines, or immune agonists within the treatment zone, reduce washout, and sustain payload release over the period in which electroporation‐induced permeability or immune permissiveness is most favorable [81, 82]. This role is particularly important in IRE‐related settings, where the ablation zone and its periphery become a transiently accessible but rapidly evolving microenvironment. Conductive hydrogel‐based electroporation in lymph nodes and hydrogel electronic microarray systems for transdermal nucleic‐acid delivery illustrate how soft materials can simultaneously localize therapy and improve electroporation function [82]. In translational terms, this category is more mature for locoregional and image‐guided applications than for systemic disease, because its main strength lies in exploiting local therapeutic windows rather than replacing whole‐body delivery platforms. This makes it a natural mechanistic bridge to the following sections on electroporation‐assisted drug delivery and antitumor immunity.

4.4. Improved In Vivo Compatibility and Translational Adaptability

Finally, biomaterials reshape electroporation by making it more physically and clinically deployable in complex in vivo environments. Many electroporation systems are optimized in simplified in vitro settings, yet in vivo use introduces curvilinear and dynamic tissue topographies, mechanical mismatch, inflammatory responses to implanted materials, and practical constraints related to access, imaging, and workflow integration. Hydrogels, soft conductors, implantable or injectable conductive interfaces, and bioresorbable electrode systems address these barriers by creating more compliant, tissue‐adaptive, and clinically viable electroporation platforms. This function becomes especially important when moving from bench‐top bulk electroporation or chip‐based micro/nano‐electroporation toward localized in vivo treatment. Soft interfaces can improve contact on irregular biological surfaces, reduce damage caused by rigid electrodes, and allow electroporation to be delivered through capillaries, catheters, postoperative cavities, or wearable/transdermal formats. Recent conductive hydrogel systems for in vivo lymph‐node editing and glioblastoma electroimmunotherapy exemplify how biomaterials can convert electroporation from a device‐centered intervention into a tissue‐adapted therapeutic interface [83]. Translational readiness also differs across electroporation modes. Bulk electroporation remains the most advanced for scalable ex vivo manufacturing, including primary human T‐cell engineering, because throughput and process integration are already being optimized [84]. Micro/nano‐electroporation is highly promising where precision and viability are critical, but scalability to macroscopic tissues, packaging, and sterilization remain bottlenecks. IRE is the most clinically mature in solid‐tumor ablation, as reflected by recent consensus‐guideline efforts, yet its next translational step will increasingly depend on biomaterial‐enabled strategies that improve selectivity, exploit post‐ablation windows, and integrate with combination therapy [85]. Overall, biomaterials do not merely add functionality to electroporation; they define whether electroporation can be made controllable, tolerable, and clinically integrable in the therapeutic settings that matter most.

Ultimately, these four mechanistic principles highlight the centrality of biomaterials in contemporary electroporation design. Instead of simply adding to the list of electroporation applications, they redefine the biological and physical parameters for safe and effective treatment. Consequently, the following sections examine key application areas ranging from gene modification and drug penetration to immune modulation and bioengineered tumor intervention. These topics are explored specifically to demonstrate how different biomaterial platforms capitalize on these performance‐shaping functions.

5. Electroporation‐Integrated Biomaterials for Gene Modification

As delineated in the preceding sections, electroporation has demonstrated exceptional utility in gene delivery, effectively circumventing the bottlenecks associated with viral vectors to facilitate CRISPR‐based editing, CAR‐T cell engineering, and DNA vaccine development. However, the widespread application of conventional bulk electroporation is constrained by inherent limitations, including compromised cell viability, heterogeneous electric field distribution, and restricted in vivo applicability. These challenges have necessitated a paradigm shift toward more systematic and innovative approaches. Consequently, contemporary research is coalescing around two primary strategies: the engineering of biomaterial‐based nanocarriers to optimize the delivery payload and interface, and the structural refinement of electroporation devices to achieve precise, high‐efficiency molecular transport. Notably, a defining breakthrough of the past half‐decade has been the shift from bulk electroporation as a stand‐alone transfection method toward integrated non‐viral gene‐modification systems in which cargo formulation, device geometry, and pulse delivery are synergistically co‐engineered.

5.1. Non‐Viral Gene Delivery Systems Based on Nanocarriers

To address the inefficiency of traditional electroporation in complex biological microenvironments, researchers have increasingly sought to synergize biomaterials with electrical stimulation. Adoptive cell therapy (ACT), which relies on the ex vivo genetic modification and expansion of autologous or allogeneic immune cells, represents a critical frontier in oncology. To optimize the pivotal gene modification step in ACT, Chakraborty et al. [86] engineered a non‐viral delivery platform utilizing polyelectrolyte‐wrapped nanocomplexes, termed “Immunoplexes” (IPs). By condensing DNA with acetylated polyethyleneimine (Ac‐PEI) into a core shielded by an anionic polyelectrolyte (PEMA), this modular system facilitates high‐efficiency transfection of both innate and adaptive immune cells, even in serum‐rich conditions. This design not only provides a targeted preconditioning interface for electroporation but also significantly enhances intracellular cargo delivery, promising to accelerate the clinical translation of immunoengineering.

In parallel, DNA vaccines offer a potent strategy for tumor immunotherapy, yet their efficacy is frequently compromised by inefficient cellular internalization. While in vivo electroporation serves as the benchmark technique for boosting uptake, it often entails adverse effects such as tissue damage and plasmid degradation. To overcome these barriers, Cui et al. [87] leveraged a microfluidic platform to systematically optimize DNA‐loaded lipid nanoparticles (LNPs), identifying a formulation with maximal transfection efficiency (TE) and minimal cytotoxicity. This optimized LNP system successfully delivered a DNA vaccine (pVAX‐hECTM) targeting the human neoantigen HER2, demonstrating the superior therapeutic potential of combining LNP technology with electroporation.

Furthermore, physiological barriers—such as the stratum corneum or corneal epithelium—pose significant challenges for gene delivery. Conventional rigid electrodes often fail to conform to these complex tissue surfaces, limiting therapeutic efficacy. To circumvent this geometric mismatch, Ren et al. [88] introduced a printable metal–polymer conductor (MPC) designed for conformal iontophoresis and electroporation. This soft, bioadaptive MPC patch enabled the localized, high‐efficiency delivery of siRNA targeting VEGF‐A to ocular tissues, achieving an approximately 16‐fold enhancement in delivery efficiency and up to 95% gene silencing. This work provides a novel materials‐based solution for overcoming robust physiological barriers in localized gene therapy.

Nevertheless, the interaction between electroporation pulses and synthetic carriers merits rigorous scrutiny. Beyond permeabilizing the target cell membrane, high‐voltage pulses may alter the physicochemical stability of some nanocarriers. Lipid‐based nanocarriers may undergo membrane destabilization or premature leakage under strong pulsed electric fields, whereas some polymeric nanoparticles may experience structural deformation, altered colloidal stability, or pulse‐dependent changes in release kinetics [79, 80]. In addition, conductive or metal‐containing nanomaterials may amplify local redox activity during pulse exposure, which can be therapeutically useful in selected settings but may also increase off‐target oxidative stress [89, 90]. Accordingly, future biomaterial‐assisted electroporation systems should be designed on the basis of pulse–material compatibility, with explicit attention to carrier robustness, electrochemical stability, and whether the material is intended to remain structurally inert or to function as a responsive component during pulse exposure.

5.2. Engineering Optimization Based on Novel Electroporation Devices

Beyond the use of biomaterials as carriers, substantial progress has been made in the rational design of the electroporation interface itself, focusing on electrode geometry, spatial configuration, and pulse parameter modulation. Liu et al. [91] pioneered the study of Nanopore Electroporation (NPE), coupling experimental data and COMSOL simulations to elucidate that NPE efficiency is intrinsically linked to cell spreading (Figure 4A). They demonstrated that well‐spread adherent cells, which form intimate contacts with nanopores, achieve localized electric field confinement and high transfection rates (>58%), whereas suspended cells exhibit significantly lower efficiency (<19%). This finding provides a critical theoretical framework for the cell‐type‐specific application of NPE.

FIGURE 4.

(A) Schematic diagrams of the model for simultaneous electroporation of a large number of cells in the NPE device, as well as the adherent cell model and suspension cell model cultured in the NPE cell culture chamber. Reproduced with permission [91]. Copyright 2023, American Chemical Society. (B) Schematic diagram of the large‐capacity continuous electroporation chip (LaViE‐Chip) for CRISPR/Cas9 gene editing. Reproduced with permission [93]. Copyright 2025, Bioelectrochemistry. (C) Schematic illustration of the TENG‐driven high‐throughput electroporation system. Reproduced with permission [94]. Copyright 2020, American Chemical Society. (D) Schematic diagram of the porous localized electroporation device (LEPD) system for delivering large functional proteins and protein‐nucleic acid constructs. Reproduced with permission [76]. Copyright 2023, American Chemical Society.

Building on these principles, NPE is being adapted for increasingly complex cellular reprogramming tasks. Ren et al. [92] reported a Nano‐electroporation system capable of efficiently delivering circular RNA (circRNA) to induce cellular reprogramming. Operating at low voltages, this system achieved transfection efficiencies exceeding 90%, rapidly reprogramming primary fibroblasts into human iPSCs while maintaining high cell viability and minimizing the release of pro‐inflammatory cytokines (IL‐6 and IL‐18).

To further push the limits of delivery precision, Duckert et al. [75] introduced High‐Definition Electroporation (HD‐EP) based on CMOS microelectrode arrays (MEAs). By generating highly focused electric fields at nanometer‐scale electrode contacts, this technology enables gene delivery with single‐cell precision. The HD‐EP platform achieved an 81% transfection efficiency for mCherry mRNA with over 90% cell survival, effectively resolving the toxicity and lack of precision inherent to bulk electroporation. However, the clinical utility of such high‐precision chips is often limited by low throughput (∼5 × 10⁴ cells). Addressing this scalability bottleneck, Li et al. [93] developed the Large‐Volume Integrated Electroporation Chip (LaVIE‐Chip). By integrating microfluidic nanostructures with parallel narrow channels to optimize field uniformity, the LaVIE‐Chip enables high‐throughput continuous processing, meeting the substantial cell manufacturing demands (10⁸–10¹⁰ cells per course) required for clinical CAR‐T therapies (Figure 4B).

Finally, addressing the formidable challenges of delivering large biomacromolecules—such as proteins and CRISPR ribonucleoprotein complexes—which are hindered by slow diffusion kinetics and structural instability, Pathak et al. [76] developed the Localized Electroporation Device (LEPD). This integrated platform combines nanoconfined electroporation (focusing fields within 400 nm interfaces), high‐throughput 96‐well plate architecture, and AI‐driven analytics (Figure 4D). This synergistic approach allows for the efficient, low‐toxicity delivery of functional proteins and gene‐editing complexes, offering a strategy that balances the competing demands of safety and efficacy.

Taken together, the studies discussed in this section indicate that the next generation of electroporation‐enabled gene modification will be defined not merely by higher transfection efficiency, but by the rational engineering of the entire delivery interface. Nanocarrier‐assisted systems contribute by protecting fragile cargos, expanding tissue accessibility, and mitigating pulse‐associated damage, whereas micro/nanoengineered electroporation devices improve field confinement, throughput, and compatibility with sensitive or hard‐to‐transfect cell types. Viewed collectively, these advances suggest a central design principle: successful non‐viral gene delivery depends on the coordinated optimization of cargo formulation, pulse parameters, and device geometry, rather than the refinement of any single component in isolation. At the same time, the field has not yet fully overcome several key translational barriers, including limited universality across cell types, industrial‐scale manufacturability, and robust in vivo adaptability. Future progress will therefore depend on the development of standardized, high‐throughput, and application‐specific electroporation platforms that can integrate biomaterial design with microfluidics, nanoengineering, and intelligent device control, ultimately bridging the gap between laboratory innovation and clinically scalable gene‐modification technologies.

6. Electroporation‐Integrated Biomaterials for Enhanced Drug Penetration

Another defining recent breakthrough is the transition from electroporation‐enhanced uptake to biomaterial‐gated drug availability. Emerging studies demonstrate that nanoliposomes, stimulus‐responsive depots, and piezoelectric platforms can synchronize payload release with the electroporation‐induced therapeutic window, thereby converting electroporation from a permeability‐enhancing tool into a spatiotemporally programmable drug‐delivery interface [95, 96]. By exploiting the enhanced permeability and retention (EPR) effect alongside active ligand‐mediated targeting, drug‐loaded nanocarriers achieve site‐specific accumulation within the tumor microenvironment. This targeted approach not only maximizes therapeutic bioavailability but also mitigates systemic toxicity by reducing dosing frequency, thereby offering a robust foundation for multimodal, synergistic, and precise oncological interventions.

6.1. Synergistic Delivery Strategies Based on Nanoliposomes

Liposomes, as a cornerstone of nanomedicine, have evolved from passive carriers into sophisticated, field‐responsive systems when integrated with electroporation. This integration has advanced from simply enhancing cellular uptake to achieving precise, electric‐field‐triggered payload release. Early efforts by Pisani et al. [97] demonstrated that subjecting cationic gentamicin‐loaded liposomes to electroporation significantly potentiated drug internalization, maximizing intracellular concentrations and promising improved clinical outcomes. However, conventional liposomes often exhibit suboptimal release kinetics under electric fields. Addressing this limitation, Silbaugh et al. [80] optimized the lipid bilayer mechanics, specifically modulating membrane fluidity to elevate electroporation‐induced release rates to over 50%. This underscores that the rational engineering of biomaterial properties is critical for amplifying the synergistic efficacy of electroporation‐mediated delivery.

To further circumvent the systemic toxicity associated with potent chemotherapeutics, Tian et al. [98] engineered a liposomal formulation of the mTOR inhibitor BEZ235 (L‐BEZ) tailored for IRE. Driven by the gradient electric fields generated during IRE, BEZ235 is efficiently liberated from the liposomal core specifically within the ablation zone. This spatiotemporally controlled release strategy not only minimized systemic adverse effects but also significantly suppressed tumor cell viability and proliferation in vivo, thereby reducing the risk of local recurrence post‐ablation.

Building on these precise delivery platforms, researchers have further explored strategies to activate specific cell death pathways. Han et al. [99] designed an “electro‐ferroptosis” nanotherapeutic (EFN) comprising iron oxide nanoassemblies (IONA) and ascorbic acid‐loaded liposomes (Lip‐AA). Upon IRE stimulation, the release of ascorbic acid triggers the reduction of IONAs, generating a surge of ferrous ions (Fe^{2 +}) that initiates ferroptosis—a regulated, iron‐dependent cell death mechanism. Concurrently, IRE enhances membrane permeability, facilitating the cytosolic accumulation of Fe^{2 +} to amplify this lethal cascade (Figure 5B). Consequently, the IRE‐triggered EFN system exhibited superior tumor growth inhibition and immune activation. Most recently, this synergistic concept has been expanded into the realm of immunotherapy. Novel cationic liposomes loaded with STING agonists have been combined with IRE to achieve a dual effect: high‐efficiency cytosolic delivery and the induction of immunogenic cell death (ICD). This combinatorial approach effectively functions as an in situ tumor vaccine, robustly orchestrating a systemic antitumor immune response.

FIGURE 5.

(A) Schematic diagram of a self‐powered controllable transdermal drug delivery system that controls drug release by collecting mechanical energy and converting it into electrical energy. Reproduced with permission [106]. Copyright 2021, Advanced Functional Materials. (B) The schematic illustration of IRE‐triggered electro‐ferroptotic nanoammunition (EFN)‐mediated ferroptosis induction for cancer therapy. Reproduced with permission [99]. Copyright 2024, Elsevier. (C) Schematic illustration of the electrically stimulated control of collagen monomer assembly within nanoliposomes to direct the formation of collagen fibrils in dermal fibroblasts. Reproduced with permission [109]. Copyright 2024, Advanced Healthcare Materials.

From a translational perspective, the combination of locally administered nanoliposomes or other injectable depots with IRE is likely to be most practical for localized and image‐accessible lesions. Although intratumoral delivery can increase drug concentration within the ablation zone while reducing systemic exposure, this strategy also adds a second local intervention to an already image‐guided and invasive ablative procedure, which may limit its practicality for deep‐seated tumors, multifocal metastatic disease, or settings requiring repeated multisite treatment [100]. Accordingly, clinically feasible next‐generation approaches may need to incorporate minimally invasive image‐guided injection workflows, catheter‐based regional delivery, or depot formulations with prolonged intratumoral retention to reduce the need for frequent re‐administration [101, 102]. In parallel, because IRE remains a fundamentally local treatment, biomaterial‐assisted IRE will likely be most effective when integrated into multimodal regimens—including systemic therapy or immunotherapy—rather than being viewed as a stand‐alone solution for disseminated cancer [103].

6.2. Intelligent Controlled Release Based on Piezoelectric Materials

Piezoelectric biomaterials introduce a qualitatively different strategy for electroporation‐enabled delivery: rather than relying exclusively on externally applied electrodes, they can convert ultrasound or biomechanical deformation into localized electrical cues at the material–tissue interface [104]. The significance of this approach lies not simply in “self‐powering,” but in the possibility of coupling three functions within a single platform—membrane sensitization, gated release, and spatially confined stimulation. In this sense, the earliest studies in the field are best understood as establishing mechanistic feasibility rather than offering direct therapeutic substitutes for conventional electroporation systems. Dolai et al. [105] provided one of the clearest foundational demonstrations by showing that a hydroxyapatite nanowire/PVDF film could generate a local potential under ultrasound sufficient to induce RE and facilitate the intracellular uptake of FITC‐dextran. By contrast, studies by Yang et al. [106], Sun et al. [107], and Wu et al. [108] shifted the emphasis from membrane permeabilization itself to electrically regulated release, showing that piezoelectric output can be used to modulate drug discharge kinetics, prolong retention, or create switchable “on/off” release behavior (Figure 5A). Even the study by Brilian et al. [109], although not strictly a piezoelectric system, is conceptually relevant here because it illustrates the same broader principle: local electric cues can be exploited not only to release cargo, but also to bias its transport across biological interfaces (Figure 5C). Taken together, these early studies suggest that the real promise of piezoelectric materials is not merely autonomous powering, but the creation of an electrically active delivery interface whose permeability and release behavior can both be tuned in situ.

Recent progress has moved this concept closer to oncology, but the field remains mechanistically heterogeneous. The most direct attempt to align piezoelectric activation with an IRE‐like therapeutic goal is the work of Pedraza et al. [110], who used ultrasound‐stimulated ferroelectric P(VDF‐TrFE) microparticles to induce cytotoxicity in cancer cells and explicitly framed the system as a possible route toward irreversible‐electroporation‐like effects. However, most of the newer tumor studies operate in a different regime: rather than replacing classical electric pulse delivery, they use piezoelectric materials to amplify membrane perturbation, ionic disequilibrium, catalytic ROS generation, or triggered drug liberation under ultrasound. This distinction is important. For example, the HER2‐targeted platform reported by Chen et al. [111] enhanced sonodynamic therapy by combining piezoelectric stimulation with membrane‐potential disturbance, RE, and Ca^{2 +} imbalance, while the pancreatic cancer systems developed by Wang et al. [112] and Song et al. [113] used piezocatalytic mechanisms to regulate NO release, chemodrug liberation, and immune modulation. Likewise, the wearable flexible ultrasound microneedle patch described by Xue et al. [114] represents a notable translational advance, but its strength lies in integrating minimally invasive local delivery, ROS amplification, and immunotherapy synergy rather than in reproducing a validated IRE‐equivalent electrical waveform. Thus, recent literature should not be overinterpreted as evidence that piezoelectric platforms are poised to supplant conventional electroporation hardware. Rather, they are emerging as sophisticated adjuncts for electro‐sensitization and release amplification, capable of rendering localized therapies highly programmable while minimizing reliance on wired devices.

In summary, piezoelectric biomaterials establish wirelessly triggered electroactive interfaces capable of locally sensitizing membranes, gating on‐demand drug release, and amplifying multimodal therapies. Early proof‐of‐concept studies demonstrated that piezoelectric films or coatings could generate localized electrical stimuli sufficient for RE‐like permeabilization or release modulation [115]. More recently, tumor‐directed systems have extended this concept to ultrasound‐driven piezoelectric nanoplatforms, coupling membrane perturbation, reactive oxygen species (ROS) generation, and ion imbalance with triggered cargo release. However, current studies remain mechanistically diverse, and genuinely electroporation‐equivalent piezoelectric systems are still uncommon.

More importantly, their translational potential is fundamentally constrained by the physics of ultrasound propagation in human tissues. Clinically, ∼1 MHz ultrasound is typically utilized for targets at depths of approximately 2.5–5 cm, whereas 3 MHz is more suitable for superficial tissues at depths of ∼0.8–1.6 cm [116, 117]. As the acoustic path length increases and tissue composition becomes more heterogeneous, energy absorption, scattering, and reflection progressively attenuate the acoustic waves reaching the implanted piezoelectric material, particularly at interfaces with significant acoustic impedance mismatches (e.g., bone or gas interfaces) [116]. Consequently, the in vivo performance of ultrasound‐triggered piezoelectric systems for deep‐seated tumors depends not only on the intrinsic material piezocoefficient but also on the acoustic frequency, target depth, coupling efficiency, device geometry, and the local tissue microenvironment. Currently, many reports still fail to quantify the electrical output generated at the tissue‐material interface in a manner directly correlative with membrane threshold phenomena, classical electroporation parameters, or deep‐tissue dosimetry. Therefore, future progress will necessitate a tighter integration of acoustic dosimetry, precise piezoelectric output profiling, release kinetics, and biological responses under clinically relevant conditions. In this sense, the most realistic near‐term role of piezoelectric biomaterials is not to serve as stand‐alone replacements for established electroporation platforms, but to function as intelligent auxiliary modules that locally lower transport barriers, gate release on demand, and enhance multimodal electroporation‐based therapy.

Taken together, biomaterial‐assisted electroporation strategies for drug penetration can be understood as efforts to address three interlinked clinical challenges: how to concentrate therapeutics at the target site, how to synchronize release with the electroporation‐induced therapeutic window, and how to minimize systemic toxicity while preserving local potency. Nanoliposomal systems are particularly effective when triggered release and local pharmacologic amplification are needed, whereas piezoelectric platforms are more promising as programmable auxiliary modules for on‐demand release and electro‐sensitization. The broader lesson from this section is that electroporation does not simply increase uptake; when coupled with rationally designed biomaterials, it can fundamentally reshape the spatiotemporal landscape of drug bioavailability.

7. Electroporation–Biomaterial Synergy for Activation of Antitumor Immune Cells

Building upon the expanding role of electroporation in drug delivery and tumor ablation, its integration with functional biomaterials has opened compelling new opportunities in cancer immunomodulation and vaccine development. Among the recent advances in this area, perhaps the most profound conceptual advancement is the recognition that IRE can create a transient immune‐permissive window following ablation, during which tumor antigen release, stromal disruption, and temporary relief of immunosuppression synergistically reverse local immune tolerance. Biomaterials can be strategically designed to exploit this window for localized checkpoint blockade, vaccine‐like antigen presentation, sustained immune‐adjuvant delivery, and phenotype‐specific reprogramming of the tumor microenvironment (TME). This emerging paradigm has substantially redefined the therapeutic scope of electroporation from a purely local ablative modality toward a more versatile platform for systemic immunotherapy design. In this context, advanced biomaterial‐assisted electroporation systems are increasingly being developed to overcome the limitations of conventional immune checkpoint inhibitors (ICIs), including delivery barriers, off‐target immune dysregulation, and acquired resistance, thereby enabling more localized, controllable, and durable antitumor immune activation (Figure 6A).

FIGURE 6.

(A) IRE induces nanoparticle drug delivery. Reproduced with permission [135]. Copyright 2024, American Chemical Society. (B) IRE combined with an oral microalgae‐based biohybrid system carrying immune checkpoint inhibitors reduces tumor burden and enhances antitumor immune responses. Reproduced with permission [122]. Copyright 2025, Advanced Science. (C) After injecting CMP nanoparticles into the tumor and performing IRE, tumor cells are killed, and damage‐associated molecular patterns (DAMPs) are released, which activate antigen‐presenting cells (APCs), promote the expression of type 1 interferon (IFN) and pro‐inflammatory cytokines, and ultimately lead to an effective anti‐tumor immune response. Reproduced with permission [123]. Copyright 2022, Small. (D) Schematic illustration of utilizing electroporation and exosome delivery for Ginsenoside Rb1 to effectively address the bottleneck of immunotherapy resistance. Reproduced with permission [128]. Copyright 2025, Journal of Nanobiotechnology.

7.1. Optimizing Delivery of Immune Checkpoint Inhibitors (ICIs)

A primary translational bottleneck in ICI therapy is the inefficient and untargeted biodistribution of monoclonal antibodies. To overcome these limitations, Li et al. [118] engineered an injectable alginate hydrogel for the sustained, localized co‐delivery of celecoxib and anti–PD‐1 antibodies. In B16‐F10 melanoma and 4T1 breast cancer models, this formulation enabled prolonged intratumoral retention while minimizing off‐target systemic exposure, ultimately enhancing checkpoint blockade efficacy. Compared with individual agents, the hydrogel‐based combination markedly elevated CD4⁺ and CD8⁺ IFN‐γ⁺ effector T‐cell populations in the TME, draining lymph nodes, and spleen, while reducing Tregs, MDSCs, and pro‐tumor cytokines. This work provided one of the first systematic demonstrations that biomaterial‐enabled codelivery of an ICI and an anti‐inflammatory agent can synergistically amplify antitumor immunity. In a complementary approach, Song et al. [119] utilized an injectable peptide hydrogel for the coordinated delivery of tumor vaccines and dual ICIs. Within this system, electroporation served as a biophysical trigger to deliver vaccine components or adjuvants and to induce immunogenic cell death, thereby generating an “in situ vaccine.” The hydrogel formed an immune‐activating depot at the tumor site, enabling sustained release of antigens and ICIs and substantially enhancing CD8⁺ T‐cell infiltration in melanoma and 4T1 models.

Recognizing the persistent issues of off‐target toxicity and drug resistance, Sun et al. [120] constructed a dual‐payload exosome system co‐delivering siRNA targeting Bruton's tyrosine kinase (BTK) and the natural anti‐tumor agent isoimperatorin (ISOIM). Following the electroporation‐mediated loading of si‐BTK and ISOIM into tumor‐derived exosomes, the resulting ISOIM/siBTK@exosome complex achieved high encapsulation efficiency and potent immunomodulation. This formulation promoted T‐cell activation, inhibited M2 macrophage polarization, suppressed tumor growth in vivo, and extended survival without causing organ damage, offering an elegant solution to overcome resistance to checkpoint blockade. Addressing IRE‐induced autophagy as a resistance mechanism in pancreatic cancer, Ye et al. [121] designed polymeric micelles (M‐R/T) co‐encapsulating inhibitors of RAGE and TLR4. When combined with IRE, these micelles blocked autophagy‐mediated cytoprotection, increased tumor cell sensitivity to electroporation, and significantly improved apoptosis, tumor suppression, and survival.

To further expand administration routes, Zeng et al. [122] developed an oral delivery platform based on Chlorella vulgaris and polydopamine for encapsulating PD‐1 inhibitors (Figure 6B). This system protected therapeutics from gastric degradation and enhanced intestinal absorption, yielding improved local drug concentrations with fewer systemic toxicities. When combined with IRE, the formulation synergistically activated antitumor immune responses, highlighting the feasibility of clinically relevant oral ICI delivery with improved patient adherence.

7.2. Reprogramming Immune Cell Phenotypes within the Tumor Microenvironment

Beyond drug delivery, electroporation–biomaterial platforms offer an additional advantage: the ability to actively reprogram the immunosuppressive TME. For example, Han et al. [123] developed metal‐phenolic network (MPN) nanoparticles through the coordination of tannic acid and manganese ions, subsequently functionalizing this Mn‐PN core with CpG‐ODNs to generate the final nanocarriers (termed CMPs). When combined with IRE, these nanoparticles were efficiently internalized by macrophages and drove polarization toward the pro‐inflammatory M1 phenotype, resulting in prolonged intratumoral retention, robust tumor suppression, and extended survival (Figure 6C).

Efficient antigen cross‐presentation plays a pivotal role in activating cytotoxic T‐cell responses; however, each step of this process constitutes a tightly regulated cascade that is difficult to modulate therapeutically [124, 125]. To enhance this cascade, Liu et al. [126] engineered liposome/CaCO₃ hybrid microgel vaccines that, in combination with IRE ablation, released FLT3L and CD40L in a pH‐responsive manner. This facilitated the migration of cDC1s to tumor‐draining lymph nodes and triggered efficient cross‐presentation. Consequently, the strategy transformed “cold” pancreatic tumors into “hot” immunogenic lesions, activated systemic antitumor immunity, suppressed metastasis, and improved survival.

Neutrophil regulation within the TME is another key determinant of IRE outcomes. In pancreatic cancer, IRE often induces neutrophil infiltration; however, these cells tend to polarize toward a tumor‐promoting N2 phenotype. Peng et al. [127] addressed this issue by designing degradable mesoporous silica nanoparticles (dMSN‐SB) for the localized release of a TGF‐β receptor inhibitor. The treatment shifted neutrophils from the N2 to the antitumor N1 phenotype, enhancing immune activation and improving IRE efficacy. This work highlights neutrophil polarization as a previously underexplored but critical target for combination electroporation immunotherapy.

Surmounting immunotherapy resistance remains another pressing challenge. Jin et al. [128] employed electroporation together with tumor‐targeting peptide–modified exosomes (T‐exo) to deliver ginsenoside Rb1. This approach improved the stability and bioavailability of Rb1, suppressed PI3K/AKT/mTOR signaling, promoted M1 macrophage polarization, and enhanced CD8⁺ T‐cell proliferation and cytotoxicity (Figure 6D). When combined with PD‐1/PD‐L1 blockade, the treatment demonstrated dual therapeutic synergy, underscoring the potential of electroporation‐enabled platforms to restore sensitivity to immunotherapy.

Highlighting the systemic potential of these combined modalities, a recent study on metastatic pancreatic ductal adenocarcinoma revealed that combining IRE with the intratumoral injection of plant virus nanoparticles (e.g., CPMV) triggered a profound abscopal effect [129]. This combination increased activated CD8⁺ T‐cell and NK cell infiltration in both primary tumors and distant liver metastases, confirming the capacity of electroporation‐biomaterial synergy to drive systemic immunity.

An important translational consideration in combining IRE with immune checkpoint inhibitors (ICIs) is the timing of immunotherapy relative to ablation. Current preclinical, translational, and early clinical evidence suggests that the most relevant opportunity for intervention likely lies in the immediate to early post‐IRE phase, rather than at a substantially delayed stage [130, 131]. This period appears to coincide with the release of tumor antigens and damage‐associated molecular patterns (DAMPs), enhanced dendritic‐cell activation, and a transient alleviation of immune suppression after ablation [131]. Available studies further indicate that these immune‐modulatory effects are time‐limited, with reduced regulatory T‐cell activity and enhanced effector T‐cell responses being most evident during the first several days to approximately 2 weeks after IRE [132, 133]. Early clinical studies in locally advanced pancreatic cancer have, notably, initiated anti–PD‐1 or PD‐1/PD‐L1 blockade at 7 days or within 2 weeks after IRE, supporting the feasibility of early post‐ablation intervention in translational settings [134]. Accordingly, ICIs or biomaterial‐based immune adjuvants may be most reasonably administered concurrently with IRE or within the first days to 1–2 weeks afterward, although a universally optimal schedule has not yet been established and will likely depend on tumor type, pulse protocol, and therapeutic mechanism.

In summary, the integration of electroporation with biomaterials is shifting cancer immunotherapy from passive drug administration toward spatially and temporally programmable immune reprogramming. The key message from this section is that electroporation contributes more than membrane permeabilization alone: it can create an immune‐permissive window through antigen release, stromal disruption, and the transient relief of immune suppression, while biomaterials determine how effectively this window is exploited through localized retention, co‐delivery, or phenotype‐specific immune modulation. The most promising future direction will therefore be to design electroporation–biomaterial combinations not only around payload choice, but also around timing, immune‐cell targets, and the dynamics of post‐ablation immune remodeling.

8. Electroporation‐Synergistic Bioengineered Materials

Conventional IRE is inherently limited by its lack of cellular selectivity, as any cell experiencing a transmembrane potential above the critical threshold undergoes structural disruption. To address this constraint, recent endeavors have focused on integrating IRE with nanomaterials to enhance targeting specificity. The central rationale is to leverage functionalized nanostructures that selectively accumulate within tumor tissues, thereby lowering the requisite electric‐field threshold by modulating cellular electrophysiology or altering the local field distribution.

8.1. Nanoadjuvants for Lowering the Electroporation Threshold

Using nanomaterials as “electrical adjuvants” to amplify local field effects has emerged as an effective strategy to reduce IRE thresholds and mitigate collateral damage to surrounding healthy tissues. For instance, Raffa et al. [136] first demonstrated that boron nitride nanotubes (BNNTs) can potentiate IRE‐induced cytotoxicity by lowering the electric field required to achieve membrane disruption. However, the biocompatibility of BNNTs is strongly influenced by their size and fabrication process, necessitating strict concentration control (<500 ng/mL) to avoid adverse effects [137, 138].

To circumvent these biocompatibility constraints, Heberlein et al. [139] introduced a non‐toxic cyclopentadecanolide (CPDL) nanoemulsion as a synergistic agent. Pre‐incubation of PANC‐1 cell suspensions with CPDL for 10–40 min followed by IRE at 1000–4500 V/cm increased acute cell‐killing efficiency by 2.2‐fold and reduced the electric field required to achieve equivalent cytotoxicity by 55%. Mechanistically, CPDL delays membrane resealing and facilitates nanoparticle‐mediated drug delivery, thereby minimizing post‐treatment sequelae in normal tissues and shifting the balance toward progressive, delayed cell death over immediate necrotic injury—an approach that may reduce inflammation triggered by high‐voltage pulses. Together, these studies underscore the promise of nanoadjuvants in lowering therapeutic thresholds and improving safety profiles.

8.2. Functionalized Nanomaterials for Enhanced Target Specificity

To further refine tumor specificity, Colacino et al. [140] developed a folate receptor (FR)–mediated targeting strategy in which folic acid (FA) was conjugated to cellulose nanocrystals (CNCs) to form CNC‐FA complexes. CNC‐FA significantly enhanced IRE‐induced cytotoxicity in FR‐positive cancer cells while exerting no effect on FR‐negative cells. Confocal imaging confirmed FA‐FR–dependent enrichment of CNC‐FA on tumor cell membranes, likely altering membrane architecture or local field distribution to potentiate electroporation. This functionalization reduced the required electric field by 50 V/cm and expanded the ablation volume (from 3 to 4 cm³), offering particular advantages at invasive tumor margins where precision is critical.

Beyond targeting, heterogeneity in intratumoral electric‐field distribution poses an additional challenge: certain tumor regions may experience only RE, enabling activation of DNA damage response (DDR) pathways and increasing recurrence risk. To overcome this, Long et al. [141] engineered a nanotherapeutic formulation (M‐TK‐OA) that disrupts RE‐induced homologous recombination (HR) and non‐homologous end‐joining (NHEJ) repair, thereby impairing clonogenic survival (Figure 7A). Combined IRE and M‐TK‐OA treatment significantly prolonged survival in both subcutaneous and orthotopic pancreatic ductal adenocarcinoma (PDAC) models and elicited CD8⁺ T cell–mediated antitumor immunity with durable memory responses.

FIGURE 7.

(A) IRE combined with the nanoformulation (M‐TK‐OA) disrupts the DDR response. Reproduced with permission [141]. Copyright 2023, American Chemical Society. (B) The flexible electronic catheter efficiently captures and kills circulating tumor cells (CTCs) through IRE. Reproduced with permission [142]. Copyright 2022, American Chemical Society. (C) The high‐voltage triboelectric nanogenerator (TENG) delivers IRE to the cavity‐side margin via a balloon catheter to prevent cancer recurrence. Reproduced with permission [143]. Copyright 2023, Elsevier.

In postoperative settings, IRE can also be applied to resected tumor margins to eradicate residual malignant cells. Along this line, Wang et al. [142] fabricated an electrospun nanofiber (NF)–based flexible catheter with high surface area and porous microstructure, readily modified with epithelial cellular adhesion molecule (EpCAM) antibodies to capture circulating tumor cells (CTCs). Integrated with MPC electrodes, the system enabled efficient CTC enrichment and complete eradication via IRE, offering a promising adjunctive strategy for mitigating metastasis and improving clinical cancer management (Figure 7B).

Taken together, these studies indicate that the most powerful role of bioengineered materials in IRE is not simply to intensify cytotoxicity, but to reintroduce selectivity into an otherwise threshold‐driven physical therapy. By lowering the effective ablation threshold, biasing field effects toward tumor cells, or suppressing survival pathways in reversibly electroporated regions, nanoadjuvants and targeted materials begin to transform IRE from a purely geometric ablation modality into a more biologically discriminating intervention. Future development in this area will depend on improving target specificity, defining dose–material–field relationships more rigorously, and validating whether these gains can be maintained in heterogeneous in vivo tumors without increasing off‐target toxicity.

9. Applications of Electroporation in Emerging Biomedical Fields

Beyond its well‐established roles in gene transfer, tumor ablation, and immunotherapy, electroporation has demonstrated growing potential across a spectrum of biomedical fields, including tissue regeneration, neural modulation, cardiovascular repair, and intracellular biosensing [144]. When combined with functional biomaterials, electroporation can further enhance nucleic acid stability and intracellular delivery efficiency, thereby expanding its therapeutic versatility. The fundamental capacity of electroporation to transiently modulate membrane permeability and cell–matrix interactions provides a biophysical basis for these diverse therapeutic innovations. In such integrated systems, electroporation functions not only as a delivery facilitator but also as a bioelectric regulator, orchestrating cellular communication and tissue remodeling across multiple biological scales. Collectively, these features confer synergistic advantages in efficacy enhancement, toxicity reduction, controlled release, and sustained therapeutic activity.

Building upon these concepts, Huysmans et al. [145] compared the intradermal expression kinetics and innate immune responses of self‐amplifying RNA (saRNA) delivered via electroporation versus lipid nanoparticles (LNPs). Intradermal electroporation‐mediated saRNA delivery exhibited a distinctive plateau‐phase expression profile, maintaining high‐level protein expression from days 3–10—significantly surpassing that achieved with either plasmid DNA (pDNA) or non‐replicating mRNA electroporation. Using IFN‐β reporter mice, the authors observed only a transient, moderate innate immune response that did not compromise saRNA translation. In contrast, saRNA‐LNP delivery produced a markedly different profile: peak expression within 24 h followed by a rapid decline, attributed to a strong, early‐onset innate immune activation (within 4 h post‐injection) that led to translational suppression. These findings underscore how electroporation‐based nucleic acid delivery can achieve a balanced interplay between immunogenicity and expression longevity.

To further expand the transdermal applications of electroporation, Xu et al. [146] developed a stretchable electronic face mask (SEFM) for noninvasive electroporation‐assisted drug delivery (Figure 8A). Fabricated with conductive ink composites to improve adhesion and conductivity on silicone substrates, the SEFM is reusable, waterproof, low‐cost, and portable. In vivo rat experiments revealed that transdermal drug uptake increased by 3–4‐fold compared with non‐electroporated controls, while human volunteer studies confirmed its ability to significantly enhance niacinamide permeation without safety concerns. This approach represents a promising paradigm for wearable, on‐demand drug delivery in personalized healthcare.

FIGURE 8.

(A) Schematic of a stretchable electronic facial mask for enhanced transdermal drug delivery via electroporation. Reproduced with permission [146]. Copyright 2023, Advanced Functional Materials. (B) The liver‐surface‐applied HGF@EV hydrogel facilitates hepatic regeneration and repair. Reproduced with permission [148]. Copyright 2025, Elsevier. (C) Schematic illustration of a bioelectronic scaffold that promotes wound healing by electrotransfecting local peri‐wound cells to induce cell proliferation, while its 3D porous structure provides a conducive microenvironment for tissue regeneration. Reproduced with permission [149]. Copyright 2024, Advanced Materials.

Hydrogels, characterized by their ECM‐mimicking 3D networks, high biocompatibility, and tunable physicochemical properties, have long been recognized as ideal scaffolds for tissue regeneration [147]. When integrated with electroporation, hydrogels not only improve the transfection efficiency of genes or bioactive molecules in encapsulated cells but also act as supportive microenvironments that sustain therapeutic effects through controlled release. The synergistic interplay between electroporation and hydrogel systems thus overcomes the inherent limitations of either modality alone, positioning this hybrid approach as a powerful platform in regenerative medicine and tissue repair.

For instance, Liang et al. [148] developed an ultrasound‐guided, liver‐targeted delivery system by combining electroporation with hepatocyte growth factor‐loaded extracellular vesicle hydrogels (HGF@EV hydrogels) to halt the rapid progression of acute liver failure (Figure 8B). The hydrogel firmly adhered to the hepatic tissue, forming a protective matrix that gradually released HGF@EVs over three days. The released vesicles fused with hepatocytes, significantly reducing reactive oxygen species (ROS) accumulation, inflammatory injury, and apoptosis during the acute phase while promoting parenchymal and vascular regeneration. Similarly, Xiao et al. [149] designed a biodegradable electrotransfection scaffold (E‐TASHI) to accelerate wound healing. High‐voltage pulses enabled the transfection of proliferative plasmids into cells at wound margins, while the hydrogel's porous structure guided cell migration and tissue ingrowth. As the hydrogel degraded, released amino acids supported dermal cell organization, facilitating synchronous epidermal–dermal regeneration and producing newly formed skin that closely resembled native tissue (Figure 8C). This strategy offers a translational pathway toward clinical wound‐healing therapies.

Together, the emerging applications discussed here suggest that electroporation is beginning to move beyond oncology and gene transfer into a broader bioelectronic therapeutic paradigm. A major lesson from these studies is that transient membrane modulation can be repurposed not only for intracellular delivery but also for orchestrating tissue‐level communication, driving regeneration, and enabling wearable or minimally invasive interventions. The most promising future directions in this area are likely to involve bioresorbable, soft, and feedback‐responsive systems that couple electroporation with controlled molecular delivery in a manner tailored to specific regenerative or physiological contexts.

10. Comparative Design of Biomaterial‐Assisted Electroporation

The major biomaterial‐assisted electroporation platforms should not be compared simply as different material categories, but as distinct solutions to different transport barriers, treatment scales, and clinical workflows. As discussed above, micro/nanoengineered interfaces, nanoparticle formulations, and hydrogel/depot systems can all improve electroporation performance, but they do so through different dominant design logics [81, 97, 150] (Figure 9). Therefore, the key question is not which biomaterial class is intrinsically superior, but which platform best matches the biological barrier, therapeutic geography, payload type, and intended mode of electroporation.

FIGURE 9.

Biomaterials reshape electroporation for programmable therapy. (A) Representative biomaterial functions in electroporation‐assisted therapy, including electric‐field modulation, cargo stabilization, local retention, and tissue‐conformal interfacing. (B) Biomaterial‐enabled electroporation can confine or redistribute the electric field, regulate local exposure, enhance membrane permeabilization, improve intracellular delivery, and reduce off‐target tissue damage. (C) These mechanisms support diverse therapeutic outcomes, including gene engineering, drug delivery, tumor ablation, and immune activation. The illustration was created with BioRender.com.

10.1. Micro/Nano‐Electrode Interfaces

Micro/nano‐electrode interfaces are governed by an interface‐centered design logic, in which the primary objective is not cargo carriage or depot formation, but precise control of the cell‐field interface. By shrinking the effective poration region and improving membrane contact, these systems enable lower‐voltage operation, better field confinement, and reduced collateral damage [151]. They are therefore particularly well suited for single‐cell manipulation, adherent‐cell delivery, and ex vivo engineering of sensitive cells. Their comparative strength lies in delivery precision and viability preservation. Conversely, their clinical translation is constrained by fabrication complexity, limited treatment volume, and the difficulty of scaling highly localized interfaces to large tissues or solid tumors [152].

10.2. Nanoparticle‐Assisted Electroporation

Nanoparticle‐assisted electroporation is most effective when the central challenge lies in cargo instability, poor intracellular trafficking, or insufficient selectivity. Nanoparticles can protect sensitive payloads, support co‐loading, enhance tissue targeting, and in some cases amplify local electric fields [95]. These features make them well suited for gene delivery, tumor sensitization, and combined chemo/gene or immune‐modulatory therapies. Their main advantage is formulation‐level control that pulse engineering alone cannot provide. However, they remain hindered by the broader bottlenecks of nanomedicine, including variable biodistribution, clearance, reproducibility, and possible carrier destabilization under strong pulsed fields [153]. Accordingly, nanoparticle‐assisted electroporation is strongest when intracellular delivery and targeting are prioritized over long‐term local depot formation.

10.3. Hydrogel‐ and Depot‐Assisted Electroporation

Hydrogel‐based electroporation platforms are most useful when the key requirement is to retain therapeutics locally and release them over time within a mechanically compliant interface. Unlike nanoparticles, hydrogels are naturally suited to tissue‐scale control. They can function as local depots, soft electrodes, or both, thereby coupling electroporation with prolonged residence time and staged delivery [154]. This makes them particularly attractive for locoregional therapy, post‐ablation immune modulation, transdermal delivery, and regenerative applications. Their main strength is the ability to prolong and spatially structure the therapeutic effect of electroporation. Their main limitation is that deep‐tissue use often requires invasive placement, and their degradation, conductivity, and electrochemical stability must be tightly controlled [155].

10.4. General Design Principles

Several design principles emerge across platforms. First, platform selection should follow the dominant barrier: micro/nano‐interfaces for precision poration, nanoparticles for cargo protection and targeting, and hydrogels for local retention and staged release. Second, the spatial scale of the material should match the spatial scale of therapy. Third, biomaterials must be co‐designed with pulse conditions and electrode geometry, because field distribution, cargo behavior, and interfacial transport are tightly coupled during electroporation. Finally, the most promising next‐generation systems will likely be hybrid rather than single‐function platforms, given that clinically relevant biological barriers are usually multifactorial.

Overall, no single biomaterial class is universally optimal for electroporation. The most effective design depends on matching material function to therapeutic barrier, treatment scale, and biological context. In this sense, the key principle of biomaterial‐assisted electroporation is not material selection alone, but the integrated co‐design of material properties, electrical input, and therapeutic objective.

11. Challenges and Future Perspectives

Although biomaterial‐assisted electroporation has generated increasingly diverse therapeutic proof‐of‐concept studies, its next major challenge is to move from empirical efficacy demonstrations toward standardized, mechanism‐correlated, and clinically reproducible systems. Beyond efficiency, selectivity, and biocompatibility, future development must address several practical barriers to clinical translation, including quantitative dosimetry, electrode/device standardization, scalable manufacturing, treatment cost, regulatory positioning, and competition with established delivery technologies.

11.1. Quantitative Dosimetry and Electrode/Device Standardization

The biological outcome of electroporation is governed not only by nominal pulse parameters, but also by electrode geometry, waveform structure, tissue conductivity, local impedance, temperature change, and dynamic changes in the treated microenvironment [156]. This complexity becomes greater when biomaterials are introduced, because hydrogels, nanocarriers, conductive depots, and electroactive interfaces can alter local conductivity, cargo release, interfacial charge transport, and immune signaling. Therefore, future studies should move beyond reporting voltage, pulse number, and pulse duration alone. They should correlate pulse delivery with material properties, electric‐field distribution, local thermal/electrochemical effects, cargo kinetics, and biological endpoints.

In addition to pulse parameters, electrode and device standardization remains a critical translational bottleneck. For IRE and related locoregional electroporation procedures, reproducible treatment requires standardized electrode geometry, inter‐electrode spacing, exposure length, insertion depth, generator output, waveform fidelity, impedance monitoring, synchronization with cardiac or respiratory motion when necessary, and image‐guided placement [157]. For biomaterial‐integrated platforms, these requirements become even more complex because soft electrodes, conductive hydrogels, microneedle arrays, or implantable bioelectronic interfaces may alter local impedance and field distribution during treatment [155, 158]. Therefore, future studies should define device‐level quality attributes together with material‐level parameters, rather than treating the electrode as a fixed background component.

The recent effort to standardize IRE protocols is instructive. For example, the 2024 Delphi consensus on pancreatic IRE proposed specific recommendations regarding electrode spacing, exposure length, test pulses, treatment pulses, pulse length, and follow‐up imaging, reflecting a broader field‐wide shift toward protocol reproducibility and treatment planning [85]. However, biomaterial‐assisted systems add further variables that conventional IRE protocols do not fully address. Thus, future biomaterial‐integrated electroporation studies should report not only electrical parameters, but also material conductivity, impedance evolution, degradation profile, release kinetics, and the spatial relationship among electrodes, materials, payloads, and target tissues.

11.2. Safety Evaluation Beyond Short‐Term Efficacy

A second major challenge is that short‐term delivery efficiency or ablation efficacy is no longer an adequate endpoint for next‐generation biomaterial‐assisted electroporation. For delivery‐oriented applications, optimization must explicitly balance membrane permeabilization efficiency with long‐term cell fitness. Pulse protocols or cargo formulations that increase intracellular delivery may compromise post‐treatment viability, proliferative capacity, differentiation potential, stemness, or immune‐cell effector function, especially in primary immune cells, stem cells, and stem‐cell‐derived products.

For tissue‐level applications, apparent selectivity at the level of cell death should be interpreted together with edema, vascular leakage, off‐target inflammation, unintended immune amplification, electrode‐associated mechanical injury, electrothermal injury, and chronic tissue remodeling [157, 159, 160]. For injectable or implantable biomaterials, additional endpoints should include biodegradation, biodistribution, material fate, local redox activity, electrochemical stability, repeated‐stimulation tolerance, and long‐term device–tissue interfacial stability [158, 161]. Thus, biocompatibility, surgical operability, manufacturing cost, inter‐patient variability, and long‐term interface stability should be treated as core translational endpoints rather than peripheral material‐science considerations.

Piezoelectric and ultrasound‐coupled systems require especially careful evaluation. Their in vivo performance depends not only on the intrinsic piezoelectric coefficient of the material, but also on acoustic frequency, tissue depth, coupling efficiency, attenuation, scattering, and local tissue composition [162]. Therefore, local ultrasound exposure should be quantitatively linked to the actual electrical output, electrochemical behavior, cargo‐release profile, and biological response at the tissue‐material interface. Without this linkage, claims of electroporation‐equivalent activity remain difficult to validate under clinically relevant conditions [163].

11.3. Scalability, Manufacturing, and Treatment Cost

For ex vivo cell engineering, manufacturability and economic viability are becoming central determinants of clinical relevance. Scalable implementation will require closed‐system processing, high‐throughput operation, batch‐to‐batch consistency, analytical comparability, rapid release testing, potency assays, and compatibility with current good manufacturing practice [164]. In this setting, future progress may depend less on marginal improvements in transfection efficiency and more on whether electroporation workflows can support reproducible and economically sustainable manufacturing of multiplex‐edited autologous or allogeneic cell therapies.

For CAR‐T and other engineered cell therapies, throughput is not only a technical parameter but also a determinant of treatment accessibility and cost. Clinical‐scale manufacturing often requires processing large numbers of cells under closed and reproducible conditions, while preserving post‐electroporation viability, expansion capacity, phenotype, and cytotoxic function [84]. Therefore, scalable electroporation platforms must be evaluated not only by editing efficiency, but also by processing capacity, automation compatibility, reagent consumption, batch failure rate, labor intensity, quality‐control burden, and cost per manufacturable therapeutic dose.

For in vivo biomaterial‐assisted electroporation, treatment cost is shaped by a different set of factors, including device cost, disposable electrode or interface cost, imaging guidance, procedural time, need for anesthesia, catheter‐ or injection‐based material placement, repeat‐treatment feasibility, and postprocedural monitoring [165]. Accordingly, clinical translation will require not only demonstration of therapeutic superiority, but also evidence that added biomaterial complexity produces a meaningful gain in safety, efficacy, local control, or quality of life relative to the additional procedural and manufacturing burden [166].

11.4. Regulatory Positioning and Combination‐Product Complexity

For in vivo or implantable biomaterial‐assisted systems, the regulatory challenge is more complex than that of stand‐alone electroporation devices. Many such platforms may function as combination products that integrate device, drug, biologic, and material components within a single therapeutic strategy. Consequently, early regulatory positioning, product‐specific CMC planning, device–material co‐validation, and definition of the primary therapeutic mode of action will be essential. Future studies should therefore align biological efficacy with quality attributes such as material composition, sterility, electrical reproducibility, release behavior, degradation products, potency, and safety‐relevant interfacial properties [158].

These requirements are particularly important for gene‐ and cell‐therapy‐related platforms. Regulatory expectations for human gene therapy products emphasize sufficient chemistry, manufacturing, and control information to support product safety, identity, quality, purity, and potency. When electroporation is combined with biomaterials, devices, cells, or therapeutic cargos, the resulting system should therefore be evaluated as an integrated therapeutic product rather than as a simple procedural enhancement.

11.5. Comparison with Competing Delivery Approaches

Biomaterial‐assisted electroporation should not be positioned as a universal replacement for viral vectors, lipid nanoparticles, conventional ablation, or established drug‐delivery systems. Viral vectors remain highly effective when durable transgene expression or broad in vivo transduction is required. Lipid nanoparticles have achieved substantial clinical maturity for RNA delivery and offer advantages in rapid formulation, scalable manufacturing, and transient expression [167]. Compared with these systems, biomaterial‐assisted electroporation is unlikely to dominate applications requiring systemic biodistribution or long‐term constitutive expression.

Its comparative advantage is stronger in settings where spatial control, transient non‐viral delivery, local retention, device‐guided intervention, or avoidance of permanent vector burden is clinically valuable. Three such niches appear particularly compelling [168, 169]. The first is non‐viral ex vivo cell engineering, where electroporation can enable programmable genome editing while avoiding permanent vector burden. The second is image‐guided locoregional therapy, especially in tumors where electroporation or IRE can be synchronized with local drug release, immune modulation, or post‐ablation depot formation. The third is regional or tissue‐interface therapy, including transdermal, regenerative, postoperative, and implantable applications, where soft biomaterials can improve tissue conformity, local retention, and temporally controlled bioelectric stimulation [170].

Taken together, the next phase of this field will be defined less by whether biomaterial‐assisted electroporation can produce isolated proof‐of‐concept efficacy, and more by whether it can deliver standardized, manufacturable, safe, cost‐conscious, and clinically differentiated solutions in well‐defined therapeutic settings.

12. Conclusion

Electroporation has evolved far beyond its original role as a transient membrane‐permeabilization technique and is increasingly recognized as a programmable therapeutic platform at the interface of physics, biomaterials, and biology. Across gene editing, engineered cell therapies, locoregional tumor treatment, immunomodulation, and regenerative medicine, electroporation provides a spatiotemporally controllable and non‐viral strategy for overcoming biological transport barriers. The integration of electroporation with functional biomaterials, including nanocarriers, hydrogels, conductive polymers, soft bioelectronic interfaces, and piezoelectric systems, further expands its capabilities from simple intracellular delivery toward localized drug depots, immune‐programming platforms, and adaptive tissue‐interfacing interventions.

Looking ahead, the most clinically meaningful advances over the next 5–10 years are likely to emerge in areas where spatial precision, transient delivery, and local therapeutic control are particularly valuable. In oncology, biomaterial‐assisted electroporation may support more personalized and image‐guided cancer therapies by combining local ablation, controlled drug release, and temporally coordinated immune modulation according to tumor type, anatomical location, and treatment response. Rather than functioning only as an ablative tool, electroporation may increasingly serve as a patient‐specific multimodal platform for local tumor control, reduced systemic toxicity, and enhanced antitumor immunity.

In cell and gene therapy, standardized ex vivo electroporation workflows may become increasingly important for non‐viral genome editing, immune‐cell engineering, and next‐generation autologous or allogeneic cell products. At the same time, biomaterial‐assisted and tissue‐localized electroporation strategies may open new possibilities for regional gene delivery or local immune‐cell programming directly within target tissues. In regenerative medicine, soft conductive biomaterials, bioresorbable electronics, and feedback‐responsive interfaces may enable transient, spatially restricted bioelectric stimulation combined with controlled molecular delivery, with potential applications in wound healing, musculoskeletal regeneration, vascular repair, and tissue‐interface reconstruction.

Overall, the long‐term significance of biomaterial‐integrated electroporation lies not merely in membrane permeabilization itself, but in the emergence of therapeutic systems that co‐design pulse parameters, material properties, device geometry, and biological context. With continued progress in imaging guidance, computational treatment planning, smart biomaterials, and bioelectronic control, biomaterial‐assisted electroporation is poised to become an enabling technology for spatially controlled precision medicine in the coming decade.

Ethics Statement

No written informed consent was obtained from the patient because no identifiable patient information is included in this review.

Conflicts of Interest

The authors declare no conflicts of interest.

Data Availability Statement

Data sharing not applicable to this article as no datasets were generated or analyzed during the current study.