Advanced Materials

Scaling neural interfaces to ever-higher channel counts has accelerated rapidly with advances in thin-film fabrication, lithography, and connectorization, enabling passive arrays to reach thousands of channels and chart credible pathways to much larger formats. Integrating active electronics directly at the sensing sites offers a complementary route to higher channel density by reducing the number of interconnects required to access large arrays. Here we introduce a monolithic flexible thin-film integrated circuit platform for active neural sensing, inspired by active-matrix display technology. The system integrates dual-gate amorphous indium gallium zinc oxide transistors on polyimide substrates to implement in-pixel transconductance amplification and row-column time-division multiplexing, improving scability for high-channel-count applications. Co-optimization of device architecture, contact engineering, and a hybrid ceramic-polymer thin-film encapsulation yields stable operation with projected lifetimes exceeding 38 years under accelerated aging. In acute and chronic in vivo rat studies, the platform exhibits negligible thermal burden, robust sensory-evoked recordings, and stable functionality over 30 days despite tissue encapsulation. These results establish display-inspired flexible thin-film electronics as a scalable building block for next-generation neural interfaces.

Scaling engineered living materials to clinically relevant dimensions is limited by diffusion-dependent depletion of oxygen and nutrients, which rapidly induces metabolic failure. We introduce glycogen as nutritional nanoparticle that provides cell-mediated, autonomous nutrient release to support long-term survival under extreme metabolic stress. We demonstrate that human mesenchymal stromal cells (hMSCs) survive for weeks in anoxia and serum deprivation when provided extracellular glycogen. Contrary to long-held assumptions, hMSCs secrete glycogen-degrading enzymes, enabling cell-density controlled extracellular glycogenolysis and sustained release of glucose and metabolic intermediates, positioning glycogen as the first-of-its-kind metabolic battery. This cell-responsive process maintains metabolic activity, limits glycolytic acidosis, and enhances pro-angiogenic signaling. To translate this mechanism into a versatile materials platform, we engineered core-shell dextran-tyramine microcapsules that stably encapsulate glycogen while permitting diffusion of enzymes and degradation products. Integrated into centimeter-scale GelMA constructs, these microcapsules maintained hMSC viability and function for at least one month under anoxia. In vivo, glycogen-loaded implants promote deep cellular infiltration, enhanced matrix remodeling, increased M2 macrophage polarization, and orchestrated accelerated vascularization. This work establishes the novel concept of glycogen-based nutritional nanoparticles as metabolic batteries to endow engineered tissues with autonomous self-feeding capacity, enabling scalable and functional living materials for regenerative medicine and related technologies.

Controlled biomineralization is central to engineering physiologically relevant hard-tissue models, yet achieving spatially organized, three-dimensional (3D) mineral deposition in microfluidic on-chip systems remains challenging. While cell-based bone-on-chip platforms offer biological complexity, they intrinsically couple mineral initiation to confounding factors such as matrix remodeling and paracrine signaling, obscuring the earliest biochemical drivers of nucleation. Drawing inspiration from bottom-up synthetic biology, we engineered an enzymatically active granular matrix that recapitulates a key osteogenic function within a perfusable 3D microenvironment. Alkaline phosphatase (ALP), the key driver of native bone formation, was covalently conjugated to poly(ethylene glycol)-based (PEG) microgels via thiol-ene photochemistry, retaining over 90% enzymatic activity after 48 h. These monodisperse microgels were assembled into a jammed, perfusable matrix within an on-chip chamber, enabling independent control over enzyme loading and substrate delivery. The system supported rapid in situ mineralization (24-48 h), yielding a carbonated, calcium-deficient, apatite-like phase characteristic of early-stage bone mineral. We demonstrate that the spatial 3D localization of enzymatic activity to discrete microscale compartments, coupled with interstitial perfusion, enables localized and near-physiological mineral formation. This mechanistically defined, acellular platform provides a programmable foundation for investigating ALP-driven 3D mineralization and establishes a modular route toward hybrid biosynthetic models of (patho)physiological tissue mineralization.

Barrier membranes govern transport and mechanochemical coupling in lung-on-chip systems but typically exhibit low open porosity, limited pore interconnectivity, and diffusion distances exceeding native thin septal regions. An interstitium-mimicking, alveolus-shaped poly({varepsilon}-caprolactone) membrane is developed using dual-templated nonsolvent-induced phase separation followed by controlled enzymatic pore enlargement. The resulting architecture achieves [~]40% total porosity with 97% pore interconnectivity and incorporates a locally thinned dome region ([~]2.5 {micro}m). This structure sustains cyclic deformation while increasing oxygen diffusivity fivefold compared with conventional Transwell(R) membranes under both acellular and epithelial-endothelial co-culture conditions. Integrated into an air-liquid interface platform, the membrane enables direct aerosol deposition and quantitative interrogation of cross-barrier mass transfer. Using carbonaceous nanoscale particulate matter as a model inhaled aerosol, controlled exposure induces dose-dependent oxidative, inflammatory, and genotoxic responses. Matched cumulative dose studies reveal distinct biological trajectories: acute high-dose exposure produces rapid cytotoxic stress and barrier disruption, whereas chronic low-dose exposure preserves viability yet promotes sustained DNA repair and genome-maintenance programs. Compartment-resolved analysis and therapeutic intervention further demonstrate the platforms utility for spatial and translational interrogation of lung injury. By restoring physiologically relevant diffusion distance, interconnectivity, and strain responsiveness, the interstitium-mimicking membrane advances lung-on-chip design toward functional replication of alveolar transport dynamics for studying lung injury and barrier dysfunction.

Implant-associated infections are driven by bacterial biofilm formation and remain difficult to eradicate using conventional antibiotic-based strategies. Here, we present a dynamically actuated reconfigurable topographical surface (DARTS) that integrates intrinsically bactericidal nanoscale surface topography with programmable mechanical actuation to achieve durable, antibiotic-free infection control. Using a scalable bottom-up nanofabrication strategy, we generate tunable wrinkled MXene topographies that exhibit contact-mediated bactericidal activity against both Gram-positive and Gram-negative bacteria without chemical leaching. Integration with a soft robotic actuator enables reversible modulation of surface geometry, which synergistically enhances bacterial removal and killing, resulting in near-complete disruption of mature biofilms. Dynamic actuation further sensitizes released bacteria to antibiotic treatment. In a mouse subcutaneous implant infection model, DARTS with actuation achieves sustained suppression of bacterial burden and markedly improves host tissue outcomes. Remote, noninvasive actuation using near-infrared laser stimulation further highlights the translational potential of this platform for implantable antibacterial applications.

Cerebral cortex organoids are powerful in vitro models that recapitulate key features of human development. However, conventional methods produce cortical organoids with spontaneous, spatially disorganized cortical regions due to limited control over morphogen distribution within local environments. Here, we present a spatially engineered hydrogel platform that drives localized posterior organizer formation in cortical organoids through controlled, localized exposure to morphogens. Using a combination of bulk photopolymerization, thermal crosslinking, and digital light processing (DLP) approaches, we fabricated hydrogels with stiffness-controlled layers that preferentially deliver morphogens to one side of the organoid, selectively inducing posterior organizer formation on the exposed face. We further validated this platform by delivering fluorescently tagged dextran, used as molecular weight-matched model morphogens, to visualize spatiotemporal delivery dynamics at the organoid interface. As a proof of principle, we also demonstrated that DLP fabrication enables the printing of dual morphogen hubs, serving as a model for establishing two opposing gradients within a single organoid. Together, this hydrogel platform enables systematic spatial patterning of cell populations in organoids, more faithfully recapitulating the spatial organization and cellular diversity of native tissues and advancing higher-fidelity models for studying human development and disease.

Three-dimensional (3D) in vitro tissue models are emerging as powerful platforms for studying development, disease, and therapeutic responses, where close monitoring of electrophysiological activity is essential. However, existing probing methods remain limited in accessibility or spatiotemporal resolution for comprehensive electrophysiological mapping of suspended 3D tissues that closely mimic the native environments. Here we introduce a Venus flytrap-inspired bioelectronic mesh system that enables the full spherical enclosure of 3D tissues in a suspended configuration. The system consists of two hemispherical meshes that envelop the tissue, constructed from highly flexible, stretchable, cell-scale ribbons interconnected into a tissue-compliant network with integrated recording electrode arrays. This architecture enables intimate, conformal tissue integration and supports stable electrophysiological recordings over 300 days. The high-resolution recordings allow precise tracking of local dynamics and correlated global signaling, enabling comprehensive assessment of tissue development as well as detailed evaluation of drug responses for disease modeling. Beyond single tissues, the mesh architecture is extended to fully enclose assembloids composed of multiple tissues, enabling characterization of cross-tissue signaling relevant to advanced heterogenous tissue modeling. Furthermore, the system is translated into array-based platforms, demonstrated by a 4x4 array integrating 1024 electrodes, for high-throughput tissue sampling and cross-study analysis. The developed bioelectronic system and integration method provide a broadly applicable platform to advance electrophysiological studies across diverse tissues and organoids.

Engineered epithelial models require three-dimensional extracellular matrix environments that support organized cell growth and allow independent access to luminal and basal compartments. However, many organ-on-chip (OoC) fabrication strategies rely on planar geometries, non-native materials, or multi-step assembly workflows that limit architectural complexity and experimental control. Here, we report a direct in-chip volumetric printing strategy for fabricating stretchable and perfusable collagen-I scaffolds inside custom OoC devices. A vitamin C-regulated ruthenium/sodium persulfate photocrosslinking system enabled high-fidelity printing of collagen-I into open-lumen architectures with ductal- and alveolar-inspired features. By generating scaffolds directly within the final culture device, this workflow eliminates post-print transfer and integrates defined collagen architectures with compartmentalized fluidic access and a mechanically actuable chip format. To support chip-based culture, printed collagen constructs were stabilized after fabrication using EDC/NHS chemistry, which limited thermally induced collagen densification, improved shape retention, and maintained scaffold anchorage during perfusion. The chip design provided separate access to the printed lumen and surrounding basal compartment, which enabled compartment-specific fluid handling while preserving scaffold integrity during inflation, stretching, and perfusion of the printed construct. On the collagen-I scaffolds, human milk-derived mammary epithelial cells formed epithelial layers with tight junctions and lactation associated markers. The platform further supported perfusion culture, in situ staining, and whole-chip volumetric imaging. Together, this work establishes direct in-chip collagen-I volumetric printing as a biofabrication strategy for creating perfusable epithelial tissue chips with native matrix architecture and compartmentalized fluidic control.

Virus-like particles represent an emerging and promising vaccine platform. However, these particles are inherently mechanically soft and have limited control over particle surface architecture, thereby constraining their immunological control. Herein, we report the rational design of bioinspired virus-like porous silica (VLPSi) nanoparticles (NPs) with tunable and mechanically rigid spike architectures that function dually as antigen delivery carriers and immune adjuvants. Using ovalbumin (OVA) as a model antigen, we systematically elucidate the spiky structure-function relationship in antigen delivery and immune response. VLPSi NPs exhibit good biocompatibility, sustained antigen release, and markedly enhanced cellular uptake and endosomal escape compared with soft spike and spherical counterparts. Mechanistic investigations combining molecular dynamics simulations and proteomic analyses reveal that rigid spike architectures reduce the energetic barrier for cellular internalization and concurrently activate dual pathways involving endosomal Toll like receptors and calcium signaling. Consequently, VLPSi with long spikes elicit significantly enhanced humoral and cellular immune responses, outperforming the particles with shorter spikes, spherical shape as well as clinically used alum adjuvant. To demonstrate translational potential, bioinspired antibacterial vaccines were produced by loading Staphylococcus aureus surface protein rEsxB. The VLPSi-based vaccine elicited robust protective immunity to achieve complete (100%) survival following lethal challenge without detectable adverse effects, whereas traditional Alum-adjuvanted formulation conferred only minimal protection, with a survival rate of 10%. Collectively, this work establishes VLPSi with tunable spikes as a mechanistically controlled platform for next generation vaccines. Graphic Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=145 SRC="FIGDIR/small/720861v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@1f6c13eorg.highwire.dtl.DTLVardef@1090d07org.highwire.dtl.DTLVardef@1364926org.highwire.dtl.DTLVardef@fc68ab_HPS_FORMAT_FIGEXP M_FIG C_FIG

Implanted brain stimulators play a crucial role in treating various neurological disorders, including Parkinsons disease (PD), Alzheimers disease, epilepsy, and depression. However, none of the existing implanted brain stimulators can realize specific neuromodulation due to fundamental disparities in signal transmission between electrical signal-induced neuronal responses and neurotransmitter-evoked neural signalling in natural neural circuits, leading to persistent challenges in biosafety and therapeutic effectiveness. Inspired by the dopaminergic neural circuit, we report a biointegrated living brain stimulator (BBS) that integrates ferroelectric bioelectronics, dopaminergic cells, and a gelatin hydrogel matrix, enabling dopamine neurotransmitter-evoked neural signalling. In contrast to conventional brain stimulators, the BBS are capable of programmed secretion of physiological-level dopamine, specifically activating nigral dopamine pathways and restoring motor function in a rodent PD model. By integrating the advantages of both bioelectronics and medicine, this lifelike BBS offers a great promise for next-generation bioelectronics, medicine, and brain-machine interfaces.

Microstructures created with flow lithography exhibit distinct functionality depending on the shape and composition of the precursor fluids, enabling applications from tissue engineering to anti-counterfeiting. However, current techniques rely on static nozzle geometries or passive hydrodynamic focusing, which commit to a fixed structure and limit dynamic reconfiguration of material architecture during fabrication. Here, we introduce ActiSculpt, an acoustofluidic platform that replaces in-channel physical structures with programmable, electronically driven acoustic streaming. By exploiting the interplay between laminar stability and acoustic streaming, we decouple deterministic fluid deformation from chaotic mixing, achieving a continuous cross-sectional displacement sensitivity of ~15 m/V. We demonstrate the generation of a diverse library of hydrogel particles whose cross-sectional moments of inertia are tunable up to 5.5-fold, establishing a direct, geometry-mediated link between acoustic parameters and the moments that govern bending and torsional rigidity. We further demonstrate continuous fiber fabrication in which acoustic parameters are varied in real time, encoding structural variation along the fibers length. The result is a platform that overcomes the one-device, one-geometry constraint of existing techniques, enabling not only on-demand reconfiguration between fabrication runs but also real-time control of material architecture. This spatiotemporal control establishes a new design axis for soft-material manufacturing.

The ability to encode and reliably read nanoscale information is increasingly important for multiplexed biomolecular detection and super-resolution imaging. DNA origami provides a uniquely programmable platform for arranging structural and functional elements with nanometer precision, enabling the creation of identifiable nanoscale patterns. In this context, DNA origami-based barcodes that incorporate gold nanoparticles (AuNPs) to encode either origami geometry or the identity of specific biological targets within defined nanoparticle patterns have been paired with transmission electron microscopy imaging for decoding. However, surface-bond AuNPs may detach during handling, purification, or biological incubation, leading to misidentification or decoding errors in barcode analysis. Here we report a rational design for the controlled encapsulation of AuNPs within DNA origami tubes to enhance nanoparticle retention and structural integrity. We engineered curvature-inducing modifications in a flat rectangular DNA origami scaffold to promote inward folding and confinement of AuNPs. These barcodes can be further functionalized on the outer surface with bioactive aptamers and/or fluorescence dyes, enabling targeted interactions with cells and optical readout. Programable dimerization further expands multiplexing capacity. This design provides a robust framework for structurally stable origami barcodes and advances the development of high-resolution, multiplexed labeling and diagnostic platforms. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=60 SRC="FIGDIR/small/725969v1_ufig1.gif" ALT="Figure 1"> View larger version (23K): org.highwire.dtl.DTLVardef@686c1aorg.highwire.dtl.DTLVardef@1914c4eorg.highwire.dtl.DTLVardef@28ad47org.highwire.dtl.DTLVardef@8847ca_HPS_FORMAT_FIGEXP M_FIG C_FIG

Bioprinting increasingly requires biomaterials that are not only printable, but structurally adaptive and biologically instructive. Here we establish an ovoprotein-derived microgel platform that couples intrinsic protein bioactivity with orthogonal interparticle photocrosslinking for modular bioprinting. Methacrylated ovoproteins yielded a photoresponsive protein-rich hydrogel matrix with retained proteomic complexity, tunable mechanics, and cell-regulatory biofunction. Endogenous tyrosine chemistry drove interparticle dityrosine coupling between ovoprotein microgels, producing cohesive, microporous, and intrinsically autofluorescent granular networks. The resulting systems displayed programmable rheology and broad compatibility across digital light processing, extrusion-based and aspiration-assisted bioprinting. Functionally, the ovoprotein microgel matrices attenuated sustained pro-inflammatory macrophage activation, promoted endothelial organization and host angiogenic invasion, and supported spheroid-mediated vascular morphogenesis with progressive sprouting, lumenization, branching and inosculation. It further enabled bioprinted osteogenic constructs with long-term maturation into bone-like mineralized tissues in vitro. These findings establish ovoprotein microgel-spheroid bioassembly as an adaptive, bioinstructive strategy for engineering vascularized and mineralized tissue constructs.

Extracellular vesicles (EVs) carry molecular signatures of their originating cells and have thus emerged as promising biomarkers. However, their clinical utility remains limited due to their low abundance and the modest sensitivity of current EV detection methods in complex biological environments. Here, we present a quantum well defect functionalized carbon nanotube sensor coupled with integrin-recognition RGD tripeptide for EV detection in human plasma. Leveraging the abundance of integrins on EV surfaces, we targeted 5{beta}1, V{beta}1, and V{beta}3 subtypes. The nanosensor exhibited robust hypsochromic shifts in defect emission upon integrin binding, achieving sub-picomolar detection limits for integrin subunits and quantifying EVs at concentrations as low as 104 EVs{middle dot}mL-1 for glioblastoma, ovarian cancer, and fibroblast cell-derived EV types. Molecular dynamics simulation indicated that integrin docking at the RGD-coupled quantum defect can substantially reshape the interfacial environments of the quantum defects, explaining the high sensitivity in EV detection in complex biological media. Finally, transmembrane protein analysis validated the expression of surface integrins across the tested EV types. The modular nanosensor construct can be targeted to detect disease-associated EV subpopulations, advancing EV-based diagnostics.

In vivo genome editing with CRISPR-Cas9 ribonucleoproteins (RNPs) holds substantial therapeutic promise, yet rapid bloodstream clearance and the absence of delivery systems capable of systemic tumor targeting have hindered its clinical translation. Herein, a supramolecular ternary complex platform is reported in which Cas9/sgRNA RNPs are co-assembled with tannic acid (TA) and phenylboronic acid (PBA)-conjugated polymers through sequential self-assembly, producing [~]30 nm core-shell ternary complexes that protect RNPs from enzymatic degradation and dissociate selectively at endosomal pH. Upon intravenous administration in subcutaneous tumor-bearing mice, these ternary complexes exhibit prolonged blood circulation and preferential tumor accumulation, achieving 37.2% gene editing at tumor sites compared with only 1.5% for free RNPs. The platform successfully knocks out previously undruggable oncogenes including mutant KRAS and polo-like kinase 1 (PLK1), markedly suppressing tumor growth in vivo. By integrating sequential supramolecular self-assembly with stimuli-responsive cargo release, this strategy establishes a generalizable framework for systemically administered in vivo CRISPR therapeutics.

Colorectal cancer (CRC) remains a major cause of cancer death, and advanced disease is still limited by resistance and systemic toxicity. We studied intrinsically active, biomimetic cerium oxide nanoparticles (CeNPs) functionalized with D- or L-cysteine (D-Cys@CeNPs and L-Cys@CeNPs) in three CRC cell lines (COLO-201, DLD-1, and LoVo) and healthy colon fibroblasts (CCD-18Co). We propose these materials act as enantioselective functional keys: cysteine stereochemistry shapes recognition at the nano-bio interface, while productive interactions allow the Ce3-rich surface to drive localized redox exchange. We measured viability, ROS as a downstream phenotypic readout, Annexin V/PI-defined cell fate, and expression of the NF-{kappa}B regulatory genes TNFAIP3 (A20), IKBKG (NEMO), and NFKBIA (I{kappa}B). Across the CRC panel, D-Cys@CeNPs caused earlier and stronger loss of viability, with the clearest effect in COLO-201, and shifted cells toward late apoptosis and necrosis. In contrast, L-Cys@CeNPs produced slower and more heterogeneous fate changes. Gene expression showed enantiomer-dependent differences in NF-{kappa}B feedback, consistent with differential pathway engagement. CCD-18Co fibroblasts were comparatively resistant to both enantiomers. Together, these findings link chiral CeNP surface design to redox-linked pathway regulation and support a materials-based route to selective anticancer activity. INTRODUCTION

The inaccessibility of intracellular bacteria has long rendered the treatment of Staphylococcus aureus infections an challenge. Studies have demonstrated that the extracellular injection system PVC can accurately deliver proteins into cells, which would not need small molecules, and enables effective intracellular delivery of antimicrobial peptides for treatment. Accordingly, we selected antimicrobial peptides including Cecropin, LL37 and Indolicidin that possess potent bactericidal activity, and established the Directed Antimicrobial Assault platform (DAAT) by leveraging the intracellular delivery capacity of PVC. DAAT Cecropin, DAAT LL37 and DAAT Indolicidin inhibited intracellular bacteria in a dose-dependent manner, with DAAT LL37 reaching 86.76% inhibition; after 72 h of treatment, viable-cell numbers reduse to 66--82-fold those of the control. Tail-fibre retargeting enabled direct extracellular S. aureus killing, while combined DAAT therapy promoted wound healing in mice. These findings expand the utility of PVC-derived nanosyringes and establish DAAT as a modular platform for intracellular antimicrobial peptide therapy.

Summary ParagraphTissues and organs in living organisms represent centimeter-scale hierarchical architectures comprising nano-to microscale, uniaxially aligned extracellular matrix (ECM) fibres with high mechanical strength, integrated with cellular components, as exemplified in tendon, skin, cartilage, bone, and blood vessels1. Here, we present a liquid-liquid interfacial spinning method to produce highly uniaxially aligned, centimeter-scale collagen fibres. The dried fibres exhibit exceptional mechanical properties, with fracture strength of 280 MPa, Youngs modulus of 6 GPa, and toughness of 17 MJ m-3, comparable to spider silk and tendon collagen, and exceeding supramolecular and double-network hydrogels1. Incorporating living cells into the collagen solution yielded centimeter-scale, cell-laden aligned fibres, with densely adherent, uniaxially aligned cells and over 80% viability. Myoblast-laden fibres recapitulate biological features of fibrotic muscle tissues, as observed in type II diabetes2. Interfacial collagen assembly further enables fabrication of dimension-controlled constructs, like 2D sheets, 0D capsules, and 1D tubes, thus providing modular building blocks for centimeter-scale 3D tissues and organ-like structures. This approach offers a versatile platform to engineer mechanically robust, cell-laden tissues with controlled hierarchical architecture.

Synthetic biology has advanced microorganisms to be programmed as production hosts, but its application to bacteria that inherently assemble extracellular materials remains limited. Komagataeibacter spp., natively synthesizes cellulose at the bacterial cell surface, creating a material-forming interface that has not been used as a programmable recruitment platform. Here we establish cell-surface display in Komagataeibacter intermedius and show that this interface can recruit defined proteins, making functionalization part of cellulose formation. By engineering LppOmpA, we displayed a fluorescent protein and genetically encoded capture modules (SpyTag and SilkTag) to selectively capture catcher-fused protein cargos onto K. intermedius cell surface. Recruitment of silk-derived structural protein before cellulose production generated silk-associated fibrous structures within the pellicles, with retained cargo signal after washing. The resulting biocomposite showed reorganized fibre-network morphology, increased surface hydrophobicity, mesoscale ordering, and improved wet-state compressive strength. Wild-type cells exposed to same conditions did not reproduce these changes, demonstrating that material properties arise from surface-directed recruitment rather than protein exposure alone. This work demonstrates the material-forming bacterial surface as a programmable engineering interface for organizing extracellular proteins, providing a general strategy for engineering living materials.

Artificial skin substitutes that simultaneously achieve mechanical robustness, regenerative bioactivity, and transplantation-scale tissue integration remain challenging to engineer. Here we report a mechanically adaptive bilayer artificial skin based on entanglement-mediated protein networks. By integrating protein chain entanglement, flexible molecular linkers, and photo-triggered intermolecular crosslinking, we establish a hierarchically organized protein matrix with enhanced toughness, structural adaptability, and regenerative compatibility. Spatial biofunctionalization further enables integration of an antibacterial Zn{superscript 2}-coordinated epidermal layer and a regenerative CLP-EGF-functionalized dermal layer within a unified construct. The engineered skin promotes cellular proliferation through PI3K-AKT-mTOR activation, exhibits sustained antibacterial activity, and supports large-area full-thickness skin replacement covering approximately 40% of the dorsal skin surface in mice. The construct further accelerates diabetic wound repair and extracellular matrix remodeling in vivo. These findings establish entanglement-mediated protein engineering as a strategy for mechanically adaptive regenerative biomaterials and provide a platform for transplantation-scale skin regeneration.