Advanced Functional Materials

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.

Most antimicrobial drug candidates currently in development are derivatives of established antibiotic classes. In contrast, antimicrobial heteroatom-doped carbon quantum dot (CQD) nanoparticles vastly differ from their chemical antibiotic counterparts and exhibit potent antibacterial activity and favourable biocompatibility, representing a promising alternative strategy, particularly for topical applications. Here, we report the incorporation of cobalt-doped carbon quantum dots (Co-CQDs) into injectable, biocompatible hydrogels capable of both sensing pH and eliminating bacteria. Ultrasmall Co-CQDs demonstrated broad-spectrum activity against gram-positive Methicillin-resistant Staphylococcus aureus (MRSA) and Gram-negative Pseudomonas aeruginosa (PAO1), mediated by membrane hyperpolarisation and reactive oxygen species (ROS) induced membrane damage. The particles showed negligible effect on primary fibroblast and endothelial cell viability at concentrations that were bactericidal to MRSA. Polymeric hydrogels were fabricated via electrospinning of chitosan, polyvinylpyrrolidone (PVP), and polyvinyl alcohol (PVA) polymer blends incorporating Co-CQD and pH-responsive HPTS particles. This approach provided accurate measurement of environmental pH within the physiological range observed across healthy and chronic wounds. In vivo, the injectable hydrogels exhibited robust antimicrobial efficacy against MRSA without impairing wound closure relative to untreated controls, while also reducing inflammatory immune responses in infected tissues. Collectively, these findings demonstrate the potential of ultrasmall metal-doped CQDs for infection control and their integration into 3D matrices as multifunctional theragnostic platforms.

While defined synthetic substrates can replace Matrigel for human induced pluripotent stem cell (hiPSC) culture and hiPSC-derived cardiomyocyte (hiPSC-CM) production, existing approaches culture cells on two-dimensional surfaces and yield structurally immature cardiomyocytes, limiting their use in disease modeling and regenerative medicine. Here, we developed a xeno-free, fully-defined cyclic RGD (cRGD)-functionalized alginate platform in which we encapsulated hiPSCs to support their expansion and in situ cardiac differentiation. cRGD functionalization was essential for hiPSC survival and pluripotency, with maximal support achieved at a low ligand density (25 M). In the presence of cRGD, hiPSC encapsulation into softer gels made from lower molecular weight alginates led to enhanced hiPSC expansion and improved cardiogenesis. Strikingly, differentiation in situ with 3D gels led to hiPSC-CM with higher structural maturity, including a markedly increased proportion of Desmin positive cardiomyocytes. Finally, after enzymatic retrieval from hydrogels, cardiomyocytes derived from softer gels formed tissue-engineered myocardium with superior contractile force compared to tissue fashioned from hiPSC-CM derived from more rigid gels. Together, these results demonstrate the promise of this defined, tunable platform for biomanufacturing of structurally mature cardiomyocytes from hiPSC.

With the push towards accessible benchtop models to capture biological events, many researchers are reaching for hydrogel platforms for 3D tissue engineering ex vivo. Recapitulating the dynamic mechanical environment cells experience in vivo requires dynamic hydrogel scaffolds whose mechanical properties can be reprogrammed with spatiotemporal precision. Here we describe a chemically simple hydrogel platform that undergoes visible-light photosoftening via a ruthenium-based photocleavable crosslinker, leveraging tetrazine-norbornene inverse electron demand Diels Alder (iEDDA) click chemistry between RuTetrazine crosslinker and norbornene-modified hyaluronic acid (NorHA). Nitrogen gas evolved during this reaction is repurposed as an intrinsic porogen, nucleating macropores (55-175 {micro}m) directly during gelation. Initial stiffness (1.5-10 kPa) and softening extent (from 50%-100% drop in storage modulus) are independently tunable through polymer and crosslinker composition. We have found RuTetrazine to be non-mutagenic and non-toxic (>80% live cell populations) once network-bound (IC50 = 0.27 mM). In a cell-instructive network co-crosslinked with an MMP-RGD-bearing peptide, human mesenchymal stromal cells (hMSCs) photosoftened in situ (2.27[-&gt;]0.54 kPa, [~]76%) spread approximately six-fold relative to stiff controls ([~]6,500 vs. [~]1,100 {micro}m2, p < 0.0001). This work demonstrates a synthetically accessible photocleavable crosslinker and a simple, macroporous hydrogel for modulating dynamic mechanical cues in three dimensions.

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.

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.

Low response rates to immune checkpoint inhibitors (ICIs) in solid tumors are often driven by insufficient tumor-infiltrating CD8 T cells and immunosuppressive tumor microenvironment (TME). Although interleukin-2 (IL-2) potently expands and activates CD8 T cells, its clinical use is limited by rapid clearance, dose-limiting toxicity, and regulatory T cell (Treg) stimulation. Engineered IL-2 variants have not yet achieved meaningful clinical efficacy. Here, polymer-modified mesoporous silica nanoparticles displaying dense, unmodified wild-type IL-2 on their surface (IL2-NP) are developed, conferring proteolytic stability and tumor retention. IL2-NP enables avidity-mediated CD8 T cell binding and enhances proliferation and effector function without increased Treg binding or proliferation. Intratumoral IL2-NP expands CD8 T cells, increases CD8/Treg ratios, and reprograms TME through dendritic cell activation and M1-like macrophage polarization. IL2-NP induces regression of both treated and untreated distant colorectal tumors in a CD8 T cell-dependent manner. IL2-NP synergizes with ICIs and leads to complete tumor regression and immunological memory that protect against rechallenge. Treatment is well tolerated, with strong efficacy also observed in triple-negative breast and metastatic ovarian cancer models. Overall, intratumoral IL2-NP elicits robust systemic antitumor immunity, offering a promising strategy to enhance ICIs, cancer vaccines, and adoptive T-cell therapies. Graphical abstractThis work introduces a nanoparticle platform that overcomes major shortcomings of IL-2 immunotherapy by presenting wild-type IL-2 at high density on the nanoparticle surface, thereby increasing binding avidity to effector T cells. The resulting IL-2 nanoparticles enhance cytotoxic T cell expansion, reprogram the tumor microenvironment, and augment responses to immune checkpoint blockade to achieve robust ant-tumor immune response in mouse tumor models. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=182 SRC="FIGDIR/small/738558v1_ufig1.gif" ALT="Figure 1"> View larger version (82K): org.highwire.dtl.DTLVardef@12f8c8corg.highwire.dtl.DTLVardef@b46b1forg.highwire.dtl.DTLVardef@e4efc5org.highwire.dtl.DTLVardef@3993e6_HPS_FORMAT_FIGEXP M_FIG C_FIG

Potent antineoplastics, such as afatinib and freebase doxorubicin, are associated with systemic toxicity. To address this issue, we developed a carrier that releases drugs, including afatinib and doxorubicin, specifically at the focus of low-intensity ultrasound. This remotely triggered and focal approach enables the release of drugs specifically at the ultrasound focus, thus mitigating undesirable off-target effects, and at concentrations governed by the duration of the applied ultrasound. We produced ultrasound-sensitive microdroplets with high encapsulation efficiencies (39.6% for afatinib and 46.6% for doxorubicin). The microdroplets consist of an ultrasound-sensitive drug delivery system based on a methoxy poly(ethylene glycol)-poly(D, L-lactide) diblock copolymer (mPEG-PDLLA) and perfluorooctyl bromide (PFOB). Antineoplastic agents were encapsulated within these microdroplets via co-evaporation during particle synthesis. The microdroplets released doxorubicin and afatinib in an ultrasound-pressure-dependent manner, with fitted half-maximal release pressures (P50) of 0.61 MPa and 0.72 MPa, respectively. Together, the effective encapsulation of hydrophobic antineoplastic agents and the dose-dependent ultrasound-triggered release provide a new method for targeted drug delivery and a foundation for future targeted chemotherapies.

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.

Lipid nanoparticles (LNPs) have become a clinical standard for systemically-administered nucleic acid drugs and vaccines, but the LNP pipeline for locally-delivered LNP therapies remains much less mature. Local delivery in lung represents a particularly compelling application space for DNA-LNP therapeutics, as local gene therapies could support sustained epithelial recovery and functional restoration in various lung diseases. However, locally-delivered DNA-LNPs face multiple barriers, including the limited availability of serum components that often support conventional LNP activity, and the additional delivery barriers posed by the nucleus. We generated hybrid peptide-lipid nanoparticles (hpLNPs) for pulmonary DNA delivery by using a core-shell assembly strategy to incorporate short histone-derived peptides, selected for their DNA-transport capacity, into a clinically inspired LNP formulation. In parallel, we evaluated serum pre-coating of LNPs as a strategy to boost hpLNP activity in the serum-poor airway environment. A peptide:DNA amine/phosphate (N/P) ratio of 0.9 was identified as the highest feasible ratio to permit peptide incorporation into hpLNPs while preserving DNA encapsulation efficiency at >90%, retaining hpLNP colloidal stability, and preserving the overall pKa for LNPs. At N/P = 0.9, hpLNPs showed markedly enhanced DNA delivery in alveolar lung cells, achieving up to a 17-fold increase in transgene expression compared to peptide-free LNPs. Transgene expression levels varied depending on serum concentration, with expression peaking in the presence of 6% serum. Furthermore, serum pre-coating was necessary to enable in vivo hpLNP activity following intratracheal administration in mice, yielding robust local GFP expression. Mechanistic studies in exosome-free serum revealed that peptide incorporation in the hpLNPs enhanced transgene delivery by increasing nanoparticle interactions with serum exosome components, resulting in significant enhancements to hpLNP uptake. Together, these findings identify nanoparticle- serum interactions as a critical determinant of LNP-mediated pulmonary DNA delivery and establish peptide incorporation and serum pre-coating as promising strategies to enable robust, localized gene transfer in serum-free environments.

Biomaterial scaffolds for repairing traumatic muscle injuries require restoration of both the anisotropic architecture and basement membrane extracellular matrix cues critical to normal muscle function. To address this need, we establish a collagen-glycosaminoglycan (CG) scaffold platform pairing an aligned pore microstructure, produced via directional freeze-drying, with basement membrane protein functionalization via carbodiimide crosslinking. Laminin and/or collagen IV are successfully tethered and retained within CG scaffolds over 7 days without significantly altering pore size or alignment, confirming stable protein functionalization and preservation of scaffold architecture. Human muscle progenitor cells show excellent viability and metabolic activity in all scaffold groups, with collagen IV functionalization significantly enhancing myotube number and fusion index. Toward establishing scaffold compatibility with non-myogenic support cells, we show that neural stem cells remain viable and metabolically active across all scaffold conditions. Overall, these findings highlight the combination of aligned scaffold architecture and collagen IV functionalization as potentially impactful for skeletal muscle tissue engineering.

Interleukin-6 (IL-6) is a pleiotropic cytokine whose aberrant elevation drives life-threatening conditions, including sepsis, cytokine storm, and autoimmune disorders, yet existing clinical detection methods demand centralized laboratory infrastructure and multi-hour assay times incompatible with rapid point-of-care decision-making. Here, we report an impedimetric aptasensor built on a programmable tetrahedral DNA nanostructure (TDN) interface anchored to a disposable gold screen-printed electrode (Au-SPE) for the ultrasensitive, label-free detection of IL-6. By systematically varying the number of thiolated base vertices from zero to three, we establish a clear and previously unreported structure-function relationship between multipodal anchoring geometry and charge-transfer resistance modulation: tripodal thiolation yields the most rigid, upright, and electrochemically responsive interface, producing the steepest analytical signal gain upon IL-6 binding at the apex-localised aptamer. Under optimised conditions (pH 7.0, 0.05 uM TDN, MCH passivation), the aptasensor exhibits a linear dynamic range of 0.0001-0.001 pg/mL, a limit of detection of 55 ag/mL, and a sensitivity of 1.55x107 ohm (pg mL-1)-1. Selectivity evaluation against seven physiologically relevant interferents such as TNF-, BSA, glucose, urea, ascorbic acid, glycine, and cysteine confirms negligible cross-reactivity, with relative responses ranging from 0.57% to 14.35% of the IL-6 signal. Spike-recovery experiments in human serum yield recoveries of 74.0-87.6% (%RSD < 4.5%), and the sensor retains functional activity for at least 21 days under refrigerated storage. This work demonstrates that thiolated vertex number is a critical and tunable design parameter for TDN-based biosensors, offering a modular, disposable platform for sub-femtogram cytokine detection with direct applicability to early sepsis diagnosis and inflammatory disease monitoring.

Governing thermal transport in engineered materials creates opportunities to redirect and recover the excess heat generated in electronic and energy-conversion devices. Materials that pair low cross-plane thermal conductivity with high in-plane thermal conductivity are particularly valuable because they confine heat and channel it away from sensitive regions, preventing localized device failure. Two-dimensional crystals are efficient building blocks for such anisotropic thermal conductors, but they are brittle, and the polymer composites used to toughen them usually forfeit much of the intrinsic anisotropy: in conventional percolation-based design, filler fraction is the only handle available, and it governs both in-plane and cross-plane conduction. Here we report a composite of Ti3C2Tx (MXene) nanosheets and squid ring teeth (SRT) inspired recombinant tandem-repeat (TR) proteins in which the protein serves as a molecular template and bridge, setting the spacing between nanosheets with angstrom-level precision through the number of tandem-repeat units and independently of the filler fraction. This structural handle provides a second, independent design parameter. At a fixed MXene loading, the number of repeats tunes the cross-plane conductivity (0.30 to 0.93 W/mK) and, with it, the thermal anisotropy ratio over a wide range (from about 70 down to 17), while the in-plane conductivity stays high (16 to 21 W/mK). We rationalize these trends with a Gaussian Network Model (GNM) of the protein embedded in a two-phase layered medium, which reproduces the measured directional conductivities from a single structural parameter and identifies the protein gallery as the cross-plane bottleneck. Extending the model to a mechanically loaded five-period stack, we find that the anisotropy is robust to reversible compression and twist, changing by only a few percent, so the number of tandem repeats, not the applied strain, is the dominant design handle. Because anisotropy is tuned structurally rather than volumetrically, these protein-MXene composites decouple thermal anisotropy from filler content, pointing toward flexible thermal materials that are not bound by the rules of mixture and percolation.

The early detection of breast cancer currently relies on expensive mammography, followed by pathology that uses biopsied, fixed, and immunohistochemically stained tissues. A live-cell detection approach could be highly beneficial as a supportive diagnostic and research tool to better understand and resolve the dynamic nature of breast cancer cells and their response to treatment in real time. Here, we present a single-walled carbon nanotube (SWCNT) near-infrared fluorescence spectral fingerprinting approach combined with machine learning to precisely detect the heterogeneity of breast cancer cells in live culture. We introduced DNA-functionalized SWCNTs to MCF-10A (a non-tumorigenic healthy control) and cancer cell lines spanning known extrinsic disease subtypes: MCF-7 (luminal A), HCC1954 (HER2+), MDA-MB-231, and MDA-MB-468 (both triple-negative). The NIR fluorescence spectra of DNA-SWCNTs across 600 individual cells within each type showed significant differences in emission peak intensities, center wavelengths, and peak intensity ratios, attributable to variations in cellular uptake and biomolecular interactions. These spectral changes likely arise from complex SWCNT cellular interaction fingerprint that includes redox-mediated modulation of the local nanotube environment, rather than from a single biomarker response. The extracted spectral features were used to train an ensemble machine learning model. The model achieved 98% classification accuracy for breast cancer detection and 95% classification accuracy for breast cancer cell subtyping. Moreover, Raman microscopy further showed that MDA-MB-468 cells exhibited the highest SWCNT uptake, whereas MCF-10A cells showed greater SWCNT aggregation, consistent with their lower broadband NIR fluorescence intensity. These results demonstrate that SWCNT NIR fluorescence fingerprints can capture cell line-specific optical signatures. This platform provides a foundation for nanomaterial-enabled biosensing strategies aimed at real-time monitoring of cancer-associated cellular states.

Modulation of immune cells as therapeutic tools has gained significant clinical relevance in the treatment of cancer. Among them, macrophages represent a promising immunotherapeutic platform not only because they can internalize tumor material, but also because they profoundly shape the tumor microenvironment through cytokine production, antigen presentation, metabolic regulation, and modulation of other immune and stromal populations. Lipid Nanoparticles (LNPs) have enabled RNA therapies to the bedside and are thus considered the gold standard for gene delivery. However, optimizing LNPs for RNA delivery to macrophages remains an active area of investigation. Here, we propose the surface modification of unPEGylated LNPs using the Layer-by-Layer (LbL) approach for enhanced RNA delivery to macrophages. Specifically, we show that fucoidan, a sulfated polysaccharide, when at the outermost layer in the LbL process provides two physicochemical advantages to unPEGylated LNPs: (1) stability in PBS and (2) resistance to lyophilization in the presence of cryoprotectant. Additionally, fucoidan improves macrophage targeting and RNA transfection efficiency compared to previously synthesized hyaluronan-decorated LbL LNPs. Fucoidan LbL LNPs (Fuc-LNPs) preferentially accumulated in CD11b+ macrophages when co-cultured with U87 glioblastoma cells, which was not observed for control PEGylated LNPs. Furthermore, Fuc-LNPs induced a higher transfection of mRNA in primary human macrophages when compared to PEGylated control LNPs. Using the model mRNA encoding CAR@CD19, Fuc-LNPs generated CAR macrophages which mediated CD19 cell ablation in vitro. Altogether, these findings highlight the potential of the LbL strategy to modulate the targeting properties of LNPs, improving RNA delivery to human macrophages and encouraging further studies using LbL LNPs for the generation of CAR-Macrophages in the context of solid tumors.

Modelling human cortical microcircuitry in vitro requires platforms that recapitulate both the compositional complexity and spatial architecture of developing neural tissue. Current organoid and assembloid models often rely on the bulk fusion of pre-differentiated, region-specific cells, lacking the capacity for emergent spatial co-differentiation and microenvironment-driven multiscale organisation. There is also a lack of neural and neuronal-glial models with photo-architectured network geometries. To address these limitations, we present a volumetric in situ differentiation system using a triculture of precision reprogrammed human iPSC-derived glutamatergic neurons, GABAergic neurons and astrocytes embedded throughout ultra-soft photocrosslinkable hydrogel microenvironments. The deterministic and spatially controlled method allows us to engineer macro-scale, interconnected human neural networks directly onto functional microelectrode array interfaces using projection photopatterning for high-throughput screening. Unlike fusion-based organoids and assembloids, our platform enables simultaneous, spatially distributed lineage differentiation and maturation, and extensive topography-guided neurite outgrowth bridging localised cellular hubs to recapitulate various aspects of neurodevelopmental patterning and synaptic integration in 3D. The model enables topographic patterning of neuronal-glial networks as well as 3D cell-embedded bioprinting with the developed triculture system. Both modes of cellular growth are studied and demonstrated here. Longitudinal electrophysiological tracking over a month of culture reveals a transition from immature, quiescent states to asynchronous, information-dense microcircuits characterised by an expanded state-space manifold and physiological excitatory-inhibitory balance. By replicating the mechanics of native brain parenchyma, the model presents a highly reproducible, scalable and flexible platform for the study of cortical microcircuitry development, neurodegenerative decline, and inter-regional network assembly.

Avascular meniscus tears exhibit minimal intrinsic healing and often progress to joint degeneration due to restricted biological repair capacity and inadequate restoration of tissue-level structure and function. Here, we report a hydrophilic polydopamine (hPDA) fueled bioglue platform that overcomes the solubility limitations of conventional polydopamine (PDA) and enables functional repair of avascular meniscus injuries. Water-soluble hPDA was synthesized via controlled depolymerization and recrystallization, yielding monomeric and oligomeric species rich in catechol, amine, and hydroxyl functionalities. Incorporation of hPDA into fibrin bioglues markedly enhanced mechanical performance, producing 520-525% increases in lap-shear modulus, 165-190% increases in adhesive strength, and a 160% increase in compressive modulus relative to fibrin controls, while degradation was markedly attenuated over 14 days. hPDA exhibited excellent cytocompatibility in both 2D and 3D cultures. In a bovine avascular meniscus explant model, hPDA fueled bioglues promoted tissue integration and aligned collagen remodeling, restoring interfacial mechanics with a 488% increase in tensile modulus and up to 150% higher pull-out strength after 6 weeks. These findings establish hPDA as a versatile bioadhesive building block with strong potential for repairing avascular meniscus tears and other mechanically demanding connective tissues. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=110 SRC="FIGDIR/small/738365v1_ufig1.gif" ALT="Figure 1"> View larger version (40K): org.highwire.dtl.DTLVardef@1f376dborg.highwire.dtl.DTLVardef@10029feorg.highwire.dtl.DTLVardef@1c4f191org.highwire.dtl.DTLVardef@cfa2fe_HPS_FORMAT_FIGEXP M_FIG C_FIG

Tissue engineered skin models are important tools for the in vitro study of physiological and pathophysiological processes as well as the valuation of therapeutic strategies and the efficacy of pharmaceutical and cosmetic compounds. Replicating the functional anatomy of cutaneous tissue is a crucial aspect in ensuring that observations made using these models are translatable to the actual situation in native skin. However, most contemporary full-thickness skin models neglect the reconstruction of the undulated microtopography of the dermal-epidermal junction (DEJ), which not only contributes to the biological functionality of the skin (e.g. stem cell niches), but also affects tissue mechanics and drug diffusion. Herein, we fabricated bilayer skin models with DEJ-like microtopographies introduced by interfacial wrinkling between a hydrogel and a nanofibrous membrane through a controllable swelling-deswelling approach. The interfacial wrinkles contributed to the structural integrity of the bilayer models. Their formation could be induced in the presence of living cells through mechanical stress-driven buckling instabilities, thus differentiating the process from commonly used pre-patterning techniques. Bilayer models supported the co-culture of human dermal fibroblasts and human epidermal keratinocytes, and the formation of stratified epithelia. Our findings provide a potential alternative method to introduce DEJ-like anatomical features into full-thickness skin tissue models.

Innate immune agonists are promising therapeutic agents to induce immune responses against cancer. However, these agents have been limited by toxicity associated with systemic accumulation and activity in off-target cells. In this work, a targeted nanoparticle (NP) platform to encapsulate and protect the Toll-like receptor 3 (TLR3) agonist polyinosinic-polycytidylic acid (poly(I:C)) and promote its specific delivery to antigen presenting cells (APCs), macrophages and dendritic cells, for activation of this cell population was designed. To determine NP physiochemical properties that promote APC delivery, we developed a library of NP surface chemistries formed by electrostatic adsorption of polyanion coatings onto liposomes using layer-by-layer (LbL) assembly and screened the particles on APCs and off-target cells. Dextran sulfate was identified as a promising coating to enhance specific APC delivery. We applied these design parameters to develop a poly(I:C)-loaded NP for an APC-targeted immunotherapy. In a model of metastatic ovarian cancer, the LbL NP prolonged poly(I:C) retention in the peritoneal space--with 2-fold remaining 24-48hr after administration compared to free poly(I:C)--ultimately reducing systemic accumulation and associated toxicities. Compared to free drug, the NP reduced the increase in serum levels of TNF, IL-6, and CXCL10 by 9-, 4-, and 31-fold respectively. NP-treated mice experienced lower weight loss and recovered more quickly at a higher poly(I:C) dose, indicating a widening of the therapeutic window. The NP formulation enhanced accumulation of poly(I:C) in the tumor 2-fold and activation of the target APC population compared to free drug, and ultimately slowed tumor growth and extended survival in combination with doxorubicin chemotherapy. Overall, this work demonstrates a modular NP delivery strategy to improve the delivery, safety, and therapeutic window of a TLR3 agonist.

Although microplastics and nanoplastics (MP/NP) are pervasive environmental contaminants, our understanding of cellular toxicity remains incomplete, as adverse effects are often attributed to long-term intracellular accumulation, while the spatiotemporal onset of cellular damage remains poorly defined. Here, we employ chemical-bond-selective stimulated Raman scattering (SRS) microscopy and cell models that decouple continuous exposure from intracellular retention to directly visualize clinically derived MP/NP-cell interactions. Cellular stress occurs primarily during MP/NP exposure, accompanied by alterations in lipid droplet (LD) composition. In contrast, following extracellular removal, intracellularly retained MP/NP become largely inert, with recovery of lipid metabolism and cellular functions. Lipidomics identifies arachidonic acid (AA) as a key dysregulated metabolite, and SRS imaging further reveals transient, spatially confined AA enrichment in MP/NP-proximal LDs during uptake. Importantly, phospholipid coating of MP/NP attenuates LD alterations and cytotoxicity while preserving particle internalization, establishing uptake-driven metabolic stress, rather than long-term intracellular retention, as primary source of MP/NP-induced damage.