Biomaterials Advances

Critical-sized bone defects represent a challenge in bone tissue engineering, due to insufficient vascularization that results in implant failure. Scaffold pre-vascularization is a promising strategy to create a functional microvascular network that integrates with host vasculature. In this study, we present a hybrid 3D construct comprising a hyaluronic acid-based hydrogel and a 3D printed polycaprolactone/{beta}-tricalcium phosphate scaffold, to support vascular network formation and osteogenic differentiation. Peptide-functionalized (i.e. RGD, YIGSR, IKVAV, QK) hydrogels were obtained via thiol-ene chemistry, using two crosslinkers (PEG-diSH or MMP-diSH). Preliminary biological experiments assessed human mesenchymal stromal cells (hMSCs), endothelial cells (hUVECs), and their co-culture, on different gel formulations. All cell conditions displayed enhanced spreading and metabolic activity on gel formulations comprising RGD; thus these (i.e. RGD only and a combination of RGD/YIGSR) were selected for further studies. Cells were then mixed with the hydrogel precursor solutions, which were injected to embed the scaffolds and crosslinked using a UV lamp. After 7 days, tubule formation was observed only in co-culture conditions, highlighting the importance of cellular crosstalk for the formation of a vascular network. Significant differences were found across the tested formulations. In the RGD-PEG constructs, hUVECs formed tubule-like structures, surrounded by hMSCs, exhibiting pericyte-like behavior, supported by the upregulation of SMA gene. Conversely, in the RGD/YIGSR-MMP conditions, hMSCs were mostly located on the scaffold fibers, and showed the highest expression of early osteogenic markers (RUNX2 and ALP). Overall, we demonstrated that the hybrid system with tailored hydrogel chemistry can support simultaneous microvascular organization and osteogenic commitment, offering a promising platform for bone tissue engineering applications. However, further studies involving longer culture periods will aim at clarifying the complex interplay between material composition, cell crosstalk and spatial organization and their influence on the maturation and stability of the vascular network.

Curcumin is a naturally occurring polyphenol that demonstrates considerable anti-cancer activity, however the aqueous insolubility, rapid metabolism and relatively low bioavailability are limiting to its clinical application. As such, a curcumin-magnesium (Cur-Mg) coordination complex was synthesized and subsequently encapsulated within DNA hydrogels (Cur-Mg-Hgel). The Cur-Mg complex was fully characterized using UV-Vis spectroscopy, FTIR and X-ray diffraction (XRD). UV-Vis, FTIR and XRD all support the formation of a coordination complex and suggest a decreased level of crystallinity compared to free curcumin. DNA hydrogels were formed and characterized using atomic force microscopy, rheology and swelling kinetic studies. In vitro cytotoxicity studies utilizing an MTT assay demonstrate dose dependent inhibition of HeLa cell proliferation and a slightly better retention of RPE-1 viability at low concentrations (suggesting some difference in sensitivity) though significant cell death is seen at higher concentrations and both cells. Intracellular production of ROS was measured using the DCFH-DA assay and is seen to increase when HeLa cells are treated with Cur-Mg-Hgel in comparison to un-treated controls. Annexin V/PI staining demonstrates primarily late or early apoptotic activity with minimal necrosis following treatment with Cur-Mg-Hgel. The evidence presented strongly supports the notion that Cur-Mg-Hgel is a ROS-modulating, pro-apoptotic Hydrogel suitable for cancer treatment. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=102 SRC="FIGDIR/small/724072v1_ufig1.gif" ALT="Figure 1"> View larger version (42K): org.highwire.dtl.DTLVardef@18727aeorg.highwire.dtl.DTLVardef@3e20adorg.highwire.dtl.DTLVardef@d3703eorg.highwire.dtl.DTLVardef@16e260e_HPS_FORMAT_FIGEXP M_FIG C_FIG

This study focuses on the development of 3D culture model dedicated to liquid cancers drug screening. The challenge addressed was to effectively retain non adherent small cells within a 3D-scaffold with tailorable mechanical properties, while proposing a fast and effective tool for drug screening. To that aim, we developed a macroporous alginate-chitosan polyelectrolyte complex (PEC) scaffold combined with a low-viscosity alginate (LVA) cell seeding solution. We hypothesized that LVA could undergo in situ pore gelation via calcium ions retained from the PEC fabrication process, enabling effective retention and homogeneous cell distribution, leading to an improved platform for drug screening and personalized medicine. First, we evaluated scaffold suitability for LVA infiltration and gelation. Microtomography revealed a highly porous architecture (98%) enabling LVA homogeneous penetration and complete gelation within 30 min, as confirmed by SEM, microscopy, rheology, and micro-rheology. Next, we assessed cell retention and biocompatibility using primary human chronic lymphocytic leukemia (CLL) cells. LVA-assisted seeding increased cell density 2.6-fold compared to medium alone, with homogeneous distribution, >80% viability over 7 days, and preserved differentiation into nurse-like cells. Finally, we demonstrated a proof of concept for drug screening. The Alginate-PEC scaffold (A-PEC scaffold) supported both qualitative live/dead imaging and rapid quantitative viability measurement with the Alamar Blue assay. Drug responses reproduced microenvironment-dependent protection effects observed in vivo. This integrated scaffold and seeding method provides a promising 3D platform for in vitro liquid cancer studies and drug screening on patient-derived hematological cancer cells. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=67 SRC="FIGDIR/small/722037v1_ufig1.gif" ALT="Figure 1"> View larger version (38K): org.highwire.dtl.DTLVardef@9b71d4org.highwire.dtl.DTLVardef@14e1dd0org.highwire.dtl.DTLVardef@1876a56org.highwire.dtl.DTLVardef@15656bc_HPS_FORMAT_FIGEXP M_FIG C_FIG

The combination of synthetic biology and additive manufacturing has driven major changes in production of biomaterials, especially through the use of three-dimensional (3D) bioprinting to create engineered living materials. However, current fabrication methods can be limited by prohibitive hardware costs and the inability to maintain structural fidelity in complex, free-form living architectures. This work demonstrates how to build a low-cost, open-source 3D bioprinting platform that can make complicated bacterial structures with complex geometry and high dimensional accuracy. A commercially available, conventional fused deposition modeling 3D printer was modified to create a bioprinting system that is simple to build. The modified bioprinter, which costs around $450, is less expensive than many commercial bioprinters. This 3D-printing technology uses slurry-based support bath methods featuring low-cost gelatin and agarose microparticles, resulting in structures with a high aspect ratio (>8:1) and feature sizes as small as 260 m. The optimization of critical printing settings, including the ability of the bioink to retract during non-print movements, resulted in a reduction of unwanted bacterial deposition by nearly two orders of magnitude. Long-term viability experiments showed that bacteria in the bioprints could survive for at least 28 days with nutrient supplementation. Additionally, 3D-printed engineered biofilms revealed that incubation conditions and extracellular matrix composition significantly impacted the mechanical properties of printed constructs, with tradeoffs between matrix production and mechanical integrity. This study showcases an accessible 3D bioprinting platform for advanced bioprinting technologies, enabling development of engineered living materials with potential applications in synthetic biology, biotechnology, and tissue engineering.

BackgroundBone graft biomaterials play a critical role in bone regeneration by influencing osteoblast differentiation and mineralization. However, comparative data regarding the osteogenic potential of commonly used graft materials under standardized conditions remain limited. Method and materialIn this in vitro experimental study, osteoblast-like cells (MG-63) were cultured with four bone graft materials, including Bio-Oss, Cerasorb, Bio-Tiss Cerabone, and Pro Osteon. The relative mRNA expression of osteogenic markers (COL1 and OPN) was evaluated at 1, 7, 14, and 21 days using real-time PCR. Alkaline phosphatase (ALP) activity and mineralization capacity were also assessed using colorimetric assay and Alizarin Red staining. Data were analyzed using one-way ANOVA and Tukey post hoc test (P < 0.05). ResultsSignificant differences were observed among the tested materials across all evaluated parameters. Bio-Oss and Cerasorb demonstrated higher gene expression levels and ALP activity compared to Bio-Tiss Cerabone and Pro Osteon (P < 0.05). Mineralization analysis showed significantly greater calcium deposition in the Bio-Oss and Cerasorb groups, whereas Pro Osteon consistently exhibited the lowest osteogenic performance. ConclusionBone graft biomaterials significantly influence osteogenic activity in osteoblast-like cells. Bio-Oss and Cerasorb showed superior osteogenic potential, while Pro Osteon demonstrated weaker performance. These findings highlight the importance of material properties in optimizing bone regeneration.

Biodegradable electrospun nanofiber (NF) scaffolds have emerged as promising materials for tissue engineering applications, including vascular grafts, because their mechanical properties and degradability can be tuned. However, their in vivo degradation behavior remains poorly understood. In this study, we characterized the in vivo degradation profiles of representative biodegradable NF materials widely used in small-caliber vascular graft research, namely polycaprolactone (PCL), poly(D,L-lactide) (PLA), polyglycolic acid (PGA), and a PCL/PLA blend, by monitoring molecular weight changes in subcutaneous and vascular environments. Electrospun NF sheets were implanted subcutaneously in mice, and tubular NF grafts were implanted into the abdominal aorta of rats. Samples were harvested for up to 48 weeks after implantation and analyzed primarily by size-exclusion chromatography (SEC) to assess time-dependent changes in molecular weight. Scanning electron microscopy (SEM) and solid-state 13C nuclear magnetic resonance (NMR) were additionally performed to evaluate ultrastructural and chemical changes associated with degradation. SEC analysis revealed distinct material-specific degradation patterns. PCL showed the slowest degradation and retained a relatively high weight-average molecular weight (Mw) in both environments. PLA exhibited marked environment dependence, with near-complete degradation in the subcutaneous environment by 48 weeks, whereas scaffold structure was maintained in the vascular environment. The PCL/PLA blend showed earlier reduction in the high-molecular-weight fraction than PCL, indicating faster scaffold breakdown. PGA degraded most rapidly and could not be evaluated beyond 2 weeks in the subcutaneous model or in the vascular model because of early graft rupture. SEM analysis further demonstrated that progressive loss of fibrous ultrastructure over time was a common feature across all materials. In addition, NF scaffolds became resistant to organic solvent after implantation in vivo, and solid-state 13C NMR analysis of the solvent-insoluble fractions detected polymer-derived signals together with additional signals consistent with biological constituents. These findings indicate that in vivo degradation of biodegradable NF scaffolds is material dependent, environment dependent, and more complex than simple hydrolytic chain cleavage alone. This study provides a quantitative framework for evaluating NF degradability and offers new insight into the design of biodegradable vascular grafts. HighlightsO_LISEC quantified long-term in vivo degradation of PCL, PLA, PGA, and PCL/PLA. C_LIO_LIDegradation was both material dependent and implantation environment dependent. C_LIO_LIIn vivo nanofiber degradation involved structural and chemical changes beyond hydrolysis. C_LI

Surface functionalization of biomaterials enables the immobilization of proteins and other molecules and can be utilized to direct the biological response to devices and implants. Fetuin-A is a blood plasma protein involved in numerous physiological processes, including the regulation of mineralization. Notably, many investigations of fetuin-A have explored its cellular interaction when in solution, but limited studies report the role of fetuin-A when used as a surface modifier. The present investigation explores the response elicited by fetuin-A on Saos-2 cells when it is immobilized on a model gold surface through the covalent reaction with dithiobis(succinimdyl propionate) (DSP). Comparative surface characterization using x-ray photoelectron spectroscopy (XPS), atomic force microscopy - infrared spectroscopy (AFM-IR) and surface plasmon resonance (SPR) confirmed the surface modifications but indicate partial inhomogeneity in the functionalizer surface coverage. The interaction of albumin and fetuin-A with the surface was quantified by radiolabeling, quartz crystal microbalance with dissipation (QCM-D) and SPR, demonstrating a higher mass of fetuin-A bound to the surface in comparison to serum albumin. Over 7 days, cells bound to the surfaces with immobilized fetuin-A showed significantly hindered proliferation of osteoblast-like cells compared to the positive control (fibronectin), presumably due to a decrease in cell metabolism. This study provides new insights into the role of fetuin-A in regulating Saos2 cell response and elucidates its potential use in combination with chemical functionalizers for biomedical applications requiring surface modification.

Sustainable, biodegradable elastomers are needed to replace fossil-based alternatives and reduce the environmental impact of traditional vibration damping materials. We investigate agarose-based hydrogels as eco-friendly vibration absorbers, examining the combined effects of polymer concentration (1-7 wt%), relative humidity (55-98%), and mechanical pre-stress on their dynamic mechanical properties. Frequency-dependent viscoelastic and vibration transmissibility tests, supported by Gaussian Process Regression (GPR), reveal that increasing agarose concentration enhances the storage modulus (E') by over an order of magnitude, reaching[~] 5 MPa depending on humidity and applied prestress. Remarkably, the damping efficiency--characterised by the loss factor (tan(d))--exhibits a highly non-monotonic trend. Maximum energy dissipation is observed at intermediate network densities, with tan(d) up to 0.21 and a loss modulus of[~] 515 kPa at 5 w% and 75% relative humidity, comparable to synthetic elastomers. GPR analysis shows that prestress controls nonlinear stiffening and transmissibility resonance behavior, while shifting peak damping from 5 wt% to 1 wt% agarose as prestress increases. These findings underscore the mechanical tunability and sustainability of agarose hydrogels, providing potential design guidance for biodegradable vibration mitigation materials.

Hydrogels are widely used as three-dimensional cell culture systems to understand the impact of cellular mechanotransduction for tissue engineering applications. Photoinitiated thiol-ene click chemistry is a commonly utilized hydrogel crosslinking mechanism that provides spatial and temporal control over hydrogel network formation and resulting mesh size and compressive properties. Despite historically documented efficiency as step-growth reactions, these reactions do not always proceed as predicted. To understand the impact of cell confinement and microenvironmental mechanics on cellular function, thiol-ene network formation must be thoroughly characterized. To this end, the objective of this work was to investigate the crosslinking dynamics to determine hydrogel network formation as assessed via mesh size and mechanical properties using a pentenoate-functionalized hyaluronic acid thiol-ene reaction. Hydrogel parameters including polymer concentration and thiol:-ene crosslinker molar ratio were modulated (4, 6, or 8 polymer weight percent and 0.15:1, 0.5:1, or 1:1 molar ratio of thiol groups to reactive -ene groups) to tune network properties including shear storage modulus and relative mesh size. Molecular Dynamics (MD) simulations were used to simulate the thiol-ene crosslinking reaction and establish a method for predicting thiol-ene reaction efficiency. Lastly, the feasibility of this hydrogel system for in vitro modeling was confirmed via assessment of metabolic activity of encapsulated primary human meniscal cells.

Cultivated meat is an emerging biotechnology that aims to produce edible tissues in an ethical and sustainable manner. However, the recreation of skeletal muscle tissue that replicates the protein composition and sensory characteristics of traditional meat is a major challenge. Skeletal muscle tissue engineering requires non-animal-based scaffolds which are inexpensive and food-safe, while meeting specific mechanical requirements with respect to viscosity, stress-relaxation and stiffness. While many of these characteristics can be fulfilled by alginate-based biomaterials, a key limitation of alginate is its lack of intrinsic attachment sites for animal cells, preventing efficient adhesion, differentiation and tissue formation. Here, we established a screening platform to evaluate extracellular matrix (ECM)-mimicking peptides as functionalisations of alginate scaffolds in 2D. Our platform enables high-throughput assessment of cell/peptide interactions, serving as a predictive tool for 3D tissue constructs. Our screen identified two RGD-containing sequences (vitronectin- and fibronectin-mimicking peptides) as most effective in promoting attachment and myogenic fusion of bovine satellite cells. Notably, these peptides outperformed more complex mixtures containing up to seven different ECM-mimicking peptides. Our findings provide a streamlined approach for optimising biomaterial functionalisations for cultivated meat applications, and lay the groundwork for future advancements in scalable, sustainable skeletal muscle tissue engineering.

Hydrogels are the preferred materials for applications mimicking soft tissues due to their high water content and tunable mechanical properties. The state of the water in these hydrated networks governs their response to mechanical loading through coupled interstitial flow and large deformations of the solid network. Reliable experimental methods for quantifying the fraction of mobile fluid during mechanical deformation remain limited. Within the theoretical framework of mixture theory, we describe hydrogels as hydrated biphasic media consisting of a deformable incompressible solid matrix and a mobile fluid phase. We developed a mechanical testing protocol that enables the experimental separation of solid and fluid contributions under loading. The method is demonstrated using biocompatible and highly versatile hydrogel phantoms of varying compositions. Controlled, incremental drained confined compression of the hydrogel samples results in free-water fractions of approximately 40%, 60%, and 77%, reflecting the systematic influence of the polymer content on the porosity and fluid mobility. Comparison with cryo-SEM-derived surface porosity reveals statistically significant differences and highlights the scale-dependent sensitivity of surface measurements compared to bulk measurements. This study introduces a new mechanical method for quantifying the free-water fraction in macroporous, ultrasoft, highly hydrated biomaterials. Furthermore, the multi-step protocols enable the separation of dissipative, fluid-related relaxation from the equilibrium response of the solid skeleton, allowing direct calibration of constitutive models for macroporous soft solids. The proposed method provides a reliable basis for the development and optimization of hydrogels for applications where fluid transport is critical, such as neural interfaces, bioelectronic platforms, and tissue-engineered constructs.

Tissue engineered constructs are increasingly used for both modeling organs and disease in vitro as well as for therapeutic intervention. In addition to collagen, these constructs commonly include native extracellular matrix proteins (ECM), such as fibronectin and laminin. Given the critical role of inflammatory pathways in disease and in response to implanted materials, it is important to understand the role these proteins play in regulating the inflammatory environment. Fibronectin and laminin influence neutrophil function and endothelial activation in 2D, but their regulation of the inflammatory environment in 3D engineered constructs is not clear. For this study, we used an inflammation-on-a-chip device that includes a model blood vessel surrounded by a collagen I hydrogel with fibronectin and/or laminin. We investigated the additive effects of both proteins and a range of concentrations for each protein to determine concentration dependence. Both fibronectin and laminin have concertation dependent effects on neutrophils and the endothelium. High concentrations (50 {micro}g/mL) of fibronectin reduced neutrophil migration, while 20 {micro}g/mL laminin reduced neutrophil extravasation and migration, potentially due to lower ICAM-1 expression by the endothelium. Interestingly, 50 {micro}g/mL of laminin significantly disrupted endothelial vessel formation and reduced ICAM-1 and VE-cadherin expression, likely due to significant changes in the collagen architecture. The inclusion of fibronectin and laminin, even at physiological levels, results in significant effects on neutrophil behavior, endothelial vessel formation, and collagen architecture. These proteins impact the inflammatory environment and thus need to be considered when modeling diseases and designing therapeutics, especially when neutrophils or an endothelium are involved. Translational Impact StatementThis work uses an inflammation-on-a-chip device to study how fibronectin and laminin impact neutrophil behavior and vascular inflammation as these proteins are commonly used in engineered constructs. We found that fibronectin impairs neutrophil migration, while laminin decreases neutrophil extravasation and migration and at higher concentrations also prevents endothelial vessel formation. Therefore, researchers should be aware that these proteins will alter the inflammatory environment when including them in engineered constructs.

Matrix-bound nanovesicles (MBVs) are a type of small extracellular vesicle (EV) embedded in the extracellular matrix (ECM) throughout the body. MBVs have been previously isolated from various tissues and in vitro-cultured cell sheets, demonstrating remarkable attributes in regenerative medicine. However, differences between MBVs and conditioned culture medium-derived EVs (liquid-EVs) have yet to be characterized, and the field currently lacks specific protein markers that can identify MBVs from other EV subtypes. Here, we isolate MBVs and liquid-EVs from bone marrow mesenchymal stem cell (MSC) sheets and define differences in size, protein, and zeta potential between these EVs. We show that there is a correlation between cell-driven ECM deposition and MBV and liquid-EV production. We also find that MBVs are smaller, contain less protein per particle, and possess lower zeta potential than liquid-EVs. Interestingly, MBVs also comprise a distinct tetraspanin profile compared to liquid-EVs, with MBVs containing more CD63 and little to no CD81. Finally, we define that CD63, LAMP1, Alix, ITG{beta}1, and GRP94 and their abundance, may be markers specifically used to identify MBVs from liquid-EVs. Our study paves the way for the characteristic differentiation between MBVs from liquid-EVs, elucidates their differences in biogenesis, and reveals a potential connection between EV and ECM production.

Bio-based materials are known for their excellent biodegradability and, in some cases, their potential to fix carbon dioxide. Owing to these properties, they are increasingly being utilized as environmentally friendly alternatives across various applications. In this study, we focused on using living cells themselves as material components, aiming to evaluate their potential as substitutes for conventional plastic-based thermal insulators. We selected two types of cells, photosynthetic purple non-sulfur bacterium Rhodovulum sulfidophilum and tobacco BY-2 plant suspension cells. After optimizing solidification conditions through the addition of pectin and cellulose nanofibers, we measured the thermal conductivity of the solidified cells under atmospheric pressure. The results showed that R. sulfidophilum exhibited 0.0553 W/m{middle dot}K, while BY-2 exhibited a thermal conductivity of 0.043 W/m{middle dot}K. Both values indicate relatively low thermal conductivity compared to existing bio-based materials, suggesting high insulation performance. Among the solidified cells, the solidified BY-2 cells showed minimal variation in thermal insulation performance under pressure changes, and had a low thermal emissivity as revealed by FT-IR analysis. Based on these findings, we propose that cell-derived materials can serve as potentially biodegradable bio-based thermal insulation materials.

Nanoclay-based biomaterials offer promise for localised growth factor presentation, yet their in vivo degradation, clearance, and systemic fate remain poorly defined. Here, we investigate the fate of a synthetic nanoclay-BMP-2 gel during ectopic bone induction using a combination of in vivo imaging, histology, and component-resolved elemental analysis. Fluorescent tracking confirmed prolonged localisation of BMP-2 within the nanoclay gel and robust bone induction despite negligible growth-factor release. Inductively coupled plasma mass spectrometry (ICP-MS) revealed divergent clearance kinetics for lithium and silicon, structurally distinct components of the clay crystalline lattice, indicating decoupled ionic and particulate degradation pathways. Early clearance was dominated by cell-mediated fragmentation and the transport of clay particulates, while later stages involved preferential lithium release associated with local clay dissolution as well as integration within newly formed bone. Systemic biodistribution analysis demonstrated rapid, transient lithium release into circulation with renal clearance, contrasted with initial hepatic and then later-phase renal handling of silicon species. Together, these findings define a multiphasic in vivo clearance model for nanoclay biomaterials consistent with progressive remodelling, localised BMP-2 activity and, importantly, safe systemic handling. This work provides mechanistic insight into the activity and clearance of nanoclay-based regenerative therapies and establishes the importance of component-resolved tracking for evaluating the biodistribution of degradable inorganic biomaterials.

BackgroundPhenome-wide association studies (PheWAS) can reveal novel associations between variants in drug-target genes and disease and, as such, can be used to predict new drug-indication pairs for repurposing drugs with a known mechanism of action. A platelet-derived growth factor receptor beta (PDGFR{beta}) PheWAS demonstrated that patients with a single nucleotide variant that reduces PDGFR{beta} expression exhibit a higher prevalence of chronic skin ulcers, skin grafts, and reconstructive surgeries. Recombinant human platelet derived growth factor BB (rhPDGF) is a therapeutic that binds to and activates PDGFR{beta} and has received FDA approval for multiple indications, including improving healing of lower extremity diabetic neuropathic ulcers, augmenting periodontal bone and soft tissue reconstruction, and stimulating orthopedic bone regeneration. Leveraging a drug-repurposing methodology informed by PheWAS, we hypothesize that rhPDGF will provide therapeutic benefit in the treatment of other complex wounds, like full-thickness surgical wounds of the head or neck that cannot heal by primary intention following skin cancer excision. MethodsThis prospective, double-blinded, single-site study aims to enroll 40 participants, randomized at a ratio of 1:1, comparing the efficacy of an advanced wound matrix saturated with rhPDGF or saline. Comparisons will be stratified by anatomical location (scalp/forehead versus face/neck) and maximum surgical defect dimensions (< 3cm versus > 3cm). The primary outcome of this study will evaluate the time in days to 81-100% granulation of the wound bed by expert clinical assessment of daily photographs. Secondary outcomes will assess the superiority of the rhPDGF-enhanced wound matrix relative to control with respect to wound granulation rate, epithelialization, complete wound healing, and patient reported outcomes (PROMs). DiscussionAlthough reconstructive techniques are available for healing complex head and neck wounds following skin cancer excision, these procedures are invasive, and older, frail patients are often suboptimal candidates. There remains a need for less invasive therapeutic approaches that reduce the healing time and mitigate the morbidity associated with chronic wounds. A PheWAS analysis identified complex wounds requiring reconstructive surgery as a novel drug-indication pair for repurposing rhPDGF. This protocol is designed to evaluate the efficacy of an rhPDGF-enhanced advanced wound matrix for healing complex head and neck wounds post skin cancer excision that cannot heal by primary intention. Clinical trial registrationThis trial is registered at ClinicalTrials.gov (NCT06634030).

Osteoporotic bone degeneration involves progressive deterioration of trabecular microarchitecture, yet most scaffold-based bone tissue engineering studies evaluate osteogenesis in structurally favorable architectures that poorly represent compromised bone environments. Here, we establish a degeneration-inspired Voronoi scaffold platform in which point spacing serves as a single tunable architectural parameter to model transitions from dense mechanically integrated to severely deteriorated trabecular-like microenvironments. Increasing point spacing from 1.25 to 2.5 mm progressively reduced scaffold connectivity and stiffness while shifting deformation behavior from distributed load transfer to localized stress concentration, as confirmed by finite element analysis and mechanical testing. Benchmarking against clinically reported HR-pQCT datasets from postmenopausal women demonstrated that the intermediate 1.75 mm point spacing scaffold represents a clinically relevant compromised trabecular-like state, whereas the 2.5 mm scaffold represents a more severely deteriorated architectural condition. These architecture-dependent mechanical and structural transitions directly regulated hMSC behavior, where high point spacing scaffolds reduced cytoskeletal organization, stress fiber density, and osteogenic mineralization, establishing an architecture-associated osteogenic dysfunction regime. Polydopamine (PDA) coating progressively enhanced cytoskeletal organization and mineralization within architecturally compromised scaffolds without altering scaffold geometry. To quantitatively assess biointerface-mediated functional recovery, a Mineralization Rescue Percentage (MRP) framework was introduced, demonstrating up to 43% restoration of architecture-associated mineralization loss following PDA coating. Collectively, this work establishes a clinically contextualized degeneration-to-rescue biomaterials framework that shifts current scaffold design paradigms beyond structurally favorable architectures toward systematic investigation and functional rescue of architecture-associated osteogenic dysfunction within compromised bone-like microenvironments. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=79 SRC="FIGDIR/small/725650v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@26833forg.highwire.dtl.DTLVardef@72b2b7org.highwire.dtl.DTLVardef@333083org.highwire.dtl.DTLVardef@b5f2d1_HPS_FORMAT_FIGEXP M_FIG C_FIG Statement of SignificanceMost scaffold-based bone tissue engineering studies evaluate osteogenesis in structurally favorable architectures that poorly represent compromised bone microenvironments associated with osteoporosis. Here, a clinically contextualized Voronoi scaffold platform is established in which point spacing serves as a single tunable architectural parameter to model transitions from mechanically integrated to structurally deteriorated trabecular-like states. By decoupling architectural and surface biointerface effects, the study demonstrates that architectural deterioration alone can drive cytoskeletal disruption and osteogenic failure. Importantly, polydopamine-mediated surface engineering partially restored cytoskeletal organization and mineralization within architecturally compromised scaffolds without altering bulk geometry. A Mineralization Rescue Percentage (MRP) framework was further introduced to quantitatively assess biointerface-mediated functional recovery within degeneration-inspired scaffold microenvironments.

Tetrahedral DNA nanostructures (TDNs) are promising nanocarriers due to their structural precision, biocompatibility, and efficient cellular uptake. However, their stability under physiological conditions remains a key challenge. In this study, TDNs were synthesized via a one-pot thermal annealing method and characterized using native PAGE, dynamic light scattering (DLS), and zeta potential analysis, confirming uniform size ([~]13 nm) and negative surface charge. Their stability was systematically evaluated across different biological media (DMEM complete, serum-free DMEM, and E3), temperatures (4 {degrees}C, 25 {degrees}C, and 37 {degrees}C), and pH conditions (4.0, 7.0, and 8.5) over 24 h. Results revealed rapid degradation in serum-containing medium, increased instability at higher temperatures, and reduced stability under acidic conditions, while serum-free, lower-temperature, and neutral to mildly basic environments enhanced structural integrity. These findings highlight the strong environmental dependence of TDN stability and provide insights for optimizing their design for biomedical applications.

Interleukin-4 (IL-4) is a key immunoregulatory cytokine that promotes type 2 inflammation, drives macrophage polarization toward an anti-inflammatory M2 phenotype, and supports tissue repair. However, clinical translation of IL-4 therapies to modulate the immune response is limited by the need for precise control over its delivery to avoid immune dysregulation. Here, we report an affinity-based strategy to modulate IL-4 delivery and bioactivity using engineered affibody proteins. A yeast surface display library was screened via magnetic- and fluorescence-activated cell sorting to identify two IL-4-specific affibodies with moderate binding affinities (dissociation constants, KD = 459 and 141 nM). Circular dichroism confirmed expected alpha-helical folding, and biolayer interferometry characterized the kinetics of IL-4 binding. Structural modeling using AlphaFold3 and RosettaDock and molecular dynamics simulations using GROMACS predicted distinct binding sites for each IL-4-specific affibody on the IL-4 protein and suggested potential interference with receptor complex formation. Bioactivity studies using murine bone marrow-derived macrophages demonstrated that IL-4 complexed with affibodies maintained Ym1 gene expression but significantly reduced Ym1 protein levels, indicating partial inhibition of IL-4 signaling. To enable controlled cytokine delivery via affinity interactions, affibodies were conjugated to polyethylene glycol maleimide (PEG-mal) hydrogels, which were loaded with IL-4. Affibody-conjugated hydrogels achieved high IL-4 loading efficiency (>90%) and exhibited sustained release over 7 days. Increasing affibody-to-IL-4 ratios significantly reduced both the rate and total amount of cytokine release. Overall, this work establishes IL-4-specific affibodies as versatile tools for tuning cytokine presentation and modulating bioactivity and provides a promising approach for regulating inflammatory responses and advancing cytokine-based therapies with improved temporal control. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=163 SRC="FIGDIR/small/723637v1_ufig1.gif" ALT="Figure 1"> View larger version (46K): org.highwire.dtl.DTLVardef@12bdb14org.highwire.dtl.DTLVardef@3c09eeorg.highwire.dtl.DTLVardef@1b00934org.highwire.dtl.DTLVardef@2c4840_HPS_FORMAT_FIGEXP M_FIG C_FIG

Patellar osteochondral allograft (OCA) transplantation is widely used to treat large full-thickness cartilage defects, yet long-term failure and reoperation rates remain high. Although surface congruity and osseous integration are emphasized clinically, cartilage thickness and mechanical compatibility between donor and recipient are not considered. Our previous work suggests that cartilage thickness mismatch can amplify local deformation at the graft boundary, potentially compromising graft longevity. This study investigates how combined mismatches in cartilage thickness and mechanical properties influence the local strain environment at the patellar OCA interface. Simplified two-dimensional axisymmetric finite element models of patellar OCA repair were developed in ABAQUS. Donor-to-recipient cartilage thickness ratios ranging from 0.33 to 3.25 were evaluated together with donor-recipient Youngs modulus mismatches (2.5-7.0 MPa). Cartilage was modeled using homogeneous linear elastic and functionally graded material formulations to account for depth-dependent stiffness. A compressive pressure of 1.0 MPa was applied to represent patellofemoral joint loading, and peak compressive and shear strains were quantified at the graft boundary. Cartilage thickness mismatch produced localized high-strain regions (HSR) of compressive and shear strain at the donor-recipient interface that were absent in thickness-matched constructs. Strain amplification increased with both thickness and mechanical property mismatch. Compressive strain exhibited directional asymmetry, with donor-side-thicker configurations producing greater amplification than recipient-side-thicker configurations. Incorporating depth-dependent cartilage stiffness reduced peak strain magnitudes but did not eliminate mismatch-driven strain amplification. These findings demonstrate that cartilage thickness and mechanical disparity can create HSR at the patellar OCA graft boundary that may predispose grafts to impaired integration and long-term failure.