Biomaterials Science

Nanomaterials have been proposed as drug delivery vehicles to enhance targeting and efficiency of traditional and novel therapeutics and have subsequently been studied for potential ecotoxicity. Previous studies have identified size, surface charge, and volume exclusion as factors that influence nanomaterial diffusion and retention. However, there is little accepted or successful quantification of how these parameters influence nanomaterial penetration relative to biological adaptation and biological response. Part of the challenge is the response of living biological interfaces to many of these nanomaterial delivery vehicles and nanosized drugs. This study aimed to emulate key physicochemical barriers to diffusion found in living biomaterials by developing a tunable, synthetic hydrogel. Through the controlled exposure of 150 kDa and 2 MDa nanodextrans with neutral and negative surface charge, we evaluated the systems ability to emulate three core physicochemical features often implicated in biofilm-associated transport resistance: size exclusion, charge interactions, and volume exclusion. We demonstrated a 30% statistically significant decrease in partition coefficients for 2 MDa nanodextran from 150 kDa nanodextran, confirming the ability of the nanocellulose-based microcaps to mimic the permeability of hydrated biomaterial matrices. These findings reflect patterns observed in, for example, living biofilm studies, where size-based diffusion hinderance is commonly reported, but charge-based interaction and volume exclusion are more context-dependent. This controllable system can be coupled with in silico modeling to understand interfacial transport phenomena for nanomaterial-biomaterial interactions. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=91 SRC="FIGDIR/small/703274v1_ufig1.gif" ALT="Figure 1"> View larger version (21K): org.highwire.dtl.DTLVardef@13c1a34org.highwire.dtl.DTLVardef@dc6c5borg.highwire.dtl.DTLVardef@14dcbd4org.highwire.dtl.DTLVardef@80f70c_HPS_FORMAT_FIGEXP M_FIG C_FIG

Tissue engineering scaffolds such as collagen-based biomaterials have long been used to mimic native extracellular matrix in a wide range of regenerative applications. Their high porosity, tunable degradation and mechanics, and cell adhesion sites provide a structure upon which cells can grow and differentiate, while they also have the potential to act as carriers for loading and release of biomolecules to aid in healing. Here we describe the inclusion of a second lyophilization step in the fabrication process to enable improved loading efficiency of bone morphogenic protein 2 as well as increased ease of end-user handling. We report mineralized collagen scaffolds demonstrate maintained microarchitecture and mechanical properties post-relyophilization with reduced variability in biomolecule loading. Relyophilization allows consistent loading and release profiles and suggests the potential to improve the translational potential of collagen scaffold biomaterials for regenerative medicine applications.

Three-dimensional (3D) bioprinted liver scaffolds offer a promising platform for drug screening, disease modelling, and regenerative medicine, yet their broader adoption is limited by the absence of robust post-fabrication preservation strategies. This study aimed to evaluate the impact of -80{degrees}C (deep freezer) preservation and evaluate the structural integrity and hepatic functionality of GelMA-decellularized liver extra cellular matrix (dECM)-based 3D bioprinted liver scaffolds. Bioinks were formulated using synthesized GelMA and solubilized rat liver dECM, and 3D scaffolds were fabricated via extrusion bioprinting into rectilinear grid scaffolds. The 3D scaffold preservations was performed by immersion into two different medium (the culture DMEM media and the other FBS-DMSO cocktail) was evaluated using MTT viability assay, and albumin assay. Preserved 3D bioprinted scaffolds retained overall architecture and cell distribution in the FBS-DMSO cocktail demonstrated by the live dead assay. Together, the data demonstrate that -80{degrees}C storage can maintain the basic cell viability ([~]80%) and a substantial fraction of liver-specific functionality in 3D bioprinted scaffolds but also highlight sensitivity to preservation-induced injury. These findings underscore the need for further optimization of cryoprotectant formulations and freezing protocols tailored to 3D bioprinted liver scaffolds, and provide a foundational framework for developing ready-to-use, cryopreserved 3D liver models for translational applications.

Polysaccharides are often used to mimic physiological environments such as for cancer research models. However, established polysaccharides can display limited long-term stability and high batch-to-batch variability. To overcome this, biomanufactured polysaccharides are increasingly utilized in biomaterials. Here, we produced and characterized Rhodotorula toruloides yeast exopolysaccharides (EPS) and used it to engineer hydrogel for culturing cancer cells. Yeast fermentation of glucose, mannose, and xylose yielded varying EPS amounts (1.68, 1.44, and 0.48 g/L, respectively) with similar compositions, suggesting a common biosynthetic pathway. The glucose-derived EPS characterization identified multiple linkage types and three molecular weight fractions (1.75, 30.0, and 1000 kDa), and its solutions exhibited Newtonian behavior, indicating minimal chain-chain interactions. Solubilizing this polydisperse EPS with polyethylene glycol diacrylate and UV-crosslinking it enabled the engineering of semi-interpenetrating polymer network hydrogel that efficiently embedded cancer spheroids. Our study introduces an integrated biomanufacturing strategy to generate stable and consistent biomaterials, applicable for tissue engineering. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=104 SRC="FIGDIR/small/703759v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@110d079org.highwire.dtl.DTLVardef@e6e390org.highwire.dtl.DTLVardef@662540org.highwire.dtl.DTLVardef@17afd5_HPS_FORMAT_FIGEXP M_FIG C_FIG

Nanotechnology is rapidly transforming medicine by enabling versatile platforms for targeted delivery, controlled release, and intracellular transport of therapeutic payloads. Polymeric mesoscale nanoparticles (MNPs) are 300 to 500 nm in diameter with a PEGylated surface that exhibit unique renal tropism, specifically toward renal tubular epithelial cells. Despite their well-described therapeutic applications and route of localization to the tubules, we do not yet understand their physicochemical stability and cellular internalization mechanisms. In this study, we investigated the stability of MNPs under stress conditions by subjecting them to repeated freeze-thaw cycles and varying storage conditions to evaluate the effects on particle size and polydispersity index. MNPs demonstrated negligible changes in size and PDI up to 4 freeze-thaw cycles. We found that both empty and dye-loaded MNPs demonstrated negligible change in size under standard -20{degrees}C storage conditions. While empty MNPs were only stable at room temperature for one day, and not at 37{degrees}C, dye-loaded nanoparticles were stable for at least eight days under both storage conditions. We then performed in vitro studies to evaluate MNP cellular uptake mechanisms using the human renal cell carcinoma cell line 786-O treated with pharmacological inhibitors of uptake pathways. We found that MNP internalization is almost entirely prevented by dynamin inhibitors, while macropinocytosis inhibition also reduced uptake, suggesting that such standard nanoparticle uptake pathways are robust to the mesoscale size range. These findings provide key insights into the stability profile and endocytosis mechanisms of MNPs, which are critical for materials scale-up and translation of novel kidney-targeted drug and gene therapies.

Buccal delivery offers a promising alternative to oral drug administration by enabling direct systemic absorption and avoiding first-pass metabolism. Multilayer polymeric films represent a promising strategy for the sequential delivery of drug and absorption enhancer in the oral cavity. Here, dual- and triple-layer films were fabricated via slot-die coating, incorporating a GLP-1 receptor agonist (GLP-1-RA) and the penetration enhancer sodium glycodeoxycholate (GDC). These were co-loaded in dual-layer films or compartmentalized in triple-layer films. Scanning electron microscopy and optical coherence tomography confirmed well-defined, distinct layers with thicknesses suitable for buccal administration (339 {+/-} 10.24 {micro}m and 487 {+/-} 36.5 {micro}m for dual- and triple-layer films, respectively). Both designs exhibited good mucoadhesion and mucosal compatibility, and preserved the secondary structure of GLP-1-RA. In vitro release studies showed rapid diffusion of GDC and GLP-1-RA from dual-layer films, whereas triple-layer films enabled sustained, sequential release of GDC and GLP-1-RA. Ex vivo porcine buccal mucosa studies showed higher GLP-1-RA and GDC flux from triple-layer films compared to dual-layer films. The films also did not compromise epithelial integrity, in contrast to the direct application of GLP-1-RA and GDC, which caused significant epithelial disruption. These results demonstrate that multilayer film architecture and spatial layering can be harnessed to control release kinetics, maximize peptide penetration, and minimize tissue stress, offering a versatile platform for safe and effective peptide delivery. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=93 SRC="FIGDIR/small/700335v1_ufig1.gif" ALT="Figure 1"> View larger version (25K): org.highwire.dtl.DTLVardef@76858borg.highwire.dtl.DTLVardef@1397e74org.highwire.dtl.DTLVardef@19d1841org.highwire.dtl.DTLVardef@a369c5_HPS_FORMAT_FIGEXP M_FIG C_FIG

Treating extensive full-thickness burn wounds remains difficult in clinical practice because available donor skin is often limited, the risk of infection is high, and many standard dressings do not perform well when defects are large or structurally complex. These limitations have shifted attention to decellularized extracellular matrix (dECM) scaffolds, which can provide physical coverage while preserving biochemical cues that may support tissue repair. Based on this rationale, we designed a decellularization method that improves reagent penetration to produce a full-thickness porcine decellularized small intestine (dSI) scaffold for use in burn wound coverage. The protocol removed most cellular material while leaving low levels of detergent residue, and it maintained the native three-layer structure of the intestinal wall. Most key ECM components, such as collagen and glycosaminoglycans, were also retained. In this study, the dSI showed several properties relevant to burn care, capacity to absorb large amounts of fluid, water vapor transmission rates similar to those reported for skin, and resisted microbial penetration in vitro. From a mechanical standpoint, the scaffold retained anisotropic behaviour and remained stable under cyclic loading. This pattern indicates that it could withstand repeated deformation instead of acting like a fragile membrane. Degradation tests under enzymatic and oxidative conditions indicate that the material breaks down in a controlled way over a period that appears consistent with typical wound-healing timelines. In vitro assays indicated that the scaffold was cytocompatible, as human dermal fibroblasts and keratinocytes both attached to its surface and continued to proliferate. Cell responses differed depending on surface orientation, suggesting that preserved intestinal layers may shape cell behaviour in ways that are often missing in thinner or more uniform matrices. Overall, full-thickness dSI appears to act as a biologically active scaffold and shows mechanical properties that exceed those of many currently used burn dressings.

Stretch is an important biomechanical stimulus facilitating tissue development in the respiratory system by programming the epithelium, endothelium, and extracellular matrix (ECM). Lung tissue undergoes stretch induced lung differentiation under normal prenatal and postnatal development. Furthermore, supraphysiological and aberrant stretch responses are known mechanisms of acute lung injury and ECM disruption. Current in vitro human tissue cyclic mechanical stretch (CMS) models suffer from significant, well-recognized disadvantages and are poorly validated in vivo for longer-term study. In vitro precision-cut lung slice (PCLS) models are commonly used to study the complex structural arrangement and cellular interactions of human tissue, as well as various lung diseases, including BPD.3 PCLS maintain lung tissue architecture and the variety of cell types present in the lung, allowing for a more realistic imitation of the lung microenvironment.3 Existing agarose-inflated PCLS models are hindered by retention of agarose media in the tissue, affecting material properties and complicating stretch studies. Our novel PCLS approach utilizes several technical innovations including a removable hydrogel for inflation and uses supportive poly(ethylene glycol) (PEG) hydrogels enable improved viability and phenotype retention during cyclic mechanical stretch (CMS). This platform will induce PCLS CMS for biochemical assays (e.g. transcriptomics, proteomics) after exposure.

Biomimetic tubular scaffolds hold great promise for tackling unmet clinical needs thanks to their biocompatibility and recapitulation of cellular microenvironments, conferring the ability to promote regeneration. Potential applications include small-diameter vascular implants and grafts for airway repair, for which no viable off-the-shelf solutions currently exist. The tubular materials (4 and 8 mm internal and external diameters) presented here consist purely of type I collagen, contain no chemical crosslinkers, and reproduce the multi-scale architecture of the native tissue including the presence of collagen fibrils. A novel two-step protocol provides materials with distinct concentric layers. A porous external structure, obtained by means of ice templating combined with collagen topotactic fibrillogenesis, favours oriented cell colonization. A smooth and much less porous internal layer provides mechanical and water-tightness properties relevant for in vivo implantation and promotes the formation of an endothelial monolayer under both static and flow conditions. The compliance of the double-layered materials under physiological pressure is close to that of piglet carotid arteries. The materials are also determined to be sufficiently flexible to provide the ability to perform ex vivo anastomosis with bronchi, although the relatively low value of suture retention strength remains a limitation for in vivo suturing.

Human hair is a keratin-based fiber with mechanical properties relevant to load-bearing biomaterials; however, its smooth cuticle limits fiber-fiber cohesion during textile-style processing. This study examines how controlled chemical decuticularization influences surface morphology and tensile behavior of intact human hair assembled into continuous one-dimensional (1D) strands. Hair was treated with oxidative bleach, sodium hydroxide (NaOH), or formic acid (FA), carded, and spun using a standardized protocol. SEM imaging showed treatment-dependent surface disruption, from minimal cuticle modification (bleach) to partial scale lifting (NaOH) and extensive cuticle removal (FA). Tensile testing revealed significant differences in Youngs modulus, ultimate tensile strength (UTS), and elongation at break (EAB) across treatments (ANOVA, p < 0.05). NaOH-treated strands exhibited the highest modulus (207 MPa), UTS (34 MPa), and moderate extensibility (28%), whereas bleach- and FA-treated strands showed reduced stiffness and strength. Compared with reference yarns, NaOH-treated strands approached the stiffness of wool and retained greater extensibility than cotton. These findings support a processing window in which partial decuticularization enhances fiber cohesion while preserving mechanical integrity. The resulting 1D strands provide a potential building block for woven biomesh structures, motivating further evaluation of durability, cyclic behavior, multi-ply configurations, and computational modeling.

Extracellular matrix (ECM) proteins assemble to form a heterogeneous connective scaffold that supports cells. Physical interactions between cells and the matrix regulate cellular behaviors and influence subsequent tissue construction. However, there is a lack of fundamental understanding regarding the contributions of individual native ECM proteins to the matrix. This gap arises from the need for nanoscopic characterization, which operates on a much smaller length scale than typical assessments in cell and tissue cultures, as well as in tissue reconstruction and clinical implantation. This study aims to systematically investigate how individual ECM proteins affect lipid membranes structurally and mechanically, and how these influences regulate cell migration. Results from Langmuir isotherm analysis, X-ray reflectivity measurements, and cell scratch assays demonstrate that strong collagen adsorption on the membrane surface disrupts lipid packing. However, its rigid network provides a sturdy scaffold for cell adhesion, thereby enhancing cell attachment and promoting cell migration. In contrast, elastin has a minimal structural or mechanical impact on the membrane during both adsorption and compression, but it benefits cells by facilitating migration and reducing the risk of infection. Fibronectin, on the other hand, exhibits complex mechanical responses to compression, characterized by significant structural rearrangements that occur during adsorption. This strong interaction with the membrane can result in excessively high adhesion forces, ultimately limiting cell motility. These findings lay the foundation for the design of artificial scaffolds that can manipulate cellular responses, a critical step toward advancing regenerative medicine and tissue engineering. SignificanceFabricating extracellular matrix (ECM) scaffolds from cells offers advantages over traditional approaches, such as decellularized tissues, which face donor limitations, and artificial scaffolds, which may hinder cellular communication. However, the slow harvesting process of cell-derived ECM has limited its clinical applications. This research is part of a larger mission to engineer ECM prescaffolds on lipid carriers tailored to cell requirements, enhancing ECM production and regulating cell behavior. The first step involves systematically analyzing the structural and mechanical effects of ECM on lipid membranes and how these effects regulate cellular behavior. This work confirms distinct characteristics of ECM proteins, advancing fundamental understanding of cell-matrix interactions and paving the way for scaffold engineering.

Alginate hydrogels are widely used for biocompatible encapsulation due to their low cost, mild gelation conditions, and scalability; however, their limited mechanical strength and poor chemical stability under physiological conditions restrict their utility for oral delivery applications. In particular, the development of robust alginate formulations capable of surviving gastrointestinal salt and pH exposures is critical for advancing encapsulated microbial therapeutics for chronic kidney disease (CKD). In this study, we investigated the incorporation of ferric iron into calcium alginate networks as a strategy to enhance gel stability while maintaining biocompatibility. Using a three-ion competition approach, we achieved controlled introduction of ferric ions into calcium alginate gels without significantly altering bulk mechanical properties relative to standard calcium alginate. Although the initial ferric-containing gels displayed comparable modulus and structure, post-treatment with chitosan under mildly acidic conditions produced a dramatic increase in gel stability in physiological salt concentrations across both acidic and neutral pH environments. Ferric-containing gels formed at pH 4.6 absorbed negligible chitosan, in contrast to iron-free alginate gels, which incorporated substantial chitosan under identical conditions. These results support the formation of a thin, dense interfacial complex between chitosan, ferric ions, and alginate at the gel surface, which reinforces the matrix and inhibits dissolution. The resulting hybrid ferric-calcium alginate formulation enabled the production of sub-millimeter beads capable of encapsulating live Thauera aminoaromatica while preserving anaerobic p-cresol degradation activity at 37 {degrees}C using nitrate as an electron acceptor. Collectively, these findings establish ferric-modified alginate hydrogels as a promising, scalable platform for stable oral delivery of encapsulated microbial therapeutics.

Electrospun fiber meshes have long served as biomaterials in a wide range of biomedical applications due to their functional similarities to extracellular matrix and highly tunable properties. Altering the mechanical behaviors of individual fibers and their microarchitecture (e.g.; diameter, crimp, orientation, density) can in principle be used to control bulk level behaviors. Moreover, electrospun meshes are often combined with softer coatings and hydrogels to control surface interactions with body tissues. Yet, fully optimizing their behaviors for specific applications remains an elusive target due to a continued lack of understanding of the micromechanical mechanisms and their relation to bulk mechanical behaviors. Our goal herein was to understand how actual nanoCT-generated 3D microfiber geometry can be used to predict bulk mechanical properties of hydrogel-mesh composites. Electrospun polyurethane meshes were fabricated with a random fiber orientation and coated with a PEG-based hydrogel. The fiber-hydrogel composite was then imaged with a nanoCT scanner at a voxel resolution of 180 nm. From these images, custom Python programs were written to segment, refine, and tesselate a high-resolution finite element of the fiber mesh and hydrogel volumes into a single integrated bi-material finite element model. The resulting mesh was used to run simulations of the planar biaxial mechanical tests used to characterize the bulk mechanical behaviors. Our framework thus enabled systematic investigations of both the macroscopic bulk mechanical response of the overall fiber mesh and the microscopic localized mechanical response of fibers under various stages of loading. The resultant simulations were accurate and predictive of the bulk mechanical responses. It is interesting to note that the fiber-hydrogel composite material experienced the largest stresses within the fiber phase and the largest strains within the hydrogel. This key result underscores that while the previous analytical model assumed local affine deformations, at the microscale this assumption does not hold. We also found very different effective fiber stress-strain responses in each model. It is likely these differences are due to the substantial heterogeneous non-affine local deformations present in the actual fiber-hydrogel composite. This finding further reveals the need for more rigorous approaches to better understand how electrospun-based materials function in order to improve their use in modern medical devices and implants.

Precision-cut lung slices (PCLS) retain the native cells and extracellular matrix that contribute to the structural and functional integrity of lung tissue. This technique enables the study of cell-matrix interactions and is particularly useful for pre-clinical pharmacological studies. More specifically, PCLS are widely used to model the complex pathophysiology of pulmonary fibrosis, an uncurable and progressive interstitial lung disease. Current ex vivo pulmonary fibrosis models expose PCLS to pro-fibrotic biochemical cues over a short timeframe (hours to days) and quickly collect samples for analysis due to viability concerns. This condensed timeline is a limitation to understanding chronic disease mechanisms. To extend the utility of ex vivo pulmonary fibrosis models, PCLS were embedded in engineered hydrogels and exposed to pro-fibrotic biochemical and biophysical cues. Hydrogel-embedded PCLS maintained greater than 80% total cell viability over 3 weeks in culture. Gene expression patterns in samples exposed to pro-fibrotic cues matched trends measured in human fibrotic lung tissue. Finally, treatment with Nintedanib, a Food and Drug Administration approved pulmonary fibrosis drug, moderately reduced fibroblast activation and influenced epithelial cell differentiation. Collectively, these results show that hydrogel-embedded PCLS models of pulmonary fibrosis extend our ability to study fibrotic processes ex vivo and, when applied to human tissues, present a new approach methodology for studying lung disease and treatment.

Articular cartilage repair is limited by the poor regenerative capacity of chondrocytes and their rapid dedifferentiation during in vitro expansion. This study investigated whether a decellularized and lyophilized cell-secreted matrix (CSM) could function as a bioactive material to regulate cell behavior, promote chondrogenic differentiation, and attenuate or reverse chondrocyte dedifferentiation without exogenous growth factor supplementation. CSM was generated from rabbit auricular perichondrial cells, decellularized, lyophilized, and characterized by histology, biochemical assays, and proteomic analysis. The resulting matrix was enriched in structurally and functionally relevant extracellular matrix proteins, including collagens, fibronectin, fibrillin, proteoglycans, and matricellular regulators, with minimal intracellular contamination and good batch-to-batch reproducibility. Functionally, CSM supported robust adhesion and proliferation of allogeneic and xenogeneic cells. Human articular chondrocytes cultured on CSM exhibited enhanced proliferation, sustained expression of cartilage-specific markers, and preserved type II collagen production over serial passages compared with standard plastic culture. CSM also promoted chondrogenic differentiation of human progenitor cells and partially reversed established chondrocyte dedifferentiation, as evidenced by increased expression of COL2A1, ACAN, SOX9, and COMP, with reduced COL1 expression and no induction of hypertrophic markers. These findings demonstrate that lyophilized CSM is a stable, off-the-shelf biomaterial capable of directing chondrocyte fate through intrinsic matrix-derived cues, highlighting its potential for cartilage tissue engineering and cell manufacturing applications.

Pancreatic intraepithelial neoplasia (PanIN) is a precursor of pancreatic adenocarcinoma (PDAC) and therefore critical to understand for identifying early-stage diagnostic and therapeutic targets. During PanIN, epithelial-to-mesenchymal transition (EMT) of pancreatic epithelial cancer cells is a crucial event which promotes invasion and early dissemination of cells into circulation before the full development of PDAC tumours. Changes in tissue mechanics are apparent during progression from PanIN to PDAC and increased local and global elasticity has been mathematically modelled in PanIN tissue as a predictive tool for diagnostics and development of personalized therapies. Aside from elasticity, viscoelasticity is emerging as a key feature of cancer which affects tissue mechanics through a combination of elastic and viscous components. Viscoelasticity has recently been shown to drive mechanosensitive cell behaviour and is known to change dramatically in PDAC progression. Hydrogels, as water-swollen polymer networks, are effective extracellular matrix (ECM) models that can recapitulate the viscoelastic properties of natural tissue. Despite this, hydrogels developed for studying cell behaviour in PanIN use purely elastic materials or have neglected the viscous component. Here, using PDAC mouse models, we show that viscoelasticity dynamically alters between healthy and PanIN-bearing tissue and have decoupled the role of elasticity and viscosity during EMT of pancreatic epithelial cancer cells using two-dimensional (2D) polyacrylamide (PAAm) hydrogels. Our work shows viscosity is critical in driving phenotypic changes associated with EMT in a pancreatic epithelial cancer cell line. These findings identify viscosity as an integral component of cell mechanosensing as PanIN develops, which may contribute to initial metastatic events via dissemination from the developing primary tumour. This should be explored further to potentially reveal novel diagnostic and therapeutic targets.

The growing prevalence of obesity necessitates innovative treatments. This study investigates a spray-dried konjac glucomannan-montmorillonite (KGM-MMT) hybrid designed to combine the fermentable, satiety-promoting effects of KGM with the lipid-binding and anti-inflammatory properties of MMT. In HFD-fed mice treated for 42 days with 2% w/w KGM-MMT, body weight gain was reduced by 7.6%, with an AUC of 5094[{+/-}[52.95, compared to 5513[{+/-}[81.35 in HFD controls (p < 0.0001). Serum IL-6 concentrations were reduced by 97% (p = 0.0002), while blood glucose decreased by 46% (p < 0.0001), outperforming reductions seen with MMT (24%, p = 0.0271) and KGM (16%, ns). Gut microbiota profiling demonstrated a significant 6.2-log[ fold increase in Lactobacillaceae (p = 0.023) and a 2.4-log[ fold increase in Enterococcaceae (p = 0.015) with KGM-MMT treatment. Predicted functional shifts revealed a 1.9-fold increase in short-chain fatty acid synthesis pathways and a 5.4-fold increase in bile acid deconjugation. Although the KGM-MMT hybrid did not consistently outperform its individual components in all measurements within the current study, it generally consolidated their metabolic benefits within a single dosage form. These findings support the utility of spray-dried KGM-MMT as a gut-targeted dietary strategy with additive effects on metabolic health. Future studies should explore underlying mechanisms and dosage effects of the hybrid formulation. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=110 SRC="FIGDIR/small/701163v1_ufig1.gif" ALT="Figure 1"> View larger version (34K): org.highwire.dtl.DTLVardef@738445org.highwire.dtl.DTLVardef@1f0d465org.highwire.dtl.DTLVardef@86e5aorg.highwire.dtl.DTLVardef@184fba8_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LISpray-dried KGM-MMT reduced HFD-induced weight gain by 7.6% in obese mice C_LIO_LISerum IL-6 and glucose levels decreased by 97% and 46%, respectively C_LIO_LI6.2-log[J and 2.4-log[J increases in Lactobacillaceae & Enterococcaceae relative abundance C_LIO_LIBile acid deconjugation and SCFA pathways increased 5.4- and 1.9-fold C_LIO_LIKGM-MMT microparticles offer additive gut-targeted benefits in metabolic disease C_LI

In vitro reconstruction of human tissue microenvironments that integrate native biochemical and biomechanical cues is essential for disease modelling, regenerative medicine, and personalized therapeutic approaches. However, most currently available engineered matrices fail to recapitulate the complexity and tissue specificity of the human extracellular matrix (ECM). To address this limitation, we developed a novel hydrogel derived from decellularized human adipose tissue (atdECM) designed to support three-dimensional culture of human cells. The decellularization and delipidation processes were first validated, and the biochemical composition and biomechanical properties of atdECM were comprehensively characterized. Human pancreatic organoids were then cultured within atdECM hydrogel, and their structural organization and transcriptional profiles were analyzed and compared with those obtained in Matrigel, the current gold-standard matrix for organoid culture. Proteomic and cytokine analyses demonstrated efficient decellularization while preserving collagen-rich ECM architecture and a diverse repertoire of soluble bioactive factors. AtdECM exhibited physiological stiffness and retained tissue-specific extracellular cues. Pancreatic organoids cultured in atdECM displayed morphological similarities with those grown in Matrigel but exhibited transcriptional profiles more consistent with physiological epithelial homeostasis, with reduced activation of inflammatory and stress-related pathways. Altogether, these findings indicate that atdECM provides a human-derived, tissue-relevant, and permissive microenvironment for human organoid generation. This platform represents a promising alternative to Matrigel for studying human tissue biology and for developing physiologically relevant in vitro models.

Proteins orchestrate essential cellular processes, including metabolism, communication, survival, and regeneration, making proteomic profiling a powerful strategy to elucidate complex biological responses. Snail slime (SnS) has emerged as a bioactive material with documented pro-healing, antioxidant, and anti-inflammatory properties; however, its effects at the proteome level in normal human dermal fibroblasts (NHDFs) remain unexplored. In this study, an LC-MS-based proteomic approach (Data are available via ProteomeXchange with identifier PXD075292) combined with network and Gene Ontology enrichment analyses was employed to investigate SnS-induced molecular reprogramming in NHDFs, followed by functional assays. Results show that SnS is well tolerated for up to 72 h, confirming its cytocompatibility, followed by proteomic analysis revealing enrichment of biological processes related to apoptosis regulation, oxidative stress response, wound healing, cell migration, and anti-aging. Network analysis identified AKT, PI3K, SRC, and KRAS family members as key hub proteins, indicating convergence on central signaling pathways controlling survival, redox balance, and migratory activity. Functional assays demonstrated a time-dependent, controlled modulation of apoptosis consistent with cellular turnover, alongside a hormetic redox response characterized by transient ROS signaling followed by enhanced antioxidant capacity. Importantly, SnS significantly accelerated fibroblast migration, achieving complete wound closure within 24 h. Collectively, these findings demonstrate that SnS induces coordinated proteomic and functional reprogramming that integrates redox modulation, controlled apoptosis, and enhanced migration, providing a mechanistic basis for its pro-healing and anti-aging effects and supporting its potential as a regenerative biomaterial.

Central nervous system (CNS) disease or injury might be treated by implanted devices, tissue regenerative scaffolds, or drug delivery platforms. However, inflammatory CNS responses limit these interventions and may worsen outcomes following damage to the CNS. Via the foreign body reaction (FBR), macrophages and glial cells trigger a "glial scar" around implants, reducing device performance, scaffold regenerative ability, or drug delivery potential. Previous studies have shown that stiffness of CNS implants significantly affects glial encapsulation, but few studies have investigated materials that truly match brain tissue stiffness. Porous precision-templated scaffolds (PTS) with uniform, interconnected, 40 {micro}m pores have shown favorable healing outcomes and a reduced FBR in numerous soft and hard tissue applications. To quantify the effects of both hydrogel compliance (stiffness) and pore size on glial encapsulation, we implanted poly(2-hydroxyethyl methacrylate-co-glycerol methacrylate) (pHEMA/GMA) PTS of varying stiffness and pore size for 4 weeks in rat brain. We observed reduced astrocyte encapsulation around PTS compared to solid hydrogel rods, reduced pro-inflammatory macrophage polarization for softer hydrogels versus stiffer hydrogels, and the presence of neuronal markers and neurogenesis within the pores. Utilizing soft, precision-porous hydrogels could provide a strategy for mitigating glial scarring and improving implant-based CNS treatments.