Search Results (2,437)

Background/Objectives: The development of multifunctional biomaterials for bone regeneration remains a key challenge in additive manufacturing. Although polylactic acid (PLA) is widely used in fused deposition modeling (FDM), its limited bioactivity and lack of intrinsic antibacterial functionality restrict its application in implantable constructs. This study aimed to develop a PLA-based composite filament combining ion-mediated bioactive potential and local antibacterial functionality using a Quality by Design (QbD) approach. Methods: PLA-based composite filaments incorporating a mollusk shell-derived biogenic calcium-containing filler (20 wt.%) and gentamicin (5 wt.%) were fabricated by solvent-free melt extrusion. A QbD framework was applied to define the Quality Target Product Profile (QTPP), identify critical quality attributes (CQAs), and assess critical material attributes (CMAs) and critical process parameters (CPPs). The material was characterized by SEM–EDS combined with ImageJ-based quantitative image analysis, TGA/DSC, mechanical testing, ICP-AES analysis of aqueous extracts, agar diffusion antibacterial assays, FDM printability assessment, and in vivo biocompatibility testing in a rat subcutaneous implantation model. Results: The developed PLA–Gen–MS material was obtained as a continuous filament with a diameter of 1.75 ± 0.05 mm and was successfully used for FDM printing of model scaffold structures. SEM–EDS confirmed matrix continuity and distribution of the calcium-containing mineral phase. ICP-AES revealed a calcium-dominant multicomponent ion release profile, with Ca as the predominant element and measurable levels of Sr, Mg, P, Mn, and Fe. TGA/DSC confirmed thermal compatibility of the components under melt-processing conditions. PLA–Gen–MS demonstrated antibacterial activity against all tested strains, with inhibition zones of approximately 20–21 mm. In vivo, the material showed a favorable preliminary tissue response compared with TiLOOP^®, including faster reduction of inflammatory infiltration and absence of foreign body giant cells by day 14. Conclusions: The QbD-guided strategy enabled the development of a multifunctional PLA-based filament integrating melt processability, structural integrity, ion-mediated bioactive potential, antibacterial functionality, printability, and favorable preliminary biocompatibility. PLA–Gen–MS can be considered a promising platform for further development of personalized bioactive and antibacterial scaffold constructs for bone regeneration. Full article

Excess fluoride originating from phosphate mining activities poses a significant environmental and public health challenge, particularly in arid and semi-arid regions where water resources are limited. This study presents a comparative evaluation of four conventional defluoridation technologies for the treatment of real phosphate mine waters collected from two major Moroccan phosphate mining sites (Youssoufia and Khouribga). The investigated processes included coagulation–flocculation using aluminum sulfate and ferric chloride, chemical precipitation with calcium hydroxide and calcium chloride, adsorption on aluminum oxide (Al₂O₃) and zirconium oxide (ZrO₂), and fluoride removal using calcined bovine bone apatite under both batch and continuous-flow conditions. Adsorption equilibrium was interpreted using Langmuir and Freundlich isotherm models, while the effects of adsorbent dosage, contact time, and water matrix composition were systematically investigated. Among the coagulation processes, aluminum sulfate achieved fluoride removal of up to approximately 82.5% in phosphate washing water and approximately 76.3% in mine drainage water, whereas ferric chloride removed about 52% of the dissolved fluoride under the reported conditions. Lime and calcium chloride exhibited moderate removal efficiencies of 66% and 61%, respectively. Aluminum oxide showed the highest equilibrium adsorption capacity (q_m = 7.14 mg g⁻¹), while zirconium oxide displayed faster fluoride uptake because of its higher surface affinity for fluoride ions. The presence of competing ions in real mine waters was associated with lower adsorption performance compared with synthetic fluoride solutions. Calcined bone apatite proved to be the most effective material, achieving approximately 83% fluoride removal within 20 min under batch conditions and maintaining good performance during continuous fixed-bed operation, producing treated water with fluoride concentrations below the World Health Organization guideline value. Overall, the results demonstrate that calcined bone apatite provides the highest fluoride-removal performance among the investigated materials under the tested conditions. Its waste-derived origin, rapid adsorption kinetics, and effective fluoride removal make it a promising material for the treatment of fluoride-rich phosphate mine waters. The comparative evaluation further indicates that integrating chemical pretreatment with adsorption may represent a promising strategy for the treatment and potential reuse of mining effluents, although the performance of such a combined treatment train should be validated experimentally. Full article

The development of sustainable, biocompatible scaffolds with enhanced structural stability remains a primary challenge in bone tissue engineering. In this study, structurally reinforced nanocomposite scaffolds were successfully fabricated by integrating green-synthesized carbon quantum dots (CQDs) into a gelatin (G) matrix, offering an innovative platform that mimics the organic–inorganic interfaces of natural bone tissue. The CQDs were derived from oatmeal via a sustainable, green hydrothermal route, serving simultaneously as zero-dimensional reinforcing fillers and bioactive agents within the biopolymer network. To ensure an additive-free fabrication process that avoids toxic chemical cross-linkers, dehydrothermal (DHT) treatment was employed, successfully modulating the interfacial and chemical cross-linking interactions between the gelatin chains and the oxygen-rich surface groups of the CQDs. Structural characterization confirmed the uniform dispersion of CQDs (average diameter 7–8 nm) within the porous gelatin framework. The incorporation of CQDs significantly improved the physicochemical properties of the scaffolds; the G/CQD 5% formulation emerged as the optimal composition, exhibiting a 118% increase in compression modulus compared to pristine gelatin. The composite demonstrated tuned swelling kinetics and a significantly reduced degradation rate, restricting mass loss after 14 days of incubation to approximately 24% compared to 40% for pristine gelatin, which is essential for maintaining a structural template during the initial stages of tissue formation. Bioactivity assays in simulated body fluid (SBF) confirmed the rapid, biomimetic induction of a crystalline hydroxyapatite layer with a natural Ca/P ratio of 1.61 within 14 days. Furthermore, preliminary in vitro assessments using Human Skin Fibroblasts (HSFs) confirmed excellent general cytocompatibility, with cell viability exceeding 90%. This study highlights the unique potential of utilizing biomass-derived carbon nanostructures and clean manufacturing processing to engineer multifunctional scaffolds with enhanced structural stability and intrinsic bioactivity for potential bone defect repairs. Full article

Tissue engineering and regenerative medicine are shifting from passive tissue replacement toward instructive platforms that regulate cellular behavior, immune responses, vascularization, and extracellular matrix remodeling. This review examines recent advances in natural, synthetic, composite, and stimuli-responsive biomaterials, biofabrication and 3D bioprinting, stem and progenitor cell therapies, extracellular vesicles and other cell-free products, immunomodulatory scaffolds, skin organoids and organ-on-a-chip systems, nanotechnology, and artificial intelligence-assisted design. Particular emphasis is placed on plastic, reconstructive, and aesthetic applications, including skin and wound repair, craniofacial bone and cartilage regeneration, peripheral nerve reconstruction, vascularization, and dental and periodontal repair. The review also considers biomodulators and skinboosters as emerging regenerative-aesthetic interventions that aim to improve dermal hydration, fibroblast activity, collagen remodeling, and skin quality rather than provide volume replacement alone. Importantly, these technologies differ substantially in translational maturity, ranging from in vitro and preclinical platforms to early clinical interventions, established clinical products, and commercially available treatments for which durable regenerative efficacy remains incompletely validated. Throughout this review, biological plausibility and preclinical efficacy are therefore distinguished from human clinical evidence, regulatory or established clinical use, and commercial availability. Progress will require standardized characterization, mechanism-linked potency assays, clinically relevant models, and outcome measures that capture functional integration, durability, safety, and aesthetic performance. Full article

Skeleton-based action recognition has attracted considerable research interest because skeleton data are inherently robust to illumination changes, viewpoint variation, background clutter, and camera motion. Nevertheless, extracting informative representations from skeleton sequences remains a challenging problem, as it requires capturing both the spatial co-occurrence patterns among body joints and the fine-grained kinematic cues that distinguish different actions. In this paper, we propose a single-stream architecture that constructs an action-specific skeleton graph directly from motion and processes it with a kinematic-aware Transformer. Rather than relying on a fixed skeleton topology, a motion-guided dynamic-graph construction module infers a per-frame adjacency matrix from short-term motion cues through a differentiable edge predictor and Gumbel-Softmax sparsification, allowing the model to discover action-driven connections between distant joints that lack direct bone connectivity (e.g., coordinated hand motion during clapping). Each joint is described by kinematic node features that combine its 3D position, instantaneous velocity, and limb-angle encodings within a single descriptor, so that both motion dynamics and higher-order limb configurations are available to the spatial encoder from the outset. A graph-attention network (GAT) encodes the spatial configuration of every frame over the learned graph, and the resulting sequence of frame descriptors is processed by a Transformer encoder that models long-range temporal dependencies; a learnable classification token aggregates the sequence, and a multi-layer perceptron (MLP) produces the final action classification. The entire model is trained end-to-end from action labels alone. We conduct a comprehensive ablation study and evaluate the proposed method on the large-scale NTU RGB+D 60 and NTU RGB+D 120 benchmarks, where the results demonstrate that our approach achieves competitive performance compared to state-of-the-art architectures. Full article

Background/Objectives: Achondroplasia is the most common genetic skeletal dysplasia associated with disproportionate short stature and is primarily caused by gain-of-function variants in the fibroblast growth factor receptor 3 (FGFR3) gene. Constitutive FGFR3 activation disrupts growth plate homeostasis and endochondral ossification through complex alterations in chondrocyte proliferation, differentiation, hypertrophy, extracellular matrix organization, and intracellular signaling. The increasing understanding of these mechanisms has enabled the transition from exclusively supportive management toward disease-modifying and precision-based therapeutic strategies. This narrative review aimed to critically synthesize current evidence on the genetic basis, molecular pathogenesis, growth plate abnormalities, and current and emerging targeted therapies in achondroplasia. Methods: A narrative literature review was conducted using PubMed/MEDLINE, Scopus, and Web of Science Core Collection, with Google Scholar used as a supplementary source, together with manual screening of the reference lists of relevant original studies, clinical trials, reviews, consensus documents, and clinical guidelines. The principal literature search covered publications from January 2010 to March 2026, while selected seminal primary studies published before 2010 were included when necessary to document the original identification of pathogenic FGFR3 variants and foundational mechanisms of FGFR3-mediated growth plate regulation. Particular emphasis was placed on FGFR3 variants, receptor activation mechanisms, growth plate dysfunction, intracellular signaling pathways, vosoritide, C-type natriuretic peptide-based therapies, FGFR3 inhibitors, ligand–receptor blockade, drug repurposing, Wnt/β-catenin modulation, and gene-based therapeutic approaches. Results: Achondroplasia is characterized by marked molecular homogeneity, with the recurrent p.Gly380Arg substitution accounting for most cases. Mutant FGFR3 displays sustained activity through partial ligand independence, enhanced receptor dimerization and kinase activation, increased receptor stability, and reduced degradation. Excessive signaling through MAPK/ERK, STAT, PI3K/AKT, IHH/PTHrP, and related pathways impairs chondrocyte proliferation and hypertrophic differentiation, alters extracellular matrix turnover, disrupts primary cilium function, and reduces longitudinal bone growth. Vosoritide provides clinical proof that pharmacological modulation of FGFR3-related signaling can improve growth velocity. Additional therapeutic strategies under clinical or preclinical investigation include long-acting CNP analogues, selective FGFR inhibitors, decoy receptors, RNA aptamers, repurposed drugs, Wnt/DKK1 pathway modulation, and gene- or enhancer-targeted interventions. Conclusions: Achondroplasia is increasingly understood as a disorder of dysregulated growth plate signaling rather than solely a condition of reduced stature. Although vosoritide has established the feasibility of disease-modifying treatment, substantial uncertainty remains regarding final adult height, skeletal proportionality, cranio-spinal development, orthopedic outcomes, and long-term safety. Future progress will depend on mechanistically informed therapeutic combinations, improved biomarkers, advanced cellular and animal models, and long-term clinical and real-world evidence. Full article

The knee is a mechanically demanding synovial organ in which joint loading, tissue deformation, cellular mechanotransduction and matrix turnover are coupled. This narrative review critically links those scales and asks where the evidence is sufficiently mature for mechanistic or clinical inference. PubMed/MEDLINE and Europe PMC were searched from database inception to 20 July 2026 using structured terms for knee biomechanics, cartilage and osteochondral mechanobiology, finite element modelling, mechanosensitive channels, osteoarthritis, machine learning and digital twins. Landmark studies were selected for foundational models, while recent studies were prioritised for causal experiments, validation and translation. Instrumented implants show that common activities generate tibiofemoral forces of several times body weight, but tissue-level exposure also depends on muscle co-contraction, geometry and material properties. Biphasic and fibril-reinforced models explain how those loads become stress, strain, fluid pressure and osmotic signals. At the cell scale, TRPV4 and PIEZO1/2 participate in overlapping, context-dependent calcium signalling rather than a universal protective–pathological binary; most causal evidence remains preclinical. Osteoarthritis is therefore framed as a mechanically amplified feedback process involving cartilage, bone, synovium and systemic modifiers. Computational degeneration models and machine-learning surrogates are increasingly informative, although prospective validation, parameter identifiability and uncertainty propagation remain limiting. The review’s added value is an explicit transmission-and-validation framework that connects whole-joint observables to molecular responses while labelling the evidence source and translational readiness at every step. Full article

Background: Homeobox transcription factors regulate developmental programs, cellular plasticity, and tumor progression, yet the role of H2.0-like homeobox (HLX) in prostate cancer (PCa) remains poorly defined. We investigated the clinical significance of HLX and its relationship to the pro-metastatic transcription factor TBX2. Methods: Transcriptomic and clinical datasets from TCGA, MET500, and SU2C/PCF cohorts were analyzed to assess HLX expression, clinicopathologic associations, and its relationship with TBX2. Functional studies in human PCa cell lines included TBX2 gain- and loss-of-function, HLX knockdown, chromatin immunoprecipitation (ChIP), and expression analyses. Shared HLX- and TBX2-associated pathways were evaluated by Reactome enrichment analysis, and Hallmark Gene Set Enrichment Analysis compared castration-resistant prostate cancer (CRPC) bone metastases with high versus low HLX expression (GSE77930; n = 5/group). In vivo relevance was assessed in an orthotopic TBX2 dominant-negative PCa xenograft model. Results: Human PCa datasets showed that HLX expression was elevated in PCa versus normal prostate tissue and associated with higher Gleason grade, lymph node involvement, aggressive molecular subtypes, and shorter disease-free survival. HLX expression also positively correlated with TBX2 across human PCa cohorts. HLX- and TBX2-associated transcriptional programs converged on extracellular matrix organization, cell adhesion, NOTCH, and VEGF-MAPK signaling pathways. Furthermore, HLX-high CRPC bone metastases were enriched for epithelial–mesenchymal transition, NOTCH, TGF-β, inflammatory, angiogenic, hypoxic, and KRAS signaling pathways. Mechanistic studies showed that HLX knockdown suppressed extracellular matrix-associated genes and key NOTCH pathway components. ChIP demonstrated direct TBX2 binding to the HLX promoter, and genetic modulation of TBX2 expression established HLX as a downstream target of TBX2. Consistent with these findings, reduced HLX expression in orthotopic TBX2 dominant-negative xenografts was associated with loss of metastatic progression. Conclusions: HLX is a candidate biomarker of aggressive PCa and a direct transcriptional target of TBX2. These findings identify a previously unrecognized TBX2–HLX regulatory axis associated with metastatic transcriptional programs and aggressive disease in advanced PCa. Full article

Growth and body conformation traits are key determinants of meat production efficiency and economic performance in sheep. However, the genetic architecture underlying these complex traits remains incompletely understood, particularly across multi-breed populations. In this study, we performed a genome-wide association study (GWAS) for seven growth and developmental traits in a combined population of 401 sheep, including Qira Black, Kyrgyz, Dorset × Hu crossbred, and Suffolk × Karakul crossbred sheep. After genotype harmonization and quality control, 47,674 autosomal SNPs were retained for analysis. Population structure was assessed using principal component analysis, and association testing was conducted using a mixed linear model incorporating breed, principal components, and a kinship matrix. A total of 44 independent loci were detected at a nominal significance threshold, encompassing 112 candidate genes. The strongest association was identified for cannon bone circumference near RPS6KA5 (Chr7; p = 1.40 × 10⁻⁷). Several biologically relevant genes involved in osteogenesis, cartilage development, and metabolic regulation were detected, including STEAP3, SLC26A2, PPARGC1B, COL11A1, CALN1, and CITED2. Two genomic regions exhibited pleiotropic effects, which were identified as being associated with multiple traits, suggesting shared genetic regulation of correlated skeletal characteristics. These findings are consistent with a polygenic architecture underlying growth trait in sheep and highlight candidate genomic regions potentially involved in skeletal development and body conformation. Although further validation is required, the identified loci provide preliminary evidence for regions that may influence growth-related phenotypes and offer a reference for future molecular breeding efforts in indigenous and crossbred sheep populations. Full article

Background/Objectives: Aluminium is a widespread environmental element and food contaminant to which the general population is continuously exposed, primarily through diet and drinking water. Although gastrointestinal absorption is generally low, bioavailability varies according to chemical form, food matrix, and interactions with dietary components. Prolonged exposure can nevertheless result in gradual tissue accumulation. This review summarises current evidence on dietary aluminium exposure, factors influencing its bioavailability, toxicokinetics, biological effects, gut microbiota interactions, and population-level health risk. Methods: A comprehensive narrative literature review was conducted using publications retrieved from PubMed/MEDLINE, Scopus, Web of Science, and Google Scholar. Original research articles, review papers, and reports issued by international organisations were critically evaluated with particular emphasis on dietary sources, drinking water, food additives, food contact materials, gastrointestinal absorption, toxicokinetics, biological mechanisms, gut microbiota, and health risk assessment. Results: Food represents the principal source of aluminium exposure in the general population, while drinking water usually contributes a smaller but continuous fraction of total oral intake. Dietary exposure varies substantially between populations and is influenced by food composition, processing practices, the use of aluminium-containing additives, and migration from food contact materials. Aluminium bioavailability is modified by chemical speciation and dietary constituents, including citrate, phosphates, silicates, phytates, polyphenols, and essential minerals. Despite limited absorption, prolonged exposure can lead to gradual aluminium accumulation, particularly in bone tissue and the central nervous system. Proposed biological mechanisms include oxidative stress, mitochondrial dysfunction, disruption of mineral homeostasis, and inflammatory signalling. Emerging evidence also indicates that aluminium may alter the gut microbiota, impair intestinal barrier integrity, and influence the gut–brain axis. Population exposure assessments show considerable regional variation, with some groups approaching or exceeding established tolerable weekly intake values. Conclusions: Dietary aluminium exposure represents a relevant issue in nutritional toxicology and food safety. Although current evidence does not establish that typical dietary exposure directly causes chronic disease, long-term exposure, differences in bioavailability, and the possibility of elevated intake in selected population groups justify continued monitoring and further prospective human studies. Future research should integrate dietary intake, aluminium speciation, nutritional status, biomarkers of internal exposure, and long-term health outcomes to improve risk assessment and support effective exposure-reduction strategies. Full article

Background: Population aging is driving a convergent rise in three disorders historically managed in isolation: cardiovascular disease, neurocognitive decline, and osteoporotic bone loss. Mechanistic data indicate that these systems are coupled through shared regulators of calcium trafficking, inflammation resolution, vascular integrity, and inflammaging. On this basis, a “Heart–Brain–Bone” axis has been proposed; it should be understood as an integrative conceptual framework that organizes evidence drawn from three separate studies, not as a validated physiological entity with agreed diagnostic criteria or demonstrated modifiability. Three lipid-soluble nutrients—long-chain omega-3 polyunsaturated fatty acids (EPA/DHA), vitamin D3 (cholecalciferol), and vitamin K2 (menaquinone-7 [MK-7])—act on overlapping nodes of this network. Methods: We performed a structured narrative review. PubMed/MEDLINE, Embase, the Cochrane Library, and Web of Science were searched from database inception to 25 June 2026 using predefined term blocks for each nutrient, each organ domain, and each candidate mechanism, and the search was updated on 7 August 2026. Records were screened against prespecified inclusion and exclusion criteria by two authors independently, with disagreements resolved by a third. The strength of evidence for each nutrient–organ relationship was graded with an explicitly defined four-level scheme ((−) to (+++)) applied separately to preclinical, observational, randomized and meta-analytic evidence. Results: Vitamin K2-dependent gamma-carboxylation of matrix Gla protein (MGP) and osteocalcin has been proposed to influence whether calcium is incorporated into the bone matrix or deposited in the arterial wall, offering a candidate mechanistic account of the “calcium paradox” associated with isolated vitamin D3 supplementation; EPA/DHA-derived specialized pro-resolving mediators may support resolution of endothelial and neuronal inflammation; and bone-, vascular- and brain-derived signals (osteocalcin, FGF23, the neurovascular unit) interconnect the three organs. These mechanisms are biologically plausible but remain insufficiently confirmed in humans. The clinical evidence is heterogeneous, formulation- and population-dependent, and comprises positive, neutral and null results: cardiovascular omega-3 trials are discordant (REDUCE-IT, which used icosapent ethyl [an EPA ethyl ester], positive; VITAL/STRENGTH/ASCEND null, predominantly in lower-risk or replete cohorts); cognitive trials are largely null or subgroup-dependent (MAPT, DO-HEALTH, VITAL); and MK-7 improves surrogate bone and calcification biomarkers and slowed coronary artery calcification in one recent randomized imaging trial (VitaK-CAC), whereas combined MK-7 plus vitamin D3 did not slow aortic valve or coronary calcification in AVADEC and MK-7 did not reduce bone loss in early menopausal women. Recognized safety signals include a dose-dependent increase in atrial fibrillation with high-dose omega-3, adverse skeletal effects of high-dose or bolus vitamin D, and clinically relevant interference of even low-dose MK-7 with vitamin K antagonist therapy. Conclusions: No adequately powered randomized trial has demonstrated that the combination of long-chain omega-3, vitamin D3 and MK-7 is superior to its individual components or to placebo for any clinical endpoint. The combined regimen is therefore mechanistically rational and hypothesis-generating rather than clinically established; benefit appears most plausible in individuals with elevated risk or demonstrable nutritional insufficiency, and least in replete, low-risk populations. Findings should be interpreted within a broader healthy-aging context that includes lifestyle and psychosocial factors. Adequately powered factorial randomized controlled trials stratified by baseline Omega-3 index, 25(OH)D and vitamin K status, with prespecified mechanistic biomarkers and hard endpoints, are required. Full article

Background: Physical activity is widely recommended for preserving musculoskeletal health and managing osteoarthritis-related pain, although its benefits are commonly framed in terms of muscle strengthening, weight control, and physical performance. This narrative review aimed to examine movement more broadly as a physiological regulator of synovial joint homeostasis, sensory calibration, and functional adaptation. Methods: A structured literature search was performed in PubMed/MEDLINE, Scopus, and Web of Science from database inception to 1 February 2026. Experimental studies, observational studies, clinical trials, systematic reviews, meta-analyses, and selected narrative reviews addressing movement-responsive joint biology or pain regulation were considered. Evidence was synthesized across four interrelated domains: mechanical, fluidic, immune-metabolic, and sensory regulation. Results: The narrative synthesis indicates that the concept of the synovial joint as a dynamic mechano-fluidic organ in which cartilage, synovium, synovial fluid, capsule, subchondral bone, periarticular tissues, and sensory pathways interact continuously. Repeated physiological movement may promote synovial fluid exchange, lubrication, cartilage nutrition, hyaluronic acid and lubricin function, matrix turnover, anti-inflammatory signaling, proprioceptive control, and exercise-induced hypoalgesia. In contrast, inactivity and unloading may impair fluid dynamics, promote muscle inhibition, stiffness, inflammatory persistence, sensory deconditioning, and loss of function. Excessive or poorly distributed loading may also disrupt homeostasis through matrix injury, inflammation, fatigue, and nociceptive sensitization. These findings informed the proposed adaptive loading window, a hypothesis-generating conceptual framework rather than a clinically validated threshold, describing the dynamic range of movement within which joint function and pain regulation may be supported without sustained tissue or symptom aggravation. Conclusions: Movement should be viewed not only as a therapeutic intervention, but also as a continuous regulator of joint biology and perception. Its clinical value may depend on identifying an individualized loading range that supports adaptation, function, and confidence in movement while avoiding both underloading and overload. Full article

The bone marrow stromal microenvironment is damaged in patients with acute leukemia. The aim of this study was to analyze changes associated with the extracellular matrix, mitochondrial function, and vesicular transport in the proteome of multipotent mesenchymal stromal cells (MSCs) in patients at the onset, in remission, before, and 1–3 months after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The study included paired MSCs samples from the bone marrow of 12 patients at the onset and in remission of acute leukemia (4 ALL, 8 AML) and eight patients before and after allo-HSCT (4 ALL, 4 AML). MSCs from eight healthy donors were used as a control. The growth characteristics and the proteome subsets describing extracellular matrix, mitochondria, and vesicular formation were studied. The proteome of the patients’ MSCs differed significantly from that of the donor MSCs, both at the onset and in remission. Changes noted in the composition of extracellular matrix proteins may affect cell adhesion and access to growth factors. Significant changes were revealed in proteins affecting mitochondrial function. Vesicular transport proteins also differed between the donor and patient groups. Unexpectedly, no differences were found between the MSCs of donors and patients before and after allo-HSCT. Full article

Extracellular vesicles (EVs) derived from osteoblasts are emerging as key regulators of bone formation, yet functional differences between vesicles from distinct extracellular compartments remain unclear. In this study, we compared media-derived extracellular vesicles (MEs) and matrix-bound extracellular vesicle-enriched fractions (MBEs) isolated from mineralized osteoblasts (MOBs) and evaluated their effects on human bone marrow-derived mesenchymal stromal cells (hBMSCs). Both preparations, characterized by nanoparticle tracking analysis and transmission electron microscopy, displayed similar size distributions (30–200 nm) and concentrations. Moreover, both preparations showed enrichment of the EV-associated marker CD63, with no detectable GAPDH and only minimal Grp94 signals in a subset of samples. mRNA profiling revealed that MBE-enriched fractions were selectively enriched in RUNX2, whereas other osteogenesis-related transcripts were comparable between them. Functional analyses demonstrated that both EV populations promoted osteogenic differentiation of hBMSCs, although with distinct biological profiles. MBE-enriched fractions were associated with higher alkaline phosphatase activity under control conditions, whereas MEs induced greater osteocalcin expression and showed a numerical tendency toward increased matrix mineralization, particularly under osteogenic conditions. These findings suggest that extracellular vesicles associated with different extracellular compartments exhibit distinct osteogenic activities rather than a uniform biological effect. Although the matrix-bound preparation likely contains extracellular matrix-associated components co-isolated during the extraction procedure, the present study highlights the importance of extracellular compartmentalization in shaping EV-associated bioactivity and provides a foundation for future studies aimed at optimizing EV-based strategies for bone regeneration. Full article

Demineralized bone matrix (DBM)-derived allografts retain extracellular matrix (ECM)-associated factors involved in bone repair, but biochemical protein recovery alone may not predict functional pathway activation. We evaluated the HEK-Blue™ TGF-β Reporter Assay as a targeted method for assessing Smad-dependent reporter activity in DBM-derived materials. ECM proteins were extracted from cortical demineralized bone granules (DBG) using guanidine hydrochloride (GuHCl) or urea and quantified after extraction, dialysis, and sterile filtration. Reporter cells were stimulated with extracted ECM proteins or directly with processed cortical and cancellous products, including DBG, wet heat-treated DBG formulated as Putty (PHT), and gamma-irradiated PHT (PGI). Non-pooled DBM sponge samples were also tested. Secreted embryonic alkaline phosphatase (SEAP) activity served as the functional reporter readout. Material-only no-cell controls assessed material-derived background. PrestoBlue™ readouts served as exploratory quality controls. The urea-derived extract yielded a higher apparent BCA-detectable protein concentration than the GuHCl-derived extract. Only the GuHCl-derived extract induced increasing SEAP activity at matched protein input. Direct stimulation showed reporter activation above the TNF-α pathway-negative cytokine control for cortical DBG, PHT, PGI, and cancellous DBG. DBM sponges also induced detectable SEAP activity. These findings support targeted functional screening of Smad-dependent TGF-β reporter activation in DBM-derived materials, but not total osteoinductivity assessment. Full article