Bone disorders and skeletal defects represent a significant clinical challenge, often requiring transplantation techniques limited by donor site morbidity and insufficient regenerative potential. Tissue engineering and regenerative medicine (TERM) strategies using 3D bioprinting have emerged as promising alternatives, but their efficacy is limited by the difficulty of directing stem cell differentiation in a controlled and reproducible manner. To address this limitation, we are proposing 3D bone printing via ultrasound-mediated osteogenic differentiation of stem cells (referred to here as '3DBonUS'). This biofabrication approach integrates low-intensity pulsed ultrasound (LIPUS) with a microfluidic-assisted 3D bioprinting system. This unprecedented approach enables biophysical stimulation of human bone marrow stromal cells (HBMSCs) during the fabrication of scaffolds, promoting osteogenic differentiation without the need for extensive post-fabrication treatments. In addition, the incorporation of microbubbles (MBs) enhanced the effects of LIPUS by amplifying mechanical signals at the cellular level. Our results revealed that the 3DBonUS system significantly upregulated key osteogenic markers (RUNX-2, ALP, COL1A1, BMP-2, OCN and OPN) as confirmed by immunofluorescence and RT-qPCR analysis. Moreover, the LIPUS-treated constructs showed a significant (p<0.05) increase in alkaline phosphatase (ALP) activity and calcium deposition, indicating enhanced mineralisation. The biofabricated constructs maintained high cell viability while exhibiting improved osteogenic differentiation, surpassing traditional 3D bioprinting approaches in both efficiency and efficacy. The 3DBonUS strategy represents a new modality in skeletal TERM, combining biofabrication with targeted mechanical stimulation, with potential for scalability of scaffold manufacturing and clinical application. Future studies will aim to validate the functional skeletal scaffolds in vivo to assess their regenerative potential, with the goal of advancing patient-specific bone implants with enhanced osteogenic properties.

The development of rapid, on-site analytical methods and test strips for alkaline phosphatase activity (APA) is crucial for addressing drinking water safety crises, as it enables eutrophication assessment and algal bloom warnings under phosphorus-limited conditions. In this study, we incorporate aggregation-induced emission luminogen (AIEgen) guest TPE-4PA into bimetallic lanthanide-based infinite coordination polymers (Tb/Eu-GMP ICPs). TPE-4PA exhibits unprecedented binding preference for Tb3+ over Eu3+, triggering coordination-induced emission (CIE) at 450 nm and modulating the antenna effect (AE) and the tandem energy transfer (TER) from Tb3+ to Eu3+. Subsequently, acetylacetone (acac) was further introduced to yield TPE-4PA@Tb/Eu-GMP-acac ICPs with an appropriate full-color fluoresce response capability. The addition of alkaline phosphatase (ALP) hydrolyzes GMP and TPE-4PA dual substrates, resulting in reduced characteristic emission of the bimetallic Ln-ICP host at 493, 548, 593, 619 and 703 nm, increased monomer emission of TPE-4PA at 392 nm, as well as the solvent-driven AIE of the enzymatic product TPE-4OH at 430 nm, which constitutes a novel ratiometric ALP sensing mechanism with pale-purple to blue fluorescence color shifts. Featuring high sensitivity, doubly assured selectivity, and operational simplicity, this "all-in-one" probe enables real-time APA monitoring via dual-substrate kinetics and is suitable for on-site ALP test strip development. Moreover, the unique color transition point of the dual-substrate-based TPE-4PA@Tb/Eu-GMP-acac ICPs probe can be designed to signal an APA surge prior to algal bloom onset, which holds significant potential for the development of microalgae pollution early-warning systems, thereby safeguarding drinking water safety.

Four-dimensional (4D) printing offers a promising strategy for fabricating shape-adaptive bone scaffolds capable of conforming to complex defect geometries; however, achieving stable fabrication, homogeneous dispersion of multifunctional components, and predictable osteogenic performance remains challenging. In this study, a solvent-free resonant acoustic mixing (RAM)-assisted strategy was introduced to fabricate fused deposition modeling (FDM)-printed shape memory P(DLLA-TMC)/β-tricalcium phosphate (β-TCP)/polydopamine (PDA) composite scaffolds. RAM enabled uniform dispersion of microscale β-TCP and nanoscale PDA within the polymer matrix under low-shear conditions, providing a robust basis for continuous filament extrusion and FDM-based 4D printing. The resulting ternary composite scaffolds exhibited suitable rheological behavior, improved compressive strength, enhanced surface hydrophilicity, and stable near-infrared-triggered photothermal shape memory performance. In vitro experiments demonstrated excellent cytocompatibility and significantly enhanced osteogenic differentiation, as evidenced by increased alkaline phosphatase activity, mineralized matrix deposition, and upregulated osteogenesis-related gene and protein expression. Furthermore, in vivo rat calvarial defect models confirmed that the composite scaffolds effectively promoted new bone formation and structural maturation. Collectively, this work establishes an integrated material-process-function framework for the scalable fabrication of multifunctional 4D-printed bone scaffolds and highlights the translational potential of RAM-assisted manufacturing for advanced bone tissue engineering applications.

A key knowledge gap exists in understanding how the decomposition of litter from different halophyte species influences microbial community dynamics in soils. This study addressed this gap through a 180-day laboratory microcosm experiment investigating the effects of leaf litter decomposition from three halophytes (Kalidium cuspidatum, Nitraria tangutorum, and Reaumuria songarica) on soil biogeochemical properties, microbial dynamics, and community compositional constancy. The main research results indicate that at 180 days, the leaf mass loss (Mm) of the three halophytes reached 40.09%-42.89%, and the decomposition constants (k) were all < 0.2. Leaf total nitrogen, lignin, and carbon/nitrogen ratio directly regulated the decomposition process. Decomposition significantly increased soil nutrient pools, including total organic carbon (57.64%-100.12%), total nitrogen (51.92%-129.80%), dissolved organic carbon (44.35%-224.40%), and dissolved organic nitrogen (24.15%-238.58%), relative to bulk soil. Microbial carbon limitation increased by 21.81%-37.99%, while nitrogen limitation was alleviated, as evidenced by a 67.86%-92.28% increase in the vector angle of enzyme stoichiometry. These changes were driven by soil chemistry (explaining 45.47% of the variance) and microbial traits (42.31%-65.77%). Plant litter decomposition reshaped the structure of bacterial and fungal communities while reshaped the structure, which was linked to microbial biomass carbon, β-glucosidase, and alkaline phosphatase (p < 0.05). Furthermore, partial least squares path modeling revealed that plant litter decomposition increased soil organic resources, thereby exacerbating microbial carbon limitation; yet, along with microbial biomass, it also influenced microbial community composition. These results underscore species-specific litter effects on soil-microbe feedbacks in a controlled microcosm, emphasizing the role of resource stoichiometry and enzymatic activity in shaping microbial community in saline ecosystems.

Cardiac calcification is an age-associated pathological process that contributes to cardiac dysfunction, arrhythmia, and sudden cardiac death, yet its underlying mechanisms remain unclear. Cardiac fibroblasts (CFs) have emerged as key mediators of ectopic calcification through osteogenic differentiation. Proprotein convertase subtilisin/kexin type 9 (PCSK9), a key regulator of cholesterol metabolism, has been implicated in cardiovascular pathology beyond its canonical role, but its involvement in cardiac calcification is unknown. In this study, aged mice exhibited cardiac dysfunction, interstitial fibrosis, and myocardial calcium deposition, accompanied by upregulation of osteogenic markers, including Runx2, OCN, and Osx. PCSK9 expression was increased in aged hearts and enriched in DDR2-positive cells. In vitro, senescent CFs displayed enhanced osteogenic differentiation, characterized by increased calcium deposition, alkaline phosphatase activity, and elevated expression of osteogenic markers. Recombinant PCSK9 promoted osteogenic differentiation in young CFs, whereas genetic deletion of PCSK9 attenuated these effects in senescent CFs. Pharmacological experiments suggest that PCSK9-mediated osteogenic differentiation is associated with activation of the ATF4 pathway and upregulation of Runx2 expression. These findings support a role for the PCSK9-ATF4-Runx2 signaling axis in osteogenic differentiation of CFs, providing new insights into age‑related cardiac calcification and identifying this pathway as a hypothesis‑generating candidate for future investigation.

This study investigated the biocompatibility and osteogenic potential of premixed bioceramic cements (PBCs)-CeraPutty, EndoCEM MTA Premixed, Well-Root PT, and One-Fil PT Injectable-as alternatives to mineral trioxide aggregate for retrograde filling in endodontic microsurgery. Setting time was measured in saline and fetal bovine serum (FBS). Cytocompatibility was assessed using MTT assays on human periodontal ligament fibroblasts with serial extract dilutions at multiple time points. Osteogenic differentiation was evaluated by alkaline phosphatase (ALP) activity and Alizarin Red S staining for mineralized nodule formation. One-way ANOVA was used for biological data and two-way ANOVA for setting time analysis (p < 0.05). All PBCs demonstrated over 90% cell viability. CeraPutty showed significantly higher ALP activity, indicating enhanced early osteogenic differentiation. All materials promoted mineral deposition. Setting time varied by material and medium, generally increasing in FBS. Overall, PBCs exhibited favourable biological and setting properties, supporting their clinical use as reliable MTA alternatives.

Alkaline phosphatases (ALPs) are enzymes involved in bone mineralisation, liver function, and intestinal health. In cattle, the activity of ALPs varies with age, nutrition, and physiological or pathological status. Despite decades of research, the understanding of isozyme-specific roles and diagnostic applications remains fragmented. This bibliometric and narrative review synthesised 473 peer-reviewed studies that measured ALPs in cattle, published between 1948 and 2025, retrieved from the Scopus database. Bibliometric analysis revealed a marked increase in publications over the past decade, with most studies focused on dairy cattle during pregnancy, lactation, and metabolic adaptation. Bone-associated ALP is consistently associated with skeletal growth and mineral metabolism, whereas hepatic- and placental-associated ALPs are linked to metabolic stress and reproductive processes. Altered activity of ALPs in serum or milk has been reported in conditions such as mastitis, milk fever, ketosis, parasitic infections, and respiratory disease, supporting its potential as a biomarker of health and disease. However, major limitations remain across the literature, including the underrepresentation of beef cattle and the frequent lack of isozyme and isoform differentiation. These limitations constrain biological interpretation and clinical applicability of measuring ALPs. Overall, ALPs have potential as tools for monitoring cattle health, supporting nutritional management and veterinary care. Future research should prioritise isozyme and isoform specific analyses and expand investigations across diverse breeds and production systems to improve the diagnostic utility of ALPs and drive its potential use in cattle systems.

This study evaluated the effects of poultry manure and chemical fertilizer on soil chemical and biological properties in pearl millet (Pennisetum glaucum), sorghum (Sorghum bicolor), and maize (Zea mays) at the Research Farm of the University of Kurdistan, Iran, during 2022 and 2023. The experiment was conducted as a split-plot arrangement based on a randomized complete block design with three replications. The main plot factor included three crops (pearl millet, sorghum, and maize), while the subplot factor consisted of five fertilizer treatments, including control, poultry manure, chemical fertilizer, and combinations of poultry manure with urea and triple superphosphate at varying rates. The results showed that, among the fertilizer treatments, the highest soil organic carbon levels across all three crops were observed in the F2 treatment (poultry manure). Soil nitrate levels were higher under pearl millet (11.75 mg kg[-][1]) and maize (11.23 mg kg[-][1]) than under sorghum (10.53 mg kg[-][1]). The highest soil potassium concentration (139.9 mg kg[-][1]) was recorded in the full chemical fertilizer treatment (F5), whereas the lowest value (118 mg kg[-][1]) was observed in the control treatment. Poultry manure significantly improved soil nutrient content, increased acid and alkaline phosphatase activities. The highest acid phosphatase activity (132.83 μg PNP g[-][1] h[-][1]) was observed with poultry manure application, while the lowest activities were recorded in the control (79.45 μg PNP g[-][1] h[-][1]) and chemical fertilizer (88.2 μg PNP g[-][1] h[-][1]) treatments. Although poultry manure increased soil EC, its benefits for soil fertility and biological activity indicate that it may be suitable under controlled application rates and regular EC monitoring.