ACS Biomaterials Science & Engineering最新文献

{"title":"Glucocorticoid Receptor Signaling Mediates Resistance to Therapy in Matrix Rigidity-Induced Dormant Brain Metastatic Breast Cancer Spheroids.","authors":"Sofia N Luna, Venu Yakati, Lalita A Shevde, Shreyas S Rao","doi":"10.1021/acsbiomaterials.5c00644","DOIUrl":"https://doi.org/10.1021/acsbiomaterials.5c00644","url":null,"abstract":"<p><p>Breast cancer is the most commonly diagnosed cancer and the leading cause of cancer mortality in females. Approximately 20-30% of patients with advanced breast cancer develop brain metastasis. Often, brain metastatic breast cancer (BMBC) exhibits a nonproliferative (dormant) phenotype and therapy resistance due to the unfavorable organ microenvironment. However, the mechanisms by which dormant BMBC micrometastases develop resistance to treatment remain unknown. In the current work, we utilized hyaluronic acid (HA) hydrogels to study the relationship between matrix rigidity-induced dormancy and the drug resistance of BMBC spheroids. BMBC spheroids were cultured on soft (∼0.4 kPa) or stiff (∼4.5 kPa) HA hydrogels, known to induce dormant versus proliferative states, and their response to Paclitaxel (PTX) or Lapatinib (LAP) treatment was measured. Spheroids on soft HA hydrogels were resistant to PTX or LAP treatment. Conversely, spheroids on stiff HA hydrogels were responsive to PTX or LAP treatment. Moreover, the resistance to therapy was mediated by glucocorticoid receptor (GR) signaling via serum/glucocorticoid-regulated kinase 1 (SGK-1) and RANBP1 in triple-negative BMBC cells and β-catenin and GSK-3β in human epidermal growth factor receptor 2 positive (HER2+) BMBC cells. Further, SGK1 inhibition alleviated drug resistance and resulted in response to treatment. Overall, this work provides evidence for dormancy associated drug resistance through GR signaling in BMBC spheroids.</p>","PeriodicalId":8,"journal":{"name":"ACS Biomaterials Science & Engineering","volume":" ","pages":""},"PeriodicalIF":5.5,"publicationDate":"2025-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144999251","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Unveiling the Interaction of Bowl-Shaped Polydopamine on Ti40Zr for Corrosion Resistance and Augmented Biomineralization Capacity.","authors":"Cheranmadevi Pugalendhi, Sudhisha Vasudevan, Vicente Amigó Borrás, Nallaiyan Rajendran","doi":"10.1021/acsbiomaterials.5c00410","DOIUrl":"https://doi.org/10.1021/acsbiomaterials.5c00410","url":null,"abstract":"<p><p>The surface chemistry of Ti40Zr alloys plays a significant role in the formation of the bowl-shaped morphology of polydopamine (PDA). The bowl-shaped PDA morphology formation mechanism on the metal surface and its potential application in biomineralization and long-term stability as an implant material were investigated systematically. A novel nonsacrificial template-assisted bowl-shaped hollow capsule PDA formation was formed on alkali-treated Ti40Zr. Our research hypothesizes that the bowl-shaped hollow structure of PDA is a result of its ion exchange activity. The surface characterization confirms the presence of bowl-shaped hollow capsule PDA and nano-PDA through HRSEM with EDS, contact angle measurement, and XRD for the crystalline phase. The corrosion resistance behavior of PDA-Ti40Zr shows high polarization resistance and enhanced coating stability in SBF which is confirmed in corrosion assessment. The potential surface biocompatibility for bone augmentation was investigated through immersion of PDA-Ti40Zr in Hank's solution for 28 days, and the morphology of the apatite layer indicated dense compact growth and coverage on the surface. The calcium ions increase as the immersion period increases in vitro biomineralization. The quantitative assessment of calcium concentration was performed by alizarin red and ALP activity up to the seventh day. The seventh day of the alizarin red assay results demonstrate that the quantity of calcified nodules increases. The osteogenic marker suggested that the differentiation and mineralization process was accelerated on PDA-Ti40Zr starting from the third day. The results of corrosion studies, bone mineralization assay, and cytocompatibility show promise that the PDA-coated Ti40Zr is a choice material for patients undergoing orthopedic surgery and permanent implantation.</p>","PeriodicalId":8,"journal":{"name":"ACS Biomaterials Science & Engineering","volume":" ","pages":""},"PeriodicalIF":5.5,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144990900","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Design of Printable Gelatin Composite-PVA Bioink for Self-Supported Fabrication of 3D Thick Porous Constructs.","authors":"Renjing Wang, Yunxia Chen, Kyle R Phillips, Chuanshen Zhou, John-Thomas T Robinson, David B Iten, Dylan Z Ver Helst, Yong Huang","doi":"10.1021/acsbiomaterials.5c00888","DOIUrl":"10.1021/acsbiomaterials.5c00888","url":null,"abstract":"<p><p>The development of thick, permeable, three-dimensional (3D) constructs is essential for advancing tissue engineering applications that require efficient mass transport and prolonged cell viability. In this study, a printable gelatin composite-poly(vinyl alcohol) (PVA) bioink is designed and evaluated for the self-supported fabrication of 3D thick porous constructs with satisfactory permeability. The proposed bioink incorporates gelatin solution, gelatin microgels, and PVA, which is utilized as a sacrificial porogen to facilitate postprinting pore formation. The rheological properties of the bioink (the PVA-to-gelatin composite v/w ratio of 1:5) demonstrate suitable shear-thinning behavior and yield-stress fluid property for extrusion-based 3D printing, and the latter enables the jamming-based physical cross-linking mechanism during printing, which works with the thermal cross-linking of gelatin solution to retain the print shape for permanent enzymatic cross-linking. Printed constructs exhibit good print fidelity and structural integrity across both two-dimensional (2D) lattice and 3D tubular geometries. After PVA removal, the freeze-dried samples show large pores formed by removed PVA, as confirmed by scanning electron microscopy and pore size analysis. Permeability tests reveal that constructs fabricated with PVA porogen removal achieve a higher permeation rate of 1.39 mm/h. NIH 3T3 fibroblast-based cell viability studies demonstrate sustained cell survival in the 10.00 mm-thick porous constructs with cell viability above 75% over 7 days with the 2D cell viability control effect considered. Despite residual PVA detected postremoval, the porous network formed by PVA removal remains effective in supporting permeability and cellular function. These findings demonstrate the potential of the printable gelatin composite-PVA bioink for fabricating thick, permeable constructs suitable for cell culture and tissue engineering applications.</p>","PeriodicalId":8,"journal":{"name":"ACS Biomaterials Science & Engineering","volume":" ","pages":""},"PeriodicalIF":5.5,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144935868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Electrospun Nanofibers Loaded with Concentrated Growth Factors and Nanohydroxyapatite for the Healing of Alveolar Bone in Tooth Extraction Wounds.","authors":"Qihang Huang, Linzi Han, Ruishi Wang, Qunli Guan, Hang Wang, Yinhui Yang, Dejiang Du, Yujia Wang, Yichen Shi, Jiaxuan Chai, Yaozhong Wang, Dechao Li","doi":"10.1021/acsbiomaterials.5c00159","DOIUrl":"https://doi.org/10.1021/acsbiomaterials.5c00159","url":null,"abstract":"<p><p>The healing of tooth extraction wounds is significantly influenced by various factors, including interference from the oral microenvironment, invasion of gingival tissue, and inflammation of the alveolar socket, all of which contribute to the substantial loss of bone tissue in the alveolar socket. This study employed electrospinning technology to fabricate polyvinyl alcohol (PVA)/sodium alginate (SA) nanofiber scaffolds infused with freeze-dried concentrated growth factor (CGF) and nanohydroxyapatite (nHA). The objective was to investigate the repair mechanism of the PVA/SA/CGF/nHA nanofibers for oral alveolar bone defects, thereby offering novel treatment strategies for bone defect repair. <i>In vitro</i> experimental results demonstrate that the addition of nHA significantly enhances both the degradation rate and swelling ratio of the PVA/SA/CGF/nHA fiber membrane. Additionally, the scaffold exhibits favorable microstructural properties and biocompatibility. The sustained release of fibrin in CGF suggests that the fibrous membrane maintains a stable three-dimensional structure, facilitating the slow and sustained release of CGF. Alkaline phosphatase (ALP) and alizarin red staining indicate a significant promotion of osteogenic differentiation of human bone marrow stem cells (hBMSCs). Furthermore, qRT-PCR results reveal increased expression levels of the ALP, collagen type-1 (Col1), runt-related transcription factor 2 (Runx2), osteocalcin (OCN), and osteopontin (OPN) genes. To further investigate the impact of the local application of the nanofiber membrane on bone regeneration, a rat tooth extraction wound model was established. These results confirm that the electrospun PVA/SA/CGF/nHA nanofiber membrane significantly promotes the proliferation, migration, and osteogenic differentiation of hBMSCs. This finding offers a novel treatment for oral alveolar bone defects and has potential clinical applications in the oral cavity.</p>","PeriodicalId":8,"journal":{"name":"ACS Biomaterials Science & Engineering","volume":" ","pages":""},"PeriodicalIF":5.5,"publicationDate":"2025-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144935825","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chitosan-Based Antibacterial Bioceramic Materials for Dental Pulp Capping.","authors":"Zhi-Yi Huang, Glemarie C Hermosa, Jyun-Sain Wu, Tung-Lin Wu, Chih-Ching Chien, Chien-Shiun Liao, Yu-Tzu Huang, Hui-Min David Wang, An-Cheng Aidan Sun","doi":"10.1021/acsbiomaterials.5c00466","DOIUrl":"https://doi.org/10.1021/acsbiomaterials.5c00466","url":null,"abstract":"<p><p>Conventional clinical approaches for regenerative endodontic procedures, root canal therapy, and vital pulp therapy often lack sufficient antimicrobial efficacy, thereby increasing the risk of post-treatment apical periodontitis. To overcome this limitation, a series of antimicrobial powders (referred to as the AC_<i>S</i> series) was synthesized through a chemical reaction between tricalcium silicate (C₃S) powder and chitosan solution pretreated with acetic acid. Following this, the AC_<i>S</i> powders were subsequently physically blended with additional C₃S to enhance the mechanical properties, thereby developing a chitosan-based bioceramic composite, named the AC_<i>S</i>-C series. The antibacterial properties of the AC_<i>S</i>-C materials were systematically evaluated using minimum inhibitory concentration (MIC) assays, inhibition zone tests, and antibacterial assessments against <i>Escherichia coli</i> and <i>Streptococcus mutans</i> as well as biofilm testing with <i>Porphyromonas gingivalis</i>. In addition, biocompatibility was assessed through cytotoxicity tests using L929 fibroblast cells. The results revealed that the AC₂₀-C formulation exhibited good antibacterial efficacy (exceeding 90%), maintained over 80% cell viability, exhibited a clear inhibition zone, and effectively inhibited biofilm formation. Regarding physical properties, the AC_<i>S</i>-C materials were able to set within 30 min and possessed sufficient compressive strength. Further structural analysis using energy-dispersive X-ray spectroscopy, X-ray diffraction, and Fourier transform infrared spectroscopy verified the successful synthesis and structural integrity of the material. The AC_<i>S</i>-C samples exhibited key functional groups, including amino, amide, Si-O, CaO, and PO₄^3-. In this study, the AC_<i>S</i>-C series represent promising antimicrobial bioceramic pulp-capping materials that combine effective antibacterial activity with favorable biocompatibility.</p>","PeriodicalId":8,"journal":{"name":"ACS Biomaterials Science & Engineering","volume":" ","pages":""},"PeriodicalIF":5.5,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144935778","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Carnosine Biofunctionalized Hydroxyapatite Induces Copper-Driven Osteogenesis and Angiogenesis, Strengthening Its Bone Regenerative Capacities.","authors":"Irina Naletova, Francesco Attanasio, Teresa Sibillano, Barbara Tomasello, Valeria Lanza, Valeria Ciaffaglione, Rita Tosto, Antonio Mio, Warren Cairns, Cinzia Giannini, Enrico Rizzarelli","doi":"10.1021/acsbiomaterials.5c00823","DOIUrl":"https://doi.org/10.1021/acsbiomaterials.5c00823","url":null,"abstract":"<p><p>Hydroxyapatite (Hap) is a prominent biomaterial used as an effective implant material in bone tissue engineering, but its use presents some points of weakness in bone regeneration efficiency. Different biofunctionalization strategies have been utilized to increase the regenerative Hap capacities. Carnosine (Car) or β-alanyl-l-histidine dipeptide has received much attention due to its beneficial effects in osteoarticular diseases and bone tissue healing. Hap functionalized in noncovalent mode with Car at a nominal Ca:Car molar ratio (10:1, 2:1, and 1:1) was synthesized. The Hap-Car composites were characterized by using X-ray diffraction, scanning electron microscopy, and Fourier transform infrared spectroscopies. The structural and morphological feature comparisons indicate a similarity between Hap-Car10:1 and Hap. The Hap-Car composites and Hap bind copper present at submicromolar concentration in the complete culture medium, determined by inductively coupled plasma-optical emission spectroscopy. Hap-Car composites enhance the biological properties of Hap in in vitro assays and promote the mineralization process and the expression of alkaline phosphatase, osteocalcin, vascular endothelial growth factor, brain derived neurotrophic factor, and bone morphogenetic protein-2 in hFOB1.19 cells. The protective and regenerative activities of the metal ion are also related to the intracellular chaperone copper chaperones for superoxide dismutase.</p>","PeriodicalId":8,"journal":{"name":"ACS Biomaterials Science & Engineering","volume":" ","pages":""},"PeriodicalIF":5.5,"publicationDate":"2025-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144935728","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction to \"Functionalized α-Helical Peptide Hydrogels for Neural Tissue Engineering\".","authors":"Nazia Mehrban, Bangfu Zhu, Francesco Tamagnini, Fraser I Young, Alexandra Wasmuth, Kieran L Hudson, Andrew R Thomson, Martin A Birchall, Andrew D Randall, Bing Song, Derek N Woolfson","doi":"10.1021/acsbiomaterials.5c00792","DOIUrl":"https://doi.org/10.1021/acsbiomaterials.5c00792","url":null,"abstract":"","PeriodicalId":8,"journal":{"name":"ACS Biomaterials Science & Engineering","volume":" ","pages":""},"PeriodicalIF":5.5,"publicationDate":"2025-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144935729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a User-Friendly Dynamic Culture System for Decellularized Scaffold-Based Tissue Engineering.","authors":"Shujuan Fan, Yerong Qian, Dan Li, Lu Lu, Jie Lian, Xin Liu, Junxi Xiang, Aihua Shi, Shasha Wei, Yi Lyu, Lifei Yang, Peng Liu","doi":"10.1021/acsbiomaterials.5c01129","DOIUrl":"https://doi.org/10.1021/acsbiomaterials.5c01129","url":null,"abstract":"<p><strong>Background: </strong>The engineering of large-scale tissues is frequently hampered by complex, inefficient perfusion bioreactors and the inherent diffusion limits of static culture. To overcome these operational and biological barriers, we developed and validated a novel, user-friendly dynamic culture platform.</p><p><strong>Methods: </strong>Our miniaturized platform seamlessly integrates a peristaltic pump, an oxygenator, and perfusion circuits into a single, easy-to-assemble unit with smart control. We demonstrated its efficacy by culturing AML12 hepatocytes on decellularized rat liver scaffolds for 7 days and comparing its performance against static culture controls.</p><p><strong>Results: </strong>Dynamic culture dramatically enhanced cell viability and functional maturity. Compared to static controls, constructs cultured in our platform exhibited a > 5-fold increase in proliferating (Ki67+) cells and a 32-fold decrease in apoptotic (TUNEL+) cells. Hepatic functions were also significantly enhanced, with increased urea production and markedly higher expression of albumin (∼1.5-fold) and the key metabolic enzymes UGT1 (∼1.4-fold) and CYP2D6 (∼2.7-fold).</p><p><strong>Conclusion: </strong>Our integrated dynamic culture platform provides a simple, reliable, and effective solution for constructing large, viable 3D tissue constructs. By overcoming critical usability and mass transport challenges, this platform represents a powerful and scalable tool for advancing tissue engineering applications, from regenerative medicine to disease modeling.</p>","PeriodicalId":8,"journal":{"name":"ACS Biomaterials Science & Engineering","volume":" ","pages":""},"PeriodicalIF":5.5,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144935843","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impact of Modified Triple Salt Monolayer Coating on Osseointegration of Endosteal Implants.","authors":"Vasudev Vivekanand Nayak, Justin E Herbert, Bruno Luís Graciliano Silva, Sophie Kelly, Camila Suarez, Maria Castellon, Pawan Pathagamage, Estevam A Bonfante, Lukasz Witek, Paulo G Coelho","doi":"10.1021/acsbiomaterials.5c00249","DOIUrl":"https://doi.org/10.1021/acsbiomaterials.5c00249","url":null,"abstract":"<p><p><b>Background:</b> Improvements in osseointegration and bone healing as a result of surface modifications indicate that the time frame following implantation necessary to achieve biomechanical capacity for functional load-bearing may be reduced. In this context, a potassium peroxymonosulfate-based modified triple salt monolayer could potentially serve as a viable surface coating to further augment bone regenerative capabilities of endosteal implants. <b>Methods:</b> Implants with resorbable blast media textured surface [Tapered Pro 3DS RBT (Laser-Lok), BioHorizons] (CTRL) were treated with a potassium peroxymonosulfate-based modified triple salt coating process to generate a stabilized monolayer (Oxion). Prior to surgical intervention, implants were subjected to surface characterization. Subsequently, implants were evaluated in a large, preclinical sheep model (<i>n</i> = 14 sheep). A total of 12 implants were placed bilaterally in the submandibular ramus (3 implants per group per sheep per side) and allowed to heal for 3- and 12-weeks (7 sheep per time point). Following the allocated healing time, the animals were euthanized, mandibles harvested, and samples isolated for histomorphometric and nanoindentation analysis, along with biomechanical assessment through implant lateral load testing. <b>Results:</b> The Oxion coated implant's surfaces yielded lower contact angle (<i>p</i> < 0.001) and higher surface free energy values (<i>p</i> < 0.001) relative to the CTRL surface. Bone-to-Implant Contact (BIC) and Bone Area Fractional Occupancy (BAFO), which were used to quantify degrees of osseointegration, were statistically homogeneous at both healing times between Oxion and CTRL surfaces. Biomechanical testing, i.e. nanoindentation and lateral loading, demonstrated improved values for Oxion implants at both early and advanced healing time points compared to CTRL (<i>p</i> = 0.001). <b>Conclusion:</b> Implant failures continue to manifest during the initial months following implant insertion due to a variety of reasons, including inadequate osseointegration, or in cases involving clinical diseases and comorbidities. These findings suggest that the time frame following implantation necessary to achieve biomechanical capacity for functional load-bearing can be further reduced due to the Oxion surface coating in addition to the potential for enhanced early biomechanical integration relative to CTRL.</p>","PeriodicalId":8,"journal":{"name":"ACS Biomaterials Science & Engineering","volume":" ","pages":""},"PeriodicalIF":5.5,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144935877","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Harvest Time-Dependent Changes in the Composition and Function of Microglia-Derived Small Extracellular Vesicles.","authors":"Muhammad Waqas Salim, Wei Zhang, Su Su Thae Hnit, Karthik S Kamath, Lyndsey E Collins-Praino, Andrew Care, Yuling Wang","doi":"10.1021/acsbiomaterials.5c00791","DOIUrl":"https://doi.org/10.1021/acsbiomaterials.5c00791","url":null,"abstract":"<p><p>Microglia are dynamic macrophage-like cells that survey the central nervous system (CNS) and mediate neuroinflammatory responses. Even under homeostatic conditions, they exhibit phenotypic plasticity in response to environmental cues. Microglia-derived small extracellular vesicles (sEVs), which are lipid bilayer vesicles encapsulating proteins, lipids, and nucleic acids, reflect the physiological state of their parent cells and influence recipient cell function. Although increasing attention has been given to their roles in neuroinflammation, neurodegeneration, and intercellular communication, the impact of experimental variables, particularly harvest timing, on microglial sEV composition remains poorly understood. This study investigated how different harvest durations (24, 48, and 72 h) influence the molecular and functional properties of sEVs released by BV2 microglial cells. sEVs were isolated via ultracentrifugation and characterized for particle size, yield, RNA and protein content, surface marker expression, and proteomic profile. While sEV size and morphology remained consistent across time points, particle yield and RNA content increased significantly with prolonged harvest, peaking at 72 h. Classical sEV markers (CD9, CD81, TSG101) were consistently expressed, suggesting stable vesicle identity. However, proteomic profiling revealed a time-dependent shift in cargo, with an increase in unique proteins and enrichment of stress- and neurodegeneration-related pathways at 72 h. Functional assays further demonstrated that sEVs collected at this time point reduced viability in both microglial and neuronal cells. Together, these findings highlight harvest time as a critical determinant of microglial sEV composition and bioactivity, with implications for their use in diagnostics, therapeutics, and drug delivery applications.</p>","PeriodicalId":8,"journal":{"name":"ACS Biomaterials Science & Engineering","volume":" ","pages":""},"PeriodicalIF":5.5,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144935871","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}