Frontiers in Bioengineering and Biotechnology最新文献

{"title":"A novel cryoprecipitate-enriched PRP (Cryo-PRP) gel with enhanced mechanical strength and regenerative capacity accelerates enterocutaneous fistula healing.","authors":"Zhang-Sheng Zhao, Zhen-Zhen Wang, Hua Ye, Xian-Lei Cai, Bin Hu, Qiu Han, You-Li Ma","doi":"10.3389/fbioe.2025.1668608","DOIUrl":"https://doi.org/10.3389/fbioe.2025.1668608","url":null,"abstract":"<p><strong>Introduction: </strong>Enterocutaneous fistula (ECF) remains a major clinical challenge due to its complex pathophysiology, high recurrence, and limited non-surgical options. Platelet-rich plasma (PRP) has demonstrated regenerative potential, but its limited mechanical strength restricts its application in high-output fistulas. To overcome this limitation, we developed a cryoprecipitate-enriched PRP (Cryo-PRP) with enhanced fibrinogen content and gel stability.</p><p><strong>Methods: </strong>Cryo-PRP was prepared by cryoprecipitation of conventional PRP. Characterization included fibrinogen quantification, thromboelastography (TEG), and scanning electron microscopy (SEM). Therapeutic efficacy was assessed in a rat ECF model by evaluating fistula closure, histological changes, and expression of angiogenic and inflammatory markers.</p><p><strong>Results: </strong>Cryo-PRP exhibited significantly higher fibrinogen levels (5.26 ± 0.78 g/L vs. 2.58 ± 0.49 g/L, P < 0.001) and greater clot firmness (TEG-MA: 37.8 ± 2.2 mm vs. 28.7 ± 1.3 mm, P < 0.001) compared with standard PRP. SEM revealed a denser and more organized fibrin network in Cryo-PRP gels. In vivo, Cryo-PRP accelerated wound healing, enhanced epithelialization, preserved crypt architecture, and reduced inflammation. Immunostaining demonstrated increased neovascularization (CD34), upregulation of regenerative markers (α-SMA, CD31, VEGF, PCNA), and suppression of TNF-α expression.</p><p><strong>Discussion: </strong>Cryo-PRP demonstrates superior mechanical and biological properties over conventional PRP, effectively promoting tissue regeneration and reducing inflammation in an ECF model. These findings support Cryo-PRP as a safe, autologous, and minimally invasive therapeutic strategy for ECF management.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1668608"},"PeriodicalIF":4.8,"publicationDate":"2025-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12542567/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145354226","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biomechanical and biological features of hyaluronic acid in combination with chondroitin and platelet rich plasma for regenerative medicine applications.","authors":"Valentina Vassallo, Celeste Di Meo, Antonella D'Agostino, Annalisa La Gatta, Donatella Cimini, Giuseppe Toro, Giovanni Iolascon, Maddalena Mastrogiacomo, Chiara Schiraldi","doi":"10.3389/fbioe.2025.1607469","DOIUrl":"10.3389/fbioe.2025.1607469","url":null,"abstract":"<p><p>Currently, one of the most common treatments for osteoarthritis (OA) is viscosupplementation using intra-articular injectable gels, often based on glycosaminoglycans (GAGs), specifically hyaluronic acid (HA) and, in some cases, chondroitin sulfate (CS). Recently, the potential benefits of pharma-grade biofermentative unsulfated chondroitin (BC) have been established, particularly when combined with high molecular weight hyaluronan (HHA). Beyond GAGs, platelet-rich plasma (PRP) has also been reported to have beneficial effects, although many clinical studies lack proper control groups. The aim of this study was to perform a comparative analysis of injectable formulations based on BC combined with HHA (HHA/BC), both alone and in combination with PRP, to evaluate their rheological and biological properties. Flow curves and mechanical spectra of HHA/BC and HHA/BC+PRP were obtained to assess their viscoelastic behavior in relation to synovial fluid characteristics. Then, these two formulations were tested on human chondrocytes isolated from OA joints to investigate their functional role <i>in vitro</i> on specific biochemical pathways. Additionally, a chondrocyte monolayer scratch assay was performed to evaluate their repair potential using time-lapse video-microscopy. Finally, chondrocytes were cultured in GAG-based gels on transwell inserts for 14 days to mimic a 3D-like <i>in vitro</i> environment. HHA/BC+PRP exhibited a consistent rheological profile, supporting its potential application in intra-articular injections. Furthermore, the maintenance of cell phenotype was confirmed through the analysis of collagen type 2A1 (COL2A1) and aggrecan (ACAN) expression. The addition of PRP further enhanced the ability of GAGs to reduce specific pro-inflammatory and degradative OA-related markers (e.g., interleukin IL-6, NF-κB, metalloprotease MMP-13, and cartilage oligomeric matrix protein COMP-2). Both HHA/BC and HHA/BC+PRP similarly prompted scratch repair. Overall, these outcomes provide deeper insights into the biochemical and biological properties of these innovative injectable formulations, highlighting their potential application in OA management.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1607469"},"PeriodicalIF":4.8,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12537726/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145344514","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Casein-assisted biomineralization of calcium carbonate microspheres for enhanced surface and adsorption properties.","authors":"Aniket Gade, Julia Nadrowska, Joanna Trzcińska-Wencel, Marek Wiśniewski, Rajesh Raut, Mahendra Rai, Patrycja Golińska","doi":"10.3389/fbioe.2025.1654712","DOIUrl":"10.3389/fbioe.2025.1654712","url":null,"abstract":"<p><strong>Introduction: </strong>Biomineralization is a key biological process by which organisms form mineralized structures, with calcium carbonate being one of the most abundant naturally occurring biominerals. The development of synthetic analogs, particularly calcium carbonate microspheres (CaCO₃-MS), holds potential for various applications, including as carrier materials.</p><p><strong>Methods: </strong>In this study, CaCO₃-MS were synthesized using a precipitation method, both with and without casein. Ammonium, sodium, and potassium carbonate were evaluated as precipitating agents to optimize microsphere formation. The physical properties of the resulting microspheres were characterized using nitrogen adsorption analysis, Brunauer-Emmett-Teller (BET) analysis, diffuse reflectance infrared Fourier transform spectroscopy (DRIFT), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and X-ray diffraction (XRD) analysis.</p><p><strong>Results: </strong>Ammonium carbonate was the most effective precipitating agent, yielding well-formed microspheres. Casein-assisted CaCO₃-MS exhibited a higher specific surface area (65 m²/g) than CaCO₃-MS synthesized without casein (47 m²/g). The casein-containing microspheres also demonstrated a more uniform spherical morphology, increased pore volume, higher surface energy, enhanced hydrophilicity, and approximately double the water adsorption capacity. However, both variants showed similar adsorption-desorption kinetics.</p><p><strong>Discussion: </strong>The presence of casein significantly improved the structural and functional properties of CaCO₃-MS, making them more suitable for use as carrier materials. Furthermore, the described method enables the large-scale, surfactant-free synthesis of uniformly sized microspheres, enhancing its practical applicability.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1654712"},"PeriodicalIF":4.8,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12537877/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145344446","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Epigallocatechin-gallate loaded BSA nanoparticles as innovative anti-inflammatory agents in immature macrophages.","authors":"Simona Martano, Mariafrancesca Cascione, Livia Giotta, Loris Rizzello, Riccardo Di Corato, Stefano Leporatti, Rosaria Rinaldi, Valeria De Matteis","doi":"10.3389/fbioe.2025.1666492","DOIUrl":"10.3389/fbioe.2025.1666492","url":null,"abstract":"<p><strong>Introduction: </strong>The development of innovative anti-inflammatory therapies is critical for addressing chronic inflammatory diseases and cancer. Epigallocatechin gallate (EGCG), a polyphenolic compound with strong antioxidant and anti-inflammatory properties, suffers from limited stability and bioavailability. Bovine Serum Albumin Nanoparticles (BSA-NPs), due to their biodegradability, non-toxicity, and high binding capacity, represent a powerful delivery system for bioactive compounds.</p><p><strong>Methods: </strong>EGCG-loaded BSA nanoparticles (EGCG@BSA-NPs) were synthesized via the desolvation method. The nanoparticles were characterized by Transmission Electron Microscopy (TEM), Dynamic Light Scattering (DLS), ζ-potential analysis, Fourier-transform infrared spectroscopy (FTIR), and UV-Vis spectroscopy. Encapsulation efficiency and antioxidant capacity were assessed by Trolox equivalent antioxidant capacity (TEAC) assays. The anti-inflammatory potential was evaluated in immature macrophages (THP-1 cells) by assessing NF-κB nuclear translocation and the stimulation of proinflammatory cytokines IL-8 and TNF-α.</p><p><strong>Results: </strong>Morphological and physicochemical analyses confirmed the successful formation of spherical EGCG@BSA-NPs with improved size uniformity and controlled surface charge. Antioxidant assays demonstrated enhanced radical scavenging activity compared with unloaded BSA-NPs and free EGCG. Cellular studies showed that EGCG@BSA-NPs reduced NF-κB nuclear translocation and decreased IL-8/TNF-α secretion, highlighting their anti-inflammatory efficacy.</p><p><strong>Discussion: </strong>These findings suggest that EGCG@BSA-NPs are an effective nanoplatform for the controlled delivery of polyphenolic compounds. By improving stability and enhancing bioactivity, they hold significant promise in modulating macrophage function and reducing inflammation, thereby supporting their potential use in chronic inflammatory disease and cancer therapy.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1666492"},"PeriodicalIF":4.8,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12537887/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145344466","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and validation of a qPCR assay for the detection of residual host cell DNA in rabies vaccines produced in Vero cells.","authors":"Danhua Zhao, Weiying Zong, Wanxin Wu, Yuhua Li, Zongsong Wu, Zhixing Yang, Shouchun Cao","doi":"10.3389/fbioe.2025.1611428","DOIUrl":"10.3389/fbioe.2025.1611428","url":null,"abstract":"<p><p>Residual host cell DNA in biological products, including rabies vaccines, poses potential health risks such as tumorigenesis and infectivity. Regulatory authorities have set limits for residual DNA levels to ensure product safety. Among various detection methods for residual DNA, quantitative PCR (qPCR) is recognized for its high sensitivity and efficiency. This study developed and validated a qPCR assay for detecting residual Vero DNA in rabies vaccines produced in Vero cells. The assay targeted two highly repetitive Vero genomic DNA sequences: the \"172bp\" sequence and the Alu repetitive sequence. The method was optimized and validated for linearity, range, quantitation limit, detection limit and specificity, etc. The qPCR assay for the \"172bp\" sequence exhibited excellent linearity, with a quantification limit of 0.03pg/reaction and a detection limit of 0.003pg/reaction. The relative standard deviation (RSD) across samples ranged from 12.4% to 18.3%, and the recovery rate was between 87.7% and 98.5%. No cross-reactivity was observed with common bacterial and cell strains, indicating a high specificity of the assay. These findings suggest that the qPCR method is a reliable approach for quantifying residual Vero DNA in pharmaceuticals and for regulatory compliance monitoring. The assay method has been adopted by local vaccine manufacturers, and has been included in the Chinese Pharmacopoeia, thus will help to enhance vaccine quality and safety.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1611428"},"PeriodicalIF":4.8,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12536431/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145344458","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Stromal cells unifying the pathology of acute and chronic skeletal muscle injury - clue for novel biomaterials.","authors":"Xianglong Liu","doi":"10.3389/fbioe.2025.1684310","DOIUrl":"10.3389/fbioe.2025.1684310","url":null,"abstract":"","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1684310"},"PeriodicalIF":4.8,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12531245/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145328448","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>OSR1</i> and <i>SIX2</i> drive divergent transcriptional programs in human kidney cells: implications for regeneration and tumorigenesis.","authors":"Naomi Pode-Shakked, Osnat Cohen-Zontag, Dorit Omer, Orit Harari-Steinberg, Einav Vax, Oren Pleniceanu, Benjamin Dekel","doi":"10.3389/fbioe.2025.1645499","DOIUrl":"10.3389/fbioe.2025.1645499","url":null,"abstract":"<p><strong>Background: </strong>The nephron progenitor cells generate approximately one million nephrons during human nephrogenesis. At 34-36 weeks of human genstation, silencing of the key kidney progenitor genes results in depletion of this progenitor pool, limiting the regeneration capacity of the mature kidney. Concurrently, the increasing incidence of end-stage kidney disease underscores the urgent need for innovative regenerative strategies.</p><p><strong>Methods: </strong>We employed lentiviral vectors to ectopically induce two key kidney progenitor genes <i>OSR1</i> and <i>SIX2</i> individually or together in primary human adult kidney (hAK) cells. We then analyzed the cellular and molecular consequences through morphological assessments, functional assays, <i>in vivo</i> transplantation studies, and comprehensive transcriptional profiling.</p><p><strong>Results: </strong><i>OSR1</i> and <i>SIX2</i> induced distinct reprogramming processes with differential functional outcomes; <i>SIX2</i> overexpression was found to maintain epithelial morphology while significantly enhancing proliferation and clonogenic efficiency. Transcriptionally, <i>SIX2</i> established epithelialization and cell-cycle networks by downregulating proximal tubule markers while upregulating distal nephron markers and proliferation genes. <i>In vivo</i>, <i>SIX2</i>-expressing cells formed organized tubular structures with a distinct luminal architecture in a proof-of-concept model. In contrast, <i>OSR1</i> overexpression was found to induce morphological changes and activate developmental morphogenetic pathways, including epithelial tube morphogenesis and canonical <i>Wnt</i> signaling; however, it did not enhance proliferation and showed minimal tubulogenic capacity <i>in vivo</i>. Unexpectedly, <i>OSR1</i> overexpression led to malignant transformation in one clone and exhibited Wilms'-tumor-like features, including expression of kidney developmental markers (i.e., SIX2, NCAM1, and WT1) and blastemal phenotype.</p><p><strong>Conclusion: </strong>Our findings suggest that <i>SIX2</i> overexpression in primary hAK cells functionally confers enhanced self-renewal and tubulogenic capacity while transcriptionally inducing a proximal-to-distal tubular cell diversion with maintained proliferative programs. In contrast, <i>OSR1</i> activates the broader developmental morphogenetic networks but poses potential oncogenic risks. The malignant transformation observed with <i>OSR1</i> overexpression provides insights into the potential cellular origins of Wilms' tumor and raises important safety considerations for regenerative medicine approaches involving developmental gene induction in adult kidney cells.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1645499"},"PeriodicalIF":4.8,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12531215/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145328144","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>In vitro</i> and <i>in vivo</i> biodegradation and biocompatibility assessment of magnesium composites for bone implants.","authors":"Samantha Gmitro, Andres Larraza, Pedram Sotoudehbagha, Andres Alayon Mata, Andrew Romero, John Lovejoy, Mehdi Razavi","doi":"10.3389/fbioe.2025.1685918","DOIUrl":"10.3389/fbioe.2025.1685918","url":null,"abstract":"<p><strong>Introduction: </strong>With an average of 6.8 million fractures per year in the United States, the current method for surgical intervention involves bioinert materials that do not promote osteogenesis and can have inflammatory reactions that hinder bone healing. Magnesium has emerged in research due to the material's biodegradability and biocompatibility; however, it has a low corrosion resistance, which can lead to hydrogen gas evolution and tissue necrosis. Therefore, magnesium is typically alloyed with rare earth elements (REEs) to increase corrosion resistance. The main goal of this study involves fabricating a magnesium (Mg)-based metal matrix nanocomposite (MMNC) containing scandium (Sc) and strontium (Sr) as alloying elements, as well as diopside (CaMgSi2O6) -based bioactive glass-ceramic nanoparticles for reinforcement.</p><p><strong>Methods: </strong>MMNCs were processed using ultrasonic melt processing and hot rolling to disperse nanoparticles and refine their microstructure. These MMNCs have undergone detailed tests to determine microstructure and degradation properties, followed by <i>in vitro</i> and <i>in vivo</i> tests to determine the MMNC's biodegradation and biocompatibility characteristics.</p><p><strong>Results: </strong>Through cell culture with human bone marrow-derived mesenchymal stem cells (hBM-MSCs) we determined that MMNCs provide <i>in vitro</i> cytocompatibility of >80%. Next, MMNC pins were implanted into rat femoral defects and monitored for 3 months post-implantation with the WE43 Mg alloy used as a control. Utilizing <i>in vivo</i> and <i>ex vivo</i> X-ray imaging and histology of these defects implanted with MMNC or WE43 pins, we found that our composite allows for no or minimal hydrogen gas evolution and fibrotic body response with osteointegration and new bone formation.</p><p><strong>Discussion: </strong>This allows for an understanding of potential applications of our composite as a biomaterial.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1685918"},"PeriodicalIF":4.8,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12531167/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145328233","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Animal experiment on osseointegration of porous titanium root analogue implants with composite CSn-TAK242 coating.","authors":"Hui Li, Dan Luo, Yudong Gao, Dashan Wang, Jianjun Yang, Zexian Xu","doi":"10.3389/fbioe.2025.1673758","DOIUrl":"10.3389/fbioe.2025.1673758","url":null,"abstract":"<p><strong>Objective: </strong>Chitosan nanoparticles loaded with TLR4 inhibitors (TAK242) were coated on porous titanium root analogue implants and placeded into beagles to investigate the role of TLR4 inhibitors in inhibiting inflammatory reactions and promoting osseointegration <i>in vivo</i>.</p><p><strong>Methods: </strong>The control group consisted of porous titanium root analogue implants fabricated via digital medical technology and 3D printing, while the experimental group included porous titanium root analogue implants with CSn and CSn-TAK242 bioactive coatings. Three groups of implants were inserted into the jaws of dogs, with their stability coefficients immediately measured upon implantation. After 3 months, samples were collected, and the bone integration and gingival attachment of the three groups were assessed using X-rays, Micro-CT, and histological section staining.</p><p><strong>Results: </strong>All groups of porous titanium root analogue implants were correctly placed within the alveolar sockets. The stability coefficients of the implants immediately post-implantation in the control group, CSn group, and CSn-TAK242 group were (64.29 ± 4.01), (62.55 ± 1.98), and (64.59 ± 3.28), respectively, with no significant statistical difference (P>0.05). Three months post-surgery, imaging and histological examinations revealed bone integration with the surrounding bone tissue for all implant groups. BIC results showed: control group (68.11 ± 3.63)%, CSn group (71.07 ± 2.83)%, and Csn-TAK242 group (78.6 ± 4.52)%, with the BIC being highest in the CSn-TAK242 group, followed by the CSn group, and lowest in the control group (P<0.05). More importantly, compared with the control group, the BV/TV of the CSn-TAK242 group was significantly higher. In addition, the Tb.Th of the CSn-TAK242 group was significantly higher than that of the control group and CSn group (P<0.05). The smooth structures at the upper ends of the implants had tight gingival tissue attachment.</p><p><strong>Conclusion: </strong>Porous titanium root analogue implants consistent with the target root morphology were successfully fabricated using digital medical technology and 3D printing. The composite CSn-TAK242 coating further enhanced the osseointegration effects of these implants.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1673758"},"PeriodicalIF":4.8,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12531250/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145328260","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Neural growth patterns: how random and aligned fibers guide 3D cell organization and pseudospheroid formation.","authors":"Jana Hlinkova, Karolina Dziemidowicz, Mathilde M Ullrich, Anne Eriksson Agger, Aina-Mari Lian, Janne Elin Reseland, Athina Samara","doi":"10.3389/fbioe.2025.1659965","DOIUrl":"10.3389/fbioe.2025.1659965","url":null,"abstract":"<p><strong>Background and purpose: </strong>Electrospun biomaterials replicate the structural complexity of the extracellular matrix (ECM), providing mechanical support and promoting neural cell survival and organization. Fiber orientation is a key determinant of neural cell behavior, influencing adhesion, migration, and differentiation. This study investigates how high seeding density combined with fiber directionality shapes SH-SY5Y culture morphology, gene expression, and early network formation; all critical factors for the design of next-generation scaffolds for neural tissue engineering.</p><p><strong>Methods: </strong>Polycaprolactone (PCL) scaffolds with either random or aligned fiber orientation were fabricated via monoaxial electrospinning. Human SH-SY5Y neuroblastoma cells were seeded at high density and cultured for 7 days, and cell viability was assessed by lactate dehydrogenase (LDH) activity. Neural, ECM, and differentiation markers were analyzed using quantitative PCR, Luminex cytokine profiling, and confocal immunofluorescence.</p><p><strong>Results: </strong>Hydrophobic PCL fibers supported cell adhesion, migration, and proliferation when cells were seeded in small clusters. After 7 days, cell coverage of the fiber-mat was significantly higher on random fibers compared to aligned ones (27.7% vs. 15.8%). Fiber orientation influenced both culture morphology and gene expression. Pseudospheroids formed on both substrates, that differed in perimeter (348.5 µm on random vs. 450.5 µm on aligned fibers, p < 0.05), with no significant difference in thickness (38.4 ± 7.7 µm vs. 43.2 ± 5.5 µm). mRNA expression of connexin 43 and β3-tubulin increased significantly from day 1 to day 7 on random fibers. On aligned fibers, mRNA patterns resembled cells cultured on glass (control), with elevated connexin 31 and doublecortin over time. Immunofluorescence showed early enrichment of nestin on aligned fibers (day 1), and greater expression of β3-tubulin, acetylated tubulin, and connexin 31 on aligned substrates, whereas fibronectin 1 was more prominent on random fibers.</p><p><strong>Conclusion: </strong>Fiber orientation significantly affected SH-SY5Y cell behaviour, including adhesion, formation of pseudospheroids, and differentiation marker expression under high-density conditions. Random and aligned fibers elicited distinct structural patterns and molecular responses, highlighting the importance of scaffold architecture in the rational design of neuroregenerative platforms. To our knowledge, this is the first study to describe scaffold-anchored neural pseudospheroids as a distinct model from conventional suspension spheroids.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"13 ","pages":"1659965"},"PeriodicalIF":4.8,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12531216/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145328455","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}