Cellular and molecular bioengineering最新文献

{"title":"Sphingolipid Metabolism Links Morphological Signatures to Immunomodulatory Potency in Human Mesenchymal Stromal Cells.","authors":"Daniel C Shah, Bobby W Leitmann, Priyanka Priyadarshani, S'Dravious A DeVeaux, Carlos Munoz, Yuwei Zhang, Kejie Rui, Nathan F Chiappa, Lauren A Hymel, William York, Molly E Ogle, Edward A Botchwey, Luke J Mortensen","doi":"10.1007/s12195-026-00933-x","DOIUrl":"10.1007/s12195-026-00933-x","url":null,"abstract":"<p><p>Interest in human mesenchymal stromal cells (MSCs) as an immune therapeutic has been on the increase over the past two decades with preclinical research yielding promising results, but so far only a singular MSC-based immunotherapy has been approved by the U.S. Food and Drug Administration (FDA). A major barrier for MSC clinical translation is the lack of reliable potency metrics and sufficient understanding of the mechanisms of action, as reported by the National Cell Manufacturing Consortium (NCMC). We integrated high-content morphological profiling and targeted sphingolipid analysis across MSC donors exhibiting a range of IFN-γ induced indoleamine-2,3-deoxygenase (IDO) activities to investigate the role of cell membrane components in MSC immunomodulatory function. High- and low IDO potency cells have distinct morphological signatures that are also reflected in the sphingolipid (SL) activity, with low IDO potency cell lines having low sphingomyelinase activity and high IDO potency cell lines having high sphingomyelinase activity. Perturbation of SL metabolism through the addition of exogenous neutral sphingomyelinase not only shifted morphological signatures to mimic those of high potency, but also significantly increased IDO activity within both high and low IDO potency donors. Additionally, these perturbations resulted in increased extracellular vesicle production with rich SL content and a stark reduction of TNF-α following EV treatment of activated macrophages. Together, these findings link morphology features from potent MSCs to sphingolipid remodeling and suggest a tractable strategy to tune MSC immunomodulatory potency.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at https://doi.org/10.1007/s12195-026-00933-x.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 4","pages":"535-548"},"PeriodicalIF":3.5,"publicationDate":"2026-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13562489/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148954962","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Kindlin-3 is a Regulator of Force-Regulated Activation of Integrin α₄β₇ by CCL25 During Lymphocyte Adhesion.","authors":"Paimin Zhuo, Zhiqing Luo, Xianwen Luo, Hefeng Wang, Cheng Dong, Quhuan Li","doi":"10.1007/s12195-026-00935-9","DOIUrl":"10.1007/s12195-026-00935-9","url":null,"abstract":"<p><strong>Purpose: </strong>Lymphocyte recruitment to inflamed intestines in inflammatory bowel disease (IBD) critically depends on shearstress-regulated integrin α₄β₇/MAdCAM-1 interactions. However, the mechano-chemical coupling between the chemokine CCL25 and the intracellular adaptor Kindlin-3 in activating α₄β₇ under physiological flow remains unclear.</p><p><strong>Methods: </strong>Utilizing parallel-plate flow chamber assays combined with fluorescence microscopy and molecular perturbation, wedemonstrate that CCL25 triggers robust integrin α₄β₇ activation and stable lymphocyte adhesion only under physiological shear stress, a process regulated by Kindlin-3.</p><p><strong>Results: </strong>Mechanistically, Kindlin-3 acts as a force transmission hub, enabling CCL25 signaling to enhance α₄β₇-mediatedadhesion strengthening. Knockdown of Kindlin-3 specifi cally enhanced the force-sensitized, CCL25-induced activationof α₄β₇.</p><p><strong>Conclusions: </strong>This study reveals, for the first time, that Kindlin-3 is crucial for translating chemokine (CCL25) signals intobiomechanical activation of integrin α₄β₇ under shear flow. Our findings establish Kindlin-3 as a critical mechano-regulator of chemokine-induced integrin activation during lymphocyte homing, providing novel mechanistic insightsinto IBD pathogenesis and highlighting this pathway as a promising target for anti-adhesion therapeutics.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at https://doi.org/10.1007/s12195-026-00935-9.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 4","pages":"549-565"},"PeriodicalIF":3.5,"publicationDate":"2026-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13562462/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148954873","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Matrix Stiffness Modulates 3D Spheroid-Derived Extracellular Vesicle Profiles and Discovery of Piezo1 Cargo.","authors":"Maulee Sheth, Supasek Kongsomros, Manju Sharma, Maria Lehn, Takanori Takebe, Vinita Takiar, Trisha Wise-Draper, Somchai Chutipongtanate, Leyla Esfandiari","doi":"10.1007/s12195-026-00931-z","DOIUrl":"10.1007/s12195-026-00931-z","url":null,"abstract":"<p><strong>Background: </strong>Increased extracellular matrix stiffness is a defining mechanical feature of solid tumors, yet how it regulates extracellular vesicle-mediated intercellular communication remains poorly understood in three-dimensional tumor microenvironments. Here, we demonstrate that ECM stiffness mechanistically regulates extracellular vesicle (EV) cargo loading in oral squamous cell carcinoma spheroids.</p><p><strong>Methods and results: </strong>Using a tunable three-dimensional spheroid culture platform, we show that increased matrix stiffness enriches tumorigenic and metastatic non-coding RNA transcripts in EVs.  At a functional level, stiffness-primed EVs influence recipient spheroid growth by modulating proliferation and apoptosis. Notably, our study reveals that EVs are enriched in parental biomolecular cargo, including the mechanosensitive Piezo1 ion channel and adhesion and stemness molecule CD44. Protein expression and small RNA sequencing analyses confirm the incorporation of these components into spheroid-derived EVs in a stiffness-independent manner.</p><p><strong>Conclusion: </strong>Together, our findings identify ECM stiffness as a mechanistic regulator of EV composition and establish EVs as biomechanical signaling vectors that further influence cell proliferation in three-dimensional microenvironments.</p><p><strong>Graphical abstract: </strong></p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at https://doi.org/10.1007/s12195-026-00931-z.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 4","pages":"519-533"},"PeriodicalIF":3.5,"publicationDate":"2026-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13562468/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148954968","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Single-Cell RNA-Sequencing Reveals Tumor Microenvironment Composition and Prior Therapy Modulate Response to Axl Inhibition.","authors":"Anisha Datta, Kate Bridges, Remziye E Wessel, Hratch M Baghdassarian, Laura C Bahlmann, Diana N Gong, Erin N Tevonian, Brian A Joughin, Douglas A Lauffenburger","doi":"10.1007/s12195-026-00930-0","DOIUrl":"10.1007/s12195-026-00930-0","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Purpose: &lt;/strong&gt;Axl is a therapeutic target under active clinical investigation for a variety of cancer indications. While pre-clinical mouse studies have been promising, clinical trials targeting Axl in combination with standard-of-care therapies have yielded mixed results. We hypothesize that this clinical translation gap may in part be due to an incomplete understanding of the effects of Axl inhibition specifically on the myeloid compartment of the human tumor microenvironment, as Axl mediates immunoregulatory signaling in myeloid cells. Utilizing human in vitro melanoma tumor microenvironment model systems, herein we generated and analyzed single-cell RNA-sequencing data to understand the impacts of tumor microenvironment composition and tumor cell prior therapy on myeloid cell response to Axl inhibition.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;We used our previously established in vitro model system to generate single-cell RNA-sequencing samples of tumor cell-macrophage co-cultures as well as tumor cell-macrophage-dendritic cell tri-cultures from four healthy buffy coat donors. We first applied LIgand-receptor ANalysis frAmework (LIANA) to our transcriptional data to infer potential intercellular crosstalk, complemented by Tensor-cell2cell to identify prevalent patterns of potential communication across the various experimental conditions. We next probed the impact of Axl inhibition on intercellular communication with orthogonalized partial least squares discriminant modeling. Finally, we conducted gene set enrichment analysis to understand how each cell type's state and signaling activity was impacted by each axis of experimental variation in our complex experimental design.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;Tensor-cell2cell yielded eight patterns of communication in the inferred intercellular interaction data, and these eight patterns were paired based on their communicating cell types as well as their shared top ligand-receptor interactions. These pairings demonstrated the impact of including dendritic cells in the model system and highlighted coordination among the myeloid cells. The specific effects of Axl inhibition on potential intercellular crosstalk were clarified with supervised modeling, demonstrating impaired tumor cell invasion and migration programs, as expected. Intriguingly, gene set enrichment analysis revealed that tumor cell prior injury with standard-of-care therapy impacted myeloid cell interferon signaling in response to Axl inhibition.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusions: &lt;/strong&gt;In this work, we demonstrated that tumor microenvironment cellular composition as well as prior treatment with standard-of-care therapy modulate myeloid cell response to Axl inhibition in human in vitro melanoma model systems. While the inferred effects of Axl inhibition on intercellular crosstalk, cell state, and cell activity were largely consistent with prior literature, the impact of tumor cell prior injury on the myeloid interferon signaling re","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 4","pages":"497-518"},"PeriodicalIF":3.5,"publicationDate":"2026-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13562482/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148954894","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Toward a Dual-Axis Model of Microbiome Modulation in Cancer Immunotherapy: Pathobiont Elimination and Functional Ecosystem Restoration.","authors":"Diwakar Davar, Hassane M Zarour, Giorgio Trinchieri","doi":"10.1007/s12195-026-00916-y","DOIUrl":"10.1007/s12195-026-00916-y","url":null,"abstract":"<p><strong>Purpose: </strong>The gut microbiome is increasingly recognized as a modulator of cancer immunotherapy efficacy, including responses to immune checkpoint inhibitors (ICIs) and chimeric antigen receptor T-cell (CAR-T) therapy. Recent clinical trials of microbiome-targeted interventions such as fecal microbiome transplantation (FMT) and live biotherapeutic products (LBPs) suggest the potential to enhance antitumor immunity and improve clinical outcomes. Yet responses remain heterogeneous and are not fully explained by engraftment of donor taxa alone.</p><p><strong>Methods: </strong>We integrate evidence from interventional trials, observational cohort studies, and principles from gut microbial ecology to develop a model hypothesis on how microbiome-targeted therapies may shape response to immunotherapy, with potential to inform future trial design, analyses, and interpretation.</p><p><strong>Results: </strong>Drawing on the available evidence, we propose that therapeutic perturbation of the gut microbiome may augment immunotherapy efficacy through two parallel axes: (1) elimination of immunosuppressive pathobionts that restrain CD8+ T-cell activation and promote myeloid-mediated immunosuppression, and (2) functional restoration of the gut ecosystem through engraftment of taxa that provide metabolites, structural cues, and immunoregulatory signals required for effective antitumor immunity. The success of both axes appears to depend on ecological processes governed by predator-prey dynamics, including colonization resistance, resilience of the resident microbiota, and the ability of administered organisms to displace entrenched dysbiotic communities. This ecological lens may help to explain discrepancies across trial designs, donor types, and intervention modalities, and suggests that complete donor engraftment is neither necessary nor sufficient for clinical benefit.</p><p><strong>Conclusions: </strong>A dual-mechanism model of pathobiont elimination and functional microbial restoration may help explain microbiome-mediated enhancement of cancer immunotherapy, highlighting a balanced immune permissive gut ecosystem as a key determinant of therapeutic success.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2026-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13348773/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148419125","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Targeted Inhibition of Nuclear Factor-κB Pathway Using Recombinant Adeno-Associated Virus 5 in 2D and 3D Glioblastoma Models.","authors":"Ceera Manikandan, Nithin Sam Ravi, Mohankumar M Kumarasamypet, Amit Kumar Jaiswal","doi":"10.1007/s12195-026-00924-y","DOIUrl":"10.1007/s12195-026-00924-y","url":null,"abstract":"<p><strong>Purpose: </strong>Glioblastoma (GBM) is a primary brain tumor with pronounced heterogeneity, aggressive nature and resistance to standard therapies. The dysregulation of nuclear factor kappa B (NF-κB) is central to glioblastoma pathology. But the limitations of existing NF-κB inhibitors have underscored the need for gene therapy approaches capable of achieving controlled and sustained pathway suppression. In this study, a previously engineered bicistronic recombinant AAV5 (rAAV5) vector co-expressing a dominant-negative mutant of IκBα (IκBα-M) alongside a luciferase reporter was packaged into viral particles. The functionality of rAAV5 viral particles was evaluated in both two-dimensional (2D) U87-MG cells and three-dimensional (3D) glioblastoma spheroids.</p><p><strong>Methods: </strong>A bicistronic rAAV5 vector co-expressing IκBα-M and luciferase was packaged and transduced into U87-MG cells at a multiplicity of infection of 10,000 vg/cell. Transduction efficiency was assessed using a rAAV5 GFP reporter virus and evaluated by flow cytometry and confocal imaging. NF-κB activity was evaluated using luciferase reporter assay, immunofluorescence, qPCR, Western blot analysis and ELISA. Apoptosis and cell viability were measured using Annexin V/PI and Live/Dead imaging respectively. ATP-based viability assay was evaluated in both 2D culture and 3D spheroids.</p><p><strong>Results: </strong>U87-MG cells transduced with rAAV5 GFP reporter virus exhibited high transduction efficiency with 82% GFP positive cells in 2D and efficiently transduced 3D spheroids. IκBα-M overexpression resulted in cytoplasmic retention of NF-κB and significant reduction in luciferase activity. NF-κB and hypoxia-inducible factor 1 subunit alpha (HIF1A) gene expression levels were downregulated and protein analysis confirmed increased NF-κB levels in the cytoplasmic protein. Additionally, cells transduced with rAAV5 virus induced apoptosis and reduced cell viability in GBM spheroids.</p><p><strong>Conclusion: </strong>The findings of this study establish that rAAV5-mediated NF-κB responsive delivery of IκBα-M enables effective and targeted suppression of aberrant NF-κB signaling, supporting its potential for further preclinical development in GBM gene therapy.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 4","pages":"465-479"},"PeriodicalIF":3.5,"publicationDate":"2026-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13562483/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148954964","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sex Differences in Angiogenic Ligand and Receptor Protein Expression During Adipose Tissue Expansion and Regression.","authors":"Yingye Fang, Silvia Gonzalez-Nieves, Vincenza Cifarelli, Princess I Imoukhuede","doi":"10.1007/s12195-026-00925-x","DOIUrl":"10.1007/s12195-026-00925-x","url":null,"abstract":"<p><strong>Purpose: </strong>Inadequate angiogenesis in obesogenic adipose tissue (AT) has been implicated in disrupted adipogenesis and metabolic disorders. While several regulators of AT angiogenesis have been identified, our understanding of the quantitative changes in angiogenic signaling proteins during obesity progression remains incomplete, particularly regarding sex-specifi c responses. This study sought to identify the dysregulated elements within the Vascular Endothelial Growth Factor (VEGF) and Platelet-Derived Growth Factor (PDGF) systems during obesity progression.</p><p><strong>Methods: </strong>We employ a mouse model, comprising both male and female mice, to investigate the changes in the VEGF/PDGF concentration and their receptor distribution in gonadal AT during short- and long-term weight gain and weight loss.</p><p><strong>Results: </strong>Female mice preserve adipose tissue angiogenic signatures during obesity progression, including sustained upregulation of endothelial VEGFR1 protein, maintenance of VEGF-A levels and endothelial cell populations, and differential regulation of PDGF levels, compared to males. These sex-specifi c patterns correlate with improved adipose expandability and may contribute to the more metabolically healthy obesity phenotype commonly observedin females.</p><p><strong>Conclusion: </strong>These sex-specifi c patterns correlate with improved adipose expandability and may contribute to the more metabolically healthy obesity phenotype commonly observed in females. Our quantitative profi ling also lays the groundwork for developing computational models of VEGF/PDGF signaling networks in AT, allowing for the simulation of complex biological interactions and the prediction of therapeutic outcomes.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at https://doi.org/10.1007/s12195-026-00925-x.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 4","pages":"481-496"},"PeriodicalIF":3.5,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13562464/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148954946","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"New Insights into 3D Bioprinting of Cartilage: A Review on Recent Advances, Challenges, and Perspectives.","authors":"Negin Khoshnood, John P Frampton, Alireza Nouri, Ali Zamanian","doi":"10.1007/s12195-026-00926-w","DOIUrl":"10.1007/s12195-026-00926-w","url":null,"abstract":"<p><p>Cartilage is an avascular tissue with a limited capacity for self-regeneration. Traditional autologous cartilage transplantation is incapable of fulfilling the increasing demand for repair of various cartilage tissue lesions. The advent of 3D bioprinting technology provides an opportunity to repair articular cartilage defects through the construction of organized living structures composed of biomaterials and cells. This technology can mimic natural cartilage by allowing control of cell distribution, and the modulation of mechanical and chemical properties with high precision. This review provides insight into the current developments in 3D bioprinting for cartilage tissue engineering. Recent studies on 3D-bioprinted cartilage tissue constructs and various bioinks are highlighted. The advantages and limitations of commonly used natural, synthetic, and composite bioinks in terms of printability, mechanical properties, and biological performance in bioprinting of anatomically shaped constructs (nasal, auricle, knee, and tracheal cartilage) are discussed. Furthermore, outlooks and challenges of 3D bioprinting of complex constructs with variable mechanical and biological properties are provided. Based on recent progress, it is expected that 3D bioprinting will lead to improved 3D tissue-engineered constructs for regeneration and repair of cartilage tissue.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 4","pages":"427-463"},"PeriodicalIF":3.5,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13562497/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148954990","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Non-linear Characterization of Commercial and Decellularized Hydrogels: Statistical Framework Enhanced by Bayesian Optimization.","authors":"D E García-García, D Marques, H Amaveda, M Mora, J Asín, I Villaoslada, P M Baptista, M A Pérez, J M García-Aznar","doi":"10.1007/s12195-026-00913-1","DOIUrl":"10.1007/s12195-026-00913-1","url":null,"abstract":"<p><strong>Scope: </strong>Hydrogels are widely used in the design of tissue substitutes because of their ability to mimic the extracellular matrix (ECM). Their mechanical cues critically influence the cellular response, making accurate characterization essential. However, it remains challenging due to their intricate nature. This study computationally evaluates the hyperelastic properties of next-generation hydrogels of high biomedical interest, including basal membrane extract and decellularized liver matrices, as well as structural proteins.</p><p><strong>Methods: </strong>We present a combined framework based on Bayesian optimization and statistical analyses that go beyond classical least-squares fitting, leveraging rheological experimental data. It defines each hyperelastic strain-energy density function, and addresses both intra- and inter-sample variability. This approach quantifies uncertainty and reveals the natural variability that deterministic models overlook, and it also enables quantification of coefficient variation with composition.</p><p><strong>Results: </strong>Validation against experimental data shows computational fits of 5% error in most cases, and low calculation time. Analyses reveal that composition-collagen addition, fibrin concentration, and decellularized extracellular matrix (dECM) age-modulate initial stiffness, nonlinearity, and overall mechanical resistance of the hydrogels.</p><p><strong>Conclusion: </strong>Hydrogels derived from basal membrane extracts exhibit comparable non-linear mechanics, while collagen addition reduces stiffness and nonlinearity. In fibrin-based hydrogels with decellularized liver matrix, mechanical behavior is concentration and age-dependent. Such insights are highly relevant for mechanobiology, enabling prediction of how cells sense mechanical cues and scaffold composition influences in vivo interactions.</p><p><strong>Supplementary information: </strong>The online version of this article (10.1007/s12195-026-00913-1) contains supplementary material, which is available to authorized users.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 3","pages":"327-344"},"PeriodicalIF":3.5,"publicationDate":"2026-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13365100/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148444536","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mechanisms and Strategies for Enhancing DNA Nuclear Entry in Gene Delivery.","authors":"Justin Sylvers, Fan Yuan","doi":"10.1007/s12195-026-00927-9","DOIUrl":"10.1007/s12195-026-00927-9","url":null,"abstract":"<p><strong>Background: </strong>Nonviral gene delivery using DNA vectors is widely used in cell engineering, vaccination, and gene therapy, but delivery efficiency remains lower than those of viral vectors and mRNA-based approaches, partly due to inefficient nuclear entry, as transfected DNA must enter the nucleus for transcription. Therefore, a mechanistic understanding of nuclear entry pathways is essential for developing strategies to improve the efficiency.</p><p><strong>Methods: </strong>This review evaluated mechanistic studies of DNA nuclear entry in mammalian cells, mathematical models of intracellular DNA trafficking, quantitative analyses of DNA nuclear accumulation and transgene expression, and strategies to enhance nuclear delivery of DNA.</p><p><strong>Results: </strong>Two mechanistically distinct pathways for DNA nuclear entry have been reported: enclosure upon nuclear envelope reformation in dividing cells, and active transport through nuclear pore complexes (NPCs). Various strategies have been developed to enhance nuclear import through these pathways; however, their effectiveness depends on multiple factors, including cell type, delivery methods, and cell cycle status. Although DNA vectors are significantly larger than the nominal inner diameter of NPCs, they may traverse NPCs through deformation and interactions with nuclear transport proteins. Quantitative studies show that DNA nuclear accumulation is time dependent and heterogeneous among individual cells within the same population.</p><p><strong>Conclusions: </strong>Nuclear entry plays a key role in determining efficiency of nonviral gene delivery. Advances in mechanistic studies, quantitative modeling, and imaging-based analyses have improved our understanding of intracellular DNA trafficking and nuclear accumulation. Integrating these insights with delivery strategies that enhance nuclear access while preserving the cellular machinery required for transgene expression will be critical for developing more efficient and reliable nonviral DNA delivery systems for therapeutic and biotechnological applications.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2026-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13431081/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148668296","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}