Cellular and molecular bioengineering最新文献

{"title":"Nanoparticle-Host Interactions: The Impact of Physiological and Pathological Factors on Biodistribution, Immune Processes, and Translational Challenges.","authors":"Louay Abo Qoura, Dmitry Kostyushev, Alessandro Parodi, Daniel I Boyarintsev, Vladimir Chulanov, Vadim S Pokrovsky","doi":"10.1007/s12195-026-00888-z","DOIUrl":"10.1007/s12195-026-00888-z","url":null,"abstract":"<p><strong>Background: </strong>The clinical translation of nanoparticles (NPs) for therapeutic applications is hindered significantly by unpredictable biodistribution in diseased patients, which results from the complex interplay between engineered physicochemical properties of NPs and specific changes in the function of biological barriers.</p><p><strong>Methods: </strong>This review delves into the multifaceted factors that govern NP biodistribution, highlighting the critical roles of intrinsic NP design, including size, shape, and surface chemistry, along with host-specific physiological and pathological conditions. We overview how these properties can change systemic circulation, organ-specific accumulation, and clearance pathways. The role of surface functionalization in targeted delivery is examined through the lens of altered serum composition, which affects protein corona formation. Particular attention is given to immune response, whether pathogen/antigen-primed, macrophage-monocyte-mediated clearance, compromised biological barriers, and host-specific factors such as sex, age, drug exposure, and gut microbiome. Disease contexts, including cancer and viral infections, are considered to evaluate translational challenges.</p><p><strong>Results: </strong>NP biodistribution is shaped by the interplay between engineered physicochemical properties and disease-associated biological changes. Intrinsic NP characteristics influence systemic circulation, organ-specific accumulation, and clearance pathways. Altered serum composition affects protein corona formation and immune responses, including overactivated or suppressed immune states. Compromised biological barriers, including increased permeability of the blood-brain barrier, enhanced renal excretion, and reduced liver retention, further modify pharmacokinetic profiles. Host-specific variability and pathological conditions introduce additional complexity, creating unique barriers that passive targeting strategies often fail to overcome.</p><p><strong>Conclusion: </strong>Developing a comprehensive grasp of these interrelated mechanisms is crucial for engineering NPs that can translate preclinical results into expected clinical outcomes, ultimately closing the gap between research innovation and patient care.</p><p><strong>Graphical abstract: </strong></p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s12195-026-00888-z.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 1","pages":"1-27"},"PeriodicalIF":5.0,"publicationDate":"2026-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13031575/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147572377","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":"Multimodal Profiling Reveals Distinct Endothelial Activation Pathways Regulated by Flow and Heparan Sulfate.","authors":"Ian C Harding, Nicholas R O'Hare, Ira M Herman, Eno E Ebong","doi":"10.1007/s12195-026-00884-3","DOIUrl":"10.1007/s12195-026-00884-3","url":null,"abstract":"<p><strong>Purpose: </strong>Atherosclerotic cardiovascular disease originates from endothelial dysfunction, characterized by a shift toward a pro-inflammatory state and increased production of reactive oxygen species (ROS). This dysfunction occurs under adverse mechanical conditions, such as blood flow oscillation, multi-directionality, recirculation, shear stress gradients, and low or stagnation flows. This study investigates how degradation of heparan sulfate (HS), a major component of the endothelial glycocalyx, drives the transition of endothelial cells from a functional, anti-inflammatory, and antioxidant phenotype under streamlined flow conditions to a dysfunctional, pro-inflammatory, and pro-oxidant phenotype when flow is stagnant. Pro-inflammatory and pro-oxidant endothelial behavior precedes atherosclerosis development.</p><p><strong>Methods: </strong>Human aortic endothelial cells were exposed to uniform shear stress (14 dynes/cm²) to model healthy endothelium. Unhealthy conditions were simulated via static conditions (0 dynes/cm²) or enzymatic HS degradation using heparinase III. Endothelial cell phenotype was assessed using fluorescent labeling, confocal microscopy, Western blotting, and RNA sequencing.</p><p><strong>Results: </strong>Endothelial cells conditioned by 14 dynes/cm² shear stress without heparinase III exhibited low expression of pro-inflammatory genes (HIF1A, VCAM1, and IL1B), minimal ROS production, and up-regulation of Kruppel-like transcription factors. Under the same flow conditions, HS degradation via heparinase III induced an inflammatory phenotype, resembling responses observed at 0 dynes/cm² shear stress, while ROS levels remained largely unaffected.</p><p><strong>Conclusions: </strong>The endothelial glycocalyx is a protective, dynamic, and complex structure, with HS as a key component. This study demonstrates that intact HS mitigates endothelial dysfunction by suppressing inflammation linked to flow-dependent atherosclerosis, but not ROS production. Future research will focus on translating these findings into HS-targeted therapies for atherosclerotic cardiovascular disease.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s12195-026-00884-3.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 1","pages":"89-110"},"PeriodicalIF":5.0,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13031597/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147572398","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":"Barrier Function of the Extracellular Matrix in AAV Gene Therapy.","authors":"Yahya Cheema, Devorah Cahn, Sahana Kumar, Matthew Wolf, Gregg A Duncan","doi":"10.1007/s12195-026-00886-1","DOIUrl":"10.1007/s12195-026-00886-1","url":null,"abstract":"<p><strong>Purpose: </strong>The extracellular matrix (ECM) is a major component of the tissue microenvironment which may pose a barrier to the distribution of AAV in target organs, preventing delivery of therapeutic cargo. We sought to address this potential barrier to AAV gene therapy by furthering our understanding of AAV-ECM interactions. We hypothesized that both the AAV serotype and ECM composition will impact AAV transport and gene delivery.</p><p><strong>Methods: </strong>AAV2, AAV6, and AAV8 viral vectors were fluorescently labeled to allow for visualization of their diffusion through the ECM. Lung, liver, and small intestinal submucosal dECM hydrogels were formulated as models of the ECM with tissue-specific biomolecular content. We then characterized AAV and nanoparticle diffusion within decellularized ECM using fluorescent video microscopy and multiple particle tracking. Additionally, we evaluated AAV transduction in dECM-incorporated 2D and 3D spheroid tissue culture models.</p><p><strong>Results: </strong>All AAV displayed reduced diffusivity through ECM as compared to similarly sized nanoparticles. AAV2 diffusion was least affected by the presence of ECM across tissue types as compared to AAV6 and AAV8. AAV transduction in dECM incorporated <i>in vitro</i> models was significantly reduced in both a 2D and 3D setting.</p><p><strong>Conclusions: </strong>These results suggest binding of AAV to the ECM may decrease their therapeutic effect in target tissues throughout the body. The barrier function of the ECM should be considered in development of AAV for gene therapy applications.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 2","pages":"163-174"},"PeriodicalIF":5.0,"publicationDate":"2026-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13129159/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147811635","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":"Development of a 3D In Vitro Model of Dupuytren's Disease as a Platform for Drug Screening.","authors":"Jarmila Knitlova, Adam Eckhardt, Daniel Hadraba, David Vondrasek, Roman Stachon, Elena Filova, Vera Jencova, Kristyna Havlickova, Tatyana Kobets, Martin Ostadal, Lucie Bacakova","doi":"10.1007/s12195-026-00885-2","DOIUrl":"10.1007/s12195-026-00885-2","url":null,"abstract":"<p><strong>Background: </strong>Dupuytren's disease (DD) is a common fibrotic disorder of the hand, characterized by progressive thickening and contracture of the palmar and digital fascia. Surgical excision remains the primary treatment; however, there are currently no therapies to prevent disease progression or recurrence. This study aims to develop a 3D in vitro model to test novel antifibrotic therapies. The model is based on decellularized pathological DD tissue seeded with patient-derived fibroblasts, capturing the role of both cellular and extracellular matrix components in disease progression.</p><p><strong>Methods: </strong>Fibrotic DD tissues were obtained from surgical excisions, sectioned, and decellularized. In parallel, primary fibroblasts were isolated from patient samples. The decellularized extracellular matrices (dECMs) were characterized with respect to biochemical composition, collagen structure, and mechanical properties. Fibroblasts were seeded onto the dECMs and cultured stepwise to initially promote proliferation, followed by differentiation into myofibroblasts. Secretomes of cells cultivated on the established 3D model were compared to those from conventional 2D cultivations. To evaluate the model´s relevance and effectiveness we tested the antifibrotic drug minoxidil.</p><p><strong>Results: </strong>The dECMs retained the pathological architecture and mechanical properties of native DD tissue, although individual ECM components were reduced after decellularization. Fibroblasts successfully adhered, proliferated, and repopulated the scaffold. The relevance of the 3D model was demonstrated by the presence of myofibroblasts with disease-relevant secretome. The responsiveness to the drug minoxidil was significantly more complex in the 3D model than in conventional 2D cultures.</p><p><strong>Conclusion: </strong>We demonstrated that dECM seeded with DD fibroblasts represents a relevant 3D in vitro model of Dupuytren's disease. The model enables antifibrotic drug screening, as demonstrated by the testing of minoxidil. Our model provides a reproducible platform also suitable for the investigation of cells and ECM contributions to palmar fascial fibrosis.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s12195-026-00885-2.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 1","pages":"111-127"},"PeriodicalIF":5.0,"publicationDate":"2026-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13031596/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147572291","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":"The Interplay Between Stiffness and Hyperglycemia on Diabetic Foot Ulcer Wound Closure.","authors":"Nourhan Albeltagy, Jennifer Patten, Karin Wang","doi":"10.1007/s12195-025-00877-8","DOIUrl":"10.1007/s12195-025-00877-8","url":null,"abstract":"<p><strong>Introduction: </strong>Diabetic foot ulcers are open wounds with impaired wound closure at the bottom of the foot. Although diabetic plantar skin is stiffer, which should enhance fibroblast mechanotransduction, fibroblasts still fail to migrate effectively. This suggests impaired wound closure is driven by another factor; hyperglycemia (≥11.1 mM glucose), which alters fibroblast mechanotransduction.</p><p><strong>Purpose: </strong>To mimic diabetic foot ulcers by developing a 2D circular in vitro wound closure model system to investigate fibroblast mechanoresponses under diabetic plantar skin stiffness and hyperglycemia.</p><p><strong>Methods: </strong>Polydimethylsiloxane was used as a substrate, fabricated at 57 kPa and 90 kPa for normal and diabetic plantar skin stiffnesses, respectively. Cell culture media contained a 5.5 mM glucose concentration simulating normal blood glucose or an altered 11.1 mM glucose concentration simulating hyperglycemia.</p><p><strong>Results: </strong>Time-lapse fluorescent imaging of wound assays reveals a restrictive effect of higher stiffness on migrating fibroblasts under normal glucose conditions, and a biphasic response to hyperglycemic conditions. Fibroblasts migrating on softer substrates mimicking normal plantar skin stiffness and under hyperglycemia have decreased velocity as predicted. Whereas cells migrating on stiffer substrates mimicking diabetic plantar skin stiffness and under hyperglycemia demonstrate increased cell velocity, overcoming the higher stiffness's restrictive effect. Despite faster cell velocities on higher stiffness, wounds under normal glucose conditions still close faster than those under hyperglycemic conditions.</p><p><strong>Conclusion: </strong>This research establishes a wound closure model demonstrating significantly slower wound closure in diabetic plantar skin with higher stiffness and hyperglycemic glucose levels compared to normal plantar skin with softer stiffness and normal glucose levels.</p><p><strong>Supplementary information: </strong>The online version of this article (10.1007/s12195-025-00877-8) contains supplementary material, which is available to authorized users.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 1","pages":"29-42"},"PeriodicalIF":5.0,"publicationDate":"2026-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13031594/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147572405","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":"FGF-2-Overexpressing Adipose-Derived Stem Cells as a Paracrine Platform for Angiogenesis-Driven Tissue Regeneration.","authors":"Daisuke Seki, Michiyo Honda","doi":"10.1007/s12195-025-00883-w","DOIUrl":"10.1007/s12195-025-00883-w","url":null,"abstract":"<p><strong>Background: </strong>The survival and function of three-dimensional tissues critically depend on the establishment of a functional vascular network that ensures oxygen and nutrient supply and waste removal. Insufficient vascularization leads to hypoxia, metabolic stress, and cell death, making angiogenesis a fundamental requirement for successful tissue regeneration. This requirement is particularly evident in highly vascularized tissues such as bone, where vascular networks closely regulate tissue metabolism and repair.</p><p><strong>Methods: </strong>Human adipose-derived mesenchymal stem cells (ASCs) were genetically modified to overexpress fibroblast growth factor 2 (FGF-2), a key regulator of angiogenesis. The angiogenic potential of these cells and the paracrine effects of their conditioned medium were subsequently evaluated, together with their effects on osteogenic differentiation to assess functional specificity.</p><p><strong>Results: </strong>Overexpression of FGF-2 in ASCs enhanced endothelial cell migration and tube formation via paracrine mechanisms, in which elevated secretion of VEGFA and other angiogenic factors acted synergistically to promote angiogenesis. In contrast, osteogenic differentiation of ASCs was significantly inhibited by FGF-2 overexpression. Notably, <i>FGFR2</i> expression, the receptor for FGF-2, was selectively downregulated during osteogenic induction, suggesting that sustained FGF-2 signaling preferentially interferes with FGFR-mediated pathways associated with osteogenic maturation rather than with early proliferative responses.</p><p><strong>Conclusion: </strong>Our results demonstrate that FGF-2-overexpressing ASCs function not as osteogenic effector cells but as a potent paracrine platform for angiogenesis. Their conditioned medium, enriched with FGF-2 and synergistic angiogenic factors, supports vascular network formation and indirectly enhances the regenerative microenvironment, highlighting its potential as a cell-free strategy for tissue regeneration.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 1","pages":"73-87"},"PeriodicalIF":5.0,"publicationDate":"2025-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13031578/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147572395","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":"Melanoma/CSPG4-Enhanced Collagen-Mediated Contact Guidance Requires Mutant Active BRAF and the CSPG4 Core Protein Cytoplasmic Domain.","authors":"G Thrivikraman, J Yang, M Price, A Giubellino, J B McCarthy, R T Tranquillo","doi":"10.1007/s12195-025-00882-x","DOIUrl":"10.1007/s12195-025-00882-x","url":null,"abstract":"<p><strong>Introduction: </strong>Chondroitin sulfate proteoglycan-4 (CSPG4) is a transmembrane cell surface proteoglycan that promotes malignant progression in melanoma. Elevated CSPG4 expression in melanoma cells is associated with several malignant phenotypic properties, including increased tumor cell invasion, tumorigenic potential, and metastasis.</p><p><strong>Methods: </strong>Magnetically aligned collagen gels with entrapped cells were used to model the aligned extracellular matrix in the tumor microenvironment and to identify the key role of CSPG4 in sensing contact guidance.</p><p><strong>Results: </strong>The data show that CSPG4-expressing WM1552C Radial Growth Phase (RGP) melanoma cells exhibit enhanced contact guidance along with increased migration speed in contrast to paired counterparts that lack CSPG4. This required the presence of a pERK 1,2 phospho-acceptor site on the cytoplasmic tail of the core protein. Furthermore, short-term treatment of CSPG4-expressing cells with the clinically used mutant active BRAF inhibitor vemurafenib reduced both guidance and speed.</p><p><strong>Conclusions: </strong>These findings support the role of CSPG4 overexpression and mutant active BRAF-in promoting increased contact guidance. The results are discussed in terms of expanding what is known about the potential tumor biology and clinical implications of CSPG4-related impact on malignant invasion during early phases of melanoma progression.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s12195-025-00882-x.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"19 1","pages":"61-72"},"PeriodicalIF":5.0,"publicationDate":"2025-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13031561/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147572460","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":"Endocytic Pathways and Actin Remodeling Mediate Everolimus-Induced VE-Cadherin Disorganization and Barrier Dysfunction.","authors":"Ken D Brandon, Yoshi Chettri, Azkah Anjum, Kimberly M Stroka","doi":"10.1007/s12195-025-00881-y","DOIUrl":"10.1007/s12195-025-00881-y","url":null,"abstract":"<p><strong>Purpose: </strong>VE-cadherin is a key component of endothelial adherens junctions, and its disorganization contributes to vascular dysfunction. While rapamycin analogs like everolimus (EVL) are clinically linked to endothelial barrier dysfunction (EBD), the underlying molecular mechanisms remain poorly defined. This study investigates how EVL alters VE-cadherin organization, trafficking, cytoskeletal architecture, and barrier function in endothelial cells.</p><p><strong>Methods: </strong>Human umbilical vein endothelial cells (HUVECs) were treated with 500 nM EVL for 4 or 24 h. Junctional VE-cadherin organization was quantified using confocal microscopy and the Junction Analyzer Program. Cytoskeletal changes were assessed via F-actin anisotropy, and pharmacologic inhibitors (chlorpromazine, chloroquine, and brefeldin A) were used to block clathrin-mediated endocytosis, lysosomal degradation, and Golgi trafficking, respectively. Barrier function was evaluated using TEER and 4 kDa FITC-dextran transwell assays.</p><p><strong>Results: </strong>EVL reduced continuous VE-cadherin and increased punctate junctions in a time-dependent but partially reversible manner. Inhibiting endocytosis or lysosomal degradation preserved VE-cadherin continuity, while Golgi disruption blocked recovery. EVL also increased F-actin anisotropy, reflecting enhanced stress-fiber alignment within individual cells, but transiently uncoupled intracellular actin organization from coordinated cytoskeletal alignment across the monolayer. Functionally, EVL decreased TEER and increased dextran permeability by 2.24-2.63-fold, indicating significant barrier disruption.</p><p><strong>Conclusions: </strong>EVL compromises endothelial barrier integrity by promoting VE-cadherin internalization and lysosomal degradation, accompanied by cytoskeletal remodeling and a Golgi-dependent, partial restoration of junctional VE-cadherin. These findings highlight endocytic, degradative, and Golgi-mediated trafficking pathways as key modulators of EVL-induced endothelial barrier dysfunction and provide mechanistic insight into the vascular effects of rapalog-based mTOR inhibition.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":" ","pages":""},"PeriodicalIF":5.0,"publicationDate":"2025-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12915895/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146225622","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":"Disrupted Sarcomere Reorganization of Cardiomyopathy-Prone Human iPSC-Derived Cardiomyocytes on a Dynamic Mechanical Substrate.","authors":"Nhu Y Mai, Xiangjun Wu, Huiyao Liu, Ariel Ash-Shakoor, Huaiyu Shi, Zhuocheng Qu, Patrick T Mather, Xinrui Wang, James H Henderson, Zhen Ma","doi":"10.1007/s12195-025-00880-z","DOIUrl":"10.1007/s12195-025-00880-z","url":null,"abstract":"<p><strong>Introduction: </strong>Approximately 15% of dilated cardiomyopathy (DCM) cases are associated with Bcl2-associated athanogene 3 (BAG3) gene mutations, which play a crucial role in myofilament organization and contractile behavior. Previous studies have highlighted the role of dynamic mechanical stress in myofibril alignment in human-induced pluripotent stem cell-derived cardiomyocytes (hiPSCCMs). In this study, we employed thermo-responsive shape memory polymers (SMPs) to mimic the dynamic mechanical environment of the extracellular matrix (ECM) and investigated their impact on myofibril assembly in healthy wild-type (WT) and BAG3 knockout (BAG3-/-) hiPSC-CMs.</p><p><strong>Methods: </strong>We synthesized Tert-Butyl Acrylate (TBA) and Butyl acrylate (BA)-based SMP substrate. hiPSC-CMs were cultured on 30oC on 40% strained (dynamic) and un-strain (static) SMPs for two days before proceeding with polymer recovery at 37oC. Myofibril components of BAG3 knock-out (KO) and WT CMs were evaluated by immunocytochemistry fluorescent images at 5 hours and 24 hours after triggering the shape changes of the SMP substrate. We quantified Z-lines and M-lines of hiPSC-CMs to evaluate sarcomere remodeling on static and dynamic substrates.</p><p><strong>Results: </strong>Our findings revealed that BAG3-/- hiPSC-CMs exhibited persistent Z-line disruption in sarcomeres compared with WT hiPSC-CMs, but M-line structures were less sensitive to mechanical stress at 5 hours, highlighted a temporal distinction in the assembly and regulation of Z-lines over M-lines. While no significant changes were detected at 5 hours, BAG3-/- CMs exhibited similar impairments in M-line organization as seen with Z-lines. These findings suggest that M-lines in BAG3-/- CMs display heightened sensitivity to dynamic mechanical actuation, but this phenotype emerges only after prolonged culture.</p><p><strong>Conclusions: </strong>This study highlights the interplay between genetic deficiency and mechanical stress to facilitate disease progression in BAG3-associated DCM.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s12195-025-00880-z.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"18 6","pages":"661-672"},"PeriodicalIF":5.0,"publicationDate":"2025-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12664868/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145653761","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":"Rotator Cuff Repair: Lessons from Immune Strategies, 3D Biofabrication and In Vivo Testing.","authors":"Yordan Sbirkov, Atanas Valev, Murad Redzheb, Furkan Bülbül, Feza Korkusuz, Denitsa Docheva, Victoria Sarafian","doi":"10.1007/s12195-025-00879-6","DOIUrl":"https://doi.org/10.1007/s12195-025-00879-6","url":null,"abstract":"<p><strong>Background: </strong>Rotator cuff injuries are the most common type of tendinopathies affecting up to 10% of young adults and more than 60% of the elderly. Tendons have notoriously limited regenerative capacity which is attributed to their low vascularisation and low cell-to-tissue ratio. That leads to an inefficient repair process resulting in fibrotic scar tissue with poorer mechanical properties. Recent advances in tissue engineering and biofabrication techniques have been anticipated with great hope in the field of regenerative medicine.</p><p><strong>Methods: </strong>In this review, we discuss the insights gained from immune-based strategies, 3D biofabrication, and in vivo testing in the context of rotator cuff repair. Particular emphasis is placed on in vivo studies that bridge the gap between laboratory innovation and clinical translation, outlining promising avenues for future therapeutic development.</p><p><strong>Results: </strong>Regardless of the huge progress in in vitro modelling and in vivo healing of RCTs in animals, clinical translation has not yet succeeded because mechanical loading, and chronic inflammation in humans are hard to recapitulate. Biologic variability, regulatory complexity, and poor reproducibility also slow down translation to the clinic.</p><p><strong>Conclusion: </strong>With a number of encouraging results so far, multidisciplinary research will continue to elucidate the complex biological processes in terms of (1) immune and tendon cell engagement and modulation, natural matrix deposition and remodelling; (2) material mimicry regarding topography, anisotropic, gradient continuity, biomechanical properties; and (3) in vivo behaviour in structural reconstruction and functional performance in the long-term. Such integrative approach is essential to overcome today's limitations in RC repair and to outline next-generation strategies to achieve improved clinical outcomes.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"18 6","pages":"549-575"},"PeriodicalIF":5.0,"publicationDate":"2025-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12664878/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145653708","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}