Tissue Barriers最新文献

{"title":"Integrated transcriptomic and functional characterization of Claudin-1 reveals its oncogenic and immunomodulatory roles in pancreatic ductal adenocarcinoma.","authors":"Anju Surendranath, Yogain Taank, Saira Hamid, Tharini Karthikeyan, Ikhlak Ahmed, Imadeldin Elfaki, Rashid Mir, Rakesh Kumar, Sameer Mirza, Mayank Singh, Shahab Uddin, Ammira S Al-Shabeeb Akil, Muzafar A Macha, Punita Dhawan, Ajaz A Bhat","doi":"10.1080/21688370.2026.2648150","DOIUrl":"10.1080/21688370.2026.2648150","url":null,"abstract":"<p><p>Pancreatic ductal adenocarcinoma (PDAC) remains among the deadliest malignancies, driven by its invasive nature and lack of effective biomarkers. Disruption of the epithelial barrier, mediated by tight junction components, is a critical yet underexplored contributor to PDAC progression. Claudins, integral regulators of tight junction integrity, display altered expression across cancers, but their prognostic and immunomodulatory roles in PDAC remain unclear. We performed an integrative analysis of 177 RNA-Seq datasets from TCGA and GTEx to characterize Claudin family alterations in PDAC. Differential expression, copy number variation, methylation, and co-expression networks were analyzed alongside clinical and survival data. Prognostic significance was assessed using Kaplan - Meier and Cox regression analyses, while immune cell infiltration was examined using deconvolution algorithms. Functional validation of Claudin-1 was conducted in Capan-1 cells using CRISPR/Cas9 knockout, followed by proliferation, wound-healing, and Western blot assays. Ten Claudin genes were significantly dysregulated, with Claudin-1 and Claudin-4 frequently amplified and associated with advanced stage and poor survival. High Claudin-1 expression correlated with reduced immune infiltration, indicating an immune-excluded phenotype characterized by immune cells retained in the tumor stroma but largely absent from the tumor parenchyma. Claudin-1 knockout markedly inhibited proliferation, migration, and EMT, evidenced by downregulation of Snail and Slug and restoration of E-cadherin expression. This integrative transcriptomic and functional study identifies Claudin-1 as a key driver of PDAC aggressiveness and immune modulation. These findings establish Claudin-1 as a promising prognostic biomarker and therapeutic target for restoring epithelial integrity and counteracting immune evasion in pancreatic cancer.</p>","PeriodicalId":23469,"journal":{"name":"Tissue Barriers","volume":" ","pages":"2648150"},"PeriodicalIF":3.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13540116/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147677217","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of local versus systemic exosomes administration on both testes in rat model of testicular ischemic reperfusion injury: shedding light on blood testis barrier.","authors":"Manar Fouli Gaber Ibrahim, Elshymaa A Abdel-Hakeem, Heba A Shawky, Soha Abdelkawy Abdelwahab, Sahar A Mokhemer","doi":"10.1080/21688370.2025.2532195","DOIUrl":"10.1080/21688370.2025.2532195","url":null,"abstract":"<p><p>Exosomes^, (EXs) competence in reproductive medicine has been proven, particularly in cases of ischemia reperfusion (IR) injury models. Testicular torsion is a major clinical concern affecting male fertility and results in ischemic reperfusion injury. To the best of our knowledge, up to date, comparative studies using different routes of EXs application have not been investigated. Thus, we aimed to investigate the effect of local versus systemic EXs administration on both testes; ipsilaterally and contralaterally and to explore the involved underlying mechanisms. Rats were allocated into four groups; the control, the ischemia reperfusion group (IR group) in which unilateral testicular IR was performed, then rats were subdivided equally into two subgroups; IR-ipsi group; in which the ipsilateral testes were investigated and the IR-contra group in which the contralateral testes were examined. Group III and group IV in which unilateral IR was performed, then injected intravenously or intratesticularly consequently with EXs and finally divided in to two subgroups IR-ipsi and IR-contra. The results revealed that testicular IR resulted in functional and structural disorders with reduction in serum testosterone and sperm indices. Interestingly, both testes showed structural histopathological changes and defects in blood testis barrier. There was also an increase in oxidative stress, inflammatory, and apoptosis markers. EXs administration improved the affected parameters probably via modulation of TLR4/MyD88/NF-κB signaling pathway. In conclusion, EXs defended against testicular IR injury. Intratesticular administration had better effects on the ipsilateral testes, whereas Intravenous route had more obvious better effects on the contralateral ones.</p>","PeriodicalId":23469,"journal":{"name":"Tissue Barriers","volume":" ","pages":"2532195"},"PeriodicalIF":3.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12959186/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144675827","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Involvement of cellular and enzymatic aspects in the complexity of diabetic neuropathy.","authors":"Anjali Gupta, Tanya Gupta, Thakur Gurjeet Singh, Randhir Singh","doi":"10.1080/21688370.2025.2581880","DOIUrl":"10.1080/21688370.2025.2581880","url":null,"abstract":"<p><p>Diabetic neuropathy (DN) is a multifaceted and progressive complication of diabetes mellitus, characterized by functional and structural damage to peripheral, autonomic, and sensory nerves. Despite its high prevalence and debilitating consequences, current therapeutic approaches remain largely symptomatic, with limited disease-modifying strategies available. The pathogenesis of DN is driven by a complex network of molecular, cellular, and enzymatic interactions, primarily instigated by chronic hyperglycemia. This review unravels the intricate molecular and cellular crosstalk underlying DN, emphasizing the roles of specific cellular and enzymatic mediators in disease progression. Key cellular players, including neurons, Schwann cells, satellite glial cells, macrophages, and bone marrow-derived cells, orchestrate and respond to pathogenic stimuli, contributing to neuroinflammation, demyelination, and axonal degeneration. Chronic hyperglycemia activates several enzymatic pathways that exacerbate oxidative stress, mitochondrial dysfunction, and vascular impairment. Among the pivotal enzymes involved is aldose reductase, which drives the polyol pathway and sorbitol accumulation; diacylglycerol (DAG)-mediated protein kinase C (PKC), linked to vascular dysfunction; poly(ADP-ribose) polymerase (PARP), which amplifies DNA damage responses; and endogenous antioxidants, such as superoxide dismutase (SOD), catalase, and glutathione peroxidase, whose dysregulation further fuels oxidative injury. Additionally, growth factors (e.g. NGF, IGF-1, and VEGF), along with metabolic regulators (like AMPK), play pivotal roles in maintaining neuronal growth, survival, and function by modulating cellular energy homeostasis, oxidative balance, and inflammatory responses. By examining these interconnected molecular mechanisms, this review highlights potential therapeutic targets and proposes future directions for mechanism-based interventions aimed at halting or reversing the progression of diabetic neuropathy.</p>","PeriodicalId":23469,"journal":{"name":"Tissue Barriers","volume":" ","pages":"2581880"},"PeriodicalIF":3.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13228964/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145422777","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Current therapeutic strategies in the management of diabetic foot ulcers.","authors":"Aniket Gupta, Rishabh Chalotra, Kajal Sharma, Anjali Gupta, Shivani Chib, Rohini Agrawal, Thakur Gurjeet Singh, Randhir Singh","doi":"10.1080/21688370.2025.2611185","DOIUrl":"10.1080/21688370.2025.2611185","url":null,"abstract":"<p><p>Diabetic foot ulcer (DFU) is a chronic and predominantly microvascular and neuropathic complication in more severe or chronic cases of diabetes mellitus. It is characterized by chronic nonhealing wounds, vascular impairment, and delayed healing process, leading to severe complications, limb amputations, and increased mortality. With an annual incidence rate of approximately 2%, DFU poses a significant global healthcare and economic burden. Despite its prevalence, current treatment options remain limited, necessitating the urgent need for a deeper understanding of the underlying molecular pathways or mechanisms to develop effective therapeutic strategies. Present work is emphasized on molecular mechanisms involved in pathogenesis of DFU and current and emerging therapeutic interventions for the treatment of DFU. Due to its high prevalence, multifaceted pathophysiology, and significant healthcare and economic burden, a thorough understanding of molecular pathways underlying DFU is essential to develop precise therapeutic interventions to improve clinical outcome and reduce the healthcare burden associated with DFU. Several therapeutic interventions have been utilized, like modulators of key signaling pathways (Wnt/β-catenin, PI3K/Akt/mTOR, JAK/STAT, and Notch), repurposed pharmacological agents (e.g. metformin, colchicine, deferoxamine, and lithium carbonate), and advanced local treatments such as bioactive hydrogels and next-generation dressings. Furthermore, regenerative approaches like gene therapy, stem cell transplantation, therapeutic peptides, and 3D-bioprinted adipose tissue constructs provide a promising strategy for restoring tissue integrity and promoting healing.</p>","PeriodicalId":23469,"journal":{"name":"Tissue Barriers","volume":" ","pages":"2611185"},"PeriodicalIF":3.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13540221/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145865714","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Potential of exosomes for targeted therapy in neuroinfectious disease management: crossing the blood-brain barrier.","authors":"Sarad Pawar Naik Bukke, Shikha Yadav, Shatrudhan Prajapati, Satla Shobha Rani, M Sunitha Reddy, Chandrashekar Thalluri, Ananda Kumar Chettupalli, Narayana Goruntla, Bhupalam Pradeepkumar, Zohre Eftekhari, Yasodha Krishna Janapati, Alrazi Eisa Shogar, Tadele Mekuriya Yadesa","doi":"10.1080/21688370.2025.2607157","DOIUrl":"10.1080/21688370.2025.2607157","url":null,"abstract":"<p><p>Neuroinfectious diseases such as meningitis, encephalitis, and myelitis continue to be a significant health issue especially in low- and middle-income nations where the timely identification and successful treatment are mostly not yet available. There is also the complicating factor of the restrictive nature of the blood-brain barrier (BBB) which greatly limits the passage of antimicrobial and anti-inflammatory agents through into the central nervous system. Exosomes which are nano-sized extracellular vesicles released by a vast variety of cells have been proposed as a promising solution to overcome this barrier because of their natural biocompatibility, low immunogenicity and capacity to enter the BBB by receptor-mediated, adsorptive-mediated and carrier-mediated processes. This review critically discusses the structural and functional dynamics of the BBB in infection, the recent discoveries in the exosome trafficking pathways, and the diagnostic and therapeutic role of exosomes in infection, including infections related to HIV-associated neurocognitive disorders, tuberculous meningitis, cryptococcal meningitis, neurotoxoplasmosis. Special consideration will be given to engineered exosomes and how they could be used to improve targeted delivery of drugs, decrease systemic toxicity and offer minimally invasive biomarkers platforms to detect disease earlier. He/she points possible solutions to CNS infections by combining mechanistic understanding with newly acquired pre-clinical and clinical results, this review emphasizes the increasing promise of exosome-based nanomedicine as a transformational technology.</p>","PeriodicalId":23469,"journal":{"name":"Tissue Barriers","volume":" ","pages":"2607157"},"PeriodicalIF":3.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13540134/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145834836","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Hyperglycaemia perturbs blood-brain barrier integrity through its effects on endothelial cell characteristics and function.","authors":"Arshad Hashmat, Jingyuan Ya, Rais Kadir, Mansour Alwjwaj, Ulvi Bayraktutan","doi":"10.1080/21688370.2024.2350821","DOIUrl":"10.1080/21688370.2024.2350821","url":null,"abstract":"<p><p>Breakdown of blood-brain barrier (BBB) represents a key pathology in hyperglycemia-mediated cerebrovascular damage after an ischemic stroke. As changes in the level and nature of vasoactive agents released by endothelial cells (ECs) may contribute to BBB dysfunction, this study first explored the specific impact of hyperglycemia on EC characteristics and secretome. It then assessed whether secretome obtained from ECs subjected to normoglycaemia or hyperglycemia might regulate pericytic cytokine profile differently. Using a triple cell culture model of human BBB, composed of brain microvascular EC (BMEC), astrocytes and pericytes, this study showed that exposure to hyperglycemia (25 mM D-glucose) for 72 h impaired the BBB integrity and function as evidenced by decreases in transendothelial electrical resistance and increases in paracellular flux of sodium fluorescein. Dissolution of zonula occludens-1, a tight junction protein, and appearance of stress fibers appeared to play a key role in this pathology. Despite elevations in angiogenin, endothelin-1, interleukin-8 and basic fibroblast growth factor levels and a decrease in placental growth factor levels in BMEC subjected to hyperglycemia vs normoglycaemia (5.5 mM D-glucose), tubulogenic capacity of BMECs remained similar in both settings. Similarly, pericytes subjected to secretome obtained from hyperglycemic BMEC released higher quantities of macrophage migration inhibitory factor and serpin and lower quantities of monocyte chemoattractant protein-1, intercellular adhesion molecule, interleukin-6 and interleukin-8. Taken together these findings indicate the complexity of the mechanisms leading to BBB disruption in hyperglycemic settings and emphasize the importance of endothelial cell-pericyte axis in the development of novel therapeutic strategies.</p>","PeriodicalId":23469,"journal":{"name":"Tissue Barriers","volume":" ","pages":"2350821"},"PeriodicalIF":4.0,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11970753/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140871984","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Slow down my beating heart: induction of cardiac fibrosis by Iroquois homeobox 2.","authors":"Madeline Petrikas, Rebecca A Wingert","doi":"10.1080/21688370.2024.2309036","DOIUrl":"10.1080/21688370.2024.2309036","url":null,"abstract":"<p><p>Cardiovascular diseases are a significant global health challenge and pervasive cause of mortality worldwide. Heart failure due to cardiovascular disease is characterized by the inability of the heart to pump blood efficiently to meet the metabolic demands of the body. The pathophysiology of heart failure involves myocardial remodeling due to excessive deposition of extracellular matrix proteins by cardiac myofibroblasts - structural changes which impair contractility, reduce compliance, and ultimately reduce stroke volume. Now, a recent report has uncovered an essential role for Iroquois homeobox 2 in the transcriptional regulation of cardiac fibrosis, illuminating new mechanistic insights that can be applied to developing future clinical therapies.</p>","PeriodicalId":23469,"journal":{"name":"Tissue Barriers","volume":" ","pages":"2309036"},"PeriodicalIF":4.0,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11875463/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139571373","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fibroblasts/three-dimensional scaffolds complexes promote wound healing in rats with skin defects.","authors":"Ting Jiang, Qiang Liu, Er-Chang Xu, Si-Yu He, Hong-Yan Liu, Chao Tian, Lan-Fang Zhang, Ze-Long Yang","doi":"10.1080/21688370.2024.2334544","DOIUrl":"10.1080/21688370.2024.2334544","url":null,"abstract":"<p><p>We aim to construct a three-dimensional nano-skin scaffold material in vitro and study its promoting effect on wound healing in vivo. In this study, hybrid constructs of three-dimensional (3D) scaffolds were successfully fabricated by combination of type I collagen (COL-1) and polylactic-glycolic acid (PLGA). Fibroblasts and human umbilical cord mesenchymal stem cells (hUCMSCs) were used to implanted into 3D scaffolds and constructed into SD skin scaffolds in vitro. Finally, the fibroblasts/scaffolds complexes were inoculated on the surface of rat wound skin to study the promoting effect of the complex on wound healing. In our study, we successfully built a 3D scaffold, which had a certain porosity. Meanwhile, the content of COL-1 in the cell supernatant of fibroblast/scaffold complexes was increased. Furthermore, the expression of F-actin, CD105, integrin β, VEGF, and COL-1 was up-regulated in hUCMSC/scaffold complexes compared with the control group. In vivo, fibroblast/scaffold complexes promoted wound healing in rats. Our data suggested that the collagen Ⅳ and vimentin were elevated and collagen fibers were neatly arranged in the fibroblast/scaffold complex group was significantly higher than that in the scaffold group. Taken together, fibroblast/scaffold complexes were expected to be novel materials for treating skin defects.</p>","PeriodicalId":23469,"journal":{"name":"Tissue Barriers","volume":" ","pages":"2334544"},"PeriodicalIF":4.0,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11875492/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140307054","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The correlation between fecal microbiota profiles and intracellular junction genes expression in young Iranian patients with celiac disease.","authors":"Mohadeseh Mahmoudi Ghehsareh, Nastaran Asri, Fahimeh Sadat Gholam-Mostafaei, Hamidreza Houri, Flora Forouzesh, Shokoufeh Ahmadipour, Somayeh Jahani-Sherafat, Mohammad Rostami-Nejad, Pasquale Mansueto, Aurelio Seidita","doi":"10.1080/21688370.2024.2347766","DOIUrl":"10.1080/21688370.2024.2347766","url":null,"abstract":"<p><p>Celiac disease (CD) is characterized by the disruption of the intestinal barrier integrity and alterations in the microbiota composition. This study aimed to evaluate the changes in the fecal microbiota profile and mRNA expressions of intracellular junction-related genes in pediatric patients with CD compared to healthy controls (HCs). Thirty treated CD patients, 10 active CD, and 40 HCs were recruited. Peripheral blood (PB) and fecal samples were collected. Microbiota analysis was performed using quantitative real-time PCR (qPCR) test. The mRNA expressions of ZO-1, occludin, β-catenin, E-cadherin, and COX-2 were also evaluated. In active and treated CD patients, the PB expression levels of ZO-1 (<i>p</i> = 0.04 and 0.002, respectively) and β-catenin (<i>p</i> = 0.006 and 0.02, respectively) were lower than in HCs. PB Occludin's level was upregulated in both active and treated CD patients compared to HCs (<i>p</i> = 0.04 and 0.02, respectively). However, PB E-cadherin and COX-2 expression levels and fecal mRNA expressions of ZO-1, occludin, and COX-2 did not differ significantly between cases and HCs (P˃0.05). Active CD patients had a higher relative abundance of the <i>Firmicutes</i> (<i>p</i> = 0.04) and <i>Actinobacteria</i> (<i>p</i> = 0.03) phyla compared to treated subjects. The relative abundance of <i>Veillonella</i> (<i>p</i> = 0.04) and <i>Staphylococcus</i> (<i>p</i> = 0.01) genera was lower in active patients in comparison to HCs. Researchers should explore the precise impact of the gut microbiome on the molecules and mechanisms involved in intestinal damage of CD. Special attention should be given to <i>Bifidobacteria</i> and Enterobacteriaceae, as they have shown a significant correlation with the expression of tight junction-related genes.</p>","PeriodicalId":23469,"journal":{"name":"Tissue Barriers","volume":" ","pages":"2347766"},"PeriodicalIF":4.0,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11970794/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140870223","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of small extracellular vesicles derived from normoxia- and hypoxia-treated prostate cancer cells on the submandibular salivary gland epithelium <i>in vitro</i>.","authors":"Ana Špilak, Andreas Brachner, Heinz-Peter Friedl, Adrián Klepe, Christa Nöhammer, Winfried Neuhaus","doi":"10.1080/21688370.2024.2347062","DOIUrl":"10.1080/21688370.2024.2347062","url":null,"abstract":"<p><p>Small extracellular vesicles (sEVs) are an important part of intercellular communication. They are phospholipid bilayer particles that carry active biomolecules such as proteins, various nucleic acids, and lipids. In recipient cells, sEVs can alter cellular functions, including cancer development and premetastatic niche formation in distant organs. Moreover, sEVs can carry cancer-specific features, which makes them promising biomarker candidates. However, the interactions of sEVs with biological barriers and consequences thereof, are not clarified yet. The blood-saliva barrier is crucial for preventing the entry of pathogens and (in)organic substances into the bloodstream, as well as molecule filtration from blood to saliva. The effects of brain derived DU145 prostate cancer (PCa) sEVs on a human submandibular salivary gland barrier (SSGB) <i>in vitro</i> were investigated. Small EVs were harvested from normoxic (N, atmospheric O₂) or hypoxic (H, 1% O₂) conditions, fluorescently labeled with CellTracker^TM Orange and thoroughly characterized. HTB-41 B2 cells were used as SSGB model cultured on 24-well ThinCert® inserts. After model optimization indicating effects of serum and serum-sEVs on barrier properties, PCa sEVs were applied to the basolateral (blood) side in either 10% serum, or serum-free conditions, and barrier integrity was continuously monitored for 40 hours. This study found that H and N PCa sEVs were uptaken by the SSGB <i>in vitro</i> model in similar quantities regardless of the media composition in the basolateral compartment. Permeation of fluorescent PCa sEVs into the apical compartment was not detectable with the applied methods. However, treatment with H and N sEVs under different serum conditions revealed distinct molecular clusters after hierarchical analysis of mRNA data measured by high-throughput qPCR, which were partly reflected at the protein level. For example, serum-reduction dependent decrease of barrier properties was accompanied with the decrease of CDH1 or Claudin-7 expression. Interestingly, the presence of H sEVs significantly increased the number of sEV-sized particles in the apical compartment of the SSGB model compared to basolaterally added N sEVs. This functional effect on the number of particles in the saliva (apical) compartment induced by different sEVs applied in the blood (basolateral) compartment might be a new approach to understand one possible mechanism how differences of salivary EVs might occur which then could be used as biomarker.</p>","PeriodicalId":23469,"journal":{"name":"Tissue Barriers","volume":" ","pages":"2347062"},"PeriodicalIF":4.0,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11875469/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140892770","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}