American journal of physiology. Lung cellular and molecular physiology最新文献

{"title":"Correction for Railwah et al., volume 319, 2020, p. L1021-L1035.","authors":"","doi":"10.1152/ajplung.00207.2020_COR","DOIUrl":"https://doi.org/10.1152/ajplung.00207.2020_COR","url":null,"abstract":"","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":"331 4","pages":"L406-L408"},"PeriodicalIF":3.8,"publicationDate":"2026-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149102791","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The role of myeloid immune cells in lung epithelial repair.","authors":"Jorge A Masso-Silva, Avnee J Kumar","doi":"10.1152/ajplung.00045.2026","DOIUrl":"10.1152/ajplung.00045.2026","url":null,"abstract":"<p><p>The lung epithelium is continuously exposed to injury via pathogens and pollutants, and its capacity for healthy, nonpathological repair is essential to maintain structure and function. In homeostatic repair, type 2 alveolar epithelial (AT2) cells-the stem cells of the alveolar epithelium-differentiate into type 1 alveolar epithelial (AT1) cells in a timed, proportionate process restoring normal epithelial architecture. When dysregulated, this process can result in pathological remodeling, potentially resulting in disease. Myeloid immune cells are among the first responders to lung injury and remain engaged across the full arc of the repair response, yet their contribution to healthy epithelial repair remains underappreciated. In this mini-review, we evaluate current evidence on the role of myeloid immune cells-mononuclear phagocytes and granulocytes-in nonpathological lung epithelial repair. Macrophages promote epithelial proliferation, migration, and barrier restoration through release of soluble mediators including oncostatin M, trefoil factor family, interleukin-1β, and tumor necrosis factor alpha, as well as through extracellular vesicle-mediated communication. Neutrophils contribute through β-catenin signaling, efferocytosis-driven macrophage reprogramming, matrix metalloproteinase-9-dependent remodeling, and direct transfer of microRNA-223 to epithelial cells. Eosinophils, mast cells, and basophils have emerging roles through granule-derived mediators and cross talk with other myeloid cells. Understanding the mechanisms by which myeloid cells promote healthy repair and how dysregulation drives pathological remodeling may uncover therapeutic targets for restoring epithelial function after lung injury.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":"L387-L395"},"PeriodicalIF":3.8,"publicationDate":"2026-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13628908/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148944138","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Aberrant expression of the PREX2-MAGI2 axis modulates myofibroblast differentiation in pulmonary fibrosis.","authors":"Yozo Sato, Koichiro Kamio, Yasuhiro Terasaki, Keisuke Yoshida, Naoyuki Kuse, Kuniko Matsuda, Yusuke Kajimoto, Kenichiro Atsumi, Shota Kaburaki, Toru Tanaka, Takeru Kashiwada, Masaru Matsumoto, Akihiko Miyanaga, Yosuke Tanaka, Yoshinobu Saito, Kazuo Kasahara, Kazufumi Honda, Masahiro Seike","doi":"10.1152/ajplung.00168.2025","DOIUrl":"10.1152/ajplung.00168.2025","url":null,"abstract":"<p><p>Myofibroblast differentiation is a pivotal event in idiopathic pulmonary fibrosis (IPF) and is driven by transforming growth factor-β1 (TGF-β1)-dependent phosphatase and tensin homolog deleted on chromosome 10 (PTEN) destabilization; however, the intermediate regulators remain incompletely defined. We tested three lung fibroblast cell lines (1 normal and 2 IPF derived) and identified the phosphatidylinositol 3,4,5-trisphosphate-dependent Rac exchanger 2 (PREX2)-membrane-associated guanylate kinase inverted 2 (MAGI2) as a critical regulator of myofibroblast differentiation. Loss-of-function analysis of MAGI2 in normal human lung fibroblasts revealed a shift toward a myofibroblast phenotype characterized by abundant α-smooth muscle actin (α-SMA) expression. In vitro experiments revealed increased PREX2 expression and decreased MAGI2 expression in IPF-derived lung fibroblasts compared with those in normal lung fibroblasts. Consistent with this finding, in vivo experiments using bleomycin-induced murine pulmonary fibrosis revealed PREX2 upregulation in contrast to MAGI2 downregulation with the progression of lesions. Notably, PREX2 depletion in IPF-derived cells enhanced MAGI2 expression, mitigating TGF-β1-induced α-SMA expression. Conversely, PREX2 overexpression promoted myofibroblast differentiation, as evidenced by MAGI2 downregulation followed by marked α-SMA expression. The PREX2-targeting suppressive <i>miR</i>-<i>338</i>-<i>3p</i> upregulated MAGI2 expression in vitro and ameliorated bleomycin-induced murine pulmonary fibrosis in vivo. Immunohistochemical staining of PREX2 on lung samples obtained from patients with IPF delineated that PREX2 was exclusively expressed in the fibroblastic foci of IPF-affected lungs. Fluorescence immunostaining revealed PREX2 colocalization with α-SMA in myofibroblasts within fibroblastic foci, corroborating our hypothesis. These findings suggest that MAGI2 acts as a negative regulator of myofibroblast differentiation by stabilizing PTEN, whereas PREX2 is aberrantly expressed in the fibrotic milieu, negatively modulating MAGI2, thereby accelerating pulmonary fibrosis.<b>NEW & NOTEWORTHY</b> In the fibrotic milieu, upregulated phosphatidylinositol 3,4,5-trisphosphate-dependent Rac exchanger 2 (PREX2) and downregulated membrane-associated guanylate kinase inverted 2 (MAGI2) expression were observed as evidenced by in vivo and in vitro analyses. PREX2 was exclusively expressed in fibroblastic foci in lung samples derived from patients with idiopathic pulmonary fibrosis (IPF) and was colocalized with α-smooth muscle actin (α-SMA) in myofibroblasts within these foci. Thus, the aberrantly expressed PREX2-MAGI2 axis modulates myofibroblast differentiation in IPF, whereby PREX2 negatively modulates MAGI2, destabilizing phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and ultimately accelerating pulmonary fibrosis.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":"L359-L374"},"PeriodicalIF":3.8,"publicationDate":"2026-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148787160","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"HYPOXIC INTENSITY REVEALS DISTINCT CAROTID BODY O2 SENSING MECHANISMS.","authors":"Nanduri R Prabhakar, Ying-Jie Peng","doi":"10.1152/ajplung.90003.2026","DOIUrl":"10.1152/ajplung.90003.2026","url":null,"abstract":"<p><p>Carotid bodies (CBs) sense arterial blood O₂ levels. Hypoxemia activates the carotid sinus nerve (CSN) afferent activity to elicit cardiorespiratory reflexes to maintain tissue oxygenation. This brief review presents emerging evidence suggesting that CBs employ distinct O₂-sensing mechanisms depending on the severity of hypoxia. During physiologically relevant moderate hypoxia (<i>p</i>O₂ ~40 mmHg), glomus cells use heme oxygenase-2 (HO-2)/carbon monoxide (CO)/cystathionine-γ-lyase (CSE)/hydrogen sulfide (H₂S) signaling pathway to stimulate CSN activity and breathing response to hypoxia. Basal CO restrains CSN activity during normoxia, whereas hypoxia reduces CO and stimulates CSN activity through H₂S. Olfactory receptor 78 (Olfr78) is required for H₂S-mediated CSN activation. CO-H₂S pathway contributes to cardiorespiratory physiological adaptations to sustained hypoxia and to the pathophysiology of diseases characterized by carotid body (CB) hyperactivity. In contrast, glomus cell responds to severe hypoxia (<i>p</i>O₂ ~5-15 mmHg) through mitochondrial complex I (NDUFS2) and complex III. Thus, CBs appear to employ severity-dependent hypoxic sensing mechanisms wherein moderate hypoxia engages the CO-H₂S pathway to activate CSN activity and drive homeostatic cardiorespiratory responses. Severe hypoxia primarily alters mitochondrial function and glomus cell metabolism. Physiological relevance of severe-hypoxia remains uncertain, because severe low O₂ levels suppresses both CB, CSN activity and ventilation.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2026-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13673981/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149102703","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dynamics of Airway Macrophage Responses and Lung Function Changes after Ozone Exposure in Humans.","authors":"Jared Radbel, Emily R Stevenson, Raymond C Rancourt, Kathleen G Black, Adriana De Resende, Derek Ge, Aesha Jobanputra, Jessica Cervelli, Jeffrey D Laskin, Howard M Kipen, Andrew J Gow, Debra L Laskin","doi":"10.1152/ajplung.00120.2026","DOIUrl":"10.1152/ajplung.00120.2026","url":null,"abstract":"<p><p>Ozone is a ubiquitous ambient air pollutant known to cause lung injury and exacerbate pulmonary disease. While rodent studies suggest that macrophages contribute to both ozone-induced lung injury and subsequent repair, their specific roles in humans remain unclear. We hypothesized that recovery from ozone-induced lung injury in humans depends on a dynamic balance between macrophage subsets. To test this hypothesis, 44 healthy male and female adults (18-40 years old) were enrolled in a randomized, single-blind cross-over study involving paired 3 h exposures to ozone (200 ppb) and filtered air. Participants were evaluated 24, 48, or 72 h after each exposure, at which time induced sputum was collected for myeloid cell analysis. Pulmonary function testing was performed before and immediately after each exposure and repeated at the follow-up visit. Compared with filtered air, ozone exposure resulted in a significant increase in the percentage of monocytes at 24 h and monocyte-derived macrophages at 24-72 h, while neutrophil percentages decreased at 48-72 h. Immediately after ozone exposure, participants showed significant reductions in FEV₁, FVC, and the FEV₁/FVC ratio, along with increases in distal airway resistance measures (R5 and R10). These functional impairments were not evident at 24-72 h. Improvements in large airway lung function from 24-72 h post-ozone exposure correlated with a higher airway monocyte-derived macrophage-to-neutrophil ratio, while improvement in R5 associated with a higher monocyte-derived macrophage-to-monocyte ratio. These findings provide new insight into the temporal sequence of human airway macrophage responses following ozone-induced lung injury and highlight coordinated macrophage dynamics that may contribute to recovery.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2026-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149057008","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Immunomodulation of the Innate Host Response by Mesenchymal-Derived Versican during Influenza A Virus Infection.","authors":"Jourdan E Brune, Mary Y Chang, Fengying Tang, Cecilia Lopez-Martinez, Stephen R Reeves, Christina K Chan, Peter Waldron, David F Boyd, Sina A Gharib, Paul G Thomas, William A Altemeier, Charles W Frevert","doi":"10.1152/ajplung.00265.2025","DOIUrl":"10.1152/ajplung.00265.2025","url":null,"abstract":"<p><p>Viral and bacterial lung infections place a significant burden on public health. Versican, an extracellular matrix (ECM) chondroitin sulfate proteoglycan, coordinates the innate immune response in multiple experimental models. Versican's potential as an immunomodulatory molecule makes it a promising therapeutic target for controlling the host's immune response to lung infection. However, versican's contribution to lung inflammation, injury, and immune cell activity during influenza A virus (IAV) infection represents a critical knowledge gap. To address our central hypothesis that mesenchymal-derived versican is pro-inflammatory and enhances the innate immune response to IAV infection, we generated a tamoxifen-inducible mouse deficient in mesenchymal-derived versican (B6. Col1a2-Cre^ERT+/-/Vcan^tm1.1Cwf, Col1a2/Vcan^-/-). We report that mesenchymal-derived versican plays a critical role in neutrophil, monocyte, and dendritic cell migration into the lungs and airways early in IAV infection. Intriguingly, mesenchymal-derived versican deficiency had the most substantial negative impact on neutrophil emigration into the lungs. We found that neutrophils were less adhesive to the ECM of Col1a2/Vcan^-/- mouse lung fibroblasts (mLFs), which had a significant decrease in versican compared to wild-type mLFs. Additionally, Col1a2/Vcan^-/- mLFs treated with poly(I:C) <i>in vitro</i> have reduced cell-associated hyaluronan. These findings suggest that fibroblast-derived versican is necessary for adhesion to lung fibroblasts by neutrophils as they transit into the lung interstitium and airways from the pulmonary vasculature. Our findings demonstrate that mesenchymal-derived versican is a key regulator of the early host immune responses to IAV.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2026-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148975522","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Adenoviral-Mediated SIRT1 Expression Restores Lung Architecture and Reduces Fibrotic Progression in Experimental Models.","authors":"Liliana Faridi Saavedra-Salazar, Nayabei Ruiz Ramirez, Hugo Christian Monroy-Rami Rez, Ana Sandoval-Rodríguez, Víctor Ruiz, Marina Galicia-Moreno, Blanca Estela Bastidas-Ramirez, Jorge Gutiérrez-Cuevas, Luis Felipe Jave-Suarez, Juan Armendariz-Borunda, Jesus Garcia-Bañuelos","doi":"10.1152/ajplung.00111.2026","DOIUrl":"https://doi.org/10.1152/ajplung.00111.2026","url":null,"abstract":"<p><p><b>Background:</b> Pulmonary fibrosis (PF) involves remodeling of lung architecture and limited therapeutic options are available. Sirtuin 1 (SIRT1), an epigenetic regulator, has emerged as a promising antifibrotic target due to its role in apoptosis, inflammation, and chromatin remodeling. <b>Methods:</b> We evaluated the effects of a recombinant adenovirus expressing human SIRT1 (Ad-SIRT1) <i>both in vitro</i> and <i>in vivo</i>. Human alveolar epithelial cells (A549) were transduced with Ad-SIRT1 or AdGFP and subjected to bleomycin-induced injury. Transduction efficiency, cell viability (MTT assay), SIRT1 localization (immunofluorescence), and expression of acetylated p53 and histone H3K9 (Western blot) were assessed. Pharmacological modulation of SIRT1 was also tested using the activator SRT1720 and the inhibitor EX527. For the <i>in vivo</i> experiments, mice with bleomycin-induced PF received intratracheal Ad-SIRT1 on day 15 after injury. Histological, immunohistochemical and molecular analyses were performed on day 21. <b>Results:</b> Ad-SIRT1-transduced A549 cells showed increased nuclear SIRT1 overexpression which restored viability of bleomycin-injured cells and reduced p53 and H3K9 acetylation, consistent with a protective deacetylating effect. These changes mirrored those obtained with SRT1720 and were reversed by EX527 treatment. <i>In vivo</i>, Ad-SIRT1 attenuated collagen deposition, reduced α-smooth muscle actin (α-SMA) expression, and decreased p53 and H3K9 acetylation. The histological scores were significantly improved compared to those of the fibrotic control group. <b>Conclusion:</b> Adenoviral SIRT1 gene delivery exerts protective effects on the alveolar epithelium <i>in vitro</i> and reverses established PF <i>in vivo</i> through epigenetic regulation of key profibrotic pathways. This dual approach highlights the translational potential of SIRT1 as a therapeutic target for PF.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2026-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148966101","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multi-omics analysis of glucocorticoid receptor crosstalk with Type I and Type II inflammatory signaling in human airway smooth muscle cells.","authors":"Sarah K Sasse, Vineela Kadiyala, Min Chen, Michael R Weaver, Shelly C McCain, Arnav Gupta, Reynold A Panettieri, Anthony N Gerber","doi":"10.1152/ajplung.00281.2025","DOIUrl":"https://doi.org/10.1152/ajplung.00281.2025","url":null,"abstract":"<p><p>Airway smooth muscle (ASM) dysfunction in obstructive airway disease is treated with glucocorticoids. Through RNA-seq analysis of cultured human ASM, we identified repressive effects of dexamethasone, a glucocorticoid, on the baseline expression of a subset of genes that are induced by either IL1B or IL13, which model Type I and Type II inflammation, respectively. ChIP-seq analysis of glucocorticoid receptor (GR) and the p65 subunit of NFkB occupancy indicated canonical motifs for both factors occur at sites of p65 occupancy but did not provide biochemical support for significant repressive tethering between GR and p65. Instead, ATAC-seq revealed significant chromatin remodeling and increased accessibility at binding motifs for the NFkB complex in association with dex + IL1B co-treatment in comparison to IL1B treatment alone. Our data support a competition-based primary repressive effect of glucocorticoids on both IL1B and IL13 signaling and provide evidence for transcriptional cooperation between GR and NFkB on a genome-wide basis in ASM, including at regulatory elements that control expression of anti-inflammatorygenes.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2026-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148928716","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chronic Mixed-Allergen Exposure Induces Pulmonary Arterial Remodeling and Vascular Dysfunction in Mice.","authors":"Mohammad Irshad Reza, Maxwell Mathias, Venkatachalem Sathish","doi":"10.1152/ajplung.00238.2026","DOIUrl":"10.1152/ajplung.00238.2026","url":null,"abstract":"<p><p>Chronic asthma can be associated with secondary pulmonary vascular dysfunction, however, the detailed functional profiling of the asthmatic pulmonary artery remains understudied. In this study, we characterized cardiopulmonary and vascular dysfunction in a chronic mixed allergen (MA) mouse model of asthma. Wild-type mice were intranasally challenged with MA or PBS for four weeks, followed by <i>in vivo</i> respiratory mechanics (flexiVent), <i>ex vivo</i> pulmonary artery wire myography, structural morphometry, and right ventricular analysis (Fulton Index). Chronic MA exposure induced significant airway hyperresponsiveness, evidenced by elevated airway resistance (Rrs) and elastance (Ers) with reduced compliance (Crs). Lung histology revealed increased pulmonary artery medial wall thickness and smooth muscle hyperplasia. Wire myography demonstrated enhanced serotonin (5-HT)-induced vasoconstriction and endothelial dysfunction in MA mice, evidenced by blunted endothelium-dependent relaxation to acetylcholine (ACh) with preserved endothelium-independent relaxation to sodium nitroprusside (SNP). Lastly MA mice had significant right ventricular hypertrophy by Fulton index. Collectively, chronic allergic airway inflammation drives vascular remodeling, endothelial dysfunction, and smooth muscle hyperreactivity in pulmonary arteries. This study demonstrates substantial structural and functional consequences of MA exposure to the pulmonary vasculature that models asthmatic pulmonary vascular dysfunction.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2026-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13637479/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148904771","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Efficient AI-supported quantification of network structures in SEM imaged human mucus: a detailed protocol.","authors":"Yannic Kerkhoff, Hanna Rulff, Paul Wilhelm Mönch, Catharina Gutwein, Aditi Loewe, Sara Timm, Ruth M Urbantat, Kai Ludwig, Rainer Haag, Michael Gradzielski, Marcus A Mall, Britta Siegmund, Matthias Ochs","doi":"10.1152/ajplung.00009.2026","DOIUrl":"10.1152/ajplung.00009.2026","url":null,"abstract":"<p><p>We present a detailed protocol for the training and application of an artificial intelligence (AI)-assisted workflow for automated analysis of pore and filament structures in scanning electron microscopy (SEM) images of human mucus from the respiratory tract and ileum. Our method leverages machine learning within the Fiji platform to classify image regions into pores and filaments, enabling precise and efficient quantification. The workflow includes preprocessing, pixel classification using a trained multilayer perceptron classifier, feature extraction, and data pooling. This approach significantly reduces analysis time compared with manual measurements while maintaining high precision. To validate our method, we compared manual and automated measurements on SEM images of airway and ileal mucus samples. High agreement was observed between datasets, with effect sizes indicating that AI-generated data variability is even less than human-to-human interobserver variation. In addition, we showed that increasing the counting frame area in automated analysis did not alter resulting distributions notably, confirming the robustness of the method. This AI-assisted tool supports high-throughput, comparative studies of biological network structures, enhancing accuracy and efficiency. Its application has the potential to accelerate research into structural and functional alterations of mucus in diseases such as cystic fibrosis and Crohn's disease, contributing to a deeper understanding of pathophysiology and the development of targeted therapeutic strategies.<b>NEW & NOTEWORTHY</b> Our study introduces a new artificial intelligence (AI)-assisted workflow for quantitative analysis of mucus network structures in scanning electron microscopy (SEM) images. By integrating machine learning with systematic uniform random sampling, this approach reduces variability and increases throughput compared with traditional manual methods. In our setup, the automated analysis was 40-120 times faster compared with manual analysis. The workflow is intended to be adapted by other biomedical research groups, particularly valuable for large-scale respiratory research.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":"L299-L311"},"PeriodicalIF":3.8,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148597669","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}