{"title":"利用Caco-2球粒芯片微流控平台评估空肠弯曲杆菌胞外囊泡与宿主的相互作用。","authors":"Silvia Tea Calzuola, Jeanne Malet-Villemagne, Debora Pinamonti, Francesco Rizzotto, Céline Henry, Christine Péchaux, Jean Baptiste Blondé, Emmanuel Roy, Marisa Manzano, Goran Lakisic, Sandrine Truchet, Jasmina Vidic","doi":"10.1021/acsbiomaterials.5c00750","DOIUrl":null,"url":null,"abstract":"<p><p><i>Campylobacter jejuni</i> is a foodborne pathogen that adheres to and invades the epithelial cells of the human intestinal tract. The extracellular vesicles (EVs) of <i>C. jejuni</i> have an important impact during pathogenicity, but their role in invasion of host intestinal epithelial cells remains largely unknown. <i>In vitro</i> models lack the complexity of tissue and fail to accurately replicate the dynamic interactions between EVs and human intestinal epithelial cells, while animal infection models have species-specific differences that limit their translational relevance and are associated with ethical concerns. To bridge this gap, we propose a microfluidic platform integrated with an impedimetric sensor to monitor <i>C. jejuni</i> EV interactions with human intestinal epithelial Caco-2 cells. When cultured in this microfluidic device, Caco-2 epithelial cells underwent spontaneous 3D morphogenesis into spheroid-like structures with diameters ranging from 50 to 100 μm. Functional assays revealed that the <i>C. jejuni</i> secretome and EVs (multiplicity of infection, MOI 10) caused a 60% reduction in Caco-2 cell viability in 2D plate cultures, as measured by the MTT assay. In contrast, 3D Caco-2 spheroids showed significantly increased resistance to cytotoxic effects of secreted virulence factors of <i>C. jejuni</i>. By combining impedance spectroscopy and live microscopic observation, the platform allowed real-time monitoring of cellular spatial growth and sensitive detection of EV interactions with intestinal epithelial cells, highlighting the protective role of 3D cell organization. The physiological relevance of the model was confirmed by TEER measurements that suggested that <i>Campylobacter</i> EVs diffused paracellularly. The developed microfluidic device is a promising platform for investigating host-microbe interactions and may have a broad impact on biomedical research on gastroenteritis.</p>","PeriodicalId":8,"journal":{"name":"ACS Biomaterials Science & Engineering","volume":" ","pages":""},"PeriodicalIF":5.4000,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Assessing <i>Campylobacter jejuni</i> Extracellular Vesicle-Host Interaction Using a Microfluidic Platform with Caco-2 Spheroides-on-Chip.\",\"authors\":\"Silvia Tea Calzuola, Jeanne Malet-Villemagne, Debora Pinamonti, Francesco Rizzotto, Céline Henry, Christine Péchaux, Jean Baptiste Blondé, Emmanuel Roy, Marisa Manzano, Goran Lakisic, Sandrine Truchet, Jasmina Vidic\",\"doi\":\"10.1021/acsbiomaterials.5c00750\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><i>Campylobacter jejuni</i> is a foodborne pathogen that adheres to and invades the epithelial cells of the human intestinal tract. The extracellular vesicles (EVs) of <i>C. jejuni</i> have an important impact during pathogenicity, but their role in invasion of host intestinal epithelial cells remains largely unknown. <i>In vitro</i> models lack the complexity of tissue and fail to accurately replicate the dynamic interactions between EVs and human intestinal epithelial cells, while animal infection models have species-specific differences that limit their translational relevance and are associated with ethical concerns. To bridge this gap, we propose a microfluidic platform integrated with an impedimetric sensor to monitor <i>C. jejuni</i> EV interactions with human intestinal epithelial Caco-2 cells. When cultured in this microfluidic device, Caco-2 epithelial cells underwent spontaneous 3D morphogenesis into spheroid-like structures with diameters ranging from 50 to 100 μm. Functional assays revealed that the <i>C. jejuni</i> secretome and EVs (multiplicity of infection, MOI 10) caused a 60% reduction in Caco-2 cell viability in 2D plate cultures, as measured by the MTT assay. In contrast, 3D Caco-2 spheroids showed significantly increased resistance to cytotoxic effects of secreted virulence factors of <i>C. jejuni</i>. By combining impedance spectroscopy and live microscopic observation, the platform allowed real-time monitoring of cellular spatial growth and sensitive detection of EV interactions with intestinal epithelial cells, highlighting the protective role of 3D cell organization. The physiological relevance of the model was confirmed by TEER measurements that suggested that <i>Campylobacter</i> EVs diffused paracellularly. The developed microfluidic device is a promising platform for investigating host-microbe interactions and may have a broad impact on biomedical research on gastroenteritis.</p>\",\"PeriodicalId\":8,\"journal\":{\"name\":\"ACS Biomaterials Science & Engineering\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":5.4000,\"publicationDate\":\"2025-06-23\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"ACS Biomaterials Science & Engineering\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1021/acsbiomaterials.5c00750\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MATERIALS SCIENCE, BIOMATERIALS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Biomaterials Science & Engineering","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1021/acsbiomaterials.5c00750","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MATERIALS SCIENCE, BIOMATERIALS","Score":null,"Total":0}
Assessing Campylobacter jejuni Extracellular Vesicle-Host Interaction Using a Microfluidic Platform with Caco-2 Spheroides-on-Chip.
Campylobacter jejuni is a foodborne pathogen that adheres to and invades the epithelial cells of the human intestinal tract. The extracellular vesicles (EVs) of C. jejuni have an important impact during pathogenicity, but their role in invasion of host intestinal epithelial cells remains largely unknown. In vitro models lack the complexity of tissue and fail to accurately replicate the dynamic interactions between EVs and human intestinal epithelial cells, while animal infection models have species-specific differences that limit their translational relevance and are associated with ethical concerns. To bridge this gap, we propose a microfluidic platform integrated with an impedimetric sensor to monitor C. jejuni EV interactions with human intestinal epithelial Caco-2 cells. When cultured in this microfluidic device, Caco-2 epithelial cells underwent spontaneous 3D morphogenesis into spheroid-like structures with diameters ranging from 50 to 100 μm. Functional assays revealed that the C. jejuni secretome and EVs (multiplicity of infection, MOI 10) caused a 60% reduction in Caco-2 cell viability in 2D plate cultures, as measured by the MTT assay. In contrast, 3D Caco-2 spheroids showed significantly increased resistance to cytotoxic effects of secreted virulence factors of C. jejuni. By combining impedance spectroscopy and live microscopic observation, the platform allowed real-time monitoring of cellular spatial growth and sensitive detection of EV interactions with intestinal epithelial cells, highlighting the protective role of 3D cell organization. The physiological relevance of the model was confirmed by TEER measurements that suggested that Campylobacter EVs diffused paracellularly. The developed microfluidic device is a promising platform for investigating host-microbe interactions and may have a broad impact on biomedical research on gastroenteritis.
期刊介绍:
ACS Biomaterials Science & Engineering is the leading journal in the field of biomaterials, serving as an international forum for publishing cutting-edge research and innovative ideas on a broad range of topics:
Applications and Health – implantable tissues and devices, prosthesis, health risks, toxicology
Bio-interactions and Bio-compatibility – material-biology interactions, chemical/morphological/structural communication, mechanobiology, signaling and biological responses, immuno-engineering, calcification, coatings, corrosion and degradation of biomaterials and devices, biophysical regulation of cell functions
Characterization, Synthesis, and Modification – new biomaterials, bioinspired and biomimetic approaches to biomaterials, exploiting structural hierarchy and architectural control, combinatorial strategies for biomaterials discovery, genetic biomaterials design, synthetic biology, new composite systems, bionics, polymer synthesis
Controlled Release and Delivery Systems – biomaterial-based drug and gene delivery, bio-responsive delivery of regulatory molecules, pharmaceutical engineering
Healthcare Advances – clinical translation, regulatory issues, patient safety, emerging trends
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Manufacturing and Technology – 3D printing, inks, organ-on-a-chip, bioreactor/perfusion systems, microdevices, BioMEMS, optics and electronics interfaces with biomaterials, systems integration
Modeling and Informatics Tools – scaling methods to guide biomaterial design, predictive algorithms for structure-function, biomechanics, integrating bioinformatics with biomaterials discovery, metabolomics in the context of biomaterials
Tissue Engineering and Regenerative Medicine – basic and applied studies, cell therapies, scaffolds, vascularization, bioartificial organs, transplantation and functionality, cellular agriculture