Development of a physiological model of human middle ear epithelium

Laryngoscope Investigative Otolaryngology

article

Muzlifah Haniffa PhD¹  Jason Powell PhD²  Chris Ward PhD³  Michael William Mather MBBS, MRes (Dist), MRCS (ENT)¹  Bernard Verdon PhD¹  Rachel Anne Botting PhD¹  Justin Engelbert BSc (Hons)¹  Livia Delpiano BSc (Hons)¹  Xin Xu BSc (Hons)⁴  Catherine Hatton BSc (Hons)³  Tracey Davey BSc (Hons)⁵  Steven Lisgo PhD¹  Philip Yates FRCS²  Nicholas Dawe FRCS²  Colin D. Bingle PhD⁶ 

[1] Faculty of Medical Sciences, Biosciences Institute, Newcastle University;Department of Otolaryngology, Freeman Hospital;Faculty of Medical Sciences, Translational and Clinical Research Institute, Newcastle University;Newcastle Biobank, Faculty of Medical Sciences, Newcastle University;Electron Microscopy Research Services, Faculty of Medical Sciences, Newcastle University;Department of Infection, Immunity and Cardiovascular Disease, The Medical School

关键词: biological models;    otitis media;    otorhinolaryngologic diseases;    respiratory mucosa;    SARS-CoV-2;   

DOI  :  10.1002/lio2.661

学科分类：环境科学（综合）

来源: Wiley

【 摘 要 】

Introduction Otitis media is an umbrella term for middle ear inflammation; ranging from acute infection to chronic mucosal disease. It is a leading cause of antimicrobial therapy prescriptions and surgery in children. Despite this, treatments have changed little in over 50 years. Research has been limited by the lack of physiological models of middle ear epithelium. Methods We develop a novel human middle ear epithelial culture using an air-liquid interface (ALI) system; akin to the healthy ventilated middle ear in vivo. We validate this using immunohistochemistry, immunofluorescence, scanning and transmission electron microscopy, and membrane conductance studies. We also utilize this model to perform a pilot challenge of middle ear epithelial cells with SARS-CoV-2. Results We demonstrate that human middle ear epithelial cells cultured at an ALI undergo mucociliary differentiation to produce diverse epithelial subtypes including basal (p63+), goblet (MUC5AC+, MUC5B+), and ciliated (FOXJ1+) cells. Mature ciliagenesis is visualized and tight junction formation is shown with electron microscopy, and confirmed by membrane conductance. Together, these demonstrate this model reflects the complex epithelial cell types which exist in vivo. Following SARS-CoV-2 challenge, human middle ear epithelium shows positive viral uptake, as measured by polymerase chain reaction and immunohistochemistry. Conclusion We describe a novel physiological system to study the human middle ear. This can be utilized for translational research into middle ear diseases. We also demonstrate, for the first time under controlled conditions, that human middle ear epithelium is susceptible to SARS-CoV-2 infection, which has important clinical implications for safe otological surgery. Level of Evidence NA.

【 授权许可】

CC BY|CC BY-NC-ND