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Although <jats:italic>Staphylococcus aureus<\/jats:italic> is the most common pathogen associated with CSOM, its interaction with middle ear epithelial cells is not well known. In the present study, we observed that otopathogenic <jats:italic>S. aureus<\/jats:italic> has the ability to invade human middle ear epithelial cells (HMEECs) in a dose and time dependent manner. Scanning electron microscopy demonstrated time dependent increase in the number of <jats:italic>S. aureus<\/jats:italic> on the surface of HMEECs. We observed that otopathogenic <jats:italic>S. aureus<\/jats:italic> primarily employs a cholesterol dependent pathway to colonize HMEECs. In agreement with these findings, confocal microscopy showed that <jats:italic>S. aureus<\/jats:italic> colocalized with lipid rafts in HMEECs. The results of the present study provide new insights into the pathogenesis of <jats:italic>S. aureus<\/jats:italic> induced CSOM. The availability of <jats:italic>in vitro<\/jats:italic> cell culture model will pave the way to develop novel effective treatment modalities for CSOM beyond antibiotic therapy.<\/jats:p>","DOI":"10.1038\/s41598-019-47079-7","type":"journal-article","created":{"date-parts":[[2019,7,25]],"date-time":"2019-07-25T10:02:44Z","timestamp":1564048964000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Otopathogenic Staphylococcus aureus Invades Human Middle Ear Epithelial Cells Primarily through Cholesterol Dependent Pathway"],"prefix":"10.1038","volume":"9","author":[{"given":"Rahul","family":"Mittal","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luca H.","family":"Debs","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amit P.","family":"Patel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Desiree","family":"Nguyen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patricia","family":"Blackwelder","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Denise","family":"Yan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paulo H.","family":"Weckwerth","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xue Zhong","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,7,25]]},"reference":[{"key":"47079_CR1","doi-asserted-by":"publisher","first-page":"16063","DOI":"10.1038\/nrdp.2016.63","volume":"2","author":"AG Schilder","year":"2016","unstructured":"Schilder, A. 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