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Variations in membrane composition and temperature lead to phase state changes in lipid membranes, which in turn influence the structure and dynamics of the associated interfacial water. In this study, molecular dynamics simulations were performed on membranes composed of dipalmitoylphosphatidylcholine (DPPC) or palmitoyl sphingomyelin mixed with cholesterol (Chol). To elucidate the effects of Chol on interfacial water, we examined the orientation and hydrogen-bonding behavior of water molecules spanning from the membrane interior to the interface. As the Chol concentration increased, a transient slowdown in water dynamics was observed in the ripple phase at 303\u00a0K. Conversely, at higher Chol concentrations, water dynamics were accelerated relative to pure lipid membranes across all temperatures studied. In particular, at a Chol concentration of 50%, the hydrogen bond lifetime in DPPC membranes decreased to \u223c0.5\u20130.7 times that of pure lipid membranes. This nonmonotonic behavior is attributed to the combined effects of membrane packing induced by Chol and reduced density of lipid molecules in the hydrophilic region, offering key insights for modulating the dynamical properties of interfacial water.<\/jats:p>","DOI":"10.1063\/5.0293607","type":"journal-article","created":{"date-parts":[[2025,12,30]],"date-time":"2025-12-30T12:49:09Z","timestamp":1767098949000},"update-policy":"https:\/\/doi.org\/10.1063\/aip-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Dual effect of cholesterol on interfacial water dynamics in lipid membranes: Interplay between membrane packing and hydration"],"prefix":"10.1063","volume":"163","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-7910-7183","authenticated-orcid":false,"given":"Kokoro","family":"Shikata","sequence":"first","affiliation":[{"name":"Division of Chemical Engineering, Department of Materials Engineering Science, Graduate School of Engineering Science, The University of Osaka , Toyonaka, Osaka 560-8531,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7138-250X","authenticated-orcid":false,"given":"Kento","family":"Kasahara","sequence":"additional","affiliation":[{"name":"Division of Chemical Engineering, Department of Materials Engineering Science, Graduate School of Engineering Science, The University of Osaka , Toyonaka, Osaka 560-8531,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3461-3660","authenticated-orcid":false,"given":"Nozomi Morishita","family":"Watanabe","sequence":"additional","affiliation":[{"name":"Division of Chemical Engineering, Department of Materials Engineering Science, Graduate School of Engineering Science, The University of Osaka , Toyonaka, Osaka 560-8531,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9241-853X","authenticated-orcid":false,"given":"Hiroshi","family":"Umakoshi","sequence":"additional","affiliation":[{"name":"Division of Chemical Engineering, Department of Materials Engineering Science, Graduate School of Engineering Science, The University of Osaka , Toyonaka, Osaka 560-8531,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7047-3183","authenticated-orcid":false,"given":"Kang","family":"Kim","sequence":"additional","affiliation":[{"name":"Division of Chemical Engineering, Department of Materials Engineering Science, Graduate School of Engineering Science, The University of Osaka , Toyonaka, Osaka 560-8531,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7176-441X","authenticated-orcid":false,"given":"Nobuyuki","family":"Matubayasi","sequence":"additional","affiliation":[{"name":"Division of Chemical Engineering, Department of Materials Engineering Science, Graduate School of Engineering Science, The University of Osaka , Toyonaka, Osaka 560-8531,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"317","published-online":{"date-parts":[[2025,12,30]]},"reference":[{"key":"2025123007490158100_c1","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1016\/s0304-4157(00)00016-2","article-title":"Structure of lipid bilayers","volume":"1469","year":"2000","journal-title":"Biochim. 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