{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,23]],"date-time":"2026-03-23T18:22:48Z","timestamp":1774290168462,"version":"3.50.1"},"reference-count":37,"publisher":"Public Library of Science (PLoS)","issue":"3","license":[{"start":{"date-parts":[[2026,3,23]],"date-time":"2026-03-23T00:00:00Z","timestamp":1774224000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:sec id=\"sec001\">\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Periprosthetic joint and implant-associated infections remain serious complications despite highly standardized diagnostic and therapeutic protocols. Repeated revision procedures substantially increase morbidity and the risk of therapeutically uncontrollable infection scenarios. This pilot study, we evaluated the effect of HMO (human milk oligosaccharides)-containing carbohydrates on S. aureus biofilm formation using crystal violet assays and implant models.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec002\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>S. aureus EDCC 5055 biofilm formation was quantified in the presence of HMO-containing carbohydrates (HMO-C) at concentrations of 5%, 7%, 9%, and 11% and lactose-alone in 96-well plates. After processing, the plate contents were read out using a 595nm Phomo plate reader (Anthos Mikrosystems, Krefeld, Germany). Biofilm activity was further evaluated on titanium disks pre-incubated with HMO-C solutions. Bacterial growth kinetics were also analyzed in TSB with the 5% to 11% HMO-C solutions.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec003\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The results demonstrated a significant reduction in S. aureus biofilm formation with the addition of HMO-C (HMO-C-5%: mean value (\n                      <jats:inline-formula id=\"pone.0342316.e001\">\n                        <jats:alternatives>\n                          <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" id=\"pone.0342316.e001g\" mimetype=\"image\" position=\"anchor\" xlink:href=\"info:doi\/10.1371\/journal.pone.0342316.e001\" xlink:type=\"simple\"\/>\n                          <mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" display=\"inline\" id=\"M1\">\n                            <mml:mi>x\u00af<\/mml:mi>\n                          <\/mml:math>\n                        <\/jats:alternatives>\n                      <\/jats:inline-formula>\n                      )=0.271nm; p\u2009=\u20090.021; HMO-C groups 7%\u221211%:\n                      <jats:inline-formula id=\"pone.0342316.e002\">\n                        <jats:alternatives>\n                          <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" id=\"pone.0342316.e002g\" mimetype=\"image\" position=\"anchor\" xlink:href=\"info:doi\/10.1371\/journal.pone.0342316.e002\" xlink:type=\"simple\"\/>\n                          <mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" display=\"inline\" id=\"M2\">\n                            <mml:mi>x\u00af<\/mml:mi>\n                          <\/mml:math>\n                        <\/jats:alternatives>\n                      <\/jats:inline-formula>\n                      \u2009=\u20090.211nm, 0.179nm, 0.147nm; All p\u2009=\u20090.001) against the positive control EDCC 5055 (PC) (\n                      <jats:inline-formula id=\"pone.0342316.e003\">\n                        <jats:alternatives>\n                          <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" id=\"pone.0342316.e003g\" mimetype=\"image\" position=\"anchor\" xlink:href=\"info:doi\/10.1371\/journal.pone.0342316.e003\" xlink:type=\"simple\"\/>\n                          <mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" display=\"inline\" id=\"M3\">\n                            <mml:mi>x\u00af<\/mml:mi>\n                          <\/mml:math>\n                        <\/jats:alternatives>\n                      <\/jats:inline-formula>\n                      \u2009=\u20090.335nm). Lactose (L) alone did not significantly affect biofilm formation (L5%\u221211%: p\u2009=\u20091.0). No significant biofilm reduction was observed for titanium implants, though medium changes indicated bacterial inhibition at higher HMO-C solutions (9%\u221211%:\n                      <jats:inline-formula id=\"pone.0342316.e004\">\n                        <jats:alternatives>\n                          <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" id=\"pone.0342316.e004g\" mimetype=\"image\" position=\"anchor\" xlink:href=\"info:doi\/10.1371\/journal.pone.0342316.e004\" xlink:type=\"simple\"\/>\n                          <mml:math xmlns:mml=\"http:\/\/www.w3.org\/1998\/Math\/MathML\" display=\"inline\" id=\"M4\">\n                            <mml:mi>x\u00af<\/mml:mi>\n                          <\/mml:math>\n                        <\/jats:alternatives>\n                      <\/jats:inline-formula>\n                      \u2009=\u20090.844nm, 0.940nm; Both p\u2009=\u20090.001).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec004\">\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>Media supplemented with HMOS-C significantly reduced S. aureus ED CC 5055 biofilm formation in vitro in the crystal violet microtiter plate assay. Bacterial invasion on titanium could not be demonstrably changed, but S. aureus growth curve was significantly reduced. Further studies with optimized implant models and standardized HMO formulations are warranted to clarify the translational potential of HMO-C for the prevention of implant-associated infections.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1371\/journal.pone.0342316","type":"journal-article","created":{"date-parts":[[2026,3,23]],"date-time":"2026-03-23T17:26:06Z","timestamp":1774286766000},"page":"e0342316","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["The anti-infective potential of human milk oligosaccharides in carbohydrate powder in implant-associated infection with Staphylococcus aureus"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1078-4983","authenticated-orcid":true,"given":"Julian","family":"Koettnitz","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Franz","family":"Koettnitz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tobias","family":"Tiemann","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4989-2367","authenticated-orcid":true,"given":"Gael","family":"Vos","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kevin","family":"Pagel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Silke","family":"Zechel-Gran","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eugen","family":"Domann","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2026,3,23]]},"reference":[{"issue":"23","key":"pone.0342316.ref001","doi-asserted-by":"crossref","first-page":"5112","DOI":"10.3390\/nu14235112","article-title":"Human milk oligosaccharides as potential antibiofilm agents: A review","volume":"14","author":"A Bhowmik","year":"2022","journal-title":"Nutrients"},{"key":"pone.0342316.ref002","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1159\/000540139","article-title":"Human milk research, more to learn?","volume":"100","author":"N Sprenger","year":"2024","journal-title":"Nestle Nutr Inst Workshop Ser"},{"issue":"12","key":"pone.0342316.ref003","first-page":"1476","article-title":"Role of human milk microbiota in infant neurodevelopment: mechanisms and clinical implications","volume":"11","author":"V Biagioli","year":"2024","journal-title":"Children (Basel)"},{"key":"pone.0342316.ref004","doi-asserted-by":"crossref","first-page":"794441","DOI":"10.3389\/fmicb.2021.794441","article-title":"Human milk oligosaccharides exhibit biofilm eradication activity against matured biofilms formed by different pathogen species","volume":"12","author":"S Jarzynka","year":"2022","journal-title":"Front Microbiol"},{"issue":"27","key":"pone.0342316.ref005","doi-asserted-by":"crossref","first-page":"11243","DOI":"10.1074\/jbc.M117.789974","article-title":"Human milk oligosaccharides inhibit growth of group B Streptococcus","volume":"292","author":"AE Lin","year":"2017","journal-title":"J Biol Chem"},{"issue":"12","key":"pone.0342316.ref006","doi-asserted-by":"crossref","first-page":"1225","DOI":"10.3390\/antibiotics13121225","article-title":"Does a Specific Sequential Combination of Antiseptic Solutions for Chemical Debridement in Periprosthetic Joint Infection Improve Outcomes vs. Solution Alone? An In Vivo Study","volume":"13","author":"M M\u00e1rquez-G\u00f3mez","year":"2024","journal-title":"Antibiotics (Basel)"},{"issue":"4","key":"pone.0342316.ref007","doi-asserted-by":"crossref","first-page":"36","DOI":"10.31083\/j.fbe1604036","article-title":"Bacterial Communities and Their Role in Bacterial Infections","volume":"16","author":"OI Guliy","year":"2024","journal-title":"Front Biosci (Elite Ed)"},{"issue":"3","key":"pone.0342316.ref008","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1021\/acsinfecdis.7b00183","article-title":"Antimicrobial and Antibiofilm Activity of Human Milk Oligosaccharides against Streptococcus agalactiae, Staphylococcus aureus, and Acinetobacter baumannii","volume":"4","author":"DL Ackerman","year":"2018","journal-title":"ACS Infect Dis"},{"issue":"6","key":"pone.0342316.ref009","doi-asserted-by":"crossref","DOI":"10.1002\/cbic.202200643","article-title":"Anti-biofilm Activity of Human Milk Oligosaccharides in Clinical Strains of Streptococcus agalactiae with Diverse Capsular and Sequence Types","volume":"24","author":"RE Moore","year":"2023","journal-title":"Chembiochem"},{"issue":"24","key":"pone.0342316.ref010","doi-asserted-by":"crossref","first-page":"16128","DOI":"10.1021\/acs.joc.0c01958","article-title":"Leveraging stereoelectronic effects in biofilm eradication: synthetic \u03b2-amino human milk oligosaccharides impede microbial adhesion as observed by scanning electron microscopy","volume":"85","author":"RE Moore","year":"2020","journal-title":"J Org Chem"},{"issue":"1","key":"pone.0342316.ref011","first-page":"28","article-title":"3D printed antimicrobial PLA constructs functionalised with zinc- coated halloysite nanotubes-Ag-chitosan oligosaccharide lactate","volume":"37","author":"A Humayun","year":"2022","journal-title":"Mater Technol (N Y N Y)"},{"issue":"1","key":"pone.0342316.ref012","doi-asserted-by":"crossref","first-page":"43","DOI":"10.3390\/md17010043","article-title":"Synthesis and Evaluation of a Chitosan Oligosaccharide-Streptomycin Conjugate against Pseudomonas aeruginosa Biofilms","volume":"17","author":"R Li","year":"2019","journal-title":"Mar Drugs"},{"key":"pone.0342316.ref013","doi-asserted-by":"crossref","first-page":"700605","DOI":"10.3389\/fmicb.2021.700605","article-title":"Antimicrobial Activities of Alginate and Chitosan Oligosaccharides Against Staphylococcus aureus and Group B Streptococcus","volume":"12","author":"M Asadpoor","year":"2021","journal-title":"Front Microbiol"},{"issue":"8","key":"pone.0342316.ref014","doi-asserted-by":"crossref","DOI":"10.1128\/genomeA.01698-16","article-title":"Complete genome and plasmid sequences of Staphylococcus aureus EDCC 5055 (DSM 28763), used to study implant-associated infections","volume":"5","author":"GK Mannala","year":"2017","journal-title":"Genome Announc"},{"issue":"1","key":"pone.0342316.ref015","doi-asserted-by":"crossref","first-page":"19364","DOI":"10.1038\/s41598-024-70244-6","article-title":"Biofilm formation of Staphylococcus aureus on various implants used for surgical treatment of destructive spondylodiscitis","volume":"14","author":"AT Zacher","year":"2024","journal-title":"Sci Rep"},{"issue":"3","key":"pone.0342316.ref016","first-page":"225","article-title":"The composition of human milk","volume":"3","author":"R Jenness","year":"1979","journal-title":"Semin Perinatol"},{"issue":"3","key":"pone.0342316.ref017","doi-asserted-by":"crossref","first-page":"104929","DOI":"10.1016\/j.micinf.2021.104929","article-title":"The prebiotic effect of human milk oligosaccharides 3\u2019- and 6\u2019-sialyllactose on adhesion and biofilm formation by Clostridioides difficile - pilot study","volume":"24","author":"M Piotrowski","year":"2022","journal-title":"Microbes Infect"},{"issue":"1","key":"pone.0342316.ref018","doi-asserted-by":"crossref","DOI":"10.1128\/msphere.00885-21","article-title":"Human milk oligosaccharides reduce murine group B streptococcus vaginal colonization with minimal impact on the vaginal microbiota","volume":"7","author":"ME Mejia","year":"2022","journal-title":"mSphere"},{"issue":"12","key":"pone.0342316.ref019","doi-asserted-by":"crossref","first-page":"3254","DOI":"10.1021\/acsinfecdis.1c00420","article-title":"Antibiofilm Activity of Human Milk Oligosaccharides against Multidrug Resistant and Susceptible Isolates of Acinetobacter baumannii","volume":"7","author":"SK Spicer","year":"2021","journal-title":"ACS Infect Dis"},{"issue":"8","key":"pone.0342316.ref020","doi-asserted-by":"crossref","first-page":"2020","DOI":"10.1021\/acschembio.8b00661","article-title":"Human milk oligosaccharides (HMOs) sensitize group B streptococcus to clindamycin, erythromycin, gentamicin, and minocycline on a strain specific basis","volume":"13","author":"KM Craft","year":"2018","journal-title":"ACS Chem Biol"},{"issue":"5","key":"pone.0342316.ref021","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1302\/2046-3758.115.BJR-2021-0395.R1","article-title":"Rifampicin restores extracellular organic matrix formation and mineralization of osteoblasts after intracellular Staphylococcus aureus infection","volume":"11","author":"FI Alagboso","year":"2022","journal-title":"Bone Joint Res"},{"issue":"10","key":"pone.0342316.ref022","doi-asserted-by":"crossref","first-page":"4518","DOI":"10.1016\/j.actbio.2014.06.013","article-title":"Rifampicin-fosfomycin coating for cementless endoprostheses: antimicrobial effects against methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA)","volume":"10","author":"V Alt","year":"2014","journal-title":"Acta Biomater"},{"key":"pone.0342316.ref023","doi-asserted-by":"crossref","first-page":"100151","DOI":"10.1016\/j.bioflm.2023.100151","article-title":"\u03b1,\u03b1-disubstituted \u03b2-amino amides eliminate Staphylococcus aureus biofilms by membrane disruption and biomass removal","volume":"6","author":"D Ausbacher","year":"2023","journal-title":"Biofilm"},{"issue":"2","key":"pone.0342316.ref024","doi-asserted-by":"crossref","first-page":"87","DOI":"10.3390\/pathogens9020087","article-title":"Novel Cytoplasmic Bacteriocin Compounds Derived from Staphylococcus epidermidis Selectively Kill Staphylococcus aureus, Including Methicillin-Resistant Staphylococcus aureus (MRSA)","volume":"9","author":"I-T Jang","year":"2020","journal-title":"Pathogens"},{"key":"pone.0342316.ref025","doi-asserted-by":"crossref","first-page":"112446","DOI":"10.1016\/j.nut.2024.112446","article-title":"How xylitol, gluten, and lactose change human gut microbiota Escherichia coli and Lactobacillus rhamnosus GG biofilm","volume":"124","author":"J Kwieci\u0144ska-Pir\u00f3g","year":"2024","journal-title":"Nutrition"},{"key":"pone.0342316.ref026","doi-asserted-by":"crossref","first-page":"143296","DOI":"10.1016\/j.ijbiomac.2025.143296","article-title":"A promising multi-component, multi-functional, drug-releasing Cu-doped Fe3O4\/bioactive glass\/Chitosan coating, applied on stainless steel substrate for biomedical applications","volume":"310","author":"M Rajabinezhad","year":"2025","journal-title":"Int J Biol Macromol"},{"issue":"6","key":"pone.0342316.ref027","doi-asserted-by":"crossref","first-page":"994","DOI":"10.3390\/antibiotics12060994","article-title":"Bacterial Adhesion Strength on Titanium Surfaces Quantified by Atomic Force Microscopy: A Systematic Review","volume":"12","author":"JDC Tardelli","year":"2023","journal-title":"Antibiotics (Basel)"},{"issue":"2","key":"pone.0342316.ref028","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1038\/pj.2012.130","article-title":"Characteristics of bacterial biofilm associated with implant material in clinical practice","volume":"45","author":"V Nandakumar","year":"2012","journal-title":"Polym J"},{"key":"pone.0342316.ref029","doi-asserted-by":"crossref","DOI":"10.3389\/fdmed.2021.695417","article-title":"Surface characterization and assessment of biofilm formation on two titanium-based implant coating materials","volume":"2","author":"B Shokeen","year":"2021","journal-title":"Frontiers in Dental Medicine"},{"issue":"3","key":"pone.0342316.ref030","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1002\/jbm.a.37635","article-title":"Influence of titanium nanoscale surface roughness on fibrinogen and albumin protein adsorption kinetics and platelet responses","volume":"112","author":"RD Boehm","year":"2024","journal-title":"J Biomed Mater Res A"},{"issue":"9","key":"pone.0342316.ref031","doi-asserted-by":"crossref","first-page":"1333","DOI":"10.1002\/jbm.a.37531","article-title":"Effect of titanium nanostructured surface on fibroblast behavior","volume":"111","author":"K Vrchoveck\u00e1","year":"2023","journal-title":"J Biomed Mater Res A"},{"issue":"3","key":"pone.0342316.ref032","doi-asserted-by":"crossref","DOI":"10.3390\/microorganisms10030586","article-title":"Mapping bacterial biofilm on features of orthopedic implants in vitro","volume":"10","author":"K Moore","year":"2022","journal-title":"Microorganisms"},{"issue":"4","key":"pone.0342316.ref033","doi-asserted-by":"crossref","DOI":"10.1128\/mbio.00298-24","article-title":"A human milk oligosaccharide prevents intestinal inflammation in adulthood via modulating gut microbial metabolism","volume":"15","author":"KM Schalich","year":"2024","journal-title":"mBio"},{"issue":"14","key":"pone.0342316.ref034","doi-asserted-by":"crossref","first-page":"4763","DOI":"10.1128\/AEM.00477-12","article-title":"Human milk oligosaccharides promote the growth of staphylococci","volume":"78","author":"KM Hunt","year":"2012","journal-title":"Appl Environ Microbiol"},{"issue":"3","key":"pone.0342316.ref035","doi-asserted-by":"crossref","DOI":"10.1002\/cbic.202100423","article-title":"The diverse antimicrobial activities of human milk oligosaccharides against group B streptococcus","volume":"23","author":"RE Moore","year":"2022","journal-title":"Chembiochem"},{"key":"pone.0342316.ref036","doi-asserted-by":"crossref","first-page":"1379042","DOI":"10.3389\/fimmu.2024.1379042","article-title":"Human milk oligosaccharides regulate human macrophage polarization and activation in response to Staphylococcus aureus","volume":"15","author":"SD Jepsen","year":"2024","journal-title":"Front Immunol"},{"issue":"21","key":"pone.0342316.ref037","doi-asserted-by":"crossref","DOI":"10.21769\/BioProtoc.4866","article-title":"A Guideline for assessment and characterization of bacterial biofilm formation in the presence of inhibitory compounds","volume":"13","author":"BA Elgamoudi","year":"2023","journal-title":"Bio Protoc"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0342316","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,23]],"date-time":"2026-03-23T17:26:12Z","timestamp":1774286772000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0342316"}},"subtitle":[],"editor":[{"given":"Ajaya","family":"Bhattarai","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2026,3,23]]},"references-count":37,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,3,23]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0342316","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,23]]}}}