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High hydrostatic pressure (HHP) is known to devitalize tissues effectively while leaving the extracellular matrix intact. However, little is known about the inactivation of the contaminating microorganisms by HHP. This study aims to investigate the ability of high\u2010pressure decontamination and to establish a treatment protocol that is able to successfully inactivate microorganisms with the final goal to sterilize bone specimens. Using <jats:italic>Escherichia coli (E. coli)<\/jats:italic> as a model organism, HHP treatment parameters like temperature and duration, pressurization medium, and the number of treatment cycles were systematically adjusted to maximize the efficiency of inactivating logarithmic and stationary phase bacteria. Towards that we quantified colony\u2010forming units (cfu) after treatment and investigated morphological changes via Field Emission Scanning Electron Microscopy (FESEM). Additionally, we tested the decontamination efficiency of HHP in bovine cancellous bone blocks that were contaminated with bacteria. Finally, two further model organisms were evaluated, namely <jats:italic>Pseudomonas fluorescens<\/jats:italic> as a Gram\u2010negative microorganism and <jats:italic>Micrococcus luteus<\/jats:italic> as a Gram\u2010positive representative. A HHP protocol, using 350\u2009MPa, was able to sterilize a suspension of stationary phase <jats:italic>E. coli<\/jats:italic>, leading to a logarithmic reduction factor (log RF) of at least \u22127.99 (\u00b10.43). The decontamination of bone blocks was less successful, indicating a protective effect of the surrounding tissue. Sterilization of 100% of the samples was achieved when a protocol optimized in terms of treatment temperature, duration, pressurization medium, and number and\/or interval of cycles, respectively, was applied to bone blocks artificially contaminated with a suspension containing 10<jats:sup>4<\/jats:sup>\u2009cfu\/mL. Hence, we here successfully established protocols for inactivating Gram\u2010negative model microorganisms by HHP of up to 350\u2009MPa, while pressure levels of 600\u2009MPa were needed to inactivate the Gram\u2010positive model organism. Thus, this study provides a basis for further investigations on different pathogenic bacteria that could enable the use of HHP in the decontamination of bone grafts intended for transplantation.<\/jats:p>","DOI":"10.1002\/jbm.b.35383","type":"journal-article","created":{"date-parts":[[2024,2,12]],"date-time":"2024-02-12T10:53:29Z","timestamp":1707735209000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Systematic enhancement of microbial decontamination efficiency in bone graft processing by means of high hydrostatic pressure using <i>Escherichia coli<\/i> as a model organism"],"prefix":"10.1002","volume":"112","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-1959-9924","authenticated-orcid":false,"given":"Henrike","family":"Loeffler","sequence":"first","affiliation":[{"name":"Biomechanics and Implant Technology Research Laboratory, Department of Orthopedics Rostock University Medical Center  Rostock 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