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This study assessed, <jats:italic>in vitro<\/jats:italic>, the antimicrobial efficacy of tetracycline hydrochloride (TCH) adsorbed onto Bio\u2010Oss\u00ae bone grafts against a range of pathogenic bacteria. Various levels of TCH were adsorbed onto Bio\u2010Oss\u00ae granules by immersing in TCH aqueous solutions of different initial concentrations for 48 h at room temperature. TCH release was assessed in phosphate buffered saline at 37\u00b0C, and its antimicrobial efficacy, up to 96 h, was tested against two Gram\u2010negative bacteria associated with periodontal diseases: <jats:italic>Aggregatibacter<\/jats:italic> (formerly <jats:italic>Actinobacillus<\/jats:italic>) <jats:italic>actinomycetemcomitans<\/jats:italic>, and <jats:italic>Porphyromonas gingivalis<\/jats:italic>, and one Gram\u2010positive bacterium associated with soft\u2010tissue and bone infections: <jats:italic>Staphylococcus aureus<\/jats:italic>. The range of TCH concentrations studied was also assessed for cytotoxicity against osteoblast\u2010like human osteosarcoma cell lines. The amount of TCH adsorbed and released from Bio\u2010Oss\u00ae was concentration dependent. All TCH adsorbed Bio\u2010Oss\u00ae resulted in a reduction of <jats:italic>A. actinomycetemcomitans<\/jats:italic>, <jats:italic>P. gingivalis<\/jats:italic>, and <jats:italic>S. aureus<\/jats:italic> and higher concentrations were generally more effective in reducing or eliminating bacterial growth. The proliferation of HOS cells was not substantially reduced except for the maximum concentration of TCH. In addition to its osteoconductive role, TCH adsorbed Bio\u2010Oss\u00ae could also be functional in negating systemically antibiotic prophylactic treatment in the prevention of implant or biomaterial related infections. \u00a9 2010 Wiley Periodicals, Inc. 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