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Antimicrobial Agents and Chemotherapy, November 2003, p. 3580-3585, Vol. 47, No. 11
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.11.3580-3585.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

In Vitro and Ex Vivo Activities of Minocycline and EDTA against Microorganisms Embedded in Biofilm on Catheter Surfaces

Issam Raad,1* Ioannis Chatzinikolaou,1 Gassan Chaiban,1 Hend Hanna,1 Ray Hachem,1 Tanya Dvorak,1 Guy Cook,2 and William Costerton2

Department of Infectious Diseases, Infection Control, and Employee Health, The University of Texas M. D. Anderson Cancer Center, Houston, Texas,1 Center for Biofilm Engineering, Montana State University, Bozeman, Montana2

Received 11 December 2002/ Returned for modification 25 May 2003/ Accepted 20 August 2003

Minocycline-EDTA (M-EDTA) flush solution has been shown to prevent catheter-related infection and colonization in a rabbit model and in hemodialysis patients. We undertook this study in order to determine the activities of M-EDTA against organisms embedded in fresh biofilm (in vitro) and mature biofilm (ex vivo). For the experiment with the in vitro model, a modified Robbin’s device (MRD) was used whereby 25 catheter segments were flushed for 18 h with 106 CFU of biofilm-producing Staphylococcus epidermidis, Staphyloccocus aureus, and Candida albicans per ml. Subsequently, each of the catheter segments was incubated in one of the following solutions: (i) streptokinase, (ii) heparin, (iii) broth alone, (iv) vancomycin, (v) vancomycin-heparin, (vi) EDTA, (vii) minocycline (high-dose alternating with low-dose), or (viii) M-EDTA (low-dose minocycline alternating with high-dose minocycline were used to study the additive and synergistic activities of M-EDTA). All segments were cultured quantitatively by scrape sonication. For the experiment with the ex vivo model, 54 catheter tip segments removed from patients and colonized with bacterial organisms by roll plate were longitudinally cut into two equal segments and exposed to either saline, heparin, EDTA, or M-EDTA (with high-dose minocycline). Subsequently, all segments were examined by confocal laser electron microscopy. In the in vitro MRD model, M-EDTA (with a low concentration of minocycline) was significantly more effective than any other agent in reducing colonization of S. epidermidis, S. aureus, and C. albicans (P < 0.01). M-EDTA (with a high concentration of minocycline) eradicated all staphylococcal and C. albicans organisms embedded in the biofilm. In the ex vivo model, M-EDTA (with a high concentration of minocycline) reduced bacterial colonization more frequently than EDTA or heparin (P < 0.01). We concluded that M-EDTA is highly active in eradicating microorganisms embedded in fresh and mature biofilm adhering to catheter surfaces.

* Corresponding author. The University of Texas M. D. Anderson Cancer Center, Department of Infectious Diseases, Infection Control and Employee Health (Unit 402), 1515 Holcombe Blvd., Houston, TX 77030. Phone: (713)792-7943. Fax: (713)792-8233. E-mail: iraad{at}mdanderson.org.

Antimicrobial Agents and Chemotherapy, November 2003, p. 3580-3585, Vol. 47, No. 11
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.11.3580-3585.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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