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Antimicrobial Agents and Chemotherapy, August 2009, p. 3308-3316, Vol. 53, No. 8
0066-4804/09/$08.00+0     doi:10.1128/AAC.01688-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Inhibitory Effects of Lactoferrin on Growth and Biofilm Formation of Porphyromonas gingivalis and Prevotella intermedia{triangledown}

Hiroyuki Wakabayashi,1* Koji Yamauchi,1 Tetsuo Kobayashi,2,3 Tomoko Yaeshima,1 Keiji Iwatsuki,1 and Hiromasa Yoshie2

Food Science & Technology Institute, Morinaga Milk Industry Co., Ltd., Zama,1 Division of Periodontology, Niigata University Graduate School of Medical and Dental Sciences, Niigata,2 General Dentistry and Clinical Education Unit, Niigata University Medical and Dental Hospital, Niigata, Japan3

Received 22 December 2008/ Returned for modification 3 February 2009/ Accepted 8 May 2009

Lactoferrin (LF) is an iron-binding antimicrobial protein present in saliva and gingival crevicular fluids, and it is possibly associated with host defense against oral pathogens, including periodontopathic bacteria. In the present study, we evaluated the in vitro effects of LF-related agents on the growth and biofilm formation of two periodontopathic bacteria, Porphyromonas gingivalis and Prevotella intermedia, which reside as biofilms in the subgingival plaque. The planktonic growth of P. gingivalis and P. intermedia was suppressed for up to 5 h by incubation with ≥130 µg/ml of human LF (hLF), iron-free and iron-saturated bovine LF (apo-bLF and holo-bLF, respectively), and ≥6 µg/ml of bLF-derived antimicrobial peptide lactoferricin B (LFcin B); but those effects were weak after 8 h. The biofilm formation of P. gingivalis and P. intermedia over 24 h was effectively inhibited by lower concentrations (≥8 µg/ml) of various iron-bound forms (the apo, native, and holo forms) of bLF and hLF but not LFcin B. A preformed biofilm of P. gingivalis and P. intermedia was also reduced by incubation with various iron-bound bLFs, hLF, and LFcin B for 5 h. In an examination of the effectiveness of native bLF when it was used in combination with four antibiotics, it was found that treatment with ciprofloxacin, clarithromycin, and minocycline in combination with native bLF for 24 h reduced the amount of a preformed biofilm of P. gingivalis compared with the level of reduction achieved with each agent alone. These results demonstrate the antibiofilm activity of LF with lower iron dependency against P. gingivalis and P. intermedia and the potential usefulness of LF for the prevention and treatment of periodontal diseases and as adjunct therapy for periodontal diseases.

* Corresponding author. Mailing address: Food Science & Technology Institute, Morinaga Milk Industry Co., Ltd., 5-1-83 Higashihara, Zama, Kanagawa 228-8583, Japan. Phone: 81-46-252-3045. Fax: 81-46-252-3059. E-mail: h_wakaby{at}morinagamilk.co.jp

{triangledown} Published ahead of print on 18 May 2009.

Antimicrobial Agents and Chemotherapy, August 2009, p. 3308-3316, Vol. 53, No. 8
0066-4804/09/$08.00+0     doi:10.1128/AAC.01688-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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