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Antimicrobial Agents and Chemotherapy, October 1998, p. 2595-2601, Vol. 42, No. 10
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Targeted Antimicrobial Photochemotherapy

Nikolaos S. Soukos,1 Laurie Ann Ximenez-Fyvie,2 Michael R. Hamblin,1 Sigmund S. Socransky,2 and Tayyaba Hasan1,*

Wellman Laboratories of Photomedicine, Department of Dermatology, Massachusetts General Hospital, Harvard Medical School,1 and Department of Periodontology, Forsyth Dental Center,2 Boston, Massachusetts

Received 8 January 1998/Returned for modification 3 June 1998/Accepted 13 July 1998

This study explores a new approach for antimicrobial therapy with light activation of targeted poly-L-lysine (pL)-chlorin e6 (ce6) conjugates. The goal was to test the hypothesis that these conjugates between pL and ce6 would efficiently target photodestruction towards gram-positive (Actinomyces viscosus) and gram-negative (Porphyromonas gingivalis) oral species while sparing an oral epithelial cell line (HCPC-1). Conjugates of ce6 with pL (average molecular weight, 2,000) having a positive, neutral, or negative charge were prepared. Illumination with red light (lambda max = 671 nm) from a diode array produced a dose-dependent loss of CFU from the bacteria, under conditions that did not affect the viability of the epithelial cells. For P. gingivalis, the cationic conjugate produced 99% killing, while the neutral conjugate killed 91% and the anionic conjugate killed 76% after 1 min of incubation and exposure to red light for 10 min. For A. viscosus, the cationic conjugate produced >99.99% killing while HCPC-1 cells remained intact. The importance of the positive charge was shown by the effectiveness of ce6-monoethylenediamine monoamide (a monocationic derivative of ce6) in killing both bacteria. The clinically employed benzoporphyrin derivative under the same conditions killed epithelial cells while leaving P. gingivalis relatively unharmed. A mixture of ce6 with pL did not show phototoxicity comparable with that of the cationic conjugate. These results were explained by the selective uptake of the conjugates by bacteria (20- to 100-fold) compared to that by mammalian cells, while free ce6 showed much less selectivity for bacteria (5- to 20-fold). The data suggest that the cationic pL-ce6 conjugate may have an application for the photodynamic therapy of periodontal disease.

* Corresponding author. Mailing address: Department of Dermatology, Massachusetts General Hospital, 50 Blossom St., WEL 224, Boston, MA 02114-2698. Phone: (617) 726-6996. Fax: (617) 726-3192. E-mail: hasan{at}helix.mgh.harvard.edu.

Antimicrobial Agents and Chemotherapy, October 1998, p. 2595-2601, Vol. 42, No. 10
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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