Abstract
Porphyromonas gingivalis (P. gingivalis) has a strong association with the pathogenesis of periodontal disease. Recurrence of periodontal disease following therapy is attributed to numerous factors, and of growing interest is the potential problem of intracellular bacteria that are able to persist and multiply within the host cell, thereby facilitating relapse of infection. The effect of antibiotic therapy in controlling P. gingivalis is questionable. Accordingly, while metronidazole is very effective against anaerobic extracellular P. gingivalis by disrupting the DNA of anaerobic microbial cells, this antibiotic does not effectively penetrate into mammalian cells to inhibit intracellular bacteria. Therefore in the present study, a modified porphyrin-linked metronidazole adducts, developed in our laboratory, was used to kill intracellular P. gingivalis. A series of experiments were performed, including cytotoxicity assays and cellular uptake of adducts by flow cytometry coupled with live cell imaging analysis, P. gingivalis invasion and elimination assays, and the analysis of colocalization of P. gingivalis and porphyrin-linked metronidazole by confocal laser scanning microscopy. Findings indicated that P. gingivalis and porphyrin-linked metronidazole were colocalized in the cytoplasm, and this compound was able to kill P. gingivalis intracellular with a sufficient culture time. This is a novel antimicrobial approach in the elimination of P. gingivalis from the oral cavity.
Highlights
Periodontitis are communicable diseases spread by bacteria
The present study aims to determine the possibility of killing intracellular bacteria using adducts of modified porphyrin linked to metronidazole, an antibiotic developed in our laboratory shown to be very effective against this anaerobic bacterium.[22,23]
Measurement of a function of cell membrane integrity It is essential to assess the function of cell membrane integrity while presented with the porphyrin-linked metronidazole compound
Summary
Periodontitis are communicable diseases spread by bacteria. Porphyromonas gingivalis (P. gingivalis) is a keystone pathogen that has strong association with the initiation and progression of periodontitis.[1,2] The severity of the disease varies, and if allowed to progress over time can result in inflammatory destruction of alveolar bone and the subsequent loss of teeth. The treatment of periodontitis aims to reduce pathogenic bacteria load on oral tissues with resulting stability of the periodontal attachment apparatus. This can be achieved through surgical means alone or with adjunctive antimicrobial regimes.[6] Systemic antibiotics have been shown to have benefit in reducing the bacterial load and improving clinical outcomes.[7] the use of broad-spectrum antibiotics carries many side effects, including a potential destruction of beneficial bacteria species and the generation of antibiotic resistance.[8,9] A pathogen-specific antimicrobial compound would be ideal in such cases
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