Atomic force microscopy of bacteria from periodontal subgingival biofilm: Preliminary study results

ABSTRACT

Objective: Atomic force microscope (AFM) is a technology that allows analysis of the nanoscale morphology of bacteria within biofilm and provides details that may be better useful for understanding the role of bacterial interactions in the periodontal disease. Material and Methods: Five patients with periodontal ≥5 mm pockets diagnosed as generalized periodontitis and five patients with slight gingivitis were selected for the investigation. Bacteria biofilms were collected and morphologically investigated by AFM application. Results: The investigation revealed how periodontitis bacteria are characterized by specific morphologic features of the cell wall. The major representative species of bacteria causing periodontal diseases have been reproduced by a three-dimensional reconstruction showing the bacteria surface details. Conclusions: The presence of complex glycocalyx structures, bacteriophage-like vesicles, spirochetes (classic and cystic morphology) and bacterial co-aggregation has been identified by the AFM analysis. The results suggest that AFM is a reliable technique for studying bacterial morphology and for examining microbial interactions in dental plaque.

Key words

Atomic force microscopy - biofilm - microbiology - periodontitis

• REFERENCES

• 1 Gursoy UK. Periodontal bacteria and epithelial cell interactions: Role of bacterial proteins. Eur J Dent 2008; 2: 231-2
• 2 van Winkelhoff AJ, Loos BG, van der Reijden WA, van der Velden U. Porphyromonas gingivalis, bacteroides forsythus and other putative periodontal pathogens in subjects with and without periodontal destruction. J Clin Periodontol 2002; 29: 1023-8
• 3 Loesche WJ, Lopatin DE, Stoll J, van Poperin N, Hujoel PP. Comparison of various detection methods for periodontopathic bacteria: Can culture be considered the primary reference standard?. J Clin Microbiol 1992; 30: 418-26
• 4 Akin L, Herford AS, Cicciù M. Oral presentation of disseminated histoplasmosis: A case report and literature review. J Oral Maxillofac Surg 2011; 69: 535-41
• 5 Hugenholtz P, Pace NR. Identifying microbial diversity in the natural environment: A molecular phylogenetic approach. Trends Biotechnol 1996; 14: 190-7
• 6 Mohamed Saber SE, El-Hady SA. Development of an intracanal mature Enterococcus faecalis biofilm and its susceptibility to some antimicrobial intracanal medications; an in vitro study. Eur J Dent 2012; 6: 43-50
• 7 Listgarten MA, Helldén L. Relative distribution of bacteria at clinically healthy and periodontally diseased sites in humans. J Clin Periodontol 1978; 5: 115-32
• 8 Kuramitsu HK, He X, Lux R, Anderson MH, Shi W. Interspecies interactions within oral microbial communities. Microbiol Mol Biol Rev 2007; 71: 653-70
• 9 Kelstrup J, Theilade J, Fejerskov O. Surface ultrastructure of some oral bacteria. Scand J Dent Res 1979; 87: 415-23
• 10 Kigure T, Saito A, Seida K, Yamada S, Ishihara K, Okuda K. Distribution of porphyromonas gingivalis and treponema denticola in human subgingival plaque at different periodontal pocket depths examined by immunohistochemical methods. J Periodontal Res 1995; 30: 332-41
• 11 Zijnge V, van Leeuwen MB, Degener JE, Abbas F, Thurnheer T, Gmür R. et al. Oral biofilm architecture on natural teeth. PLoS One 2010; 5: 9321
• 12 Greif D, Wesner D, Regtmeier J, Anselmetti D. High resolution imaging of surface patterns of single bacterial cells. Ultramicroscopy 2010; 110: 1290-6
• 13 Dubrovin EV, Voloshin AG, Kraevsky SV, Ignatyuk TE, Abramchuk SS, Yaminsky IV. et al. Atomic force microscopy investigation of phage infection of bacteria. Langmuir 2008; 24: 13068-74
• 14 Allan EJ, Hoischen C, Gumpert J. Bacterial l-forms. Adv Appl Microbiol 2009; 68: 1-39
• 15 Bradley DE. Ultrastructure of bacteriophage and bacteriocins. Bacteriol Rev 1967; 31: 230-314
• 16 MacDonald AB. Spirochetal cyst forms in neurodegenerative disorders: Hiding in plain sight. Med Hypotheses 2006; 67: 819-32
• 17 Listgarten MA, Socransky SS. Electron microscopy of axial fibrils, outer envelope, and cell division of certain oral spirochetes. J Bacteriol 1964; 88: 1087-103
• 18 De Ciccio A, McLaughlin R, Chan EC. Factors affecting the formation of spherical bodies in the spirochete treponema denticola. Oral Microbiol Immunol 1999; 14: 384-6
• 19 Bik EM, Long CD, Armitage GC, Loomer P, Emerson J, Mongodin EF. et al. Bacterial diversity in the oral cavity of 10 healthy individuals. ISME J 2010; 4: 962-74
• 20 Quevedo B, Giertsen E, Zijnge V, Lüthi-Schaller H, Guggenheim B, Thurnheer T. et al. Phylogenetic group- and species-specific oligonucleotide probes for single-cell detection of lactic acid bacteria in oral biofilms. BMC Microbiol 2011; 11: 14
• 21 Dunne WM. Bacterial adhesion: Seen any good biofilms lately?. Clin Microbiol Rev 2002; 15: 155-66
• 22 Preus HR, Olsen I, Gjermo P. Bacteriophage infection--a possible mechanism for increased virulence of bacteria associated with rapidly destructive periodontitis. Acta Odontol Scand 1987; 45: 49-54
• 23 Hansma HG, Pietrasanta L. Atomic Force Microscopy and other scanning probe microscopies. Curr Opin Chem Biol 1998; 2: 579-84