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Micro-Raman Spectroscopy Reveals the Presence of Octacalcium Phosphate and Whitlockite in Association with Bacteria-Free Zones Within the Mineralized Dental Biofilm

Published online by Cambridge University Press:  04 February 2019

Furqan A. Shah Open the ORCID record for Furqan A. Shah [Opens in a new window]
Furqan A. Shah*
Affiliation:
Department of Biomaterials, Sahlgrenska Academy, University of Gothenburg, Göteborg, Sweden
*
*Author for correspondence: Furqan A. Shah, E-mail: furqan.ali.shah@biomaterials.gu.se
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Abstract

Through a correlative analytical approach encompassing backscattered electron scanning electron microscopy (BSE-SEM), energy dispersive X-ray spectroscopy (EDX), and micro-Raman spectroscopy, the composition of the mineralized biofilm around a dental implant, retrieved due to peri-implantitis, was investigated. The mineralized biofilm contains two morphologically distinct regions: (i) bacteria-containing zones (Bact+), characterized by aggregations of unmineralized and mineralized bacteria, and intermicrobial mineralization, and (ii) bacteria-free zones (Bact−), comprised mainly of randomly oriented mineral platelets. Intramicrobial mineralization, within Bact+, appears as smooth, solid mineral deposits resembling the morphologies of dental plaque bacteria. Bact− is associated with micrometer-sized Mg-rich mineral nodules. The Ca/P ratio of Bact+ is higher than Bact−. The inorganic phase of Bact+ is carbonated apatite (CHAp), while that of Bact− is predominantly octacalcium phosphate (OCP) and whitlockite (WL) inclusions. Compared with native bone, the inorganic phase of Bact+ (i.e., CHAp) exhibits higher mineral crystallinity, lower carbonate content, and lower Ca/P, C/Ca, Mg/Ca, and Mg/P ratios. The various CaPs found within the mineralized dental biofilm (CHAp, OCP, and WL) are related to the local presence/absence of bacteria. In combination with BSE-SEM and EDX, micro-Raman spectroscopy is a valuable analytical tool for nondestructive investigation of mineralized dental biofilm composition and development.

Keywords

Apatitedental implantmineralized biofilmoctacalcium phosphateRaman spectroscopywhitlockite
Type
Biological Science Applications
Information
Microscopy and Microanalysis , Volume 25 , Issue 1 , February 2019 , pp. 129 - 134
Copyright
Copyright © Microscopy Society of America 2019 

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References

Akcali, A & Lang, NP (2018). Dental calculus: The calcified biofilm and its role in disease development. Periodontol 2000 76(1), 109–115.Google Scholar
Albouy, JP, Abrahamsson, I, Persson, LG & Berglundh, T (2008). Spontaneous progression of peri-implantitis at different types of implants. An experimental study in dogs. I: Clinical and radiographic observations. Clin Oral Implants Res 19(10), 997–1002.Google Scholar
Bannerman, A, Williams, RL, Cox, SC & Grover, LM (2016). Visualising phase change in a brushite-based calcium phosphate ceramic. Sci Rep 6, 32671.Google Scholar
Beier, BD, Quivey, RG & Berger, AJ (2010). Identification of different bacterial species in biofilms using confocal Raman microscopy. J Biomed Opt 15(6), 066001.Google Scholar
Bjørnøy, SH, Bassett, DC, Ucar, S, Strand, BL, Andreassen, JP & Sikorski, P (2016). A correlative spatiotemporal microscale study of calcium phosphate formation and transformation within an alginate hydrogel matrix. Acta Biomater 44, 254–266.Google Scholar
Buchwald, T, Okulus, Z & Szybowicz, M (2017). Raman spectroscopy as a tool of early dental caries detection–new insights. J Raman Spectrosc 48(8), 1094–1102.Google Scholar
Cheng, PT (1987). Formation of octacalcium phosphate and subsequent transformation to hydroxyapatite at low supersaturation: A model for cartilage calcification. Calcif Tissue Int 40(6), 339–343.Google Scholar
de Aza, PN, Guitián, F, Santos, C, de Aza, S, Cuscó, R & Artús, L (1997). Vibrational properties of calcium phosphate compounds. 2. Comparison between hydroxyapatite and β-tricalcium phosphate. Chem Mater 9(4), 916–922.Google Scholar
Flemming, HC, Wingender, J, Szewzyk, U, Steinberg, P, Rice, SA & Kjelleberg, S (2016). Biofilms: An emergent form of bacterial life. Nat Rev Microbiol 14(9), 563–575.Google Scholar
Gonzales, F & Sognnaes, RF (1960). Electron microscopy of dental calculus. Science 131(3394), 156–158.Google Scholar
Holliday, R, Preshaw, PM, Bowen, L & Jakubovics, NS (2015). The ultrastructure of subgingival dental plaque, revealed by high-resolution field emission scanning electron microscopy. BDJ Open 1, 15003.Google Scholar
Ivleva, NP, Kubryk, P & Niessner, R (2017). Raman microspectroscopy, surface-enhanced Raman scattering microspectroscopy, and stable-isotope Raman microspectroscopy for biofilm characterization. Anal Bioanal Chem 409(18), 4353–4375.Google Scholar
Kakei, M, Nakahara, H, Kumegawa, M, Yoshikawa, M & Kunii, S (2000). Demonstration of the central dark line in crystals of dental calculus. Biochim Biophys Acta 1524(2), 189–195.Google Scholar
Kani, T, Kani, M, Moriwaki, Y & Doi, Y (1983). Microbeam x-ray diffraction analysis of dental calculus. J Dent Res 62(2), 92–95.Google Scholar
Kodaka, T, Debari, K & Higashi, S (1988). Magnesium-containing crystals in human dental calculus. J Electron Microsc (Tokyo) 37(2), 73–80.Google Scholar
LeGeros, RZ (1985). Preparation of octacalcium phosphate (OCP): A direct fast method. Calcif Tissue Int 37(2), 194–197.Google Scholar
LeGeros, RZ, Sakae, T, Bautista, C, Retino, M & LeGeros, JP (1996). Magnesium and carbonate in enamel and synthetic apatites. Adv Dent Res 10(2), 225–231.Google Scholar
Lustmann, J, Lewin-Epstein, J & Shteyer, A (1976). Scanning electron microscopy of dental calculus. Calcif Tissue Res 21(1), 47–55.Google Scholar
Ramakrishnaiah, R, Rehman, GU, Basavarajappa, S, Al Khuraif, AA, Durgesh, BH, Khan, AS & Rehman, IU (2015). Applications of Raman spectroscopy in dentistry: Analysis of tooth structure. Appl Spectrosc Rev 50(4), 332–350.Google Scholar
Rizzo, AA, Martin, GR, Scott, DB & Mergenhagen, SE (1962). Mineralization of bacteria. Science 135(3502), 439–441.Google Scholar
Rizzo, AA, Scott, DB & Bladen, HA (1963). Calcification of oral bacteria. Ann N Y Acad Sci 109, 14–22.Google Scholar
Schroeder, HE (1969). Formation and Inhibition of Dental Calculus. Vienna: Hans Huber.Google Scholar
Shah, FA, Lee, BEJ, Tedesco, J, Larsson Wexell, C, Persson, C, Thomsen, P, Grandfield, K & Palmquist, A (2017). Micrometer-sized magnesium whitlockite crystals in micropetrosis of bisphosphonate-exposed human alveolar bone. Nano Lett 17(10), 6210–6216.Google Scholar
Shah, SR, Tatara, AM, D'Souza, RN, Mikos, AG & Kasper, FK (2013). Evolving strategies for preventing biofilm on implantable materials. Mater Today 16(5), 177–182.Google Scholar
Shi, H, He, F & Ye, J (2016). Synthesis and structure of iron- and strontium-substituted octacalcium phosphate: Effects of ionic charge and radius. J Mater Chem B 4(9), 1712–1719.Google Scholar
Tsuda, H & Arends, J (1993). Raman spectra of human dental calculus. J Dent Res 72(12), 1609–1613.Google Scholar
Wopenka, B & Pasteris, JD (2005). A mineralogical perspective on the apatite in bone. Mater Sci Eng C Mater Biol Appl 25(2), 131–143.Google Scholar
Zander, HA, Hazen, SP & Scott, DB (1960). Mineralization of dental calculus. Proc Soc Exp Biol Med 103, 257–260.Google Scholar
Zhou, H, Yang, D, Ivleva, NP, Mircescu, NE, Schubert, S, Niessner, R, Wieser, A & Haisch, C (2015). Label-free in situ discrimination of live and dead bacteria by surface-enhanced Raman scattering. Anal Chem 87(13), 6553–6561.Google Scholar
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