Hostname: page-component-78c5997874-8bhkd Total loading time: 0 Render date: 2024-11-08T05:00:11.575Z Has data issue: false hasContentIssue false

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*
Affiliation:
Department of Biomaterials, Sahlgrenska Academy, University of Gothenburg, Göteborg, Sweden
*
*Author for correspondence: Furqan A. Shah, E-mail: [email protected]
Get access

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.

Type
Biological Science Applications
Copyright
Copyright © Microscopy Society of America 2019 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Akcali, A & Lang, NP (2018). Dental calculus: The calcified biofilm and its role in disease development. Periodontol 2000 76(1), 109115.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), 9971002.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, 254266.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), 10941102.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), 339343.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), 916922.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), 563575.Google Scholar
Gonzales, F & Sognnaes, RF (1960). Electron microscopy of dental calculus. Science 131(3394), 156158.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), 43534375.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), 189195.Google Scholar
Kani, T, Kani, M, Moriwaki, Y & Doi, Y (1983). Microbeam x-ray diffraction analysis of dental calculus. J Dent Res 62(2), 9295.Google Scholar
Kodaka, T, Debari, K & Higashi, S (1988). Magnesium-containing crystals in human dental calculus. J Electron Microsc (Tokyo) 37(2), 7380.Google Scholar
LeGeros, RZ (1985). Preparation of octacalcium phosphate (OCP): A direct fast method. Calcif Tissue Int 37(2), 194197.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), 225231.Google Scholar
Lustmann, J, Lewin-Epstein, J & Shteyer, A (1976). Scanning electron microscopy of dental calculus. Calcif Tissue Res 21(1), 4755.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), 332350.Google Scholar
Rizzo, AA, Martin, GR, Scott, DB & Mergenhagen, SE (1962). Mineralization of bacteria. Science 135(3502), 439441.Google Scholar
Rizzo, AA, Scott, DB & Bladen, HA (1963). Calcification of oral bacteria. Ann N Y Acad Sci 109, 1422.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), 62106216.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), 177182.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), 17121719.Google Scholar
Tsuda, H & Arends, J (1993). Raman spectra of human dental calculus. J Dent Res 72(12), 16091613.Google Scholar
Wopenka, B & Pasteris, JD (2005). A mineralogical perspective on the apatite in bone. Mater Sci Eng C Mater Biol Appl 25(2), 131143.Google Scholar
Zander, HA, Hazen, SP & Scott, DB (1960). Mineralization of dental calculus. Proc Soc Exp Biol Med 103, 257260.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), 65536561.Google Scholar
Supplementary material: PDF

Shah supplementary material

Figures S1-S5

Download Shah supplementary material(PDF)
PDF 4.2 MB