Skip to main content
Top
Published in: BMC Oral Health 1/2018

Open Access 01-12-2018 | Research article

The effect of chlorhexidine on dental calculus formation: an in vitro study

Authors: Yuuki Sakaue, Shoji Takenaka, Tatsuya Ohsumi, Hisanori Domon, Yutaka Terao, Yuichiro Noiri

Published in: BMC Oral Health | Issue 1/2018

Login to get access

Abstract

Background

Chlorhexidine gluconate (CHG) has been proven to be effective in preventing and controlling biofilm formation. At the same time, an increase in calculus formation is known as one of considerable side effects. The purpose of this study was to investigate whether mineral deposition preceding a calculus formation would occur at an early stage after the use of CHG using an in vitro saliva-related biofilm model.

Methods

Biofilms were developed on the MBEC™ device in brain heart infusion (BHI) broth containing 0.5% sucrose at 37 °C for 3 days under anaerobic conditions. Biofilms were periodically exposed to 1 min applications of 0.12% CHG every 12 h and incubated for up to 2 days in BHI containing a calcifying solution. Calcium and phosphate in the biofilm were measured using atomic absorption spectrophotometry and a phosphate assay kit, respectively. Morphological structure was observed using a scanning electron microscope (SEM), and chemical composition was analyzed with an electron probe microanalyzer (EPMA).

Results

The concentrations of Ca and Pi following a single exposure to CHG increased significantly compared with the control. Repeatedly exposing biofilms to CHG dose-dependently increased Ca deposition, and the amount of Ca was five times as much as that of the control. Pi levels in CHG-treated biofilms were significantly higher than those from the control group (p < 0.05); however, the influence of the number of exposures was limited. Analyses using an SEM and EPMA showed many clusters containing calcium and phosphate complexes in CHG-treated biofilms. Upon composition analysis of the clusters, calcium was detected at a greater concentration than phosphate.

Conclusions

Findings suggested that CHG may promote mineral uptake into the biofilm soon after its use. It is necessary to disrupt the biofilm prior to the start of a CHG mouthwash in order to reduce the side effects associated with this procedure. The management of patients is also important.
Literature
1.
go back to reference Lindhe J, Karring T, Lang N. Clinical periodontology and implant dentistry. 3rd ed. Blackwell Munksgaard: Copenhagen; 2003. Lindhe J, Karring T, Lang N. Clinical periodontology and implant dentistry. 3rd ed. Blackwell Munksgaard: Copenhagen; 2003.
2.
go back to reference Figuero E, Nóbrega DF, García-Gargallo M, Tenuta LMA, Herrera D, Carvalho JC. Mechanical and chemical plaque control in the simultaneous management of gingivitis and caries: a systematic review. J Clin Periodontol. 2017;44(Suppl 18):116–34.CrossRef Figuero E, Nóbrega DF, García-Gargallo M, Tenuta LMA, Herrera D, Carvalho JC. Mechanical and chemical plaque control in the simultaneous management of gingivitis and caries: a systematic review. J Clin Periodontol. 2017;44(Suppl 18):116–34.CrossRef
3.
go back to reference Van der Weijden FA, Slot DE. Efficacy of homecare regimens for mechanical plaque removal in managing gingivitis a meta review. J Clin Periodontol. 2015;42(Suppl 16):77–91.CrossRef Van der Weijden FA, Slot DE. Efficacy of homecare regimens for mechanical plaque removal in managing gingivitis a meta review. J Clin Periodontol. 2015;42(Suppl 16):77–91.CrossRef
4.
go back to reference Sälzer S, Slot DE, Van der Weijden FA, Dörfer CE. Efficacy of inter-dental mechanical plaque control in managing gingivitis - a meta-review. J Clin Periodontol. 2015;42(Suppl 16):92–105.CrossRef Sälzer S, Slot DE, Van der Weijden FA, Dörfer CE. Efficacy of inter-dental mechanical plaque control in managing gingivitis - a meta-review. J Clin Periodontol. 2015;42(Suppl 16):92–105.CrossRef
5.
go back to reference Serrano J, Escribano M, Roldán S, Martín C, Herrera D. Efficacy of adjunctive anti-plaque chemical agents in managing gingivitis: a systematic review and meta-analysis. J Clin Periodontol. 2015;42(Suppl 16):106–38.CrossRef Serrano J, Escribano M, Roldán S, Martín C, Herrera D. Efficacy of adjunctive anti-plaque chemical agents in managing gingivitis: a systematic review and meta-analysis. J Clin Periodontol. 2015;42(Suppl 16):106–38.CrossRef
6.
go back to reference Santos A. Evidence-based control of plaque and gingivitis. J Clin Periodontol. 2003;30(Suppl 5):13–6.CrossRefPubMed Santos A. Evidence-based control of plaque and gingivitis. J Clin Periodontol. 2003;30(Suppl 5):13–6.CrossRefPubMed
7.
go back to reference Jones CG. Chlorhexidine: is it still the gold standard? Periodontol. 1997;15:55–62.CrossRef Jones CG. Chlorhexidine: is it still the gold standard? Periodontol. 1997;15:55–62.CrossRef
8.
go back to reference Van Strydonck DA, Slot DE, Van der Velden U, Van der Weijden F. Effect of a chlorhexidine mouthrinse on plaque, gingival inflammation and staining in gingivitis patients: a systematic review. J Clin Periodontol. 2012;39(11):1042–55.CrossRefPubMed Van Strydonck DA, Slot DE, Van der Velden U, Van der Weijden F. Effect of a chlorhexidine mouthrinse on plaque, gingival inflammation and staining in gingivitis patients: a systematic review. J Clin Periodontol. 2012;39(11):1042–55.CrossRefPubMed
9.
go back to reference Overholser CD, Meiller TF, DePaola LG, Minah GE, Niehaus C. Comparative effects of 2 chemotherapeutic mouthrinses on the development of supragingival dental plaque and gingivitis. J Clin Periodontol. 1990;17:575–9.CrossRefPubMed Overholser CD, Meiller TF, DePaola LG, Minah GE, Niehaus C. Comparative effects of 2 chemotherapeutic mouthrinses on the development of supragingival dental plaque and gingivitis. J Clin Periodontol. 1990;17:575–9.CrossRefPubMed
10.
go back to reference Yates R, Jenkins S, Newcombe R, Wade W, Moran J, Addy M. A 6-month home usage trial of a 1% chlorhexidine toothpaste (1). Effects on plaque, gingivitis, calculus and toothstaining. J Clin Periodontol. 1993;20:130–8.CrossRefPubMed Yates R, Jenkins S, Newcombe R, Wade W, Moran J, Addy M. A 6-month home usage trial of a 1% chlorhexidine toothpaste (1). Effects on plaque, gingivitis, calculus and toothstaining. J Clin Periodontol. 1993;20:130–8.CrossRefPubMed
11.
go back to reference Charles CH, Mostler KM, Bartels LL, Mankodi SM. Comparative antiplaque and antigingivitis effectiveness of a chlorhexidine and an essential oil mouthrinse: 6-month clinical trial. J Clin Periodontol. 2004;31:878–84.CrossRefPubMed Charles CH, Mostler KM, Bartels LL, Mankodi SM. Comparative antiplaque and antigingivitis effectiveness of a chlorhexidine and an essential oil mouthrinse: 6-month clinical trial. J Clin Periodontol. 2004;31:878–84.CrossRefPubMed
12.
go back to reference Kumar S, Patel S, Tadakamadla J, Tibdewal H, Duraiswamy P, Kulkarni S. Effectiveness of a mouthrinse containing active ingredients in addition to chlorhexidine and triclosan compared with chlorhexidine and triclosan rinses on plaque, gingivitis, supragingival calculus and extrinsic staining. Int J Dent Hyg. 2013;11:35–40.CrossRefPubMed Kumar S, Patel S, Tadakamadla J, Tibdewal H, Duraiswamy P, Kulkarni S. Effectiveness of a mouthrinse containing active ingredients in addition to chlorhexidine and triclosan compared with chlorhexidine and triclosan rinses on plaque, gingivitis, supragingival calculus and extrinsic staining. Int J Dent Hyg. 2013;11:35–40.CrossRefPubMed
13.
go back to reference Jepsen S, Deschner J, Braun A, Schwarz F, Eberhard J. Calculus removal and the prevention of its formation. Periodontol. 2011;55:167–88.CrossRef Jepsen S, Deschner J, Braun A, Schwarz F, Eberhard J. Calculus removal and the prevention of its formation. Periodontol. 2011;55:167–88.CrossRef
14.
go back to reference White DJ. Dental calculus: recent insights into occurrence, formation, prevention, removal and oral health effects of supragingival and subgingival deposits. Eur J Oral Sci. 1997;105:508–22.CrossRefPubMed White DJ. Dental calculus: recent insights into occurrence, formation, prevention, removal and oral health effects of supragingival and subgingival deposits. Eur J Oral Sci. 1997;105:508–22.CrossRefPubMed
15.
go back to reference Yamaguchi M, Noiri Y, Kuboniwa M, Yamamoto R, Asahi Y, Maezono H, Hayashi M, Ebisu S. Porphyromonas gingivalis biofilms persist after chlorhexidine treatment. Eur J Oral Sci. 2013;121:162–8.CrossRefPubMed Yamaguchi M, Noiri Y, Kuboniwa M, Yamamoto R, Asahi Y, Maezono H, Hayashi M, Ebisu S. Porphyromonas gingivalis biofilms persist after chlorhexidine treatment. Eur J Oral Sci. 2013;121:162–8.CrossRefPubMed
16.
go back to reference Ohsumi T, Takenaka S, Wakamatsu R, Sakaue Y, Narisawa N, Senpuku H, Ohshima H, Terao Y, Okiji T. Residual structure of Streptococcus mutans biofilm following complete disinfection favors secondary bacterial adhesion and biofilm re-development. PLoS One. 2015;10:e0116647.CrossRefPubMedPubMedCentral Ohsumi T, Takenaka S, Wakamatsu R, Sakaue Y, Narisawa N, Senpuku H, Ohshima H, Terao Y, Okiji T. Residual structure of Streptococcus mutans biofilm following complete disinfection favors secondary bacterial adhesion and biofilm re-development. PLoS One. 2015;10:e0116647.CrossRefPubMedPubMedCentral
17.
go back to reference Pesciaroli L, Petruccioli M, Fedi S, Firrincieli A, Federici F, D’Annibale A. Characterization of Pleurotus ostreatus biofilms by using the Calgary biofilm device. Appl Environ Microbiol. 2013;79:6083–92.CrossRefPubMedPubMedCentral Pesciaroli L, Petruccioli M, Fedi S, Firrincieli A, Federici F, D’Annibale A. Characterization of Pleurotus ostreatus biofilms by using the Calgary biofilm device. Appl Environ Microbiol. 2013;79:6083–92.CrossRefPubMedPubMedCentral
18.
go back to reference Najafi MH, Taheri M, Mokhtari MR, Forouzanfar A, Farazi F, Mirzaee M, Ebrahiminik Z, Mehrara R. Comparative study of 0.2 and 0.12% digluconate chlorhexidine mouth rinses on the level of dental staining and gingival indices. Dent Res J. 2012;9:305–8. Najafi MH, Taheri M, Mokhtari MR, Forouzanfar A, Farazi F, Mirzaee M, Ebrahiminik Z, Mehrara R. Comparative study of 0.2 and 0.12% digluconate chlorhexidine mouth rinses on the level of dental staining and gingival indices. Dent Res J. 2012;9:305–8.
19.
go back to reference Matthews D. No difference between 0.12 and 0.2% chlorhexidine mouthrinse on reduction of gingivitis. Evid Based Dent. 2011;12:8–9.CrossRefPubMed Matthews D. No difference between 0.12 and 0.2% chlorhexidine mouthrinse on reduction of gingivitis. Evid Based Dent. 2011;12:8–9.CrossRefPubMed
20.
go back to reference Sidaway DA. A microbiological study of dental calculus. II. The in vitro calcification of microorganisms from dental calculus. J Periodontal Res. 1978;13:360–6.CrossRefPubMed Sidaway DA. A microbiological study of dental calculus. II. The in vitro calcification of microorganisms from dental calculus. J Periodontal Res. 1978;13:360–6.CrossRefPubMed
21.
go back to reference Wong L, Sissons CH. Human dental plaque microcosm biofilms: effect of nutrient variation on calcium phosphate deposition and growth. Arch Oral Biol. 2007;52(3):280–9.CrossRefPubMed Wong L, Sissons CH. Human dental plaque microcosm biofilms: effect of nutrient variation on calcium phosphate deposition and growth. Arch Oral Biol. 2007;52(3):280–9.CrossRefPubMed
22.
go back to reference Eilberg RG, Judy K, Lovino E, Kornfeld P, Phelan J, Ellison R. Relationship between plaque mineralization in vitro and calculus formation in vivo. J Dent Res. 1973;52(1):45–8.CrossRefPubMed Eilberg RG, Judy K, Lovino E, Kornfeld P, Phelan J, Ellison R. Relationship between plaque mineralization in vitro and calculus formation in vivo. J Dent Res. 1973;52(1):45–8.CrossRefPubMed
23.
go back to reference Karduck P. Quantitative near-surface microanalysis and depth profiling by EPMA. In: Love G, Nicholson WAP, Armigliato A. (eds) Modern developments and applications in microbeam Analysis. Vienna: Mikrochimica Acta. Splinger; 1980;15:109–23. Karduck P. Quantitative near-surface microanalysis and depth profiling by EPMA. In: Love G, Nicholson WAP, Armigliato A. (eds) Modern developments and applications in microbeam Analysis. Vienna: Mikrochimica Acta. Splinger; 1980;15:109–23.
24.
go back to reference Escribano M, Figuero E, Martín C, Tobías A, Serrano J, Roldán S, Herrera D. Efficacy of adjunctive anti-plaque chemical agents: a systematic review and network meta-analyses of the Turesky modification of the Quigley and Hein plaque index. J Clin Periodontol. 2016;43(12):1059–73.CrossRefPubMed Escribano M, Figuero E, Martín C, Tobías A, Serrano J, Roldán S, Herrera D. Efficacy of adjunctive anti-plaque chemical agents: a systematic review and network meta-analyses of the Turesky modification of the Quigley and Hein plaque index. J Clin Periodontol. 2016;43(12):1059–73.CrossRefPubMed
25.
go back to reference James P, Worthington HV, Parnell C, Harding M, Lamont T, Cheung A, Whelton H, Riley P. Chlorhexidine mouthrinse as an adjunctive treatment for gingival health. Cochrane Database Syst Rev. 2017;31(3):CD008676. James P, Worthington HV, Parnell C, Harding M, Lamont T, Cheung A, Whelton H, Riley P. Chlorhexidine mouthrinse as an adjunctive treatment for gingival health. Cochrane Database Syst Rev. 2017;31(3):CD008676.
26.
go back to reference Schroeder HE, Shanley D. Formation and inhibition of dental calculus. J Periodontol. 1969;40:643–6.CrossRefPubMed Schroeder HE, Shanley D. Formation and inhibition of dental calculus. J Periodontol. 1969;40:643–6.CrossRefPubMed
27.
go back to reference Kodaka T, Debari K, Higashi S. Magnesium-containing crystals in human dental calculus. J Electron Microsc. 1988;37:73–80. Kodaka T, Debari K, Higashi S. Magnesium-containing crystals in human dental calculus. J Electron Microsc. 1988;37:73–80.
28.
go back to reference Rose RK, Hogg SD, Shellis RP. A quantitative study of calcium binding by isolated streptococcal cell walls and lipoteichoic acid: comparison with whole cells. J Dent Res. 1994;73(11):1742–7.CrossRefPubMed Rose RK, Hogg SD, Shellis RP. A quantitative study of calcium binding by isolated streptococcal cell walls and lipoteichoic acid: comparison with whole cells. J Dent Res. 1994;73(11):1742–7.CrossRefPubMed
29.
go back to reference Wong L, Sissons CH, El P, Cutress TW. Calcium phosphate deposition in human dental plaque microcosm biofilms induced by a ureolytic pH-rise procedure. Arch Oral Biol. 2002;47(11):779–90.CrossRefPubMed Wong L, Sissons CH, El P, Cutress TW. Calcium phosphate deposition in human dental plaque microcosm biofilms induced by a ureolytic pH-rise procedure. Arch Oral Biol. 2002;47(11):779–90.CrossRefPubMed
30.
go back to reference Allen DL, Kerr DA. Tissue response in the Guinea pig to sterile and non-sterile calculus. J Periodontol. 1965;36:121–6.CrossRefPubMed Allen DL, Kerr DA. Tissue response in the Guinea pig to sterile and non-sterile calculus. J Periodontol. 1965;36:121–6.CrossRefPubMed
31.
go back to reference Nichols FC, Rojanasomsith K. Porphyromonas gingivalis lipids and diseased dental tissues. Oral Microbiol Immunol. 2006;21:84–92.CrossRefPubMed Nichols FC, Rojanasomsith K. Porphyromonas gingivalis lipids and diseased dental tissues. Oral Microbiol Immunol. 2006;21:84–92.CrossRefPubMed
Metadata
Title
The effect of chlorhexidine on dental calculus formation: an in vitro study
Authors
Yuuki Sakaue
Shoji Takenaka
Tatsuya Ohsumi
Hisanori Domon
Yutaka Terao
Yuichiro Noiri
Publication date
01-12-2018
Publisher
BioMed Central
Published in
BMC Oral Health / Issue 1/2018
Electronic ISSN: 1472-6831
DOI
https://doi.org/10.1186/s12903-018-0517-3

Other articles of this Issue 1/2018

BMC Oral Health 1/2018 Go to the issue