Skip to main content

Advertisement

SpringerLink
Log in

Effects of low-concentrated chlorhexidine on growth of Streptococcus sobrinus and primary human gingival fibroblasts

  • Original Article
  • Published:
Clinical Oral Investigations Aims and scope Submit manuscript

Abstract

The aim of this study was to investigate the in vitro effects of low concentrations of chlorhexidine (CHX) on the proliferation of Streptococcus sobrinus (ATCC 33478) and primary human gingival fibroblasts (HGF). Liquid cultures of bacteria or human gingival fibroblasts were exposed to CHX concentrations ranging from 0.07 to 40 μM in microtiter plates at 37°C for 24 h. Bacteria or cells grown without CHX served as controls. The effects of CHX were determined either by measurements of the optical density (OD) of bacterial cultures or by evaluation of cell growth with the DNA-intercalating fluorescent stain H33342 in comparison to untreated controls. Results were evaluated calculating means and standard deviations. Data were statistically analyzed by an ANOVA using Post Hoc tests (p<0.005). No growth inhibition of S. sobrinus was found at concentrations between 0.07 and 0.15 μM CHX, whereas 0.3 μM CHX led to an elongated (2 h more) lag phase and 0.6 μM CHX induced a lag phase of 4 h more and a minor inclination of the curve in the log phase. Concentrations of CHX≥1.25 μM completely inhibited growth of S. sobrinus. On the contrary, CHX showed no significant effect on growth of HGF at concentrations ≤5 μM. A slight growth inhibition was only observed at a concentration of 5 μM. CHX-concentrations of 10 and 20 μM reduced cell growth to 88 or 75% of control assays. Data analysis showed that overall treatment effects were highly significant (p<0.005). Our data reveal that chlorhexidine inhibits proliferation of S. sobrinus even at very low concentrations while concentrations of CHX≤5 μM are not cytotoxic to human gingival fibroblasts.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1.
Fig. 2.

Similar content being viewed by others

References

  1. Arweiler NB, Netuschil L, Reich E (2001) Alcohol-free mounthrinse solutions to reduce supragingival plaque regrowth and vitality. A controlled clinical study. J Clin Periodontol 28:168–174

    Article  CAS  PubMed  Google Scholar 

  2. Babich H, Wurzburger BJ, Rubin YL, Sinensky MC, Blau L (1995) An in vitro study on the cytotoxicity of chlorhexidine digluconate to human gingival cells. Cell Biol Toxicol 11:79–88

    CAS  PubMed  Google Scholar 

  3. Bonesvoll P (1977) Oral pharmacology of chlorhexidine. J Clin Periodontol 4:49–65

    CAS  PubMed  Google Scholar 

  4. Botelho MG (2000) The minimum inhibitory concentration of oral antibacterial agents against cariogenic organisms. Microbios 103:31–41

    CAS  PubMed  Google Scholar 

  5. Cline NV, Layman DL (1992) The effects of chlorhexidine on the attachment and growth of cultured human periodontal cells. J Periodontol 63:598–602

    CAS  PubMed  Google Scholar 

  6. Decker EM, Weiger R, von Ohle C, Wiech I, Brecx, M (2003) Susceptibility of planktonic versus attached Streptococcus sanguinis cells to chlorhexidine. Clin Oral Invest (in press)

  7. Grönroos L, Mättö J, Saarela M, Luoma AR, Luoma H, Jousimies-Somer H, Pyhälä L, Asikainen S, Alaluusua S (1995) Chlorhexidine susceptibilities of mutans streptococcal serotypes and ribotypes. Antimicrobial Agent and Chemotherapy 39:894–898

    Google Scholar 

  8. Hansel C, Leyhausen G, Mai UEH, Geurtsen W (1998) Effects of various resin composite (co)monomers and extracts on two caries-associated micro-organisms in vitro. J Dent Res 77:60–67

    CAS  PubMed  Google Scholar 

  9. Helgeland K, Heyden G, Rølla G (1971) Effect of chlorhexidine on animal cells in vitro. Scand J Dent Res 79:209–215

    CAS  PubMed  Google Scholar 

  10. Hidalgo E, Dominguez C (2001) Mechanisms underlying chlorhexidine-induced cytotoxicity. Toxicol In Vitro 15:271–276

    Article  CAS  PubMed  Google Scholar 

  11. Kohler B, Bjarnason S (1992) Mutans streptococci, lactobacilli and caries prevalence in 15 to 16 year olds in Goteborg. Part II. Swed Dent J 16:253–259

    CAS  PubMed  Google Scholar 

  12. Koontongkaew S, Jitpukdeebodintra S (1995) Interaction of chlorhexidine with cytoplasmatic membranes of Streptococcus mutans GS-5. Caries Res 29:413–417

    CAS  PubMed  Google Scholar 

  13. Luoma H, Murtomaa H, Nuuja T, Nyman A, Nummkoski P, Ainama J, Luoma AR (1978) A simultaneous reduction of caries and gingivitis in a group of school-children receiving chlorhexidine-fluoride applications. Results after 2 years. Caries Res. 12:290–298

    Google Scholar 

  14. Mariotti AJ, Rumpf DAH (1999) Chlorhexidine-induced changes to human gingival fibroblast collagen and non-collagen protein production. J Periodontol 70:1443–1448

    CAS  PubMed  Google Scholar 

  15. Netuschil L, Weiger R, Preisler R, Brecx M (1995) Plaque bacteria counts and vitality during chlorhexidine, meridol and listerine mouthrinses. Eur J Oral Sci 103:355–361

    CAS  PubMed  Google Scholar 

  16. Pesonon T, Holmalahti J, Pohjola J (1995) Determination of chlorhexidine in saliva using high-performance liquid chromatography. J Chromatogr B Biomed Appl 665:222–225

    Article  PubMed  Google Scholar 

  17. Portenier I, Haapasalo H, Ørstavik D, Yamauchi M, Haapasalo M (2002) Inactivation of the antibacterial activity of iodine potassium iodide and chlorhexidine digluconate against Enterococcus faecalis by dentin, dentin matrix, type-I collagen, and heat-killed microbial whole cells. J Endodont 28:634–637

    Google Scholar 

  18. Pucher JJ, Daniel JC (1993) The effects of chlorhexidine digluconate on human fibroblasts in vitro. J Periodontol 62:526–532

    Google Scholar 

  19. Rølla G, Løe H, Schiøtt CR (1971) Retention of chlorhexidine in the human oral cavity. Archs Oral Biol 16:1109–1116

    Google Scholar 

  20. Salem AM, Adams D, Newman HN, Rawle LW (1987) Antimicrobial properties of two aliphatic amines and chlorhexidine in vitro and in saliva. J Clin Periodontol 14:44–47

    CAS  PubMed  Google Scholar 

  21. Shani S, Friedman M, Steinberg D (2000) The anticariogenic effect of amine fluorides on streptococcus sobrinus and glucosyltransferase in biofilms. Caries Res 34:260–267

    Article  CAS  PubMed  Google Scholar 

  22. Shapiro S, Giertsen E, Guggenheim B (2002) An in vitro oral biofilm model for comparing the efficacy of antimicrobial mouthrinses. Caries Res 36:93–100

    Article  CAS  PubMed  Google Scholar 

  23. Spijkervet FK, van Saene JJ, van Saene HK, Panders AK, Vermey A, Fidler V (1990) Chlorhexidine inactivation by saliva. Oral Surg Oral Med Oral Pathol 69:444–449

    CAS  PubMed  Google Scholar 

  24. Steinberg D, Rothman M (1996) Antibacterial effect of chlorhexidine on bacteria adsorbed onto experimental dental plaque. Diagn Microbiol Infect Dis 26:109–115

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors would like to thank Mr. Oliver Staperfeld and Mr. Dennis DiJulio for their excellent technical assistance.

Author information

Authors and Affiliations

  1. Department of Conservative Dentistry & Periodontology, Medical University Hannover, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    S. Dogan, H. Günay & G. Leyhausen

  2. Department of Restorative Dentistry, School of Dentistry, University of Washington, Seattle, USA

    W. Geurtsen

Authors
  1. S. Dogan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. H. Günay
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G. Leyhausen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. W. Geurtsen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to S. Dogan.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dogan, S., Günay, H., Leyhausen, G. et al. Effects of low-concentrated chlorhexidine on growth of Streptococcus sobrinus and primary human gingival fibroblasts. Clin Oral Invest 7, 212–216 (2003). https://doi.org/10.1007/s00784-003-0226-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00784-003-0226-3

Keywords

Search

Navigation