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Assessment of multispecies biofilm growth on root canal dentin under different radiation therapy regimens

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Abstract

Objectives

To assess the growth of a multispecies biofilm on root canal dentin under different radiotherapy regimens.

Materials and methods

Sixty-three human root dentin cylinders were distributed into six groups. In three groups, no biofilm was formed (n = 3): NoRT) non-irradiated dentin; RT55) 55 Gy; and RT70) 70 Gy. In the other three groups (n = 18), a 21-day multispecies biofilm (Enterococcus faecalis, Streptococcus mutans, and Candida albicans) was formed in the canal: NoRT + Bio) non-irradiated + biofilm; RT55 + Bio) 55 Gy + biofilm; and RT70 + Bio) 70 Gy + biofilm. The biofilm was quantified (CFUs/mL). Biofilm microstructure was assessed under SEM. Microbial penetration into dentinal tubules was assessed under CLSM. For the biofilm biomass and dentin microhardness pre- and after biofilm growth assessments, 45 bovine dentin specimens were distributed into three groups (n = 15): NoRT) non-irradiated + biofilm; RT55 + Bio) 55 Gy + biofilm; and RT70 + Bio) 70 Gy + biofilm.

Results

Irradiated specimens (70 Gy) had higher quantity of microorganisms than non-irradiated (p = .010). There was gradual increase in biofilm biomass from non-irradiated to 55 Gy and 70 Gy (p < .001). Irradiated specimens had greater reduction in microhardness after biofilm growth. Irradiated dentin led to the growth of a more complex and irregular biofilm. There was microbial penetration into the dentinal tubules, regardless of the radiation regimen.

Conclusion

Radiotherapy increased the number of microorganisms and biofilm biomass and reduced dentin microhardness. Microbial penetration into dentinal tubules was noticeable.

Clinical relevance

Cumulative and potentially irreversible side effects of radiotherapy affect biofilm growth on root dentin. These changes could compromise the success of endodontic treatment in oncological patients undergoing head and neck radiotherapy.

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Data availability

The data that support the findings of this study are available on request from the corresponding author. The data are not publicly available due to privacy or ethical restrictions.

References

  1. van Dijk B, Lemans JVC, Hoogendoorn RM, Dadachova E, de Klerk JMH, Vogely HC, Weinans H, Lam MGEH, van der Wal BCH (2020) Treating infections with ionizing radiation: a historical perspective and emerging techniques. Antimicrob Resist Infect Control 9:121. https://doi.org/10.1186/s13756-020-00775-w

    Article  PubMed  PubMed Central  Google Scholar 

  2. De Felice F, Polimeni A, Valentini V, Brugnoletti O, Cassoni A, Greco A, de Vincentiis M, Tombolini V (2018) Radiotherapy controversies and prospective in head and neck cancer: a literature-based critical review. Neoplasia 20:227–232. https://doi.org/10.1016/j.neo.2018.01.002

    Article  PubMed  PubMed Central  Google Scholar 

  3. Velo MMAC, Farha ALH, da Silva Santos PS, Shiota A, Sansavino SZ, Souza AT, Honório HM, Wang L (2018) Radiotherapy alters the composition, structural and mechanical properties of root dentin in vitro. Clin Oral Investig 22:2871–2878. https://doi.org/10.1007/s00784-018-2373-6

    Article  PubMed  Google Scholar 

  4. Douchy L, Gauthier R, Abouelleil-Sayed H, Colon P, Grosgogeat B, Bosco J (2022) The effect of therapeutic radiation on dental enamel and dentin: a systematic review. Dent Mater 38:e181–e201. https://doi.org/10.1016/j.dental.2022.04.014

    Article  CAS  PubMed  Google Scholar 

  5. Fonseca JM, Troconis CC, Palmier NR, Gomes-Silva W, Paglioni MD, Araújo AL, Arboleda LP, Filho AJ, González-Arriagada WA, Goes MF, Lopes MA, Brandão TB, Vargas PA, Ribeiro AC, Santos-Silva AR (2020) The impact of head and neck radiotherapy on the dentine-enamel junction: a systematic review. Med Oral Patol Oral Cir Bucal 25:e96–e105. https://doi.org/10.4317/medoral.23212

    Article  PubMed  Google Scholar 

  6. Gupta N, Pal M, Rawat S, Grewal MS, Garg H, Chauhan D, Ahlawat P, Tandon S, Khurana R, Pahuja AK, Mayank M, Devnani B (2015) Radiation-induced dental caries, prevention and treatment - A systematic review. Natl J Maxillofac Surg 6:160–166. https://doi.org/10.4103/0975-5950.183870

    Article  PubMed  PubMed Central  Google Scholar 

  7. Rodrigues RB, Soares CJ, Junior PCS, Lara VC, Arana-Chavez VE, Novais VR (2018) Influence of radiotherapy on the dentin properties and bond strength. Clin Oral Investig 22:875–883. https://doi.org/10.1007/s00784-017-2165-4

    Article  PubMed  Google Scholar 

  8. Venegas SC, Palacios JM, Apella MC, Morando PJ, Blesa MA (2006) Calcium modulates interactions between bacteria and hydroxyapatite. J Dent Res 85:1124–1128. https://doi.org/10.1177/154405910608501211

    Article  CAS  PubMed  Google Scholar 

  9. Teughels W, Van Assche N, Sliepen I, Quirynen M (2006) Effect of material characteristics and/or surface topography on biofilm development. Clin Oral Implants Res 17:68–81. https://doi.org/10.1111/j.1600-0501.2006.01353.x

    Article  PubMed  Google Scholar 

  10. Parmar D, Hauman CH, Leichter JW, McNaughton A, Tompkins GR (2011) Bacterial localization and viability assessment in human ex vivo dentinal tubules by fluorescence confocal laser scanning microscopy. Int Endod J 44:644–651. https://doi.org/10.1111/j.1365-2591.2011.01867.x

    Article  CAS  PubMed  Google Scholar 

  11. de Miranda RR, Silva ACA, Dantas NO, Soares CJ, Novais VR (2018) Chemical analysis of in vivo-irradiated dentine of head and neck cancer patients by ATR-FTIR and Raman spectroscopy. Clin Oral Investig 23:3351–3358. https://doi.org/10.1007/s00784-018-2758-6

    Article  PubMed  Google Scholar 

  12. Lopes CCA, Rodrigues RB, Cenci MS, Uehara JLS, Maske TT, Limirio PHJO, Soares PBF, Novais VR (2021) Effect of ionizing radiation and cariogenic biofilm challenge on root-dentin caries. Clin Oral Investig 25:4059–4068. https://doi.org/10.1007/s00784-020-03736-0

    Article  PubMed  Google Scholar 

  13. Hillesheim LC, Hoffmann JB, Schuldt DP, Tedesco M, Bortoluzzi EA, Teixeira CS (2017) Intracanal irrigating solutions prior to calcium hydroxide medication and its effects on root dentin strength. Braz Dent J 28:46–50. https://doi.org/10.1590/0103-6440201700698

    Article  PubMed  Google Scholar 

  14. de Barros da Cunha SR, Fonseca FP, Ramos PAMM, Haddad CMK, Fregnani ER, Aranha ACC (2017) Effects of different radiation doses on the microhardness, superficial morphology, and mineral components of human enamel. Arch Oral Biol 80:130–135. https://doi.org/10.1016/j.archoralbio.2017.04.007

    Article  CAS  PubMed  Google Scholar 

  15. Yamin PA, Pereira RD, Lopes FC, Queiroz AM, Oliveira HF, Saquy PC, Sousa-Neto MD (2018) Longevity of bond strength of resin cements to root dentine after radiation therapy. Int Endod J 51:1301–1312. https://doi.org/10.1111/iej.12945

    Article  CAS  PubMed  Google Scholar 

  16. Cancelier PDA, Machado RG, Savaris JM, Bortoluzzi EA, Teixeira CDS, Minamisako MC, Rodrigues PM, Netto VR, Dutra-Horstmann KL, Garcia LDFR (2023) Effect of the timing of radiation therapy on the push-out strength of resin cement to root dentine. Aust Endod J 49:122–131. https://doi.org/10.1111/aej.12699

    Article  PubMed  Google Scholar 

  17. Coelho SM, Pandolfo MT, Bortoluzzi EA, Rossetto HL, Pereira RP, Minamisako MC, Rodrigues PM, Netto VR, Ramos TDS, Teixeira CDS, Garcia LDFR (2024) Effect of radiation therapy on fracture resistance of simulated immature teeth submitted to root reinforcement. Int J Paediatr Dent 34:3–10. https://doi.org/10.1111/ipd.13073

    Article  PubMed  Google Scholar 

  18. Cunha Neto MAD, Coêlho JA, Pinto KP, Cuellar MRC, Marcucci MC, Silva EJNL, Andrade FB, Sassone LM (2021) Antibacterial efficacy of triple antibiotic medication with macrogol (3Mix-MP), traditional triple antibiotic paste, calcium hydroxide, and ethanol extract of propolis: an intratubular dentin ex vivo confocal laser scanning microscopic study. J Endod 47:1609–1616. https://doi.org/10.1016/j.joen.2021.07.014

    Article  PubMed  Google Scholar 

  19. de Almeida J, Pimenta AL, Pereira UA, Barbosa LCA, Hoogenkamp MA, van der Waal SV, Crielaard W, Felippe WT (2018) Effects of three γ-alkylidene-γ-lactams on the formation of multispecies biofilms. Eur J Oral Sci 126:214–221. https://doi.org/10.1111/eos.12411

    Article  CAS  PubMed  Google Scholar 

  20. Albuquerque MTP, Nagata J, Bottino MC (2017) Antimicrobial efficacy of triple antibiotic-eluting polymer nanofibers against multispecies biofilm. J Endod 43:S51–S56. https://doi.org/10.1016/j.joen.2017.06.009

    Article  PubMed  PubMed Central  Google Scholar 

  21. Gil ACK, Prado MM, da Rocha LR, Benfatti C, Filho GS, de Almeida J (2023) In vitro evaluation of membranes for regenerative procedures against oral bacteria. Braz Dent J 34:57–65. https://doi.org/10.1590/0103-6440202305060

    Article  PubMed  PubMed Central  Google Scholar 

  22. Alfadda S, Alquria T, Karaismailoglu E, Aksel H, Azim AA (2021) Antibacterial effect and bioactivity of innovative and currently used intracanal medicaments in regenerative endodontics. J Endod 47:1294–1300. https://doi.org/10.1016/j.joen.2021.05.005

    Article  PubMed  Google Scholar 

  23. Deng DM, Hoogenkamp MA, Exterkate RA, Jiang LM, van der Sluis LW, Ten Cate JM, Crielaard W (2009) Influence of streptococcus mutans on enterococcus faecalis biofilm formation. J Endod 35:1249–1252. https://doi.org/10.1016/j.joen.2009.05.038

    Article  PubMed  Google Scholar 

  24. Ricucci D, Siqueira JF Jr (2010) Biofilms and apical periodontitis: study of prevalence and association with clinical and histopathologic findings. J Endod 36:1277–1288. https://doi.org/10.1016/j.joen.2010.04.007

    Article  PubMed  Google Scholar 

  25. Epstein JB, Chin EA, Jacobson JJ, Rishiraj B, Le N (1998) The relationships among fluoride, cariogenic oral flora, and salivary flow rate during radiation therapy. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 86:286–292. https://doi.org/10.1016/s1079-2104(98)90173-1

    Article  CAS  PubMed  Google Scholar 

  26. Siqueira JF Jr, Alves FR, Versiani MA, Rôças IN, Almeida BM, Neves MA, Sousa-Neto MD (2013) Correlative bacteriologic and micro-computed tomographic analysis of mandibular molar mesial canals prepared by self-adjusting file, reciproc, and twisted file systems. J Endod 39:1044–1050. https://doi.org/10.1016/j.joen.2013.04.034

    Article  PubMed  Google Scholar 

  27. Inagati CM, Scheffel DLS, Anovazzi G, Alonso JRL, Christoffoli MT, Pashley DH, De Souza Costa CA, Hebling J (2021) Proteolytic activity and degradation of bovine versus human dentin matrices. J Appl Oral Sci 29:e20210290. https://doi.org/10.1590/1678-7757-2021-0290

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Silva EJNL, Carvalho NK, Prado MC, Senna PM, Souza EM, De-Deus G (2019) Bovine teeth can reliably substitute human dentine in an intra-tooth push-out bond strength model? Int Endod J 52:1063–1069. https://doi.org/10.1111/iej.13085

    Article  CAS  PubMed  Google Scholar 

  29. de Souza GHM, Pandolfo MT, Bortoluzzi EA, da Silveira Teixeira C, Rossetto HL, da Rosa AF, Machado RG, da Fonseca Roberti Garcia L (2022) Effect of root perforation repair with mineral aggregate-based cements on the retention of customized fiberglass posts. Odontology 110:535–544. https://doi.org/10.1007/s10266-022-00686-9

    Article  CAS  PubMed  Google Scholar 

  30. Polce S, Gogineni E, Antone J, Ghaly M, Keith Frank D, Segal JD, Parashar B (2021) Dental radiation dosimetric maps from intensity-modulated radiation therapy planning for head and neck cancers. Head Neck 43:1428–1439. https://doi.org/10.1002/hed.26611

    Article  PubMed  Google Scholar 

  31. van der Waal SV, van der Sluis LW (2012) Potential of calcium to scaffold an endodontic biofilm, thus protecting the micro-organisms from disinfection. Med Hypotheses 79:1–4. https://doi.org/10.1016/j.mehy.2012.03.012

    Article  CAS  PubMed  Google Scholar 

  32. Lin S, Yang L, Chen G, Li B, Chen D, Li L, Xu Z (2017) Pathogenic features and characteristics of food borne pathogens biofilm: biomass, viability and matrix. Microb Pathog 111:285–291. https://doi.org/10.1016/j.micpath.2017.08.005

    Article  CAS  PubMed  Google Scholar 

  33. Xie J, Yang L, Peters BM, Chen L, Chen D, Li B, Li L, Yu G, Xu Z, Shirtliff ME (2017) A 16-year retrospective surveillance report on the pathogenic features and antimicrobial susceptibility of pseudomonas aeruginosa isolates from FAHJU in Guangzhou representative of Southern China. Microb Pathog 110:37–41. https://doi.org/10.1016/j.micpath.2017.06.018

    Article  CAS  PubMed  Google Scholar 

  34. Huang J, Pinder KL (1995) Effects of calcium on development of anaerobic acidogenic biofilms. Biotechnol Bioeng 45:212–218. https://doi.org/10.1002/bit.260450305

    Article  CAS  PubMed  Google Scholar 

  35. Patrauchan MA, Sarkisova S, Sauer K, Franklin MJ (2005) Calcium influences cellular and extracellular product formation during biofilm-associated growth of a marine Pseudoalteromonas sp. Microbiol (Reading) 151:2885–2897. https://doi.org/10.1099/mic.0.28041-0

    Article  CAS  Google Scholar 

  36. Yokoyama K, Takeo K (1983) Differences of asymmetrical division between the pseudomycelial and yeast forms of Candida albicans and their effect on multiplication. Arch Microbiol 134:251–253. https://doi.org/10.1007/BF00407768

    Article  CAS  PubMed  Google Scholar 

  37. Brawner DL, Cutler JE (1985) Changes in surface topography of Candida albicans during morphogenesis. Sabouraudia 23:389–393

    Article  CAS  PubMed  Google Scholar 

  38. Gow NA, Gooday GW (1984) A model for the germ tube formation and mycelial growth form of Candida albicans. Sabouraudia 22:137–144. https://doi.org/10.1080/00362178485380211

    Article  CAS  PubMed  Google Scholar 

  39. Jenkinson HF, Lala HC, Shepherd MG (1990) Coaggregation of Streptococcus Sanguis and other streptococci with Candida albicans. Infect Immun 58:1429–1436. https://doi.org/10.1128/iai.58.5.1429-1436.1990

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Jham BC, da Silva Freire AR (2006) Oral complications of radiotherapy in the head and neck. Braz J Otorhinolaryngol 72:704–708. https://doi.org/10.1016/s1808-8694(15)31029-6

    Article  PubMed  Google Scholar 

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Acknowledgements

The authors would like to thank the Laboratory of Molecular Genetics of Bacteria (GeMBac - UFSC), Laboratory of Biomechanics and Dental Materials (UFPel), Central Laboratory of Electron Microscopy (LCME - UFSC), Multi-User Laboratory for Biology Studies (LaMEB - UFSC), the Laboratory of Endodontics (UFSC), the Department of Restorative Dentistry (UFPel), the Department of Dentistry (University of Southern Santa Catarina) and the Oncological Research Center (CEPON) for the equipment support.

Funding

The authors would like to thank the “Coordination for the Improvement of Higher Education Personnel” (CAPES) and the “National Council for Scientific and Technological Development” (CNPq) for their financial support.

Author information

Authors and Affiliations

  1. Department of Dentistry - Endodontics Division, Health Sciences Center, Federal University of Santa Catarina, Campus Reitor João David Ferreira Lima, Florianópolis, CEP: 88040-900, Santa Catarina, Brazil

    Taynara Santos Goulart, Cleonice da Silveira Teixeira & Lucas da Fonseca Roberti Garcia

  2. Department of Endodontics, University of Southern Santa Catarina, Palhoça, Santa Catarina, Brazil

    Tainara Hawerroth & Josiane de Almeida

  3. Department of Chemical Engineering, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil

    Karina Cesca

  4. Department of Restorative Dentistry, Federal University of Pelotas, Pelotas, Rio Grande do Sul, Brazil

    Rodrigo Rohenkohl Silva & Rafael Ratto de Moraes

  5. Department of Radiotherapy, Oncology Research Center (CEPON), Florianópolis, SC, Brazil

    Mariana Comparotto Minamisako, Marcio Toshio Umeda Takashima & Nayara Cardoso Cábia

  6. Department of Diagnosis & Oral Health, Endodontics Division, University of Louisville, Louisville, KY, USA

    Eduardo Antunes Bortoluzzi

  7. Department of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil

    Ricardo Ruiz Mazzon

Authors
  1. Taynara Santos Goulart
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  2. Tainara Hawerroth
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  7. Mariana Comparotto Minamisako
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  9. Nayara Cardoso Cábia
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  10. Eduardo Antunes Bortoluzzi
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  12. Josiane de Almeida
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  13. Lucas da Fonseca Roberti Garcia
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Contributions

Taynara Santos Goulart: Conceptualization; Data curation; Investigation; Methodology; Writing - original draft; Writing - review & editing. Tainara Hawerroth: Formal analysis; Writing - original draft; Writing - review & editing. Cleonice da Silveira Teixeira: Formal analysis; Methodology; Writing - original draft; Writing - review & editing. Karina Cesca: Methodology; Writing - review & editing. Rodrigo Rohenkohl Silva: Methodology; Writing - review & editing. Rafael Ratto de Moraes: Methodology; Writing - review & editing. Rodrigo. Mariana Comparotto Minamisako: Methodology; Writing - review & editing. Marcio Toshio Umeda Takashima: Methodology; Writing - review & editing. Nayara Cardoso Cábia: Methodology; Writing - review & editing. Eduardo Antunes Bortoluzzi: Validation; Visualization; Methodology; Writing - review & editing. Ricardo Ruiz Mazzon: Data curation; Investigation; Methodology; Writing - review & editing. Josiane de Almeida: Data curation; Investigation; Methodology; Writing - review & editing. Lucas da Fonseca Roberti Garcia: Conceptualization; Supervision; Project administration; Visualization; Writing - review & editing.

Corresponding author

Correspondence to Lucas da Fonseca Roberti Garcia.

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Ethics approval

This in vitro experimental study was previously approved by the Research Ethics Committee of the Federal University of Santa Catarina (UFSC) (Protocol no. 5.921.189). Considering the institutional guidelines in which this study was performed, ethical approval for using bovine teeth was not required. The study design was conducted following the ethical standards laid down in the 2008 Declaration of Helsinki.

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The authors declare no competing interests.

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Goulart, T.S., Hawerroth, T., da Silveira Teixeira, C. et al. Assessment of multispecies biofilm growth on root canal dentin under different radiation therapy regimens. Clin Oral Invest 28, 324 (2024). https://doi.org/10.1007/s00784-024-05719-x

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