1887

Abstract

Purpose. In vitro analyses of virulence, pathogenicity and associated host cell responses are important components in the study of biofilm infections. The Candida-related infection, denture-associated oral candidosis, affects up to 60 % of denture wearers and manifests as inflammation of palatal tissues contacting the denture-fitting surface. Commercially available three-dimensional tissue models can be used to study infection, but their use is limited for many academic research institutions, primarily because of the substantial purchase costs. The aim of this study was to develop and evaluate the use of in vitro tissue models to assess infections by biofilms on acrylic surfaces through tissue damage and Candida albicans virulence gene expression.

Methodology. In vitro models were compared against commercially available tissue equivalents (keratinocyte-only, SkinEthic; full-thickness, MatTek Corporation). An in vitro keratinocyte-only tissue was produced using a cancer-derived cell line, TR146, and a full-thickness model incorporating primary fibroblasts and immortalised normal oral keratinocytes was also generated. The in vitro full-thickness tissues incorporated keratinocytes and fibroblasts, and have potential for future further development and analysis.

Results. Following polymicrobial infection with biofilms on acrylic surfaces, both in-house developed models were shown to provide equivalent results to the SkinEthic and MatTek models in terms of tissue damage: a significant (P<0.05) increase in LDH activity for mixed species biofilms compared to uninfected control, and no significant difference (P>0.05) in the expression of most C. albicans virulence genes when comparing tissue models of the same type.

Conclusion. Our results confirm the feasibility and suitability of using these alternative in vitro tissue models for such analyses.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000677
2018-01-11
2019-10-17
Loading full text...

Full text loading...

/deliver/fulltext/jmm/67/3/364.html?itemId=/content/journal/jmm/10.1099/jmm.0.000677&mimeType=html&fmt=ahah

References

  1. Flemming HC, Wingender J. The biofilm matrix. Nat Rev Microbiol 2010;8:623–633 [CrossRef][PubMed]
    [Google Scholar]
  2. Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: a common cause of persistent infections. Science 1999;284:1318–1322 [CrossRef][PubMed]
    [Google Scholar]
  3. Sutherland IW. The biofilm matrix—an immobilized but dynamic microbial environment. Trends Microbiol 2001;9:222–227 [CrossRef][PubMed]
    [Google Scholar]
  4. Costerton JW, Cheng KJ, Geesey GG, Ladd TI, Nickel JC et al. Bacterial biofilms in nature and disease. Annu Rev Microbiol 1987;41:435–464 [CrossRef][PubMed]
    [Google Scholar]
  5. Hall-Stoodley L, Stoodley P. Developmental regulation of microbial biofilms. Curr Opin Biotechnol 2002;13:228–233 [CrossRef][PubMed]
    [Google Scholar]
  6. Bradshaw DJ, Marsh PD, Allison C, Schilling KM. Effect of oxygen, inoculum composition and flow rate on development of mixed-culture oral biofilms. Microbiology 1996;142:623–629 [CrossRef][PubMed]
    [Google Scholar]
  7. Bradshaw DJ, Marsh PD, Watson GK, Allison C. Role of Fusobacterium nucleatum and coaggregation in anaerobe survival in planktonic and biofilm oral microbial communities during aeration. Infect Immun 1998;66:4729–4732[PubMed]
    [Google Scholar]
  8. Fux CA, Costerton JW, Stewart PS, Stoodley P. Survival strategies of infectious biofilms. Trends Microbiol 2005;13:34–40 [CrossRef][PubMed]
    [Google Scholar]
  9. Trautner BW, Darouiche RO. Role of biofilm in catheter-associated urinary tract infection. Am J Infect Control 2004;32:177–183 [CrossRef][PubMed]
    [Google Scholar]
  10. Ramage G, Martínez JP, López-Ribot JL. Candida biofilms on implanted biomaterials: a clinically significant problem. FEMS Yeast Res 2006;6:979–986 [CrossRef][PubMed]
    [Google Scholar]
  11. Wolcott RD, Rhoads DD, Bennett ME, Wolcott BM, Gogokhia L et al. Chronic wounds and the medical biofilm paradigm. J Wound Care 2010;19:45–53 [CrossRef][PubMed]
    [Google Scholar]
  12. Gendreau L, Loewy ZG. Epidemiology and etiology of denture stomatitis. J Prosthodont 2011;20:251–260 [CrossRef][PubMed]
    [Google Scholar]
  13. Djeribi R, Bouchloukh W, Jouenne T, Menaa B. Characterization of bacterial biofilms formed on urinary catheters. Am J Infect Control 2012;40:854–859 [CrossRef][PubMed]
    [Google Scholar]
  14. Percival SL, Suleman L, Vuotto C, Donelli G. Healthcare-associated infections, medical devices and biofilms: risk, tolerance and control. J Med Microbiol 2015;64:323–334 [CrossRef][PubMed]
    [Google Scholar]
  15. Aas JA, Paster BJ, Stokes LN, Olsen I, Dewhirst FE. Defining the normal bacterial flora of the oral cavity. J Clin Microbiol 2005;43:5721–5732 [CrossRef][PubMed]
    [Google Scholar]
  16. Dewhirst FE, Chen T, Izard J, Paster BJ, Tanner AC et al. The human oral microbiome. J Bacteriol 2010;192:5002–5017 [CrossRef][PubMed]
    [Google Scholar]
  17. Wade WG. The oral microbiome in health and disease. Pharmacol Res 2013;69:137–143 [CrossRef]
    [Google Scholar]
  18. Ghannoum MA, Jurevic RJ, Mukherjee PK, Cui F, Sikaroodi M et al. Characterization of the oral fungal microbiome (mycobiome) in healthy individuals. PLoS Pathog 2010;6:e10007131000718 [CrossRef][PubMed]
    [Google Scholar]
  19. Ritz HL. Microbial population shifts in developing human dental plaque. Arch Oral Biol 1967;12:1561–1568 [CrossRef][PubMed]
    [Google Scholar]
  20. Marsh PD. Dental plaque as a microbial biofilm. Caries Res 2004;38:204–211 [CrossRef][PubMed]
    [Google Scholar]
  21. Edgerton M, Levine MJ. Characterization of acquired denture pellicle from healthy and stomatitis patients. J Prosthet Dent 1992;68:683–691 [CrossRef][PubMed]
    [Google Scholar]
  22. Salerno C, Pascale M, Contaldo M, Esposito V, Busciolano M et al. Candida-associated denture stomatitis. Med Oral Patol Oral Cir Bucal 2011;16:e139e143 [CrossRef][PubMed]
    [Google Scholar]
  23. Newton AV. Denture sore mouth: a possible aetiology. Br Dent J 1962;112:357–360
    [Google Scholar]
  24. Williams DW, Kuriyama T, Silva S, Malic S, Lewis MA. Candida biofilms and oral candidosis: treatment and prevention. Periodontol 2000 2011;55:250–265 [CrossRef][PubMed]
    [Google Scholar]
  25. Shulman JD, Rivera-Hidalgo F, Beach MM. Risk factors associated with denture stomatitis in the United States. J Oral Pathol Med 2005;34:340–346 [CrossRef][PubMed]
    [Google Scholar]
  26. Rogers H, Wei X-Q, Lewis MAO, Patel V, Rees J et al. Immune response and candidal colonisation in denture associated stomatitis. J Clin Cell Immunol 2013;4:1–7
    [Google Scholar]
  27. Cavalcanti YW, Morse DJ, da Silva WJ, del-Bel-Cury AA, Wei X et al. Virulence and pathogenicity of Candida albicans is enhanced in biofilms containing oral bacteria. Biofouling 2015;31:27–38 [CrossRef][PubMed]
    [Google Scholar]
  28. Silva S, Henriques M, Oliveira R, Azeredo J, Malic S et al. Characterization of Candida parapsilosis infection of an in vitro reconstituted human oral epithelium. Eur J Oral Sci 2009;117:669–675 [CrossRef][PubMed]
    [Google Scholar]
  29. Silva S, Hooper SJ, Henriques M, Oliveira R, Azeredo J et al. The role of secreted aspartyl proteinases in Candida tropicalis invasion and damage of oral mucosa. Clin Microbiol Infect 2011;17:264–272 [CrossRef][PubMed]
    [Google Scholar]
  30. Yadev NP, Murdoch C, Saville SP, Thornhill MH. Evaluation of tissue engineered models of the oral mucosa to investigate oral candidiasis. Microb Pathog 2011;50:278–285 [CrossRef][PubMed]
    [Google Scholar]
  31. Colley HE, Hearnden V, Jones AV, Weinreb PH, Violette SM et al. Development of tissue-engineered models of oral dysplasia and early invasive oral squamous cell carcinoma. Br J Cancer 2011;105:1582–1592 [CrossRef][PubMed]
    [Google Scholar]
  32. Jennings LR, Colley HE, Ong J, Panagakos F, Masters JG et al. Development and characterization of in vitro human oral mucosal equivalents derived from immortalized oral keratinocytes. Tissue Eng Part C Methods 2016;22:1108–1117 [CrossRef][PubMed]
    [Google Scholar]
  33. Wayakanon K, Thornhill MH, Douglas CW, Lewis AL, Warren NJ et al. Polymersome-mediated intracellular delivery of antibiotics to treat Porphyromonas gingivalis-infected oral epithelial cells. FASEB J 2013;27:4455–4465 [CrossRef][PubMed]
    [Google Scholar]
  34. Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J et al. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 2009;55:611–622 [CrossRef][PubMed]
    [Google Scholar]
  35. Jayatilake JA, Samaranayake YH, Samaranayake LP. An ultrastructural and a cytochemical study of candidal invasion of reconstituted human oral epithelium. J Oral Pathol Med 2005;34:240–246 [CrossRef][PubMed]
    [Google Scholar]
  36. Dongari-Bagtzoglou A, Kashleva H. Development of a novel three-dimensional in vitro model of oral Candida infection. Microb Pathog 2006;40:271–278 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.000677
Loading
/content/journal/jmm/10.1099/jmm.0.000677
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error