The Open Dentistry Journal




ISSN: 1874-2106 ― Volume 14, 2020

Whole Saliva has a Dual Role on the Adherence of Candida albicans to Polymethylmetacrylate



N Elguezabal1, J.L. Maza2, S. Dorronsoro3, J. Pontón4, *
1 Department of Inmunología, Microbiología y Parasitología, Facultad de Medicina y Odontología, Universidad del PaísVasco, Spain
2 Department of Estomatología, Facultad de Medicina y Odontología, Universidad del PaísVasco, Spain
3 Department of Biología Oral, Facultad de Odontología, Universidad Nacional de Córdoba, República Argentina
4 Department of Inmunología, Microbiología y Parasitología, Facultad de Medicina y Odontología, Universidad del PaísVasco, Spain

Abstract

Adhesion of Candida albicans to acrylic of dental prostheses or to salivary macromolecules adsorbed on their surface is believed to be a critical event in the development of denture stomatitis. In previous studies our group has shown that adhesion of C. albicans germ tubes to polystyrene is decreased by saliva whereas C. albicans yeast cells adhesion to the same material is enhanced. The results presented in this study confirm this dual role played by whole saliva, since it decreased the adhesion of germ tubes but increased the adhesion of yeast cells to polymethylmetacrylate (PMMA). These effects mediated by whole saliva do not seem to be related to an inhibition of the germination of C. albicans, since similar levels of filamentation were observed in presence and absence of saliva. These results may give new insights into the conflicting role of saliva in the adhesion of C. albicans to acrylic resins of dental prostheses.



Article Information


Identifiers and Pagination:

Year: 2008
Volume: 2
First Page: 1
Last Page: 4
Publisher Id: TODENTJ-2-1
DOI: 10.2174/1874210600802010001

Article History:

Received Date: 21/11/2007
Acceptance Date: 12/12/2007
Electronic publication date: 8/1/2008
Collection year: 2008

2008 Bentham Science Publishers Ltd.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.5/), which permits unrestrictive use, distribution, and reproduction in any medium, provided the original work is properly cited.


* Address correspondence to this author at the Departamento de Inmunología, Microbiología y Parasitología, Facultad de Medicina y Odontología, Universidad del País Vasco, Apartado 699, E-48080 Bilbao, Vizcaya, Spain; Tel: 34-94-6012855; Fax: 34-94-6013495; E-mail: jose.ponton@ehu.es





INTRODUCTION

Candida albicans is a dimorphic fungus that is commensal in the gastrointestinal and reproductive tracts of healthy individuals. Under certain predisposing conditions, C. albicans can convert into a pathogen capable of causing a variety of oral infections including pseudomembranous candidiasis, erythematous candidiasis and hyperplasic candidiasis, as well as Candida-associated denture stomatitis, Candida-associated angular cheilitis, rhomboid glossitis and chronic mucocutaneous candidiasis [1Aguirre JM. Candidiasis orales Rev Iberoam Micol 2002; 19: 17-21.].

Candida-associated denture stomatitis is an inflammatory process that affects the oral mucosa of 25-65% of patients wearing removable dental prostheses [2Budtz-Jörgensen E. Etiology, pathogenesis, therapy and prophylaxis of oral yeast infections Acta Odontol Scand 1990; 48: 61-9.]. The etiology is multifactorial consisting of either ill-fitting prostheses leading to mechanical irritation or poor hygiene leading to chronic infection. Regardless of the initiating process C. albicans is the major cause of fungal origin in denture stomatitis [3Budtz-Jörgensen E. The significance of Candida albicans in denture stomatitis Scand J Dent Res 1974; 82: 151-90.]. The first step implicated in denture stomatitis is adherence to acrylic or to salivary pellicles adsorbed on the surface of dental prosthesis being the most important event in the ability of C. albicans to colonize dentures in the mouth [4Vasilas A, Molina L, Hoffman M, Haidaris C. The influence of morphological variation on Candida albicans adhesion to denture acrylic in vitro Arch Oral Biol 1992; 37: 613-22.].

C. albicans can produce biofilms on natural surfaces, such as teeth, and foreign surfaces, such as prostheses. These biofilms are normally resistant to common antimicrobial therapy an increasing problem in clinics [5Douglas LJ. Candida biofilms and their role in infection Trends Microbiol 2003; 11: 30-6.].

Saliva is the biologic fluid that bathes all oral surfaces and acts as a defense against microorganisms present in the oral cavity [6Marcotte H, Lavoie MC. Oral microbial ecology and the role of salivary immunoglobulin A Microbiol Mol Bio Rev 1998; 62: 71-109.]. However, contradictory results have been obtained when the effect of saliva on the adherence of C. albicans to acrylic and plastic has been assessed. Several studies have shown that saliva reduces the adherence of C. albicans to plastic [7McCourtie J, MacFarlane T, Samaranayake L. Effect of saliva and serum on the adherence of Candida spp to clorhexidine-treated denture acrylic J Med Microbiol 1986; 21: 209-13.-9San Millán R, Elguezabal N, Regúlez P, Moragues MD, Quindós G, Pontón J. Effect of salivary secretory IgA on the adhesion of Candida albicans to polysterene Microbiology 2000; 146: 2105-12.], a restorative material [10Elguezabal N, Maza JL, Pontón J. Inhibition of adherence of Candida albicans and Candida dubliniensis to a resin composite restorative dental material by salivary secretory IgA and monoclonal antibodies Oral Dis 2004; 10: 81-6.,11Maza JL, Elguezabal N, Prado C, Ellacuría J, Soler I, Pontón J. Candida albicans adherence to resin-composite restorative dental material: Influence of whole human saliva Oral Surg Oral Med Oral Pathol 2002; 94: 589-92.], PMMA [12Silva Moura J, da Silva WJ, Pereira T, Del Bel Curry AA, Rodrigues-Garcia RC. Influence of acrylic resin polymerization methods and saliva on the adherence of four Candida species J Prosthet Dent 2006; 96: 205-11.,13Karaagaclioglu L, Gulsen C, Yilmaz B, Ayham N, Semiz O, Levent H. The adherence of Candida albicans to acrylic resin reinforced with different fibers J Mater Sci Mater Med Epub ahead of print Aug1; 2007] and epithelial cells [14Umazume M, Ueta E, Osaki T. Reduced inhibition of Candida albicans adhesion by saliva from patients receiving oral cancer therapy J Clin Microbiol 1995; 33: 432-9.] but others have observed that saliva enhances the adherence of C. albicans to PMMA [15Edgerton M, Scannapieco F, Reddy M, Levine M. Human submandibular-sublingual saliva promotes adhesion of Candida albicans to polymethylmethacrylate Infect Immun 1993; 61: 2644-52.], polystyrene [4Vasilas A, Molina L, Hoffman M, Haidaris C. The influence of morphological variation on Candida albicans adhesion to denture acrylic in vitro Arch Oral Biol 1992; 37: 613-22.] and epithelial cells [16Holmes AR, Bandara BMK, Cannon RD. Saliva promotes Candida albicans adherence to human epithelial cells J Dent Res 2002; 81: 28-32.]. In a previous paper, we have shown that whole saliva plays a different role in the adhesion of C. albicans to polystyrene depending on the morphological phase of C. albicans, since it enhanced the adhesion to polystyrene of yeast cells but decreased the adhesion of germinated cells [9San Millán R, Elguezabal N, Regúlez P, Moragues MD, Quindós G, Pontón J. Effect of salivary secretory IgA on the adhesion of Candida albicans to polysterene Microbiology 2000; 146: 2105-12.]. In the present study, we have assessed the influence of whole saliva on adherence of C. albicans to PMMA in an attempt to extend our previous observation to the acrylic used to make dental prostheses.

MATERIALS AND METHODOLOGY

C. albicans serotype A (NCPF 3153), a filamentous strain obtained from the National Collection of Pathogenic Fungi (NCPF, Bristol, United Kingdom), C. albicans Ca2, a germ tube-deficient mutant of the parental strain serotype A NCPF 3153, kindly supplied by Dr. A. Cassone (Istituto Superiore di Sanita, Rome, Italy) and C. albicans UC1, an oral isolate from the Universidad de Cordoba (Argentina), were used in these experiments. The strains were maintained at 4 ºC on slants containing 20 g of glucose, 10 g of yeast extract, and 20 g of agar per liter.

The experimental protocols to obtain human saliva were approved by the Institutional Review Board of the School of Medicine and Odontology at the University of the Basque Country, Leioa, Spain, and the subjects gave their informed consent. Unstimulated whole saliva samples were collected and pooled from 5 healthy donors to eliminate sample variations. The donors had not taken any medication during the 3 months preceding the study and had no active periodontal disease or active caries. Saliva was centrifuged at 6000 g for 30 min at 4 ºC and the supernatant was stored at 4 ºC to be used the same day or stored at –80 ºC until used.

The acrylic strips for the adhesion assay were prepared as described by Samaranayake and MacFarlane [17Samaranayake LP, MacFarlane TW. An in vitro study of the adherence of Candida albicans to acrylic surfaces Arch Oral Biol 1980; 25: 603-9.], with some modifications. Briefly, transparent self-polymerizing acrylic powder (1.5 g of PMMA powder) was spread on an aluminium-foil-covered glass slide (2.5 x 7.5 cm). Monomer liquid (1ml) was poured on to the surface of the slide and immediately a second slide similar to the first was placed on top of the polymerizing mixture, and the slides were firmly secured at both ends with two binder clips. After bench curing for 30 min, the glass slides were separated. The resultant acrylic strips were cut into 5 x 5 mm squares and immersed in distilled water for 1 week to leach excess monomer. The strips were then ultrasonicated for 20 min, washed again in sterile distilled water, dried and used for the adhesion assay.

A modification of the method described by Tronchin et al. [18Tronchin G, Bouchara JP, Robert R, Senet JM. Adherence of Candida albicans germ tube to plastic: ultrastructural and molecular studies of fibrillar adhesins Infect Immun 1988; 56: 1987-93.] was used in all adhesion experiments. Briefly, yeast cells were inoculated in medium 199, pH 6.7, at a final concentration of 8 x 105 cells/mL and were incubated for 2 h at 37 ºC or 25 ºC in 24-well tissue culture polystyrene plates containing the PMMA pieces and 350 µL of the yeast cell suspension. After incubation, the pieces were removed from the wells, washed with saline solution, and germination was quantified by counting the total number of cells and the number of germinated cells in 12 fields (0.64 mm2 each) per PMMA square by means of a graticule mounted in the focus of the ocular. The percentage of germination was calculated by the following equation: % Germination = (germinated cells/ total cells) x 100. PMMA pieces were washed three times with saline, and the adhesion was quantified by counting the total number of cells in the same 12 fields. The percentage of adhesion was calculated by the following equation: % adhesion = (adhered cells/ total cells) x 100. All values quoted represent mean figures derived from at least 4 independent assays. To determine the effect on adherence of the salivary pellicles, the PMMA pieces were previously incubated in 350 µL of whole saliva for 30 min at 37 ºC.

The ANOVA test was used to assess the significance of differences between means in adherence assays. Data were considered significant at P<0.05.

RESULTS

At 37 ºC and in the absence of human saliva, the adhesion of C. albicans NCPF 3153, UC1 and Ca2 to PMMA increased with time, reaching a maximum at 120 minutes (Fig. 1). However, the levels of adhesion were not the same for the three strains since C. albicans Ca2 reached only a 25% of adhesion whereas C. albicans NCPF 3153 and UC1 exceeded 90% of adhesion. At 25 ºC (Fig. 2), adherence of all three strains increased with time, although the percentages of adhesion reached were significantly lower than the ones observed at 37 ºC for strains C. albicans NCPF 3153 and UC1 (P<0.01 at 80 and 120 min) but similar to those observed for C. albicans Ca2.

Fig. (1)

Adhesion of C. albicans NCPF 3153 (black), UC1 (white) and Ca2 (gray) to PMMA at 37ºC. Results represent the means of quadruplicate determinations ±SD.



Fig. (2)

Adhesion of C. albicans NCPF 3153 (black), UC1 (white) and Ca2 (gray) to PMMA at 25ºC. Results represent the means of quadruplicate determinations ±SD.



Fig. (3)

Adhesion of C. albicans NCPF 3153 (black), UC1 (white) and Ca2 (gray) to PMMA at 37ºC in presence of saliva. Results represent the means of quadruplicate determinations ±SD.



Fig. (4)

Germination of C. albicans NCPF 3153 (black) and UC1 (white) to PMMA at 37ºC. Results represent the means of quadruplicate determinations ±SD.



Fig. (5)

Germination of C. albicans NCPF 3153 (black) and UC1 (white) to PMMA at 25ºC. Results represent the means of quadruplicate determinations ±SD.



Fig. (6)

Germination of C. albicans NCPF 3153 (black) and UC1 (white) to PMMA at 37ºC in presence of saliva. Results represent the means of quadruplicate determinations ±SD.



Addition of saliva to the adhesion assay changed the kinetics of adhesion observed in absence of saliva (Fig. 3). At 40 min of incubation at 37ºC, the adherence observed for all the strains was higher than the adherence in the absence of saliva at the same period of time, with a rise in adhesion ranging from 11.4% (C. albicans NCPF 3153) to 4.2% (C. albicans Ca2). However, at 80 and 120 min in the presence of saliva the adhesion of C. albicans to PMMA was significantly reduced (P<0.001).

Since adhesion of C. albicans to plastic materials has been shown to be related with germination [18Tronchin G, Bouchara JP, Robert R, Senet JM. Adherence of Candida albicans germ tube to plastic: ultrastructural and molecular studies of fibrillar adhesins Infect Immun 1988; 56: 1987-93.,19San Millán R, Ezkurra PA, Quindós G, Robert R, Senet JM, Pontón J. Effect of monoclonal antibodies directed against Candida albicans cell wall antigens on the adhesion of the fungus to polystyrene Microbiology 1996; 142: 2271-7.], we assessed the filamentation in the conditions of the adhesion assay. At 37ºC and in absence of saliva, germination increased with time in C. albicans NCPF 3153 and UC1 (Fig. 4). At 40 min of incubation the majority of the C. albicans cells presented yeast morphology and only 3% showed short germ tubes. Longer periods of incubation produced an increase in the percentage of germination and an extension in the length of the germ tubes. The maximum level of germination, which was similar for both strains reaching 85% at 120 min. However, germination at 25ºC (Fig. 5) was very low and remained fairly constant through the time of incubation studied (4-7%). As expected, C. albicans Ca2 did not germinate at 25 or 37ºC in presence or absence of saliva (data not shown). In the presence of saliva (Fig. 6), germination also increased with time showing levels similar to those reached by the controls in absence of saliva.

DISCUSSION

C. albicans adheres to a variety of surfaces in the oral cavity that are constantly bathed by saliva [6Marcotte H, Lavoie MC. Oral microbial ecology and the role of salivary immunoglobulin A Microbiol Mol Bio Rev 1998; 62: 71-109.]. Saliva has a defensive role in the oral cavity that is non-specific (mucins, lysozyme, peroxydase, histatins, etc.) and specific (secretory IgA). Mucins enhance agglutination preventing colonization [20Hoffman MP, Haidaris CG. Analysis of Candida albicans adhesion to salivary mucin Infect Immun 1993; 61: 1940-9., 21Tabak LA, Levine MJ, Mandel ID, Ellison SA. Role of salivary mucin in protection of the oral cavity J Oral Pathol 1982; 11: 1-7.] whereas lysozyme, peroxydase [6Marcotte H, Lavoie MC. Oral microbial ecology and the role of salivary immunoglobulin A Microbiol Mol Bio Rev 1998; 62: 71-109.] and histatins [22Fitzgerald DH, Coleman DC, O´Connell BC. Susceptibility of Candida dubliniensis to salivary histatin 3 Antimicrob Agents Chemother 2003; 47: 70-6., 23Oppenheim FG, Xu T, McMillan FM, et al. Histatins, a novel family of histidine-rich proteins in human parotid secretions J Biol Chem 1988; 263: 7472-7.] are candidacidal. On the other hand, saliva provides water, nutrients and adherence factors [6Marcotte H, Lavoie MC. Oral microbial ecology and the role of salivary immunoglobulin A Microbiol Mol Bio Rev 1998; 62: 71-109.]. A number of studies have shown that the adherence of C. albicans to acrylic materials [4Vasilas A, Molina L, Hoffman M, Haidaris C. The influence of morphological variation on Candida albicans adhesion to denture acrylic in vitro Arch Oral Biol 1992; 37: 613-22.,7McCourtie J, MacFarlane T, Samaranayake L. Effect of saliva and serum on the adherence of Candida spp to clorhexidine-treated denture acrylic J Med Microbiol 1986; 21: 209-13.-9San Millán R, Elguezabal N, Regúlez P, Moragues MD, Quindós G, Pontón J. Effect of salivary secretory IgA on the adhesion of Candida albicans to polysterene Microbiology 2000; 146: 2105-12.,15Edgerton M, Scannapieco F, Reddy M, Levine M. Human submandibular-sublingual saliva promotes adhesion of Candida albicans to polymethylmethacrylate Infect Immun 1993; 61: 2644-52.] can be modulated by saliva. However, the precise role of saliva on the adhesion of C. albicans to acrylic is not clear, since both an increase and a decrease in adhesion have been described [4Vasilas A, Molina L, Hoffman M, Haidaris C. The influence of morphological variation on Candida albicans adhesion to denture acrylic in vitro Arch Oral Biol 1992; 37: 613-22.,7McCourtie J, MacFarlane T, Samaranayake L. Effect of saliva and serum on the adherence of Candida spp to clorhexidine-treated denture acrylic J Med Microbiol 1986; 21: 209-13.,8Samaranayake LP, McCourtie J, MacFarlane TW. Factors affecting the in vitro adherence of Candida albicans to acrylic surfaces Arch Oral Biol 1980; 25]. A possible explanation for this contradictory effect has been proposed by San Millán et al. [9San Millán R, Elguezabal N, Regúlez P, Moragues MD, Quindós G, Pontón J. Effect of salivary secretory IgA on the adhesion of Candida albicans to polysterene Microbiology 2000; 146: 2105-12.] when studying the adhesion of C. albicans to polystyrene, since whole saliva decreased or enhanced the adhesion of C. albicans to polystyrene depending on the morphological phase of C. albicans. Thus, it is possible that whole saliva plays a dual role on the adhesion of C. albicans to plastic materials used to make dental prostheses, decreasing adhesion of germinated cells and enhancing the adhesion of yeast cells.

The results presented in this study confirm this dual role played by whole saliva, since it decreased the adhesion of germ tubes to PMMA but increased the adhesion of yeast cells to the acrylic. Although germination is an important factor in the adhesion of C. albicans to plastic surfaces and epithelial cells [18Tronchin G, Bouchara JP, Robert R, Senet JM. Adherence of Candida albicans germ tube to plastic: ultrastructural and molecular studies of fibrillar adhesins Infect Immun 1988; 56: 1987-93., 24Kimura LH, Pearsall NN. Relationship between germination of Candida albicans and increased adherence to human buccal epithelial cells Infect Immun 1980; 28: 464-8.] and in this study we have observed that adhesion increases in parallel with germ tube formation, the effects mediated by whole saliva do not seem to be related to an inhibition of the germination of C. albicans, since similar levels of filamentation were observed in presence and absence of saliva. Secretory IgA and possibly other salivary components are likely to modulate the adhesion of C. albicans to PMMA by binding the cell wall surface of the fungus. In the inhibitory effect, binding of the cell wall will mask the fungal adhesins, while the enhancement of adhesion may be mediated by a reorganization of the cell wall adhesins induced by certain types of antibodies [10Elguezabal N, Maza JL, Pontón J. Inhibition of adherence of Candida albicans and Candida dubliniensis to a resin composite restorative dental material by salivary secretory IgA and monoclonal antibodies Oral Dis 2004; 10: 81-6.,11Maza JL, Elguezabal N, Prado C, Ellacuría J, Soler I, Pontón J. Candida albicans adherence to resin-composite restorative dental material: Influence of whole human saliva Oral Surg Oral Med Oral Pathol 2002; 94: 589-92.,19San Millán R, Ezkurra PA, Quindós G, Robert R, Senet JM, Pontón J. Effect of monoclonal antibodies directed against Candida albicans cell wall antigens on the adhesion of the fungus to polystyrene Microbiology 1996; 142: 2271-7.]. Interestingly, the dual effect caused by whole saliva may be specific of plastic materials since it was not observed with the composite Herculite, where only the inhibitory effect on the adhesion of C. albicanswas observed [11Maza JL, Elguezabal N, Prado C, Ellacuría J, Soler I, Pontón J. Candida albicans adherence to resin-composite restorative dental material: Influence of whole human saliva Oral Surg Oral Med Oral Pathol 2002; 94: 589-92.].

Future studies should be aimed at finding antimicrobial agents that can decrease C. albicans adhesion to PMMA or kill C. albicans adhered to PMMA surface. In this sense, more studies like the one performed recently by Manfredi et al. [25Manfredi M, Merigo E, Salati A, et al. In vitro candidacidal activity of a synthetic killer decapeptide (KP) against Candida albicans cells adhered to resin acrylic discs J Oral Pathol Med 2007; 36: 468-71.] showing that a synthetic killer peptide has potential candidacidal effect on C. albicans cells adhered to acrylic surfaces should be done.

CONCLUSIONS

In conclusion, the results presented in this study demonstrate that whole saliva decreases the adherence of C. albicans to PMMA, although our results show it plays a dual role that depends on the morphological phase of the fungus.

ABBREVIATIONS

PMMA = Polymethylmetacrylate
NCPF = National Collection of Pathogenic Fungi
SD = Standard deviation

ACKNOWLEDGEMENTS

This project has been financed with grant GIU06/56 from Universidad del País Vasco, Spain.

REFERENCES

[1] Aguirre JM. Candidiasis orales Rev Iberoam Micol 2002; 19: 17-21.
[2] Budtz-Jörgensen E. Etiology, pathogenesis, therapy and prophylaxis of oral yeast infections Acta Odontol Scand 1990; 48: 61-9.
[3] Budtz-Jörgensen E. The significance of Candida albicans in denture stomatitis Scand J Dent Res 1974; 82: 151-90.
[4] Vasilas A, Molina L, Hoffman M, Haidaris C. The influence of morphological variation on Candida albicans adhesion to denture acrylic in vitro Arch Oral Biol 1992; 37: 613-22.
[5] Douglas LJ. Candida biofilms and their role in infection Trends Microbiol 2003; 11: 30-6.
[6] Marcotte H, Lavoie MC. Oral microbial ecology and the role of salivary immunoglobulin A Microbiol Mol Bio Rev 1998; 62: 71-109.
[7] McCourtie J, MacFarlane T, Samaranayake L. Effect of saliva and serum on the adherence of Candida spp to clorhexidine-treated denture acrylic J Med Microbiol 1986; 21: 209-13.
[8] Samaranayake LP, McCourtie J, MacFarlane TW. Factors affecting the in vitro adherence of Candida albicans to acrylic surfaces Arch Oral Biol 1980; 25
[9] San Millán R, Elguezabal N, Regúlez P, Moragues MD, Quindós G, Pontón J. Effect of salivary secretory IgA on the adhesion of Candida albicans to polysterene Microbiology 2000; 146: 2105-12.
[10] Elguezabal N, Maza JL, Pontón J. Inhibition of adherence of Candida albicans and Candida dubliniensis to a resin composite restorative dental material by salivary secretory IgA and monoclonal antibodies Oral Dis 2004; 10: 81-6.
[11] Maza JL, Elguezabal N, Prado C, Ellacuría J, Soler I, Pontón J. Candida albicans adherence to resin-composite restorative dental material: Influence of whole human saliva Oral Surg Oral Med Oral Pathol 2002; 94: 589-92.
[12] Silva Moura J, da Silva WJ, Pereira T, Del Bel Curry AA, Rodrigues-Garcia RC. Influence of acrylic resin polymerization methods and saliva on the adherence of four Candida species J Prosthet Dent 2006; 96: 205-11.
[13] Karaagaclioglu L, Gulsen C, Yilmaz B, Ayham N, Semiz O, Levent H. The adherence of Candida albicans to acrylic resin reinforced with different fibers J Mater Sci Mater Med Epub ahead of print Aug1; 2007
[14] Umazume M, Ueta E, Osaki T. Reduced inhibition of Candida albicans adhesion by saliva from patients receiving oral cancer therapy J Clin Microbiol 1995; 33: 432-9.
[15] Edgerton M, Scannapieco F, Reddy M, Levine M. Human submandibular-sublingual saliva promotes adhesion of Candida albicans to polymethylmethacrylate Infect Immun 1993; 61: 2644-52.
[16] Holmes AR, Bandara BMK, Cannon RD. Saliva promotes Candida albicans adherence to human epithelial cells J Dent Res 2002; 81: 28-32.
[17] Samaranayake LP, MacFarlane TW. An in vitro study of the adherence of Candida albicans to acrylic surfaces Arch Oral Biol 1980; 25: 603-9.
[18] Tronchin G, Bouchara JP, Robert R, Senet JM. Adherence of Candida albicans germ tube to plastic: ultrastructural and molecular studies of fibrillar adhesins Infect Immun 1988; 56: 1987-93.
[19] San Millán R, Ezkurra PA, Quindós G, Robert R, Senet JM, Pontón J. Effect of monoclonal antibodies directed against Candida albicans cell wall antigens on the adhesion of the fungus to polystyrene Microbiology 1996; 142: 2271-7.
[20] Hoffman MP, Haidaris CG. Analysis of Candida albicans adhesion to salivary mucin Infect Immun 1993; 61: 1940-9.
[21] Tabak LA, Levine MJ, Mandel ID, Ellison SA. Role of salivary mucin in protection of the oral cavity J Oral Pathol 1982; 11: 1-7.
[22] Fitzgerald DH, Coleman DC, O´Connell BC. Susceptibility of Candida dubliniensis to salivary histatin 3 Antimicrob Agents Chemother 2003; 47: 70-6.
[23] Oppenheim FG, Xu T, McMillan FM, et al. Histatins, a novel family of histidine-rich proteins in human parotid secretions J Biol Chem 1988; 263: 7472-7.
[24] Kimura LH, Pearsall NN. Relationship between germination of Candida albicans and increased adherence to human buccal epithelial cells Infect Immun 1980; 28: 464-8.
[25] Manfredi M, Merigo E, Salati A, et al. In vitro candidacidal activity of a synthetic killer decapeptide (KP) against Candida albicans cells adhered to resin acrylic discs J Oral Pathol Med 2007; 36: 468-71.
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