Dental Materials Journal 2010; 29(2): 206–212

Comparative effects of two different artificial body fluids on Candida albicans

adhesion to soft lining materials
Caner VURAL1, Guven OZDEMIR1, Huseyin KURTULMUS2, Ovul KUMBULOGLU2 and Mutlu ÖZCAN3
1
Department of Biology, Microbiology Section, Faculty of Science, Ege University, Izmir, Turkey
2
Department of Prosthodontics, Faculty of Dentistry, Ege University, Izmir, Turkey
3
Dental Materials Unit, Center for Dental and Oral Medicine, Clinic for Fixed and Removable Prosthodontics and Dental Materials Science, Zurich
University, Zurich, Switzerland
Corresponding author,  Huseyin KURTULMUS;  E-mail:  h_kurtulmus@yahoo.com


This study investigated the C. albicans adhesion to cold- and heat-polymerized soft lining materials that were initially incubated in
two different artificial body fluids, namely saliva and nasal secretion, and examined the surface roughness the materials (cold and
heat polymerized soft liner) tested in vitro. Cold (Visco Gel) and heat-polymerized (Molloplast B) soft liner specimens (N=32, n=8 per
group) (10×10×1.5 mm) were randomly produced to express the relationship between surface roughness and contamination, and
influence of body fluids, and incubated in 1.5 ml contaminated solutions for 2 h. After fixation, all of materials were evaluated under
optical microscope (×400) and SEM. Surface roughness measurements were examined with profilometre for each material. Data were
analyzed using two-way ANOVA, Tukey’s HSD and Dunnett T3 tests (α=0.05). Material type (p<0.05) and contamination media
(p<0.05) showed a significant influence on the C. albicans adherence. The surface roughness of cold polymerized soft liner (Visco Gel)
was significantly higher than heat-polymerized soft liner (Molloplast B) (p<0.05).

Keywords: Adhesion, Candida albicans, Denture soft lining materials


Received Sep 4, 2009: Accepted Dec 10, 2009

Currently, soft lining materials are available in

INTRODUCTION
Human body, as a host, has many kinds of resident
microorganisms that could be classified as both
domestic and opportunistic pathogens1). These
organisms are closely related with the host systems
such as cell-host tissues or cell-biomaterial interactions
that could be harmful for the host. Some of the
remainder organisms are opportunistic pathogens. The
members of the resident microflora spread out in the
host area (i.e. mouth, skin, gut)1). One such residential
human microflora is Candida albicans (C. albicans)
which is a eukaryotic and opportunistic pathogen,
causing candidiasis known as the most common fungal
infection in human being2-4). This organism being
mainly found in oral cavity causes stomatitis. Denture
stomatitis, more commonly known as ‘denture sore
mouth’, is frequently observed in the maxilla of elderly
denture wearers5-8). Prevalence of denture stomatitis in
association with C. albicans has been reported to range
between 11 and 67% in complete denture wearers5,9-14).
Continuous denture wearing appears to facilitate
denture stomatitis by increasing the local injury and
the time of mucosal exposure to denture plaque9,15,16).
Resilient denture lining materials are used to overcome
such injury. Such materials reduce the traumatic effect
that a denture may have on patients with thin atrophic
mucosa or with normal mucosa but with resorbed
ridges, sharp alveolar ridge crest, deep anatomic
undercuts, bony protuberances, bruxomania, or where
the oral mucosa exhibits a reduced tolerance to the
load applied by the denture17-21). They also facilitate
comfort when used in obturators for acquired and
congenital cleft palate16-20).
silicone elastomers and soft acrylic compounds. Soft
denture liners are also defined as soft polymers which
may be applied to the fitting or mucosal surface of the
dentures. They reduce the occlusal forces and distribute
them more evenly on the underlying mucosal tissues22).
These materials can be classified as provisional or
definitive according to their composition being either
silicone rubber or acrylic resin. Soft denture liners can
be either cold (chemically)- or heat-polymerized23,24).
The adherence of C. albicans to host cells or
polymers such as denture acrylic resins or soft lining
materials is the first step in colonization, yielding to
development of pathogenesis and eventually causing
infection25-28). Soft lining materials have been found to
be more prone to microbial adhesion than acrylic resin
denture base materials29-31). They have demonstrated
the ability to interact with oral microorganisms because
of their surface texture and the physical and chemical
affinity to microorganisms. Verran and Maryan27)
showed more funcal adhesion was found on rough
surfaces than smooth surfaces. Differences in surface
topography affect the attachment of microorganisms to
a surface, with higher numbers of cells retained on
rougher surfaces. Surface irregularities would increase
the likelihood of microorganisms remaing on the
surface27). When soft lining materials are used for
relining the maxillofacial prosthesis, they may expose
to the nasal secretion. The relationship between the
salivary or nasal secretions on denture material
surfaces and C. albicans colonization is complex,
particularly when the surface aging of soft lining
materials are taken into account32). When wearing a
denture, the base becomes colonized with pellicles
 Dent Mater J 2010; 29(2): 206–212 207

composed of salivary or nasal secretions which may
provide receptor sites for the adherence of
microorganisms. Some studies have reported that soft
lining materials have an inhibitory effect on yeast
growth14,33,34) providing that additional variables may
contribute to yeast growth in vivo. In fact, since yeast
cells tend to colonize more in acidic conditions, it can
be hypothesized that nasal secretion, due to its low
pH35), would result in more microbial colonization.
Dental literature contains many case reports on the
construction or application silicone liners but to the
authors’ knowledge there is no information on C.
albicans adhesion in nasal secretion. Particularly in
patients who have gone tumor surgery and thereafter
acquire obturators for maxillofacial defects, post-nasal
secretion is often unavoidable. In fact, especially in
patients with postnasal flux, intra-oral prosthetic
appliances are also exposed to nasal fluid
contamination36). In such situations, oral flora of the
patients is affected. Hence, prosthesis exposed to such
environments may be indirectly affected from the
microorganisms in nasal secretion.
The objectives of this study therefore were to
investigate C. albicans adhesion and surface roughness
onto two commercial silicone liners based on different
polymerization processes in the presence of artificial
saliva and nasal secretion and to evaluate the
colonization microscopically. The tested null hypotheses
were that nasal secretion would result in more C.
albicans colonization than artificial saliva and heat-
polymerization would result in less C. albicans
adhesion.
liner (Visco Gel) was supplied as a powder, liquid and a
sealer liquid with the chemical composition based on
polyethylmethacrylate (powder) and phthalyl butyl
glyconate and ethanol (liquid), heat-polymerized soft
liner (Molloplast B) was supplied as a single paste and
adhesive (bonding liquid) based on polydimethylsiloxan
(polymer) and benzyl peroxide (initiator).
Specimen preparation
Pink modeling wax forms (10×10 mm, thickness: 1.5
mm) were punched from a sheet of wax. Dental stone
(BPB Formula, Newark Works, Bowbridge Lane,
Newark, Nottinghamshire, UK) was mixed, poured into
the shallow part of a dental flask and allowed to set.
After setting, the stone was lubricated using petroleum
gel. Consequently, specimens were flasked with
vacuum, heated for 10 min, and the wax was removed
using a hot water spray27,37). Flasks were cooled, and
then the stone was lubricated using a CO-MO sealant
(John Winter and Co. Ltd, Halifax, UK). Wet
polymerization technique was used for the heat-
polymerized liners (Fig. 1). The cold-polymerized soft
lining material (Visco Gel) was processed according to
the manufacturer’s directions and polymerized in the
flask for 15 min at the room temperature, and then
made to a uniform size (10×10×1.5 mm thickness) using
the mold (Fig. 1). After polymerization, specimens
trimmed and kept in polyethylene containers in a dry
environment until required (1 week). In order to

MATERIALS AND METHODS

Two different denture soft lining materials were used
in this study, namely Visco Gel (Dentsply Ltd., De Trey
Division, Weybridge, U.K), a cold-polymerized
transparent acrylic resin soft lining material (Batch#:
071000;1479) and a heat-polymerized one, Molloplast B
(Detax Karl Huber GmbH, Ettingen, Germany) Fig. 1 Test specimens with smooth surface (10×10×1.5
(Batch#: 17-610) (Table 1). While cold-polymerized soft mm).

Table 1 The brand names, corresponding polymerization modes, chemical compositions, shades, batch numbers and
manufacturers of the materials used in this study
Brand name Polymerization Chemical Shade Batch Manufacturer
mode composition number

Visco Gel Cold-cured acrylic Polyethyl methacrylate (powder) Translucent 071000 Dentsply Ltd.,
resin soft lining and phthalyl butyl glycolate, 1479 De Trey
material ethanol (liquid) Division,
Weybridge,
U.K.

Molloplast B Heat-cured silicone Polydimethylsiloxan (polymer) and Pink 17-610 Detax Karl Huber
soft lining material benzyl peroxide (initiator) GmbH,
Ettlingen,
Germany
208 Dent Mater J 2010; 29(2): 206–212
produce test specimens of comparable smoothness, they
were prepared against glass.
Surface roughness of soft lining materials
The surface roughness (Ra) of the test samples was
measured with a profilometre (Mitutoyo Surftest 301,
Tokyo, Japan), where a stylus traverses across the
layer of the surface, and an amplified trace of the
profile is provided4,27). The Ra value is the arithmetical
average of all departures of the profile through the
mean sample lengt. All measurements were recorded
by one operator.
Candida survey
C. albicans strain ATCC 60193 was obtained from
microorganism stock culture (Basic and Industrial
Microbiology Section, Department of Biology, Ege
University, Izmir, Turkey). For enrichment of
microorganisms, stock culture was inoculated into
Mueller Hinton Broth, Fluka® with 500mM sucrose
medium and incubated for 24 h4,34). After incubation,
cells were collected by centrifugation (Hettich Rotina
35 R Zentrifugen, Germany), harvested and washed
three times in sterile 0.1M phosphate buffered saline
(PBS) with 0.89% NaCl at pH: 7.228,37), and re-
suspended in PBS to an optical density of 0.5 at 540
nm8), corresponding to a cell concentration of 1-5×106
CFU/ml. OD of cell suspension was previously
determined in either saliva or nasal secretion for the
specimens to be inoculated in saliva or nasal secretion,
respectively8).
Adherence assay
Specimens were sterilized in an autoclave (HiClaveTM
HC-50L, Hirayama, Japan) at 121°C for 20 min. They
were then placed in separate sterile tubes and
incubated with either 1.5 ml artificial saliva (pH=7)
that was contaminated with C. albicans cells (set to 0.5
OD, 540 nm, in advance)8) or with 1.5 ml artificial nasal
secretion (pH=4.8) at 37°C with orbital shaking (100
rpm) for 2 h8). The artificial saliva was composed of
0.220 g/L calcium chloride, 1.07 g/L sodium phosphate,
1.68 g/L sodium bicarbonate, and 2 g/L sodium azide
0.2% (NaN3)38) and artificial nasal secretion consisted of
Na+ 107±4 mM, Cl– 120±6 mM, K+ 8.7±0.4 mM obtained
according to in vivo microdialysis procedure (IVMD)35).
After incubation, in order to remove the unattached
cells, the specimens were gently removed from the
tubes and rinsed by dipping them into the PBS solution
for three times for approximately 75 s. Then, for
fixation of the attached cells, specimens were treated
with 100% ethanol for 3 s and left to dry in sterile
plates. Specimens were stained using sterilized, fixated
Methylene Blue stain (Merck®) for 1 min and
subsequently, evaluated under optical light microscope
(Olympus CH20, Olympus Singapore PTE Ltd., City,
Singapore) at ×400 magnification4). On each specimen,
15 different consecutive areas were counted. Visible
measurement field was calculated in mm2 and the
obtained data were expressed in cell/mm2.
Complementary to the optical microscopy analysis,
specimens were also observed under Scanning Electron
Microscope (SEM) (JEOL JSM-5200, Kyoto, Japan)
after they were fixed with 2% gluteraldehyde,
dehydrated with ethanol (at 25, 50, 75 and 100% for 5,
5, 5 and 10 min, respectively) and coated with Au-Pd
(×750 to ×3500 magnification).
Statistical analysis
The statistical analysis was performed with the SPSS
software package (version 11.5; SPSS, Chicago, IL,
USA). Mean ranks obtained from adherence assay were
evaluated with using two-way analysis of variance
(ANOVA) and paired sample T-test. Since significant
differences were found between or within groups,
Tukey’s HSD was used to determine the differences.
The results of normality and homogeneity test
(Kolmogorov-Smirnov) indicated that the residual
values were normally distributed when plotted against
predicted values. The uniformity and normality tests
did not violate the statistical assumptions. In all
comparisons, statistical significance was declared if the
p value was less than 0.05.
RESULTS
Surface roughness
The results are presented as surface roughness values
(a Ra unit) of average surface roughness measurements
of soft lining materials in Table 2. The number of C.
albicans on smooth surfaces was low. Significantly
more cells were counted on the rough surfaces than on
smooth surfaces. The surface roughness of cold-
polimerized soft liner (Visco Gel) was significantly
higher than heat-polimerized soft liner (Molloplast B)
(p<0.05).
C. albicans adherence
Both the material type (p=0.05) and contamination
media (p=0.05) showed a significant influence on the C.
albicans adherence (two-way ANOVA, paired sample T-
test).
Cold-polimerized soft lining (Visco Gel) material
Table 2 Mean surface roughness values (Ra) and
standard deviations for surface roughness
Materials Ra / Mean - (SD)
Visco Gel (n=10) 15.67 (2.20)
Molloplast B (n=10)   0.21 (0.06)
Ra: A unit of the arithmetical average of all departures of
the profile through the mean sample length.
SD: Standard deviation
n: Number of samples
 Dent Mater J 2010; 29(2): 206–212
showed more C. albicans adherence in both saliva and
nasal secretion (mean rank: 9040.75 and 2550.55,
respectively) than that of heat-polimerized soft liner
(Molloplast B) (mean rank: 4149.18 and 1350.31,
respectively) (p<0.05, Tukey’s HSD).
C. albicans adherence onto cold-polimerized soft
liner (Visco Gel) specimens that were incubated with
nasal secretion (mean rank: 3900.86) was significantly
less than with saliva (mean rank: 13189.93) (p=0.05).
Approximately 3.5 fold difference was observed between
nasal and saliva in adhesion results (Fig. 2).
Microscopical evaluations
In general, C. albicans adherence was observed in
cluster forms on all of materials. Whole attached cells
were viewed in blastospore morphology. While cold-
polimerized soft liner (Visco Gel) specimens presented
local colonization of the cells, heat-polimerized soft
liner (Molloplast B) specimens exhibited smoothly
dispersed cells (Figs. 3a-c).
Fig. 2 Mean ranks of C. albicans adherence (cell/mm2)
according to the material and the contamination
medium.
Fig. 3 SEM micrograph of a) Visco Gel contaminated with artificial saliva having C. albicans. Note the cells in big
clusters spread on the surface (×750 original magnification), b) Visco Gel with individual cells and small clusters
in saliva (×1000). Note the crinkled surfaces, c) C. albicans on Molloplast B surface. Note the smoother surface
compared to Visco Gel (×1000).
209
DISCUSSION
Diseases or inflammations such as stomatitis may
occur as a consequence of accumulation ability of the
cells on the polymeric materials that is closely related
to the London- van der Waals (the surface energy
between particles which generates dipole interactions)
and electrostatic forces that facilitate cell adhesion39,40).
When the surface roughness is increased, it triggers
microbial adhesion and the number of adhered cells
also increases remarkably4,27,39). During microbial
colonization, cells produce acidic substances as an
outcome of their natural metabolisms that affect the
pH of the surface they are interacting with. On the
other hand, pH of the medium to which the materials
are exposed plays also a significant role on microbial
colonization41). This study was undertaken in order to
investigate the C. albicans adherence levels to
commercial denture soft liners when they were exposed
to two different human body secretions, namely saliva
and nasal secretions that were artificially created. The
high number of yeast cells presented as ‘‘per mm2’’
facilitated comparison with other studies, but is
somewhat misleading because three-dimensional
accumulations of cells, not monolayers, were observed
and counted. Regardless of the polymerization type of
the materials tested, increased C. albicans adherence
was found in artificial saliva (pH=7). Although artificial
saliva was prepared at neutral state, by-products of C.
albicans could have changed the pH of the whole
solution42,43). Artificial saliva can be served as a
screening medium in this study.
Contamination of the specimens with artificial
nasal secretion yielded to significantly less C. albicans
adherence compared to artificial saliva. Therefore the
hypothesis is rejected. The nasal secretion prepared
had a pH of 4.8. Less C. albicans adherence in this
medium could be attributed to the presence of NaNO3
in this medium that might have inhibited colonization,
created ionic bonds and thereby had repellent effect.
210 Dent Mater J 2010; 29(2): 206–212
Other reason could be due to the storage duration of
the specimens in this media (2 h) which may not be the
critical time point. Unfortunately, to the authors’
knowledge, there exists no study in the dental
literature using nasal secretion as a contamination
medium for soft liners to make comparisons of the
findings of this present study. In this study, artificial
saliva and nasal secretion were used for all specimens
separately. Interaction between organism and fluids
that are aforementioned cannot be commented on in
this situation. It was reported that in-vivo
microorganisms never attach directly to denture
materials without salivary pellicle44). In this
experiment, although no pellicle was initially formed
and all specimens were incubated directly with
artificial saliva and nasal secretion that was added to
C.albicans, the results showed that C. albicans does
not require pellicle for adhesion. Although composition
of nasal secretion has been defined earlier35), C.
albicans adherence on soft lining materials in the
presence of artificial nasal secretion was not studied.
Currently, in a case controlled clinical trial, the effect
of human nasal secretion on C. albicans adherence on
liners is being evaluated where in vivo results will be
collected and correlated to the findings of this in vitro
study. This study therefore should be considered for
screening purposes.
In this study, artificial saliva and nasal secretion
coated heat (Molloplast B) and cold (Visco Gel)-
polymerized soft liners were tested. In all materials
tested, some degree of C. albicans adherence was noted
being significantly less for heat-polymerized soft liner
(Molloplast B), confirming the hypothesis. On the other
hand, the SEM findings and surface roughness
measurement applied with profilometre presented
fairly smooth surfaces with heat-polymerized soft liner
(Molloplast B) material compared to cold-polymerized
soft liner (Visco Gel) that is most probably related to
the polymerization modes of the materials tested.
According to the Tari et al.4), Nikawa et al.30,32), in their
investigation on fungal adherence with or without a
salivary, showed there was no difference in fungal
adherence between to saliva coated and uncoated soft
lining materials. Therefore uncoated materials as
control material was not tested in this study. Two
different soft lining materials were processed against
glass slides to the direction of suggestions of
researchers’ studies in reviewed literatures4,27,30,32). Still,
there was a difference in the adhesion of C. albicans
between soft lining materials was observed. This result
essentially promoted that polymerization method was
effective. Besides, more adhesion may be observed on
the materials when processed against dental stone.
Larger cells such as yeasts are more easily
dislodged from smooth surfaces than the smaller
microorganisms. In this study, the exposure of the
specimens to either artificial saliva or nasal secretion
was 2 h due to technical reasons. Although salivary
proteins has been considered to serve as a source of
nutrients for microorganisms for growth and
reproduction requirements, as well as assisting in
microbial succession of dissimilar bacterial species in
developing plaque31,45), prolonged period of
contamination may lead to possible cell death35). In fact,
prolonged exposure of the soft liners to such media may
have more in-vivo implications regarding the functional
lifespan of liners since host surfaces in contact would
be then more heavily colonized with commencing
microorganisms, many of which are opportunist
pathogens. The synergistic effect of the microorganism
colonization with low pH together with the conditions
of the contamination media may lead to material
degradation and eventually rougher surfaces.
In this study, to determine of surface texture of the
liners, roughness measurements were undertaken. The
surface roughness of a material used for a removable
prosthesis is a importance since it affects, directly or
indirectly, retention, staining resistance, plaque
accumulation, as well as oral tissue health and patient
comfort4,28). In the present study, cold-polymerized soft
liner (Visco Gel) was rougher than heat-polymerized
soft liner (Molloplast B). The surface roughness value
for heat-polymerized soft liner (Molloplast B) and cold-
polymerized soft liner (Visco Gel) was found similar to
those reported by Tari et al 4). This may be because the
same technique was used for measurement. Separately,
microscopy analysis has shown heat-polymerized soft
liner (Molloplast B) with smooth surfaces. Surface
roughness is a significant factor in the attachment and
adhesion of microorganisms on surfaces with an
increase in roughness causing increased retention of
cells4,27,31). It is known to be a factor in the entrapment
of microorganisms on surfaces and their protection
from shear forces27,46). In this study, yeast cells were
retained on roughened surfaces in higher numbers than
on smooth. Cells were observed within surface
irregularities. The results of present study may be
explained by the effect reported previously4,27,46).
Namely, it demonstrated that increased surface
roughness increased retention of yeast.
From the clinical point of view, maxillofacial and
dental/oral prostheses mounted into patients should be
frequently controlled by the dentist. Particularly in
patients who have gone tumor surgery and thereafter
acquire obturators for maxillofacial defects, post-nasal
secretion is often unavoidable. Such prosthesis exposed
to the nasal area comprises appliances for nasal,
midfacial, large orbital defects, combined facial
prosthesis with defects in the maxilla and tracheal
stent. In fact, especially in patients with postnasal flux,
intra-oral prosthetic appliances are also exposed to
nasal fluid contamination. In such situations, oral flora
of the patients is affected. Hence, prosthesis exposed to
such environments may be indirectly affected from the
microorganisms in nasal secretion. Based on the
obtained data, maxillofacial and oral/dental prostheses
exposed to nasal secretion seems to collect less C.
albicans than the ones exposed to saliva. Nonetheless,
smooth surfaces and heat-polymerization of such
materials may delay microorganism colonization. In
 Dent Mater J 2010; 29(2): 206–212
this study, C. albicans was used as a model
microorganism. The obtained results may change with
other Candida species such as C. tropicalis, Candida
dubliniensis, Candida glabrata, C parapsilosis, C.
guilliermondii, or with other microorganisms such as
S. aureus or S. mutans31,39,46). In this study, no microbial
agents were used in the ‘‘cleaning’’ (washing) process,
but it would be of interest to compare the effectiveness
of denture cleansing agents against microorganisms
immobilized on surfaces.
CONCLUSIONS
Cold-polymerized soft-lining material tested showed
more C. albicans adherence in both saliva and nasal
secretion being widely spread onto the material than
that of the heat-polymerized one, where C. albicans
accumulation was observed more concentrated in
blastospore morphology. The surface roughness of cold-
polymerized soft liner (Visco Gel) was significantly
higher than heat-polymerized soft liner (Molloplas B).
Cold-polymerized soft liner (Visco Gel) with roughened
surface showed a greater adherence of C. albicans.
Contamination of the soft liners in artificial nasal
secretion led to less C. albicans adherence than that of
artificial saliva.
REFERENCES
1) Marsh PD. Are dental diseases examples of ecological
catastrophes? Microbiology 2003; 149: 279-294.
2) Budtz-Jörgensen E. Etiology, pathogenesis, therapy, and
prophylaxis of oral yeast infections. Acta Odontol Scand
1990; 48: 61-69.
3) Zegarelli DJ. Fungal infections of the oral cavity.
Otolaryngol Clin North America 1993; 26: 1069-1089.
4) Tari BF, Nalbant D, Al FD, Kustimur S. Surface roughness
and adherence of Candida albicans on soft lining materials
as influenced by accelerated aging. J Contemp Dent Pract
2007; 8: 18-25.
5) Budtz-Jorgensen E, Stenderup A, Brabowski M. An
epidemiologic study of yeast in elderly denture wearers.
Community Dent Oral Epidemol 1975; 3: 115-119.
6) Budtz-Jorgensen E. Clinical aspects of Candida infections in
denture wearers. J Am Dent Assoc 1978; 96: 474-479.
7) Renner RP, Andors L, Mcnamara TF. The role of Candida
albicans in denture stomatitis. Oral Surg 1979; 47: 323-328.
8) Bulad K, Taylor RL, Verran J, McCord JF. Colonization and
penetraton of denture soft lining materials by Candida
albicans. Dent Mater 2004; 20: 167-175.
9) Nyquist G Denture sore mouth. Acta Odontol Scand 1952;
10: 1-15.
10) Love WD, Goskafa FA, Mixson FJ. The aetiology of mucosal
inflammation associated with dentures. J Prosthet Dent
1967; 18: 515-527.
11) Farman AG, Nutt G. Oral Candida, debilitating disease and
atrophic lesions of the tongue. J Biol Buccale 1976; 4: 20-28.
12) Arendrof TM, Walker DM. Oral candidal populations in
health and disease. Br Dent J 1979; 147: 267-272.
13) Martin MV, Lamb DJ. Frequency of Candida albicans
serotypes in patients with denture-induced stomatitis and
in normal denture wearers. J Clin Pathol 1982; 35: 888-891.
14) Budtz-Jorgensen E, Theilade E, Theilade J. Quantitive
relationship between yeasts and bacteria in denture-induced
stomatitis. Scand J Dent Res 1983; 91: 134-142.
211
15) Budtz-Jorgensen E, Bertram U. Denture stomatitis 1. The
aetiology in relation to treatment and infection. Acta
Odontol Scand 1970; 28: 71-92.
16) Ettinger RL. The aetiology of inflammatory papillary
hyperplasia. J Prosthet Dent 1975; 34: 254-261.
17) Lammie GA, Storer R. A preliminary report on resilient
denture plastics. J Prosthet Dent 1958; 8: 8411-8424.
18) Travaglini EA, Gibbons P, Craig RG. Resilient liners for
dentures. J Prosthet Dent 1960; 10: 664-672.
19) Gonzalez JB, Laney WR. Resilient materials for denture
prostheses. J Prosthet Dent 1966; 16: 438-444.
20) Clarke DA. Resilient lining materials. Dent Tech 1975; 28:
132-137.
21) Williamson RT. Clinical application of soft denture liner: a
case report. Quintessence Int 1995; 26: 413-418.
22) Mack PJ. Denture soft lining materials: clinical indications.
Aust Dent 1989; 34: 454-458.
23) Nikawa H, Hamada T. Binding of salivary or serum proteins
to C. albicans in vitro. Arch Oral Biol 1990; 35: 571-573.
24) Garcia R, Leon BLT, Oliveira VMB, Del Bel Cury AA. Effect
of a denture cleansers on weight, surface roughness, and
tensile bond strength of two resilient denture liners. J
Prosthet Dent 2003; 89: 489-494.
25) Douglas LJ. Adhesion of Candida species to epithelial
surfaces. Crit Rev Microbiol 1987; 15: 27-43.
26) Segal E. Pathogenesis of human mycosis: role of an adhesion
to host surfaces. Microb Sci 1987; 4: 344-347.
27) Verran J, Maryan CJ. Retention of Candida albicans on
acrylic resin and silicone of different surface topography. J
Prosthet Dent 1997; 77: 535-539.
28) Waters MGJ, Williams DW, Jagger RG, Lewis MAO.
Adherence of Candida albicans to experimental denture soft
lining materials. J Prosthet Dent 1997; 77: 306-312.
29) Okita N, Orstavik D, Orstavik J, Ostby K. In vivo and in
vitro studies on soft denture materials: Microbial adhesion
and tests for antibacterial activity. Dent Mater 1991; 7: 155-
160.
30) Nikawa H, Iwanaga H, Kameda M, Hamada T. In vitro
evaluation of C. albicans adherence to soft denture-lining
materials. J Prosthet Dent 1992; 68: 804-808.
31) Radford DR, Sweet SP, Challacombe SJ, Walter JD.
Adherence of C. albicans to denture-base materials with
different surface finishes. J Dent 1998; 26: 577-583.
32) Nikawa H, Jin C, Hamada T, Makihira S, Kumagai H,
Murata H. Interactions between thermal cycled resilient
denture lining materials, salivary and serum pellicles and
C. albicans in vitro: Part II. Effects on fungal colonization. J
Oral Rehabil 2000; 27: 124-130.
33) Razek MKA, Mohamed ZM. Influence of tissue-conditioning
materials on the oral bacteriologic status of complete
denture wearers. J Prosthet Dent 1980; 44: 137-142.
34) Wright PS. The effect of soft lining materials on the growth
of Candida albicans. J Dent 1980 ; 8: 144-151.
35) Grubb BR, Chadburn JL, Boucher RC. In vivo microdialysis
for determination of nasal liquid ion composition. Am J
Physiol 2002; 282: 1423-1431.
36) Shifman A. Clinical applications of visible light-cured resin
in maxillofacial prosthetics. Part 1: Denture base and reline
material. J Prosthet Dent 1990; 64: 578-582.
37) Jin C, Nikawa H, Makihira S, Hamada T, Furukawa M,
Murata H. Changes in surface roughness and color stability
of soft lining materials caused by denture cleansers. J Oral
Rehabil 2003; 30: 125-130.
38) Rodrigues Garcia RCM, Leon BLT, Oliveira VMB, Del Bel
Cury AA. Effect of a denture cleanser on weight, surface
roughness, and tensile bond strength of two resilient
denture liners. J Prosthet Dent 2003; 89: 489-494.
39) Moura JS, da Silva WJ, Pereira T, Del Bel Cury AA,
Rodrigues Garcia RCM. Influence of acrylic resin
212 Dent Mater J 2010; 29(2): 206–212
polymerization methods and saliva on the adherence of four
Candida species. J Prosthet Dent 2006; 96: 205-211.
40) Yildirim MS, Hasanreisoglu U, Hasirci N, Sultan N.
Adherence of Candida albicans to glow-discharge modified
acrylic denture base polymers. J Oral Rehabil 2005; 32: 518-
525.
41) Karaagaclioglu L, Can G, Yilmaz B, Ayhan N, Semiz O,
Levent H. The adherence of Candida albicans to acrylic
resin reinforced with different fibers. J Mater Sci: Mater
Med 2008; 19: 959-963.
42) Verran J, Shakespeare AP, Willcox MDP, Knox KW. The
effect of pH on adhesion and hyphal formation by strains of
Candida albicans. Microbial Ecology in Health and Disease
1991; 4: 73-80.
43) Baena-Monroy T, Moreno-Maldonado V, Franco-Martínez F,
Aldape-Barrios B, Quindós G, Sánchez-Vargas LO. Candida
albicans, Staphylococcus aureus and Streptococcus mutans
colonization in patients wearing dental prosthesis. Med Oral
Pathol Oral Cir Bucal 2005; 10: E27-E39.
44) Radford DR, Challacombe SJ, Walter JD. Denture plaque
and adherence of Candida albicans to denture-base
materials in vivo and in vitro. Crit Rev Oral Biol Med 1999;
10: 99-116.
45) Verran J, Taylor RL, Lees GC. Bacterial adhesion to inert
thermoplastic surfaces. J Mater Sci Mater in Med 1996; 7:
597-601.
46) Verran J, Lees GC, Shakespeare AP. The effect of surface
roughness on the adhesion of Candida albicans to acrylic.
Biofouling 1991; 3: 183-192.