THE FUNCTION OF ENZYMES IN REMOVING CANDIDA ACCUMULATED ON DENTURE PLAQUE

Dentistry
THE FUNCTION OF ENZYMES IN
REMOVING CANDIDA ACCUMULATED
ON DENTURE PLAQUE
SENAY CANAY
SIBEL ERGÜVEN
NURAN YULUG
SUMMARY: The denture plaque that is a cause of denture stomatitis has Candida albicans to a
great extent. The plaque accumulated should be removed in order to prevent infections related to
Candida albicans. Since chemical denture cleansing agents have some inconveniences in application
and are less effective, in recent years the use of enzyme containing cleansers has become popular. The
aim of this study was to evaluate the efficacy of enzymes in removing C. albicans from the denture
plaque. The plaque was accumulated on polymethyl-methacrylate plates by the strain of C. albicans
and the effect of enzymes such as papain, trypsin and amylase were investigated by the cell-counting
procedure. Papain, a proteolytic enzyme, was found to be effective (80%).
Key Words: Candida albicans, denture plaque, enzymes.
INTRODUCTION
The hygiene of denture has been overlooked by
Mechanical denture cleansing does not always yield
patients and physicians for unknown reasons. However,
good results. The use of chemical denture cleansers
colonies of bacteria and fungi grown on denture plaque
produces more effective results especially in geriatric
give rise to denture stomatitis. According to Neill the soil-
patients and in people who have problems in wearing den-
ing of denture occurs in 3 phases (11): first mucin and
tures. An effective denture cleanser must have the ability
food debris accumulate upon the surface of denture, then
to remove microbial plaque formed and prevent its rebuild.
plaque builds up, this plaque acts as a matrix and then
In addition, it must be capable of removing mucin, food
calcification begins.
debris, calculus and stains. In the meantime, however, it
There is a close relationship between denture stomati-
should not cause harm to denture base material (4).
tis seen in most of the patients who are denture wearers
Chemical denture cleansers are classified into various
and Candida albicans (3, 6, 8,12,13). Thus, the presence
groups such as alkaline peroxides, alkaline hypochlorites,
of Candia albicans has been shown on the denture
acids, disinfectants and enzymes (4, 8,11,14,16).
plaque by studies (14). For this reason, Candida should
Alkaline peroxides are particularly effective on new
be removed from the surface of denture.
plaque and satins formed. To be effective they must
Another uneasy cosmetic concern is stains which
remain in contact with denture for a long period. These
accumulate on the surface of denture. They are formed
cleansers cause bleaching of the acrylic resin base when
as a result of various habits such as consumption of
routinely used.
colored foodstuffs and smoking. The satins adhere
Alkaline hypochlorites remove stains on denture and
biologically to the surface of the denture in the same way
decompose mucin and other organic substances. They
as plaque (1). This situation causes odor and also impairs
are effective against both bacteria and fungi and dissolve
the esthetic quality of denture.
the organic matrix of denture plaque by acting directly.
Besides, these materials cause some harmful effects
From Department of Prothodontics, Faculty of Dentistry,
such as tarnish and corrosion of the metal components of
Hacettepe University,
dentures. The routine use of alkaline hypochlorites may
From Department of Microbiyology, Faculty of Medicine,
also give rise to bleaching of the acrylic resin base.
Hacettepe University, Ankara, Türkiye.
Effective against calculus or stains on dentures, acids
Journal of Islamic Academy of Sciences 4:1, 87-89, 1991
83

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ABILITY OF ENZYMES TO REMOVE CANDIDA
CANAY, ERGÜVEN, YULUG
may corrode the metal component in dentures. Disinfec-
Figure 2: Candida species seen on the acrylic plates (x320).
tants such as dilute solution of chlorhexidine gluconate or
salicylate reduce visibly the amount of denture plaque.
However, when the treatment is discontinued, a relapse is
seen. They are effective especially against fungi. As their
daily use leads to coloring of denture, they have a limited
field of application.
Enzymes breakdown the macromolecules of glyco-
proteins, mucoproteins and mucopolysaccharides found
in denture plaques into less adhesive small units (6). They
are effective against both bacteria and fungi. Enzymes
are very suitable in removing the heavy deposits from
denture plaque and they also prevent new plaque forma-
tion. Their use has no harmful effect on the denture base
material and its other component. In this study, the
cleansing-ability of some enzyme containing solutions,
which are superior to chemical disinfectants, to remove
Candida albicans from acrylic resin denture plaque were
investigated.
Enzymes
The standards of enzymes, Papain, Trypsin and Amylase
MATERIALS AND METHODS
were obtained from Ankara Refik Sydam Hifzissihha Institute.
Preparation of test specimen
0.1 mg/ml solutions of these enzymes in PBS (pH 7.2) were pre-
Fifty specimens, 10x10x2 mm in size, were prepared from
pared. For each enzyme five 10ml tubes were prepared. Tubes
polymethyl-methacrylate (DeTrey Dentsply QC-20 Denture
having only 10 ml PBS were used as control.
Material, France) which was used as denture base material and
both of the surfaces were polished by standard polishing proce-
Removing of Candida
dures (Figure 1). Specimens were kept in distilled water to pre-
Plates having plaque as a result of incubation with Candida
vent dehydration.
albicans were placed in the solutions of enzymes. Tubes were
incubated at 37°C for 8 hours. After incubation, plates were
Preparation of fungal suspension
washed ten times in saline solution. They were fixed in methanol
The strain of Candida albicans (RSKK 81-623) used in this
and stained with crystal violet. After drying, fungal cells adhered
study was supplied by Ankara Refik Saydam Hifzissihha Institute
to the plates were counted by microscopy (Figure 2) and the
from Institute Pasteur. The cultures were grown in peptone-yeast
mean of each count was used. By comparing with the control
extract glucose (PYG) medium containing 1% peptone, 0.5% yeast
plates, the Candida-removing percentage by enzymes was
extract and 2% glucose. Ten ml suspensions of these cultures
determined.
(5x106 cfu/ml) were prepared. Acrylic resin plates were immersed
in these suspensions and incubated for 12 hours at 37°C.
RESULTS AND DISCUSSION
The Candida-removing ability of enzymes from acrylic
plates was illustrated in Table 1. According to these
Figure 1: Standard acrylic plates (10x10x2 mm).
results, amylase was ineffective for removing Candida
albicans. Proteolytic enzymes, Trypsin and Papain were
effective 5 and 80% respectively.
Candida albicans as a significant cause of denture
stomatitis can be isolated not only from the tissue surface
on which the denture fits but from the inner surface of
denture as well (5,10). During the growth and metabolism
of Candida species organic acids that decrease the pH
are formed. These acids may have a direct cytotoxic
effect on the mucosal epithelium. Moreover, the acidic pH
may activate the acid proteases or phospholipases of
Candida albicans and may result in the inflammation of
mucosa. As this type of inflammation increases, more
Candida albicans may adhere to the surface of denture.
Many studies have been conducted to evaluate the effi-
88
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ABILITY OF ENZYMES TO REMOVE CANDIDA
CANAY, ERGÜVEN, YULUG
Table 1: The enzymes effective in removing Candida albicans.
2. Augsburger RH, Jaffar ME : Evaluation of seven propri-
etary denture cleansers. J Prosthet Dent, 47:356, 1982.
3. Budtz-Jorgensen E, Knudsen AM : Chlorhexidine gel and
Enzyme
% cell
Mean of the
Control
steradent employed in cleaning dentures. Acta Odontol Scand,
removed Candida albicans-cell
counted
36:83, 1977.
?
4. Budtz-Jorgensen E : Materials and methods for cleaning
-amylase
0
100
100
dentures. J Prosthet Dent, 42:619, 1979.
Proteolytic enzyme
5
95
100
5. Burns DR, Burns DA, DiPietro GJ, Gregory RL :
Trypsin
Response of processed resilient denture liners to Candida albi-
Proteolytic enzyme
80
20
100
Papain
cans. J Prosthet Dent, 57:507, 1987.
6. Connor JNE, Schoenfeld CM, Taylor RL : An evaluation of
an enzyme denture cleanser. J Prosthet Dent, 37:147, 1977.
cacy of commercial denture cleansing agents used for
7. Ghalichebaf M, Graser GN, Zonder HA : The efficacy of
removing the denture plaque (1, 2, 7, 8,10,13,15,16).
denture-cleansing agents. J Prosthet Dent, 48:515, 1982.
However, most of them have some known disadvan-
8. Lambert JP, Kolstad R : Effect of a benzoic acid-detergent
tages. It has been reported that enzyme -containing
germicide on denture-borne Candida albicans. J Prosthet Dent,
cleansers are less harmful and more effective. In our
55:699, 1986.
country this type of study with enzymes has not yet been
9. Minagi S, Tsunoda T, Yoshida K, Tsuru H : Objective test-
done.
ing of the efficiency of denture-cleansing agents. J Prosthet
Several methods have been used in order to deter-
Dent, 58:595, 1987.
mine the plaque cleanser efficacy of denture cleansers.
10. Moore TC, Smith DE, Kenny GE : Sanitization of den-
The weight method was suggested by some of the inves-
tures by several denture hygiene methods. J Prosthet Dent,
tigators (13). However, this method is not very sensitive.
52:158, 1984.
More quantitative results can be obtained by counting the
11. Neill DJ : A study of materials and methods employed in
cells grown on plaque. In our studies, we used this
cleaning dentures. Br Dent J, 124:107, 1968.
method.
12. Santarpia RP, Pollock JJ, Renner RP, Spiechowics E :
The cells of C. albicans found diffusively on denture
An in vivo replica method for the site-specific detection of Can-
plaque adhere to the acrylic resin by means of proteins
dida albicans on the denture surface in denture stomatitis
and polysaccharides. Consequently, this plaque can be
patients: Correlation with clinical disease. J Prosthet Dent,
removed from the surface of acrylic by enzymes cleaving
63:437, 1990.
these bonds (13).
13. Sharp EW, Verran J : Denture cleansers and in vitro
The enzymes were found to be of optimum effect in
plaque. J Prosthet Dent, 53:584, 1985.
plaque removal in 8 hours as reported by the researchers
14. Tamamoto M, Hamada T, Miyake Y, Suginaka H : Ability
(6). We also used this period in our experiments. There
of enzymes to remove Candida. J Prosthet Dent, 53:214, 1985.
are studies in which various enzymes have been used in
15. Tarbet WJ, Axelrod S, Fratarcangelo PA : Denture
denture cleansing. Particularly yeast lytic enzyme group
cleansing : a comparison of two methods. J Prosthet Dent,
and proteolytic enzymes were found to be effective. In our
51:322, 1984.
study from protedytic enzyme group Trypsin was effective
16. Türköz Y, Karaagaclioglu L, Misirligil A : Muhtelif
5% whereas Papain was effective 80%. Our results are
kimyasal protez temizleyici maddelerin protez plaklarindaki Can-
consistent with the other studies (9,14).
dida albicans aktivitesine etkileri. AU Dis Hek Fak Derg, 15:47,
As a conclusion, in denture cleaning in order to pre-
1988.
vent Candida induced denture stomatitis, the use of pro-
teolytic enzyme-containing solutions such as papain
could be suitable. Patients were instructed to remove their
dentures when they retire at night. It is our opinion that if
patients could immerse their dentures in enzyme-contain-
ing solutions at night, 8 hours sleeping time would serve
well to complete the cleansing procedure.
Correspondence:
Senay Canay
REFERENCES
Hacettepe University,
1. Altman MD, Yost KG, Pitts G : A spectrofluorometric pro-
Faculty of Dentistry,
tein assay of plaque on dentures and of denture cleaning effi-
Dept. of Prosthodontics,
cacy. J Prosthet Dent, 42:502, 1979.
Ankara, TÜRKIYE.
Journal of Islamic Academy of Sciences 4:1, 87-89, 1991
89

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Document Outline

• JIAS Volume 4, No 1
• The Function of Enzymes in Removing Candida Accumulated on Denture Plaque
• Summary
• Introduction
• Materials and Methods
• Results and Discussion
• References

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