Dental Caries - A Student Project

Sharan Bisla 2000

The information on these pages is provided as an instructional tool for general biology and
introductory microbiology classes. It is not intended as a diagnostic aid, nor should it
replace consultation with medical professionals.

What is dental caries?

There are relatively few infectious diseases of the upper alimentary canal but one common
disease is dental caries. Dental caries, a pathological process of destruction of tooth
structure by microorganisms, is also known as tooth decay and is commonly called
"cavities." Dental plaque, which leads to caries, is the oral flora which adhere to teeth and
break down tooth enamel.

History of dental caries

Prehuman skeletons more than 500,000 years old indicate that very little caries was
prevalent. In the In Europe during the Medieval period less than 10% of human teeth had
cavities and they were mostly on parts of the teeth exposed by the receding gums of older
people. During the 17th century there was an increase in the incidence of tooth decay and
location of cavities changed to include the sides and biting surfaces of teeth. Young people
were also starting to get tooth decay along with older people. Then in the European diet
flour and sugar (sucrose) were introduced and this lead to an increase in dental caries. (1)

Archaeological evidence from preColumbian Mesoamerica indicates that dental caries

became prevalent among people at about the time the agriculture shifted to the triumvirate
of maize, beans, and squash, all of which have high sugar contents.
A number of factors are necessary for caries to occur. These factors include the interaction
of the host, oral flora, and the substrate which is primarily sucrose (sugar). In order for
tooth destruction to occur, there must be a susceptible host, cariogenic microorganisms and
a suitable substrate present for a sufficient length of time. Tooth brushing can reduce the
likelihood of caries by reducing number of cariogenic organisms and removing the
substrate. (2)

How dental caries occurs and the cariogenic bacteria

involved
Organisms predominant in plaque are of the genus Streptococcus, with Streptococcus
mutans being the species most important in the formation of dental caries. According to the
Bergey's Manual of Systematic Bacteriology (3), S. mutans are non-motile Gram positive
cocci. They are 0.5-0.75 micrometers in diameter, occurring in pairs, short medium length
chains, without capsules. S. mutans have several properties that are important in the carious
process. First, they colonize on tooth surfaces. Second, they synthesize insoluble
polysaccharides from sucrose. This allows adhesion to smooth surfaces and appears to be
important in the formation of smooth surface caries. Third, they ferment sucrose to form
lactic acid.

Another organism that is important in the development of caries is the Lactobacillus.

Lactobacillus are gram positive, non-spore forming rods. They normally constitute only a
small fraction of plaque flora when compared to S. mutans. Lactobacillus species are not
important in the initiation of caries but in the continuation.

Variety of microorganisms in the oral flora

We streaked samples of oral flora on blood agar plates in order to see the variety of oral
microorganisms. The teeth were brushed a couple hours before taking samples.

The blood agar plates were incubated aerobically at 37C, temperature of the mouth. These
blood agar plates were incubated for two days and we noted that white-gray colonies grew
on the surface of the agar. This gave us an idea of the various kinds of colonies that were
present in dental plaque. We saw a large variety of colonies varying in type, shape, size and
color.
This semester Dr. Jacobs and I made Gram stains of our own dental plaque from different
regions of the teeth. This Gram staining was performed not only to reveal the size and
shape of the various bacteria, but also tell us some general information about their basic
chemistry. These Gram stains were viewed through the microscope and we observed there
was great diversity in the oral flora found in the dental plaque.

This picture shows the diversity in the

oral flora found in dental plaque. The
spheres (cocci) represent the
Streptococcus and the rods show us
Lactobacillus. These two types of
bacteria were the most predominant in
dental plaque.

This picture illustrates a cheek cell

in the center covered by various
microorganisms present in dental
plaque.
There were Gram positive and Gram negative rods (bacilli), spheres (cocci), and spirals
(spirillae). There were differences in bacteria present in different samples of the mouth. We
found that even in a "clean" mouth (the teeth had recently been brushed) there were still
millions of bacteria in the dental plaque.

Corrosion chemistry of oral bacteria

Lactic acid is a strong acid that is effective in demineralization of tooth structure. S. mutans
are present in the mouth only when a solid surface such as teeth is present. Salivary
concentrations of S. mutans range from undetectable to 10,000,000 colony forming
units/ml, with a mean concentration of about 100,000 CFU/ml. (4)

Streptococcus mutans and Lactobacillus are acidogenic and aciduric meaning that they can
produce acids which can dissolve the tooth substance (calcium phosphate in the form of
hydroxyapatite crystals) and that they can survive and even produce acids in a low pH
environment. Compared to other oral flora, Streptococcus mutans and Lactobacillus are
cariogenic and have enzymes with greater resistance to acid. They also have greater
capacities of the cells to extrude protons from the cytoplasm. The most important enzyme
for extrusion of protons is ATPase. This enzyme is found in higher amounts and is working
at lower pH with S. mutans and Lactobacillus.

Cariogenic bacteria produce lactic acid when they are exposed to high concentrations of
sugar. Lactic acid has a pK of 3.8, while other carboxylic acids have a pK of 4.8. (The
lower the pK number, stronger the acid and weaker the base.) Lactic acid can chelate the
calcium phosphate and forces the chemical equilibrium to shift so the saliva, which is
supersaturated with calcium phosphate ions, can not replace the calcium phosphate as
rapidly as it is being pulled away from the teeth. Dental caries is essentially corrosion of
teeth enamel.

The bacterial cell membrane is permeable to undissociated carboxylic acids. These

undissociated acids act as carriers to bring protons back into the cytoplasm. Bacteria that
make lactic acid as a fermentation product can grow and produce acids at one pH unit lower
than bacteria that make acetic, propionic, or butyric acid as a fermentation product. For this
reason, these bacteria are especially important in formation of dental caries. (5)

Experiments we conducted confirm corrosion chemistry

of dental caries
During this semester we conducted an experiment to see how certain oral flora can destroy
the tooth enamel which is made of calcium phosphate.

We produced a minimal salts medium which consisted of calcium phosphate, sucrose, agar,
water and other chemicals. After creating this minimal salts medium it was poured into
sterile plates and solidified. Then samples of the different colonies that were grown on the
blood agar plates were taken and smeared on the solidified calcium phosphate medium
plates. These plates were also incubated at 37C, again the temperature of the mouth for
about three weeks and were observed twice a week, at the beginning and middle of the
week. As time progressed I observed there was a clearing effect being produced around
certain colonies of bacteria because of acid production.

We concluded from these experiments certain bacteria can ferment the sucrose into organic
acids such as lactic acid. The lactic acid lowers the pH enough to dissolve the calcium
phosphate in the medium as it would with tooth enamel (see pictures of results from the
experiment).

Prevention
Dental caries is a complex multifactoral disease. One of the most important factors in the
etiology of dental caries is the patient's level of oral hygiene. If thorough hygiene is
maintained there will not be a build up of plaque and bacterial colonization of tooth
structure is not initiated. Maintenance of good oral hygiene requires education, motivation
and reinforcement if compliance is to be achieved. The dietary profile also plays a
important role in dental caries. The amount of sucrose consumed, the consistency of the
food, and the frequency of feeding all affect the level of caries activity. The availability of
fluoride in water will also decrease dental caries. Fluoride is incorporated into the calcium
phosphate matrix of enamel and reduces the occurrence of dental caries.

What did I learn?

Before doing this directed study I was unaware of the vast diversity in bacteria that is
present in the mouth. Each human has its own unique oral flora and the composition of the
oral flora determines the susceptibility to tooth decay. The microorganisms that adhere to
the tooth surface are called dental plaque. The dental plaque causes tooth decay by
fermenting sucrose to lactic acid.

The most interesting part of the study was creating a calcium phosphate medium which
represented tooth enamel and growing colonies of bacteria to confirm corrosion chemistry
of dental caries. It was interesting to look at dental caries from both microbiological and
chemical perspectives and how they combined to cause tooth decay.

References
(1) Nester, E. W., C. Roberts, N. Pearsall, D. Anderson, and

M. Nester. 1998. Microbiology: A Human Perspective. WCB McGraw-Hill , Boston.

(2) http://sdm.uchc.edu/ProjectD/Knowledge/1.Texts/4.Disease/DentalCaries.HTML Back

to text

(Dental school website at the University of Connecticut)

(3) Holt, J. G., N. R. Kreig, P. Sneath, J. Staley, and S. Williams. 1994.Bergey's Manual of
Determinative Bacteriology Ninth Edition. Williams&Wilkins, Baltimore.

(4) http://sdm.uchc.edu/ProjectD/Knowledge/1.Texts/4.Disease/DentalCaries.HTML

(5) http://www.odont.lu.se/mutans/mutgen.html Back to text