1

Saliva non-
protein organic
compounds.
Besides proteins there
are other organic
compounds in human saliva:

a) Steroid hormones estrogens, and

androgens,(testosterone),
progesterone, cortisol.

b) Urea.Urea content in mixed saliva

is 75 90% of blood serum
concentration. Some oral cavity
bacteria release urease which splits urea to CO 2 and NH3.
Formed ammonia attaches proton and converted to
NH4+-ion which increases saliva pH. Oral cavity bacteria
can use urea as a source of nitrogen.

c) Uric acid and creatinine, their content depends on blood

concentration.

d) Total lipid amount is equal 60-70 mg/L. They are palmitic,

stearic, eucosapentanonic, oleic, cholesterol and its esters
and small amount of phospholipids.

e) Amino acids

f) Lactate and pyruvate. Lactate and pyruvate increasing

in dental deposit contributes pH shifting to acidic
region, activation of demineralization and caries
development.

g) Carbohydrates are present mainly in bound with

proteins condition. Free monosaccharides are formed by
microorganismsenzymes hydrolysis of di- and
oligosaccharides. Formed monosaccharides are rapidly
used as energy source by oral cavity microbes and
serve for dextran and levan building. Glucose
2

concentration in mixed saliva is 0,06-0,15 mMol/L in the norm.

In hyperglycemia glucose content is considerably increased.

Saliva biologically active compounds.

Saliva glands synthesize and secret to saliva a number of
biologically active peptides: insulin-like protein, nerve growth
factor (NGF), epithelium growth factor (EGF), parotin,
renin, erythropoietin, thymocytetransforming factor,
mesoderm growth factor, granulocytopoiesis factor.

1) Nerve growth factor (NGF) is oligomer in inactive form,

built up from 2 -, 2 - and 2 -chains. In the course of
activation oligomer dissociates, -chains are degraded by
partial proteolysis.

NGF participates in regulation of metabolism of

sympathetic nerve fibers.

Receptors are tyrosine kinases.NGF increases glucose

consumption by target these cells, activates Na+ , K+ -
ATP-ase work, synthesis of a particular enzymes
responsible for nucleotides and lipids synthesis.

NGF stimulates oral cavity tissues wound healing.

Epithelium growth factor (EGF) is polypeptide. Its found in

blood, urine, gastric and pancreatic juice, breast milk,
cerebrospinal fluid. Androgens, progestin and thyroxin
increase synthesis and rise concentration of this factor in
blood.

The targets of EGF are epithelial cells of oral cavity

mucosa,gullet, esophagus, cornea, breast gland, lung alveoli, and
also vessel endothelium chondrocytes. Formed hormone-
receptor complexes are plunged by endocytosis into the
cell and activate matrix processes replication,
transcription and translation.

Produced in salivary glands EGF is released to saliva and with

saliva enters stomach and further to intestine. EGF renders
mitogenetic effect in mucosa damage of gastro-intestinal
tract.EGF enhance proliferation and keratinization of
3

epithelium, decrease hydrochloric acid secretion in

stomach, stimulate gastric and duodenum ulcers healing,
growth of vessels.

2) Parotin is a protein nature hormone(it was found in parotid

salivary gland for the first time and named as parotin). In
mixed saliva its present by the group of structurally
similar protein: parotin S, A, B and C. One of them
parotin S is synthesized by submandibular glands.

Hormone regulate phospho-calcium metabolism and like

wise calcitonin contributes enamel mineralization. Hormone
enhances cartilage mineralization, stimulates nucleic acids and
protein synthesis in odontoblasts, dentin and bones mineralization
and decreases calcium and glucose concentration in blood
plasma.

3) Insulin-like protein(Ilp) has similar to insulin structure,

composed of 2 polypeptide chains A and B. Protein is
formed by duct cells. In diabetes mellitus it content is
considerably increased in saliva. Insulin-like protein
increases glucose consuption by these cells and
formation of energetic reserve glycogen.

4) Saliva kallikrein is specific serine-protease hydrolyzing

peptide bonds in proteins, formed mainly by cationogenic
amino acids (arginine and lysine).

Kallikreinis secreted in active and in inactive form as

prekallikrein. The main function of kallikrein is partial
proteolysis of specific proteins and formation of active
peptides bradykinin and kallidin(kinins).

Kinins possess multiple of physiological functions. In saliva

gland ducts kinins change local blood flow, increase
secretion of water and electrolytes by salivary glands.
Kinins influence microcirculation.

Excessive kinins formation contributes inflammation

development.
4

5) Renin is a proteolytic enzyme synthesized in

submandibular glands, also in kidney, adenohypophysis
and seminal glands. Angiotensin formed under renin
action causes vasoconstriction, contributes
salivation, increase reparation of oral cavity mucosa.

Oral cavity protective systems.

In healthy persons oral cavity microflora species
composition is constanct and barely quantity of microbes
can be substantially changed. This depends on salivation
intensity, food character, oral cavity hygienic state, somatic
diseases presence.

Increased quantity of microorganisms in oral cavity may

be conditioned by two factors: 1) damage of saliva
formation and salivation, impairmentof mastication and
deglutition(swallowing); 2) difficulty of washing-out of
microorganisms by saliva for example in caries, pathological
dento-gingival pockets, badly fitted fixed dental prosthesis
( denture).

Oral cavity normal microflora is composed of hundred

species among which most of all are anaerobic microflora
representatives.These microorganisms split
carbohydrates to lactate which decrease pH and by this
reduce the growth of putrefactive microorganisms which
get to oral cavity from environment.

Quantity of inhabiting microorganism in oral cavity is in

dynamic balance due to antibacterial saliva factors and
leukocytes incoming to oral cavity. Saliva possessing
antibacterial properties is able to control quantitative and
qualitative composition of microflora and thereby
maintenance of oral cavity homeostasis.

Oral cavity protective systems are divided to nonspecific

and specific.

Oral cavity nonspecific protective factors for cariesogenic and

other bacteria are conditioned by:
5

- lysozyme, lactoferrin, lactoperoxidase content;

-barrier function of mucous and submucosal tunic.

Present compliment system proteins and immunoglobulins

in saliva provide bacteriostatic and bactericidal activity of oral
fluid.

Lysozyme (muramidase from latin murus wall) is

glycosidase. Lysozyme splits glycosidic bond in murein
polysaccharide chains.

Murein is heteropolysaccharide of glycoprotein

composition of bacteria cell wall.Damage of murein structure
leads to change of membrane permeability, lysis, and
phagocytosis of damaged cells.

Oral cavity mucous tunic wounds healing(having contact

with large amount of different microorganisms including
pathogenic) depends on saliva lysozyme activity.

In humans lysozyme is found also in tears, milk, on nose mucous

tunic. The most reach source of lysozyme is chicken egg protein.

Peroxisase (lactoperoxidase).Perxidases are heme-

containing glycoproteins.

Two groups of peroxidases reveal activity in mixed saliva. They

differ in isoelectric point. They are produced by salivary glands or
enter saliva from blood granulocytes. Salivary gland
peroxidases have maximal activity at pH 5,0 - 6,0.

There is small amount of hydrogen peroxide (H2O2) in oral fluid

which is produced mainly by aerobic bacteria. H2O2 is non-
enzymatically broken down with superoxide anion
formation. Formed O2attacks and induces death of
anaerobic microorganisms which have not enzymatic
protection from superoxide anions.

There are 10 times more anaerobes than aerobes in oral

cavity. The increase of anaerobe bacterial clump (which produce
lactate) leads to mixed saliva pH reduction and peroxidase
activation.
6

Enzyme catalyzes oxidation-reduction reaction with high velocity

in with hydrogen peroxide and ions of I-, Br- or SCN- participation
(coming to saliva from blood).

H2O2O2 -

spontaneously

H2O2 + SCN - OSCN- + H2O O2 - +

SCN -

Peroxydase spontaneously

Oxygen active radicals are spontaneously arisen from formed

hypothiocyanate (OSCN-). AOS (active oxygen species) damage
cell membrane lipids of anaerobic microorganisms with high
reactivity. This contributes to restoration of equilibrium between
aerobe and anaerobe in oral cavity. This pathway of active oxygen
formation provides ten times more powerful antibacterial effect
than non-enzymatic peroxide cleavage.

Therefore saliva peroxidase biological function is

promotion of oxygen active species formation which
inhibit growth of microorganisms by damaging their
membranes. At the same time hypothiocyanate-ion and active
oxygen species do not damage oral cavity epithelial cells which
have effective enzymatic systems rapidly inactivating these ions.

Saliva immunoglobulins.
There are representatives of 5 classes of immunoglobulins in
saliva: IgA, IgD, IgE, IgG and IgM, but IgA reveal the particular
protective properties. Secretory component (SC) is
glycoprotein synthesized by glandular epithelium cells and
exposed on basolateral membrane surface. It is a receptor which
specifically interact with dimer (IgA)2-J. Formed complex enters
cell by endocytosis and migrates to apical part of membrane.
7

During exocytosis transmembrane part is cleaved from SC-

receptor by the activity of proteolytic enzymes. Immunoglobulin
composed of IgA 2 monomers combined by J-chain and
shortened secretory componen enters secret. Formation
of this complex structure increases stability of IgA to
proteolysis by mixed saliva enzymes and influence of
denaturating factors (temperature, pH and others).

Secretory immunoglubulins (aIgA):

-reduce adhesion of bacteria on oral cavity mucous tunic

by binding microorganisms;

-damage microorganism membranes and induce their

depth by activating compliment system;

-decrease adsorption and reproduction of viruses in

mucous tunic epithelial cells by binding viruses;

-prevent caries development by decrease of cariesogenic

streptococcus adhesion on tooth enamel.

Presence of some blood coagulating and anticoagulating

system factors are also concerns to saliva protective function.
These proteins provide effective protection in microtraumae of
oral cavity which arise every day during food intake.

Gingival fluid.
8

Gingival fluid or fluid of gingivalsulcus is a complex biological

medium which plays an important role in maintenance of normal
condition of periodontium tissues.

Gingival fluid fills up the fissured space between tooth

surface and gingival brim (edge). Gingival sulcus is lined by
epithelium attached to enamel cuticle and has depth of 1, 0 1, 5
mm.

In the norm sulcus bottom is settled at the level of enamel-

cement conjunction but deepened by aging.

Gingival fluid is blood serum transudate which enters

gingival sulcus through post capillary venulae.

Amoun of transudate coming to gingival sulcus is decreased in

the morning but increased in the evening. 0, 5 2, 4 ml of
transudate is secreted during twenty four hours.

Gingival fluid is moved away by gum lymthatic vessels that is

why compounds from it can penetrate not only to oral cavity but
to subjacent tissues.

The main function of gingival fluid is protective.Gingival

fluid has pH 6,3 7,93, maintains mixed saliva pH and
prevent its reduction.

Gingival fluid possesses antimicrobial activity because

contain a lot of protective proteins immunoglobulins,
complement system, and also creates barrier
amortization protection of tooth in response to
mastication load.Change in gingival fluid amount affects tooth
mobility and can lead to morphological alteration of periodontium
tissues.

Gingival fluid contains electrolytes, proteins, enzymes,

cast-off epithelium cells,leucocytes, microorganisms and
their metabolites, extracellular fluid of gum tissues.
Superficial epithelial cells participating in attachment to enamel
contain low amount of mitochondria, has reduced metabolic
processes activity and that is why the rate of their regeneration is
considerably higher than other periodontium cells.
9

Amount and composition of gingival fluid are changed in

periodontium inflammation. In patients with periodontopathy
composition and volume of gingival fluid are changed owing
tooutflow of extracellular liquid from inflammatory mucous tunic
of gums.

Gingival fluid cells are mainly polymorphonuclear

leukocytes, amount of which can be increased at different stages
of periodontopathy. Gingival fluid is the main source of leukocytes
in oral fluid. Leukocytes are absent in oral fluidtill eruptionhence
till gingivalgroove formation.

Gingival sulcus microflora is very varied,coccal bacteria

are present in large amount.

In inflammation development gum microflora composition is

changed, amount of gramm-positive bacteria become less, but
amount of spirochetes, fusobacteria, leptotrichia and
others are increased. Association of fusobacteriae settling
in gum pockets with spirochetes inhabiting dental plaque
are the main causative agent of pyo-inflammatory and
necrotizing ulcerative gingivitis processes.

Gingival fluid electrolytes. Na+ and K+ amount in gingival

fluid is lower than in blood plasma. In periodontitis these
ions amount is increased.

Gingival fluid is one of the sources of fluoride in oral

cavity and fluoride concentration in gingival fluid and
blood plasma are equal.

Gingival fluid proteins. Protein amount in gingival fluid and

blood plasma is 60-70 g/L. Albumin, IgG, IgA, IgM, sIgA,
complement system proteins coming in to gingival fluid
from blood plasma.

Complement system proteins. Complement system is

present by 20 proteins which are formed mainly in the liver and
enter gingival fluid from blood. They are inactive in blood
serum, but in antigen-antibody complex formation are
activated by interaction with each other and partial
10

proteolysis. Part of proteins reveal proteolytic activity at that

and carry out partial proteolysis of other proteins of system.

As a result of transformations the assembly of protein

structures bindsto bacterial membranes. Formed
complexes generate pores in microorganism membrane
that leads to metabolism damage and their depth.

Peptides releasing in the process of compliment system

activation induce vasodilatation and attraction of
phagocytic cells to microorganism clump spots
(chemotaxis).

Besides that complement increases the ability of

phagocytic cells to bind, engulf and break down damaged
microorganisms (phagocytosis).

Gingival fluid proteins forms sticky film which links

gingival groove with tooth surface. Cell adhesion is provided
by proteins of extracellular matrix fibronectin, lamininand fibrin.
Present in gingival fluid plasmin hydrolyzes fibrin of film and
prevents formation of more dense (stiff) film damaging entrance
of gingival fluid to gingival groove.

Gingival fluid enzymes.

Gingival fluid enzymes enter from blood plasma, cast-off

epithelium cells, leukocytes or bacterial cells. Activity of
these enzymesis not determined under normal condition, but is
increased in inflammation development in periodontium.
One of pathogenetic factors in periodonopathy is the activity of
bacterial hydrolases of aggression and invasion. They are:

-hyaluronidase which hydrolyzes hyaluronic acid and thereby

contributes penetration of microbes deep into periodontium
tissues;

-neuraminidase which detaches neuraminic acid(sialic acid)

from glycoprotein and as a result breaks down membrane
structure of epithelial cells and increases possibility of interaction
of these cells with microorganisms;
11

-collagenase catalyzes connective tissue collagen proteolysis

and facilitates microorganisms migration to periodontium tissue;

-phospholipase C (lecithinase C) hydrolyzes phosphatidylcholine

(lecithin) of cell membranes and increases membrane
permeability. This leads to damage of metabolism and cell depth;

-Plasmin (fibrinolysin) which carries out fibrin proteolysis

(formed in inflammation) and contributes microbe penetration
deep to periodontium;

-DNAasesplits DNA and causes retardation of tissue

regeneration;

-Proteases hydrolyze immunoglobulins, complement system

proteins and suppress protective system of gingival fluid.

Cast-off epithelium cells are source of enzymes of

carbohydrate, amino acids, fatty acids metabolism,
common metabolic pathways, inactive collagenase in
complex with 2 -macroglobulin inhibitor.

Elastase with wide substrate specificity enters the

gingival fluid from leucocytes. In the norm it is inhibited by 2
macroglobulin and 1-antitrypsin. Both enzymes can be activated
by the action of trypsin like proteases.

Gingival fluid low-molecular organic compounds are

products ofgingival groove bacteria: ammonia, hydrogen
sulphide, indol, butyric, lactic, acetic, propionic and formic
acids. There are not or a few amount of these compounds in
normal state. But in periodontium tissue
inflammationcaused by change of gingival groove
microbial flora, amount of these compounds are sharply
increased. Each of these compound has its own mechanism of
toxicity, can damage function and even cause necrosis of
epithelial cells first of all.

Ammonia (NH3) is produced by microorganisms in the amino

acids catabolism and also from urea by the actionof bacterial
urease. NH3 enter to gingival epithelium where is converted to
12

NH4 by proton binding. As a result pH in these cells is

increased.

Urea + H2O CO2 + 2NH3

pHconservation at optimal level in cells is provided by two

reactions: glutamine synthesis and reductive amination of
-ketoglutarate.Increases of the rate of these reactions leads to
decreased amount of NADH, -ketoglutarate and energetic
metabolism damage in epithelial cells.

ATP ADP + P i

Glu+ NH3
Gln

Glutamine synthetase

NADH + H + NAD+

-kG + NH 3

Glu + H 2 O

Glutamate dehydrogenase

- Low-molecular fatty acids are synthesized from

glucose by microorganisms. They pass through cellular
and mitochondrial membranes of epithelial cells and
uncouple oxidation and phosphorylation. Reduced
ATP/ADP ration damages synthetic processes necessary for
cell.

- Indole is a product of tryptophan metabolism by enzymes

mainly of Porphyromonasgingivalis. Indole enters gingival
fluid and further into epithelial cells. Indole is foreign
substance (xenobiotic) for cells and its accumulation
damages cell metabolism.
13

- Hydrogen sulfide (H2S)is formed by microorganisms in the

course of sulfur-containing amino acids catabolism. H2S is
an inhibitor of MRC cytochrome oxidase and reduce
energetic metabolism in epithelial cells.

pH of gingival fluid depends on microbial microflora and

content of their vital (waste) products ammonia, lactate
and low-molecular fatty acids. Presence of these
compounds define pH (6,3 7,93) of gingival fluid more
high than mixed saliva.

Acceleration of pathogenic microorganisms reproduction and

increase of their products of metabolism in gingival fluid can
occur in stress, uncontrolled antibiotics usage, oral hygiene
breach, improper feeding.

Large microorganisms clumps of normal microflora are able to

induce alterations in paradentium tissues. Under these conditions
activity of local protective mechanisms is not sufficient to reduce
their pathogenic activity. In
paradentiuminflammationgingival groove is increased
formingparadentiumpockets, volume and composition of
gingival fluid is changed.

A big role in inflammatory process development plays

mediators of inflammation such as biological amines
(histamine and serotonin), prostaglandins and others. In
paradentopathycontent mainly of PGE1 and PGE2 are increased in
gingival fluid andthis contributes vasodilation and their
permeability increasing.

For analysis of paradentium tissue state, determination of

paradentitisseverity degree and differential diagnosis of gingivitis
and paradentitis activity technics of gingival fluid examination are
used. Gingival fluid is obtained by insertion of filter paper
stripfirmly to gingival groove on 3 5 min. Fluid is collected with
micropipette, rarelysterile cottonturunda is used.
14

Obtained data analysis allows estimating:

- fluid release intensity;

- free amino acid content in gingival fluid;

- initial condition of paradentium tissues;

- epithelial and connective tissue cells population;

- activity of a number of enzymes and microorganisms vital

activity products presences.

Dental deposit formation and caries development.

Dental deposit is one of the main pathogenic reasons of
caries development.

Primaryaries lesionis arisen at places where favorable

conditions for dental deposit accumulation are created:
fossae, fissures, on aproximal surfaces and cervical
regions.

Oral cavity conditions such as moist medium, constant

temperature and food products contribute rapid
microorganism reproduction forming resistant (constant)
microflora.

Dental deposit appearanceis one of theresults of oral

cavity microorganism vital activity.
15

Dental deposit is soft sticky (adhesive) lump (generation)

on tooth cervix region or on all tooth surface. Dental
deposit is mainly formed on the surface with roughness
and in dento-gingival region.

Dental deposit is easily removed by toothbrush and

rubbed off (eliminated) during mastication of firm
food.But in two hours after tooth brushing its completely
restored.

The main components of dental deposit are (1)saliva

glycoproteins, (2)microorganisms, (3(polysaccharides of
bacterial genesis, (4)cast-off epithelium of mucous tunic.

Pellicle

Between dental deposit and enamel surface there is thin

microbe-free layer - acquired pellicle. Its formed as a
result of glycoproteins spontaneous adsorption and
16

aggregation, first of all mucins. Negatively charged groups of

these molecules interact with calcium ions, and basic groups
interact with hydroxyapatite phosphate groups.

Pellicle fulfills protective function because reduces the

possibility of undesirable isomorphic replacements in
enamel hydroxyapatites in 4-6 times. This biological
membrane can regulate different ions diffusion from
saliva to enamel or from enamel to saliva.

Pellicle formation is considerably accelerated at reduced

pH in oral cavity and presence of calcium and phosphate
ions.

Ions interact with glycoproteins of saliva and by reducing

their solubility reinforce these molecules adhesion on
tooth surface.Pellicle has the thickness of 1 to 10
micrometer, tooth deposit 5 200 micrometer.

Microorganism participation in pellicle genesis is not

necessarily but their presence accelerates the process.

Bacteria secrete lactate and other organic acids which

induce reduction of oral cavity pH.H+ increased
concentration suppresses sialic acid residues dissociation
in mucin composition.Charge reduction of macromolecules
leads to their precipitation on enamel surface.Bacterial
enzyme neuraminidase detaches N-acetylneuraminic (sialic) acid
and fucose that leads to chemical modification in mucin. That is
why mucins in pellicle composition contain 10 times less
carbohydrates than mucins soluble in saliva.

After completion of pellicle formation practically

simultaneouslycast-off epithelium cells, microorganisms
and saliva glycoproteins are adsorptedon pellicle surface.

Streptococcus mutans plays the main role of dental

deposit formation on smooth tooth surface.They synthesize
extracellular sticky (adhesive) and insoluble polysaccharide of
glucose dextran, which is an important component of
dental deposit.
17

Dextran synthesis catalyzes dextran-sucrase (dextranase -

specific glycosyltransferase). Enzyme hydrolyzes sucrose
to glucose and fructose and then attaches glucose
molecules by -1,6 and at points of branching by -1,3-
glycosiduc bond to growing dextran molecule. Dextranase is
specific to only sucrose.

From fructose another polysaccharide is synthesized

levan. It plays the role of extracellular energetic reserve
of microorganisms.

Glycoproteins, cast-off epithelium cells and dextran

form the base of dental deposit which is occupied by oral
microorganisms , first of all S.mutans.

Initially representatives of constant flora participatein

microorganisms adhesion. First microbial cells are settled
down and reproduced in crypts on the tooth surface and
then pass on smooth surface.

Some microorganisms can stimulate dental deposit generation

and its mass increasing another doesnt influence on this process,
but thirddecelerate dental formation. Microorganisms of the
last group secret glycosidases and hydrolyze extracellular
polysaccharides which necessary for other bacteria
adhesion.

Manymicrobial cells are unable to attachdirectly to pellicle

but may settle on the surface of other bacteria forming
the link cell-to-cell. For example anaerobic streptococci
possess high adhesive properties towards epithelium and
tooth enamel and also ability to aggregation with other bacteria
of oral cavity: bacteroids, fusobacteria.

All adhesion process flow very rapidly and during 5 minutes

bacterial cells amount on 1 cm2of deposit is increased from 103 to
105 106. Low CO32- , H2PO4- , HPO42- concentrations stimulate
adsorption of microorganisms on tooth enamel surface.

If at early stages of dental deposit development the main

cells are anaerobic coccus microflora and then it give way
to rod-shaped (bacillary) and thread-like forms. In the
18

following thick net layer threaded(filamentary) microorganisms

are formed with inclusion of other microorganism colonies.

Presented epithelial cells in dental deposit are degraded after

adsorption on pellicle surface.

During approximately 8 hours intensity of adhesion is

stable but after 1-2 days dental deposit turn into stage of
mature plaque. Amount of attached bacteria is again increased
reaching 107 108/cm2. Formation of the last surface layer is
completed. This layer is composed of microorganisms which able
to assimilate and oxidize an acetic, pyruvic, and lactic acid that is
neutralize acidic products of other bacteria metabolism.

Dental deposit of such structure is the most dangerous for

tooth enamel.Synthesized by bacteria of 1-st and the 2-nd
layers hydrolytic enzymes proteinases, glycosidases and
also organic acids lactic, propionic, acetic may induce
hydrolysis of pellicle and chemical modifications in enamel
hydroxyapatite structures.

The followed components enter into the dental deposit

composition:

saliva proteins, bacterial and cast-off epithelial cells;

carbohydrates glucose, hexosamines, sialic acid, acidic

glycosaminoglycans, polysaccharides - dextran, products
of soluble polysaccharide levan hydrolysis;

membrane lipids of epithelial and bacterial cells;

proteases, glycosidases, lipases and other enzymes mainly

bacterial genesis.

Dental deposit chemical compositiondepends on age, food

character, and hygienic state of oral cavity.

Its composed of 78 80 % of water.

Its distinguished the cell and cell-free fractions in dental deposit.

19

Cell fraction is represented by epithelial cells,

streptococci ( ~70%), veilonellae and neisseriae ( ~15%)
and also other bacteria and yeast fungi ( ~15%).

Cell-free fraction includes mineral compounds, proteins,

and carbohydrates. Amount of macro- and micro-elements in
dental deposit varies: 1 mg of dental deposit dry mass contains
about 3,4g of calcium, 8,4gof phosphorus, 4,2g of
potassium and 1,3g of sodium. Calcium and phosphorus
mainly enter dental deposit from saliva but it is possible
entrance from enamel, and as far as dental deposit
maturation these ions amount grows.

Dental deposit includes ions of cobalt, strontium, iron,

magnesium, manganese, fluoride and other.

Fluoride concentration may be in ten and even hundred

times more than in saliva (from 6 to 180 g/ g). Itdepends
on fluoride amount in drinking water. Fluoride inclusion to dental
deposit occurs at the expense of (1) salt formation with calcium
(CaF2), (2) complexes with deposit proteins, (3) inclusion to
apatites and also (4) by bacteriaabsorption.

Dental deposit and teeth caries.

Caries is pathological process in which demineralization
and destruction of hard tooth tissues occur with the
following dental cavity formation.

Causes of caries development may be divided to three

groups:

poor diet with sucrose predominance, drinking

water with low fluoride content, disorders and
functional changes in organism condition, extreme
influence;

damage of properties and composition of oral fluid,

carbohydrate food remnants and excessive
accumulationof dental deposit;
20

disturbance of tooth tissue resistance, defective

structure, change of chemical content, genetic
defects.

In the norm enamel is in state of dynamic balance

between slow but constant processes of de- and
remineralization.

In the process of enamel maturation it crystal structure is

thickened (consolidated) and amount of micro-spaces
isdecreased - that is full value mineralization make tooth
tissues caries resistant.

Fluoride plays the most important role in increase of

caries resistance. Relative hydroxyapatites replacement
to fluoroapatites [Ca10(PO 4)6F2] contributes increase
resistance of surface enamel layer to acids effect (caries
resistance).

But formation of carboapatites [Ca10(PO4)5(CO3)(OH)2]make

enamel fragile (brittle) and reinforce caries susceptibility.

As far as dental deposit enlargement saliva protective properties

for enamel is weakened.But effect of metabolic products and
enzymes of bacteria settling in dental deposit is increased.

Porous structure of dental deposit (tooth plaque)

permitsnutrients penetration to deep layers and provide
active vital activity of microorganisms with formation of
lactate and other organic acids. Protons formed in
acidsdissociationparticipate in calcium replacement in enamel
hydroxyapatite crystals or even induce their damage.

Ca10(PO4)5(CO3)(OH)2 + H + Ca92H(PO4)5(CO3)(OH)2 + Ca2+

Acids dissolve apatites, and especially intensively in inter-

prismspaces because penetrate under surface enamel
layer and induce it demineralization. Surface layer
structure is changed less because includes more
fluoroapatites.Active flow of this process leads to gradual
increasing of micro-spaces between crystals of enamel
21

prisms.Microorganisms dash to these spaces creating the

source of acid formation in the interior of enamel.

Prolongedexistence of demineralization center leads to

dissolution also surface (more resistant) layer of enamel.
Carious process can be stopped but only at the stage of white
spot on enamel up to formation enamel carious defect. Clinically
this is revealed by formation of pigmentedbrown or black spot.

Microorganisms which influence the processes running

in dental deposit and enamel can be divided to two
groups:

anaerobic microorganisms which products of vital

activity are lactic and other organic acids reducing
pH of saliva and internal layers of dental deposit;

aerobic microorganisms of surface layer assimilating

lactate, pyruvate, acetic acid, amino acids and
contributes pH increasing.

Relation of aerobic microorganisms colonizing oral cavity to

anaerobic is from 1:10 to 1:100. In case of violation of this
proportion composition and quantity of constant microflora is
changed. This is accompanied by reproduction of transient
microorganisms in oral cavity which induce caries and
different forms of paradentopathy.

Streptococci of oral cavity are sort first of all to

cariesogenic microorganisms. They are representatives of
healthy people oral cavity microflora, but under a particular
conditions can accelerate caries development because actively
participate in carbohydrate metabolism.

At that pH of dental deposit can be reduced to critical 5,

0 and even low. Splitting of extracellular polysaccharides
by bacterial enzymes in caries cavity and entranceof their
catabolic products to anaerobic glycolysis leads to further
lactate accumulation.

Formation of acids in dental deposit and change of pH is

especially actively occurs if carbohydrates enter oral
22

cavity and first of all sucrose. Presence of 10g of sucrose in

food induces increase of lactate content in saliva to 10 16 times.
And in pH reducing to 6,2 saliva from supersaturated of calcium
and phosphate becomes undersaturated and consequently
demineralizing.

Pathways of sucrose usage by oral cavity bacteria

depends on it amount in food. In low concentration
sucrose is rapidly hydrolyzes to glucose and fructose
which enter the glycolysis. In high content in food sucroseis
used not only as energetic substrate but converted to
extracellular polysaccharides levan and dextran by bacteria.

Appearance of different acids in dental deposit is inherited with a

particular species of microorganisms. Streptococci synthesize
mainly lactic acid;lactobacteriatogether with lactic can form
propionic, acetic and butyric acids; neisseria and veillonella
convert lactic acid to propionic and acetic, and carbon dioxide and
water to pyruvic and acetic.

Dental deposit pH depends not only on organic acids

formation but access of saliva to it. Saliva buffer systems first
of all bicarbonate are able to neutralize acids. That is why dental
deposit more connecting with saliva has higher pH.

Saliva influencescariesogenic activity of microorganisms.

Secreted proteins help to attachment of microbial cells to tooth
surface, and at the same time oral fluid wash oral cavity and
contribute their removing. To anti-caries factors it may be related
the group of microorganisms which use proteins and amino
acidsas nutrients. Ammonia forming in the course of their
metabolism binds protons and reducesH+participationin
isomorphic replacements.

Cariesogenic enzymes of dental deposit.

More than 50 different enzymes are present in dental deposit.
Most of them impact destructive effect in tooth
tissues.Microorganisms produce hydrolases: proteinases,
sulfatases, phosphatases, glycosidases which are
aggressive enzymes.
23

Sulfatases detaching SO3H groups induce damage of

protein-glycosaminoglycan complexes and that is why
breaks down organic basis of enamel and dentin.

Protein phosphatases dephosphorylating proteins disturb matrix

interaction with hydroxyapatite crystals.

The most cariesogenic enzymes are collagenase and

hyaluronidase which hydrolyze matrix of dentin and
supply bacteria with source of nutrition
therebydeepencarious cavity.

Caries development contributes the following changes:

saliva secretion in xerostomia;

protein composition (content of specific and non-

specific defense factors in saliva);

buffer capacity of saliva (possibility to maintain pH

at optimal level);

content of calcium, phosphates and other ions in

saliva which define remineralizing potential.

Tooth stone and paradental tissue inflammation.

Inorganic compounds precipitation in dental deposit (that

is it mineralization) leads to formation of dental calculus.
Partially mineralized dental calculus contains bacteria;completely
formed dental calculus is microbe-free formation.

Mineralization of dental deposit organic matrix formation

of dental calculus.

Saliva is the source of calcium, phosphate and other ions

participating in mineralization.

Intensive reproduction of pathogenic microorganisms in dental

deposit leads to increase of their metabolic products in saliva
organic acids: lactate, acetate, butyrate and others.

Dissociating acids increase proton concentration in saliva

CH3 COOH CH3 COOH + H+

24

They damage calcium phosphate micelle structure

because protonate phosphate groups of diffuse layer.

HPO42- + H + H2 PO4-

Under these conditions calcium ions are washed off surface

layer of micelle and takes part in dental deposit
mineralization.Facultative anaerobes (settling dental deposit)
secret waist metabolic products nitrogen, ammonia and urea.

NH3 interacts with phosphate groups:

NH3 + H2 PO - NH4+ + HPO42-

Anions HPO42- bind Ca2+ and form poorly soluble salt

CaHPO42H2O, which to give rise to dental calculus
formation.

At first stages formed mineral CaHPO42H2O is composed 50%

of all apatite types. Dental calculus of this composition is
easily removed but by aging it composition changed and
this lead to formation of compound of more complex
composition: octocalcitriphosphate and hydroxyapatites.

Ca10(PO4)6 2(H2O)

CaHPO42H2O Ca8H2(PO4)65H2O
Ca10(PO4)6(OH)2

Octocalcitriphosphate
hydroxyapatites

Carbonate apatite, magnesium salts are present in small amount

in dental calculus: (MgHPO46H2O) and (CaMg)3(PO4)2, fluoride in
the form of fluoroapatite, calcium fluoride (CaF 2) and as complex
with organic compounds.

Conditions for dental deposit mineralization and dental

calculus formation are:

participation of acid-forming microorganisms (H +);

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increase of Ca2+ and PO43- ions in saliva induced by

reduced stability of saliva micelle;

reproduction of microorganisms producing NH3 and

urea;

increase of amount of dead bacteria metabolites

(Glu, NH3, citrate, -ketoglutarate and others )
whichare able to keep Ca2+ and PO4 3- ;

participation of alkaline phosphates, which

catalyzesphospho-organic compounds hydrolysis
and increase PO43-content in deposit.

Mature dental calculus is composed from inorganic (70%-

90%) and organic components.The main inorganic
compounds are calcium (39%), phosphorus (19%),
magnesium (0,8%) and carbonates (1,9%). Also there are
large group of trace elements in dental calculus: sodium, zinc,
strontium, bromine, cooper, manganese, tungsten, aurum,
aluminum, iron, fluoride. Presented in dental calculus Ca-binding
glyco- and prosphoproteins are composed 0, 1 -2, 5%.

In dependence on localization its distinguished above-

gingival and sub-gingival dental calculus, they have different
chemical composition, mechanism of formation and sources of
calcium and phosphates.

Above-gingival dental calculus resides above crest of gingival

edge, has yellowy- grayish color, which depends on iron oxides,
cooper, nicotine and other compounds entering to organism. It
26

has solid or clay-like consistence and is easily separated from

tooth surface by scraping or spalling. Above-gingival dental
calculus is found more often on cheek surface of molar teeth in
front of parotid salivary gland ducts and on lingual surface of
frontal [anterior] teeth of mandibular. Dental calculus formed at
dento-gingivaledgeleads to difficulty circulation of dental fluid and
that is why to damage of it protective functions.

Sub-gingival calculus is formed under gingiva and cant be

seen in oral cavity examination. Calculus usually thick and solid,
dark-brown or greenny-black color and tightly attached to tooth
surface.

Above-gingival dental calculus usually is related to salivary

type, but sub-gingival one - to serum type because the source
of calcium and phosphate infirst case is saliva, in the second
case is gingival fluid (and it is a transudate of blood serum. First
has pathological importance in teeth caries development, the
second facilitates pathological processes in patadentium.

Paradentium pathology.

Soft dental deposit producing toxins (NH3 , H2S, lactate,

indole and cetera) induces gingiva inflammation. Gradually
space (gingival pocket) is formed between gingiva and tooth.
Accumulated deposits on the tooth root are mineralized and
converted to subgingival calculus. Dental calculus destroy tooth-
gingival link and contributes spread of the infection into the
depths of paradentium tissue.
27

Paradentitis is inflammation of paradentium tissues

accompanied by gingiva, periodontum, alveolar bone and
tooth destruction. Paradentitis is dystrophic damage of all
paradentium elements.

Factors stimulating inflammation development of paradentium

tissue are oral cavity microorganisms, dietary habits, dental
deposit, low (insufficient) load on tooth maxillomandibular
system, general condition of organism.

Tooth-gingival pockets, dental deposit and bacterial deposit on

mucous coat of gingiva are the main locality of oral cavity
microorganisms.
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Saliva contains not more than 5 mln/ml of microorganisms. But

there are several billions of microbes in 1 ml of tooth-
gingival pocket content. And much more in dental deposit.
Paradentitis development is in direct dependency on
amount of dental deposit and common microbial state of
oral cavity.

Microorganisms of dental deposit, tooth-gingival pocket and

gingival liquid produce different pathogenic compounds. They
are concentrated in liquid of tooth- gingival pockets,
penetrate to gingiva and damage periodontium.

Besides NH3, H2S, lactate, indol, low-molecular fatty acids

microorganisms of tooth-gingival pockets produce amines,
(hydroxylamine NH2OH), which is strong oxidant:

NH2OH +2e NH3 + O-

Source of electrons may be heme or Fe 2+-containing proteins:

cytochromes of ERC, antioxidant defense and other cellular
enzymes. Hydroxylamine breaks mitochondrial oxidative
phosphorylation and induces cell membrane damage by
active radicals, and activates lipid peroxidation.

Enzymes produced by microorganisms of tooth-gingival

pocket and gingival fluid has Inflammatory and toxic
effect on epithelial cells of periodontium. They easily
penetrate to gingival tissue with helping of bacterial
hyaluronidase which hydrolyzes the main GAG of
extracellular matrix. Pathological action is enforced with
joint activity of collagenase.

Bacterial neuraminidase splits the neuraminic acid residues and

this leads to changes the structure of oligosaccharide membrane
of epithelium and periodontium cells. This stimulates production
of antibodies to damaged cells; proteins of complement system
enhance cytotoxic effect of antibodies. Complement system
activation is accompanied by formation of peptides stimulating
leucocytes migration to the region of damaged cells and their
accellation.
29

Bacterial phospholipase C and DNA-hydrolase reveal pathogenic

properties. Phospholipase C catalyzes the hydrolysis of
leucocyte membranes and induces their damage, and
retards phagocytic response. Bacterial proteases have the
most aggressive impact on paradentium tissue. Collagenase
hydrolyzes gingival and alveolar bone collagen and
induces damage of protein matrix (stroma) of these
tissues.Bacteria of dental deposit produce elastase which
is able to damage elastin of vessel walls and inducing
increased hemorrhage. Microorganisms S. sanguisproduce
specific protease which can hydrolyze sIgA of saliva.
30