This document discusses various biological and chemical indicators that can be used to assess oral health and disease risk. It describes several tests and analyses that can be performed on saliva, dental plaque, subgingival plaque, and gingival crevicular fluid to evaluate factors like salivary flow rate, pH, microbial content, temperature, and inflammatory markers. Methods for collecting and analyzing samples are provided for many of the tests. The results of these analyses can provide information on periodontal disease risk and systemic conditions like cardiovascular disease and diabetes that are linked to oral infections.

1. Biological and Chemical
g
indicators of disease risk
By
y
Dr Wael Mohamed Swelam
17 March 2009

2. Background:
g
• Oral tissue functions are controlled by
different molecules/factors
diff
t
l
l /f t
– Salivary flow rate
– Salivary components modifying its pH
(acidity/alkalinity)
( idit / lk li it )
– Microorganisms
– Grenetic
• Al oral tissue functions affect its
Also,
l i
f
i
ff i
environment
– Temperature
– Enzymes and immunokines
So, they are considered indicators
for health & disease
17 March 2009
Dr. Wael M Swelam
2

3. 17 March 2009
Dr. Wael M Swelam
3

4. Microbiological evaluation
g
1. Dental plaque
a.
a Tools
• Plastic toothpick/dental instrument
• Pad of plastic test strip
• Culture medium at 37˚C/48h
• Microscope with 10X magnification
b. Advantages
 High specificity , based on microorganisms
identified = tooth risk for caries
c. Limitations
Information can’t be generalized
I f
ti
’t b
li d
Doesn’t reflect the biological ecology of oral
cavity
Inconvenient to identify colonization of organisms
y
g
from multiple sites
17 March 2009
Dr. Wael M Swelam
4

5. Microbiological evaluation
g
2. Saliva microbiological culture
a.Principle:
• Saliva is the vehicle for microorganisms
• "un-stimulated" or "stimulated" saliva
unb.Precautions
Careful
C f l recording f all medications; contraindicated
di for ll
di ti
t i di t d
during
Systemic antibiotic
Transient antihistaminic
T
i t tihi t i i
Avoid foods/drinks or disclosing tablets 4h before test
Avoid dental prophylaxis before sample collection
17 March 2009
Dr. Wael M Swelam
5

6. Test for Unstimulated Saliva
Materials needed:
• Measure-cup (graduated test tube with conical base)
• Funnel
• A watch or timer
h
How to measure.
1) The person sits in an upright position with his head inclined
forward so that the production of saliva is collected in the floor
of the mouth and then flows out over the lip.
2) Saliva formed is let to drip into the measure-cup for 15 minutes.
3) The result of this collection is expressed as milliliters per minute.
17 March 2009
Dr. Wael M Swelam
6

7. Test for Stimulated Saliva
Materials needed:
p
paraffin for chewing to stimulate saliva secretion
g
• A piece of p
• Measure-cup (graduated test tube with conical base)
• Funnel
• A watch or timer
How to measure.
1) The person chews a piece of paraffin until it becomes soft.
Before the collection is started the first portion of saliva is
swallowed.
2) Start timer and the chewing is continued for another 5 minutes
(for subjects with high secretion rate, 3 minutes may be enough).
3) The saliva is spat out at short intervals in a measure-cup during
the collection period.
4) The colleted saliva is then measured. The measurement should
not include the foam which is formed during the collection. The
result is expressed as mililiters per minute.
17 March 2009
Dr. Wael M Swelam
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8. Salivary flow rate analysis
y
y
Unstimulated Saliva
(ml/minute)
more than 0 25
th 0.25
0.1 - 0.25
less than 0.1
normal
l
low
very low
Stimulated Saliva (ml/minute)
More than 1.0
Normal
0.7 1.0
07-10
Less than 0.7
17 March 2009
Low
Very low
Dr. Wael M Swelam
8

9. 2 Laboratory analysis of cultured saliva
2.
Serially dilute the sample
Culture samples on SB20 agar for streptococci
;
in anaerobic environment; WHY?
Culture samples on Rogosa agar for
lactobacilli species
Added bacitracin, antibiotic to the cultured
samples, WHY?
Keep samples in culture at 3 ˚C/ 8
37˚C/48h
Count the colonies
17 March 2009
Dr. Wael M Swelam
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10. 3. Side chair analysis of Salivary S Mutans
a.Commercial set “Dentocult SM Strip mutans” is based on
broth,
Usage of selective culture broth and
The adherence and growth of mutans streptococci bacteria on
the test strip
b.
b Materials needed :
 Mitis salivarius broth for culture,
 Add 5µg bacitracin tablet; WHY?
 Start saliva stimulation,
 After 2 min. introduce the plastic strip provided in the kit
in-between the lips
p
 Rotate it to collect sample, remove & attach to the cap
 Close Cap onto the vial and keep for 48h/37˚C
17 March 2009
Dr. Wael M Swelam
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11. Caries risk category
High
Moderate
Low
17 March 2009
Mutans streptococci
(CFU/ml)
≥5.5
≥5 5 ×105
≥1 ×105 ~ <5.5×105
<1 ×105
1 10
Dr. Wael M Swelam
11

12. Analysis of Salivary pH as indicator for its buffering capacity
a.
a Principle:
• Salivary buffering capacity is pH dependent
• Salivary pH is directly related to
• Acidogenicity of microenvironment
• Sucrose challenge in the diet
Dentobuff® Strip System
Materials needed: Dentobuff® Strip, a kit which includes
p
• Paraffin tablet for chewing to stimulate saliva secretion
• A test strip containing acid and ph-indicator
• A standard color chart
• A disposable p p
p
pipette
• A cup or tube
• A timer
How to measure.
1) Saliva is collected in the same way as described in Tests for
Stimulated Saliva. Usually buffer capacity is taken
simultaneously with secretion rate
2) The pipette is used and one droplet of this stimulated saliva is
placed on a small test pad of a test strip
strip.
3) Wait exactly 5 minutes, as color will change with time, for the
reaction of saliva and indicator.

13. Salivary p analysis result
y pH
y
Compare the color of the test pad with the
standard color chart. This color indicator
reflects the pH on the strip.
Final ph value
6.0 or more
4.5 - 5.5
4.0
4 0 or less
17 March 2009
Buffer capacity
High
Medium
Low
Dr. Wael M Swelam
13

14. Summery of saliva testing
17 March 2009
Dr. Wael M Swelam
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15. 17 March 2009
Dr. Wael M Swelam
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16. A. Analysis of subgingival plaque
Precautions
Careful recording for all medications; contraindicated during Antimicrobial including
systemic antibiotic
Removal of heavy deposits of supragingival plaque Transient antihistaminic
1. Direct examination using microscope; have limited value only to assess motile
spirochetes
2. Culture organism and perform sensitivity test; sample preservation is critical for
experimental success
3. PCR / DNA probes
4.
4 Perioscan “office based test indicate the presence of pathogens (T denticola B
office-based test”
(T. denticola, B.
forsythus, & porphyromonas gingivalis) capable to hydrolyze BANA enzyme
17 March 2009
Dr. Wael M Swelam
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17. Analysis of subgingival Temperature
a.Principle:
• Inflammation is usually related to increase in organ
temperature; WHY?
b Tools: using either infrared thermometer OR periotemp
b.Tools:
c. Protocol:
Apply the tip similar to periodontal probe calibration
First ll t th
Fi t collect the normal healthy core temp, then measure the
l h lth
t
th
th
subgignival temp
The instrument uses light indicator system (Normal=green,
Slightly elevated=yellow, and Elevated=red)
17 March 2009
Dr. Wael M Swelam
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18. Objective: PST “Commercial” test to check two
j
interleukin-1 genes closely correlated to
advanced periodontal diseases
Tools & Procedure:
– Collect finger stick blood sample
– DNAase-free blotting paper
– PCR technique to evaluate gene polymorphism
17 March 2009
Dr. Wael M Swelam
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19. Background:
1)
2)
3)
Inflammation of the gingiva, usually referred to as gingivitis, arises in
the region of the crevicular epithelium well before any inflammation is
clinically visible
y
Vasculature of gingival tissues provide a vehicle for entire
immune process to occur in response to a bacterial challenge in
subgingival environment
GCF is a serous transudate
a)
b)
In healthy condition it provide homeostasis maintenance
mechanism
It change into inflammatory exudate during inflammation to deliver
immune cells and inflammatory exudate to local environment
Objective: Immunologic analysis (Antigens, Antibodies, and Mediators) will be a
measure of changes in host response to periodontitis
17 March 2009
Dr. Wael M Swelam
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20. Tools:
1) M k
Markers against; i t l ki 1β i t l ki 6
i t interleukin-1β, interleukin-6,
interleukin-8, tumor necrosis factor-α
2) Enzymatic activity; neutral proteases, lactate
y
g
,β g
,
dehydrogenases, β-glucouronidase, and alkaline
phosphatase
Precautions:
a) Moisture control
b) Careful removal of existing debris
Procedure:
1. Insert filter paper strip into gingival sulcus
2. Leave it in place till it appear saturated
3. Remove it and measure collected volume in periotron
4. ELISA to analyze markers and antibodies
l
k
d
b d
17 March 2009
Dr. Wael M Swelam
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21. 17 March 2009
Dr. Wael M Swelam
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22. Background:
1)
2)
3)
91%
Diagnosis of Oral cancer is dependent on clinical exam, brush biopsy,
toluidine blue, and biopsy
Cancer cells change the body metabolism which can be traced
metabolism,
by its products in body fluids including saliva
Saliva screening for oral cancer was +ve for
a)
b)
c)
d)
e)
)
HIV
IL-1β,
IL-8,
Spermidine acetyltransferase,
Ornithin decarboxylase
O i hi d
b
l
Tools: Oral Fluid Nano-Sensor Test (OFNASET)
17 March 2009
Dr. Wael M Swelam
22

23. OFNASET
Automated,
Integrated eicroelectromechanical system that
g
y
will enable simultaneous and rapid detection of
Multiple salivary protein, and
Nucleic acid targets
17 March 2009
Dr. Wael M Swelam
23

24. • Microfluidic chip “using a detection system based on
up-converting p
p
g phosphor technology” has been
p
gy
developed to detect HIV,
• TB assay to detect antibodies in saliva,
• M l i test will identify nucleic acid as well as antigens
Malaria
ill id if
l i
id
ll
i
17 March 2009
Dr. Wael M Swelam
24

25. Oral pathogens & their toxins can be transmitted through
p
g
g
blood to other organs
1) Cardiovascular diseases (CVD):
a) B th CVD & periodontal di
) Both
i d t l disease are i fl
inflammatory di d
t
disorder
b) The relationship is indirect rather than direct
Test
Total serum cholesterol
Low-density lipoprotein
cholesterol (LDL)
High-density lipoprotein
cholesterol (LDL)
C-reactive protein
17 March 2009
Normal
Increase risk of CVD
<200mg/dl
200-239mg/dl, borderline high
≥240mg/dl, high
<100mg/dl
<100 /dl
100-129 mg/dl, above optimal
130-159mg/dl, borderline high
g/ ,
g
160-189mg/dl, high
≥190mg/dl, very high
≥60mg/dl, high
<40mg/dl, low
1.5-3.0mg/dl, moderate risk
<1.0mg/dl
>3mg/dl, high risk
Dr. Wael M Swelam
25

26. 1) Diabetes mellitus:
a) Oral infections trigger inflammatory mediators = increase insulin
resistance
b) Conversely, diabetes increases risk for p
)
y
periodontal disease as
indicated by (Glycated hemoglobin) level
Test
T t
Impaired
Glucose
Gl
tolerance
Normal
N
l
Diabetes
Di b t
Casual blood glucose conc.
< 100 mg/dl
Fasting l
F ti plasma glucose (FPG)
l
< 100 mg/dl
/dl
100-125
100 125 mg/dl
/dl
≥ 126 mg/dl
/dl
Oral glucose tolerance test
(OGTT)
< 140 mg/dl,
high
> 140- <200 mg/dl
≥ 200 mg/dl
Clycated hemoglobin (A1C)
17 March 2009
≥ 200 mg/dl
<6.0
<6 0 mg/dl
/dl
Dr. Wael M Swelam
26