Here saliva as a diagnostic biomarker has been explained via this powerpoint . extraction of saliva and diagnostic equipments and techniques are explained here by Dr Harshavardhan Patwal

1. Dr Harshavardhan Patwal

2. • Saliva is “the aqua-vita” of the oral cavity.
• the principal protector of the soft and hard oral tissues.
Diminished secretion:
• oral tissues become susceptible to infection
• ability to masticate, swallow, speak and taste may be
disturbed.

3. Saliva is a clear, slightly acidic mucoserous exocrine secretion.
• Whole saliva:
“It is a complex mixture of fluids from major and minor salivary
glands and from gingival crevicular fluid, which contains oral
bacteria and food debris”. Edgar,1992.

4. Development of salivary glands
The three major sets of glands
• Parotid, submandibular,sublingual
• they originate in a uniform manner by oral ectodermal
epithelial buds invading the underlying mesenchyme

5. Classification
It is based on
• Size and location
• The histochemical nature of secretory products
On basis of size
Major salivary glands
Parotid gland
Submandibular gland
Sublingual gland
Minor salivary glands
Labial and buccal
glands
Glossopalatine glands
or lingual glands

7. Composition of saliva
(Mandel & Wotman 1976, Core 1992)

10. 99% water
1% large organic and inorganic molecules.
Large organic
molecules
• protein,
• glycoproteins
• lipids
Small organic
molecules
• glucose
• urea
Electrolytes
• sodium,
• calcium,
• chloride
• phosphates

11. • Organic constituents:
– Protein
• Comprise approx. 200mg/100ml only 3% of the protein
conc. in plasma.
– Enzymes
– Immunoglobulins
– Antibacterialproteins
– Mucousglycoproteins
– Tracesof albumin,polypeptides andoligopeptides.

12. • Inorganic constituents:
– sodium, potassium, chloride and bicarbonate are the main
contributors to the osmolarity of saliva.
– Bicarbonate is also the principal buffer in saliva.
– Thiocyanate activates antibacterial effect of
sialoperoxidase.
– Fluoride content similar to plasma

13. The known stimulus may be
• Psychological (eg. Thinking of tamarind)
• Visual (eg. Seeing delicious food)
• Taste (tasting good food)
• Others (during vomiting)
Stimulated secretion of saliva is due to reflex (salivary reflex)
• Conditional
• Unconditional
Regulation of secretion
Spontaneous Stimulated

14. Factors influencing the composition of saliva
Flow Rate
• The accepted range of normal flow for unstimulated
saliva is anything above 0.1ml/min.
• For stimulated saliva flow rate is approx 0.2ml / min.
• As the flow rate increases, the concentration of proteins,
sodium, chloride and bicarbonate rises, while the levels of
phosphate and magnesium fall.

15. Flow rate in salivary glands according to individual
constituents
• total protein, amylase, sodium, bicarbonate
Substances whose concentration increases as the
flow rate increases:
• phosphate, urea, aminoacid, uric acid, serum, albumin
Substances whose concentration decreases with
the increase in flow rate:
• potassium, fluoride
Substances whose concentration does not change
with change in flow rate:
• Chlorine, calcium, protein bound carbohydrates
Substances whose concentration decreases at
first but increase as flow rate increases:

16. Differential
• In unstimulated whole saliva, the parotid gland contributes only about 10%
• stimulated whole saliva contains lower levels of calcium
Circadian rhythm
• The levels of calcium and phosphate ions are low in early morning.
Nature of stimulus
• salt stimulates higher protein content.
• sugar stimuli give rise to a high amylase content in saliva.
Diet
• Change in phosphate and urea conc. induced by dietary alterations
may be reflected in the saliva.
Duration of stimulus

17. Salivary function can be organized into
Mastication
and
deglutition
Taste
Facilitation
of speech
Buffering
action
Excretory
function
Starch
digestion
Maintenance
of the tooth
integrity
Antibacterial
factor
Pellicle and
plaque
formation.

19. Buffering action
• bicarbonate , phosphate and amphoteric proteins: the salivary ph is
usually maintained alkaline.
• If the salivary ph falls from alkaline to acidic certain constituents of
saliva get precipitated -tartar - removing calcium from the tooth -
caries.
Mastication and deglutition
• This helps to convert food into a soft bolus which is coated with a
layer of mucous which acts as a lubricant and facilitates swallowing
(deglutition)
Taste
• has to be in solution.
• Saliva provides the water for this purpose and helps in the
appreciation of taste.

20. Starch digestion
• This is the only digestive function of saliva and is due to
ptyalin, which is a weak amylolytic enzyme
Boiled starch
Soluble starch
Erythrodextrin and maltose
Achrodextrin and maltose
Isomaltose and maltose
(Maltase converts maltose into glucose)

21. Facilitation of speech
• Saliva lubricates the oral cavity of proper activation of speech
Excretory function
• Helps in excreting certain heavy metals like lead and iodine
Maintenance of the tooth integrity
Saliva is supersaturated with calcium and phosphate ions that
provides minerals for
• post eruptive maturation.
• to counteract tooth dissolution by saliva (solubility product
principle)
• forms a film of glycoprotein on the teeth (the pellicle) that may
act as a diffusion barrier

22. Antibacterial factors: that influence bacteria.
Specific
Immunoglobulins
aggregate
specific bacteria
Non-specific
Lactoferrin
Lysozymes
Sialoperoxidase
Histidine

23. Role in oral disease
The role of saliva in oral disease is most apparent when
salivary flow is markedly reduced
• Pellicle and plaque deposition
• Plaque mineralisation to form calculus
• Dental caries

24. Pellicle and plaque deposition
• It is partly cellular, fundamentally bacterial, and partly
acellular, from bacterial, salivary and dietary sources.
It initiates plaque deposition by pellicle formation (or
cuticle) which occurs in stages
• Bathing of tooth surfaces by salivary fluid, which contains
numerous protein constituents
• Selective adsorption of certain negatively and positively
charged glycoproteins (electrostatic attraction of charged
molecules is a factor)
• Loss of solubility of the adsorbed proteins by surface
denaturation and acid precipitation
• Alteration of the glycoproteins by enzymes from bacteria and
the oral secretions

25. pellicle formation ( made up of salivary proteins and other macromolecules)
The amino-terminal segment of Proline-rich proteins adheres to the tooth,
leaving the carboxy-terminal region free to bind to the bacteria.
saliva continues to provide agglutinating substances to the intercellular matrix
and bacterial intercellular adhesion results.
Secondary colonization. Salivary proteins and carbohydrates serve as a substrate
for metabolic activity of the bacteria.
Salivary urea and ammonia have a profound effect on bacterial activity and
final plaque ph

26. Plaque mineralization and calculus formation
• Salivary calcium, phosphorus, magnesium, sodium, and
potassium become part of the gel like interstices of the plaque
and influence mineralization.
• Esterase, pyrophosphatase and possibly acid phosphatase may
play a role in plaque mineralization
• Persons who are heavy calculus formers have higher levels of
salivary glycoproteins than non-calculus formers.

27. Dental caries
Saliva can affect caries in five general ways
• To mechanically cleanse and thus lessen plaque accumulation
• To reduce enamel solubility by plaque modification through
calcium, phosphate and fluoride
• To buffer and neutralize the acids either produced by
cariogenic organisms or introduced directly through diet
• By direct anti bacterial activity
• By aggregation or clumping bacteria and reducing adherence
to tooth surface

28. SALIVA AS A DIAGNOSTIC TOOL
Analysis of saliva may be useful for the diagnosis of
• hereditary disorders
• autoimmune diseases
• malignant and endocrinal disorders
• assessment of therapeutic levels of drugs
• monitoring of illicit drug use

29. Collection of saliva
Aspects to be considered to collect saliva
Whether resting or stimulated saliva will be detected, and if
stimulated, how will it be stimulated?
The amount of saliva needed to complete the analysis
Pre-treatment of saliva before assaying and storage until assay.
If the patient may be taking medication or have a disease causing
dry mouth
Whether quantitative or qualitative assays will be seen on the
specimens

30. The main methods of whole saliva collection
Draining
Spitting
Suction
absorbent
methods
In it preweighed sponge is placed in the pt’s mouth for a set
amount of time. After collection the sponge is weighed again
and the volume of saliva is determined.
It requires the pt. to allow saliva to flow from the mouth into
a pre weighed test tube or graduated cylinder for a time
period.
Pt. allows the saliva to accumulate in the mouth and
then expectorates into a pre weighed graduated
cylinder usually every 60 sec for 2 to 5 min
It uses an saliva ejector to draw saliva from the mouth
into a test tube for defined time period.

31. Commercially available kits are
Sialometer, Salivette, Omnisal,
Orasure are used for collection
of saliva.
Paper indicator strips to
measure the pH of saliva.
Strip test to measure the
buffer capacity of saliva
Dip slide test for oral bacteria. Lab on chip Oral fluid nanosensor test
Testing of saliva

33. Advantages
• Can be collected non invasively
• more accurate reflection of the active hormone in the body
specially steroid hormones
• can be collected with devices that will be stable at room
temperature for extended periods
• the health hazards associated with blood collection such as
cross contamination among patient do not apply to saliva
• secretory leucocyte protease inhibitor (SLPI) may be another
factor contributing to the safety of saliva as a diagnostic
specimen. SLPI expresses anti virus activity against free HIV-
1 and lymphocyte derived tumor cell lines.

34. Disadvantages
• Direct spitting into a tube or absorption in cotton balls
performs most saliva collections. samples not sterile and
subjected to bacterial degradation over time
• Interpretation of saliva assays is still difficult although
diurnal and monthly patterns generally parallel serum
values
• Polar hormones such as thyroxin and the peptide hormones
are subjected to variations by flow rate, so reliable levels
cannot be obtained in saliva
• Proficiency testing programmes are not yet available for
saliva, which makes validation of laboratory tests for
certified laboratories difficult.

35. Clinical problems in which saliva contributes to diagnosis.
• Digitalis toxicity.
• Stomatitis in cancer chemotherapy
• Immuno deficiency of secretory IgA
• Cigarette usages
• Dietary nitrates, nitrites and gastric cancer
• Ovulation time

36. Salivary assays in diagnosis
• Raised Na+,K+,Ca2+ and PO4
-levels
Sialadenitis
• Raised Na+,Ca2+,Mg2+ and Cl-levels
Radiation damage
• Raised Na+,Cl-and PO4
- in parotid gland saliva
• Raise total protein and 2-microglobulin levels in parotid
gland saliva
Sjogren’s syndrome
• Raised Na+,K+,Ca2+ and PO4
-levels.
• Raised total proteins ,amylase ,lysozyme in submandibular
gland saliva and glycoproteins in parotid gland saliva .
Cystic fibrosis
The findings of Mandel (1980) include the following

37. • Raised K+ levels
• Raised total protein and amylase in parotid gland saliva
Alcoholic cirrhosis
• Raised Ca2+ levels
• Raised total protein
Hyperparathyroidism
• Raised Ca2+ levels
• Raised total protein, IgA, IgG, IgM, and raised glucose levels
Diabetes mellitus
• Depressed HCO3
-levels
Chronic pancreatitis
• Possibly raised Na+ levels
Psychiatric illness (not other wise specified)
• Raised Na+ and K+levels
Digitalis intoxication
• Depressed amylase and lysozyme levels
Sarcoidosis

38. Drug monitoring
• Drug levels in saliva reflect the free non protein bound portion in
plasma
• therapeutic drug monitoring is most effectively used when the saliva
to plasma concentration ratio is constant
The determination of drugs in saliva depends on their
• concentration in the blood
• diffusion capacity
• liposolubility and molecular size.
Examples
anti convulsant drugs such as phenytoin, primidone, ethosuximide,
carbamazepine
theophylline monitoring for asthmatic children
salivary lithium in manic depressive patient
high correlation between ethanol concentrations in saliva and in serum.

39. Screening for antiviral and viral antigens
complete concordance between salivary and serum finding
for HIV positive people
the proportion of specific to total immunoglobulins is
similar in the saliva and serum of each individuals

40. Hormones monitoring
• the liposoluble hormones with lower molecular weights can be
detected reliably (Kaufman E, 2002).
• The steroid hormones assayed in saliva includes cortisol,
testosterone, 5α dihydro testosterone, 17 beta hydroxy
progesterone, progesterone, 17beta estradiol, sterol, estrone.
• estriol measurement during pregnancy for detecting fetal
growth retardation and the estriol progesterone ratios for
preterm labor.
• significant correlation between salivary and plasma insulin and
melatonin
• Higher salivary cortisol levels detected in severe periodontitis,
a high financial strain, and high emotion-focused
coping(Genco et al. 1998).

41. Application of salivary analysis of medicine
inorganic ions
• thiocyanate ion excellent indicator of smokers
• High levels of nitrate in the saliva associated with carcinoma
of the digestive tract

42. Saliva for periodontal diagnosis
• Probing depth
• Clinical attachment level
• Bleeding on probing (BOP)
• Plaque index (PI)
• Radiographic loss of alveolar bone (Polson & Goodson 1985)
• Monitoring of the microbial infection (Listgarten 1992)
• Analysis of the host response in GCF (Lamster 1997)
• Genetic analysis (Kornman 1997)
information primarily about disease severity, and are not useful
measures of disease activity.

43. Limitations of traditional methods
• Insufficient for determining site of active disease
• Insufficient for quantitative measurement of response to
therapy
• Insufficient for measuring susceptibility to future disease
progression.
• Time consuming
• Subject to measurement error

44. Why Saliva
• Contains biomarkers for unique physiological aspects of
periodontal/peri-implant disease.
• Quantitative changes in the biomarkers can identify patients
with enhanced disease susceptibility
• Identify sites with active disease.
• Identifying sites that will have active disease in future.
• Simple and non-invasive method of collection

45. Proposed markers for disease include
• proteins of host origin (i.e. enzymes, immunoglobulins)
• phenotypic markers (epithelial keratins)
• host cells, hormones (cortisol)
• bacteria and bacterial products
• volatile compounds and ions (Mandel 1991)

46. Markers affecting the dental biofilm
marker Relationship with periodontal disease periodontal disease
Specific Immunoglobins
IgA,IgM,IgG
Interfere in adherence and bacterial
metabolism, increased conc. in saliva of
periodontal patients
Chronic and Aggressive
Non specific Mucin Interfere with colonization of Aa Aggressive
Lysozyme Regulates biofilm accumulation Chronic
Lactoferrin Inhibits microbial growth, increased
correlation with Aa
Aggressive
Histatin Neutralizes LPs and enzymes known to
affect periodontium
Chronic and Aggressive
Peroxidase Interfere with biofilm accumulation,
increased correlation with periodontal
patients
Chronic
Systemic C reactive protein Increased conc found in serum and
saliva of periodontal patients
Chronic and Aggressive

47. Specific markers:
• Ig are important specific defense factors of saliva.
• the preponderant immunoglobulin found is IgA.
• Major and minor salivary glands contribute all of the secretory
IgA (sIgA) and lesser amounts of IgG and IgM.

48. sIgA
• forms specific immune defense mechanism in saliva
• parotid gland responsible for the majority of the IgA (Nair
1986).
• important in maintaining homeostasis in the oral cavity.
• control the oral micro biota by reducing the adherence of
bacterial cells to the oral mucosa and teeth (Morcotte and
Lovice.1998)
• two subclasses IgA1 and IgA2.
• sIgA levels, undetectable in newborns, increase progressively
and reach adult values in stimulated saliva by 2–4 years of age,
and in unstimulated saliva by 6–8 years of age (Burgio1980).

49. IgG
present in low concentration.
concentration increases during inflammation of the
periodontal tissues (Wilton,1989, Shapiro, 1979).

50. Immunoglobulin isotypes in saliva
• Basu (1976) increased IgG concentration in saliva and
decreased IgA concentration before periodontal therapy as
compared to post-treatment levels. Salivary levels of IgG and
IgA found to be higher in a group of NIDDM patients with
periodontitis.
• Guven (1982) positive correlation between the severity of
inflammation and IgA concentration.
• Sandholm (1984) Salivary IgA, IgG, and IgM levels were
higher in the JP patients
• Harding (1980) found decreased levels of IgA and IgG, but
elevated sIgA concentration, in saliva of the patients with
NUG.

51. Specific immunoglobulins in saliva
• Eggert (1987) saliva from treated periodontitis patients had
higher IgA and IgG for periodontal pathogens (P.g and T.d).
• Sandholm (1987) The level of salivary IgG antibody to A.a
was significantly elevated in 55% of the patients with
untreated JP and in 28% of the treated patients and 57% in AP.
• Schenk(1993)patients with a low mean number of bleeding
gingival units demonstrated significantly higher levels of
salivary IgA antibody reactive with S.mutans, A.a, and
Eubacterium.

52. Non specific markers:
Mucins
• glycoproteins produced by salivary glands
• mucins(MG1 and MG2)
• cytoprotection
• Lubrication
• protection against dehydration
• maintenance of viscoelasticity in secretions.
• The mucin MG2 affects the aggregation and adherence of
bacteria (A.a) and decreased conc of MG2 in saliva may
increase colonization with Aa.

53. Enzymes
Enzymes found in whole saliva originate from three
main sources:
(1) the actual salivary secretions per se
(2) the GCF, stemming from PMNs and tissue
degradation
(3) bacterial cells from dental biofilms and mucosal
surfaces. (Chauncey 1961).
Ingman et al. 1993 enzyme activity in whole saliva appears to
reflect the severity of periodontal disease

54. • salivary enzymes reported in increased conc in
periodontal disease are hyaluronidase, lipase, β-
glucuronidase and chondritin sulfatase, amino acid
decarboxylases, catalase, peroxidase and collagenase.
• proteolytic enzymes in the saliva contribute to the
initiation and progression of periodontal disease
• saliva contains antiproteases that inhibit cysteine
proteases such as cathepsins, antileucoproteases that
inhibit elastase and (TIMP) to inhibit the activity of
collagen – degrading enzyme

55. • Nakamura and Slots (1983) noted higher enzyme
activity in AP patients for alkaline phosphatase,
esterase, β-glucuronidase, β-glucosidase, and cysteine
aminopeptidases in JP.
• Gibert et al. relationship between attachment loss in
the periodontal group and a drop in ALP activity in
serum.
• Zambon et al. (1985) reduced salivary levels of
caprylate esteraselipase, leucine, valine and cysteine
aminopeptidases, trypsin, b-galactosidase, b
glucuronidase and b-glucosidase
decrease in proportions of subgingival black pigmented
bacteroides and motile organisms noted after treatment,
suggesting them as potential source.

56. Lysozyme
• hydrolytic enzyme cleaves the linkage between the glycopeptide (muramic
acid) – found in the cell wall of certain bacteria’s.
• cause lysis of bacterial cells by interacting with monovalent anions and
with proteases found in saliva. leads to destabilization of the cell membrane
as a result of the activation and degranulation of endogenous bacterial
autolysins.
• Lysozyme targets Veillonella species and A.a.
• It probably repels certain transient bacterial invaders of the mouth (Pullock
et al 1985)
Jalil et al. (1993) patients with low levels in saliva are more
susceptible to plaque accumulation which is considered as risk
factor for periodontal disease.

57. Lactoperioxidase
• Peroxidase activity is derived from 2 sources.
– Human lactoperoxidase (HS-LPO) is synthesized and
secreted by salivary glands.
– Myeloperoxidase (MPO) is found in PMN leucocytes &
migrate in to oral cavity by gingival crevice.
Salivary peroxide
• removes toxic hydrogen peroxide produced by oral
microorganisms and reduces acid production in the dental
biofilm, thereby decreasing plaque accumulation.
• Guven et al. (1996)higher activity in the diabetes patients,
serve as a marker for gingival inflammation

58. The lactoperoxidase-thiocynate system in saliva
catalyses the formation of bactericidal compounds e.g.
hypothiocyanate, by peroxidation of thiocyanate
bactericidal to certain strains of lactobacillus and streptococcus
prevents the accumulation of lysine and glutamic acid, both of
which are essential for bacterial growth.
Myeloperoxidase
• released by leukocytes and is bactericidal for Actinobacillus
• inhibits the attachment of Actinomyces to hydroxyapatite.
• Increased MPO activity was found in saliva of RPP and AP.

59. Histatin
• salivary protein with antimicrobial properties
• secreted from parotid and submandibular glands.
• neutralizes the LPS located in membrane of gram negative
bacteria.
• inhibitor of host and bacterial enzymes involved in the
destruction of the periodontium
• involved in the inhibition of release of histamine.

60. Other proteins
Cysteine proteinases
• proteolytic enzymes originated from pathogenic bacteria,
inflammatory cells, osteoclasts and fibroblasts.
• collagenolytic activity, which may cause tissue destruction.
(Cutler et al. 1995).
• Cystatins are physiological inhibitors of cysteine proteinases
• Henskens,1996 After periodontal treatment, total cystatin and
cystatin C concentration decreased to control levels.
• Evren, 2008 total saliva cystatin C levels were higher in health

61. Lactoferrin
•is a iron binding glycoprotein produced by salivary glands
•inhibits microbial growth by sequestering iron from the
environment thus depriving bacteria.
•strongly upregulated in mucosal secretions during gingival
inflammation and is detected at high conc. in saliva of patients
with periodontal disease.
•effective against Actinobacillus species (Arnold, 1980)

62. Platelet activating factor (PAF)
• a potent phospholipid mediator of inflammation
• Garito,1995 A significant positive correlation was observed
between the level of PAF in saliva and measures of periodontal
inflammation

63. Serine proteinases
Elastase
• Produced by PMN leukocytes.
• Elastase is held in inactive state within cell, by inhibitors
(α1proteinase inhibitor and α2 macroglobulin).
• Elastase is able to degrade proteoglycans and can also activate
latent collagenase
• Nieminen,1993 levels correlate with bleeding sites
• significantly higher in patients group from gingivitis to
periodontitis.
• Ingman,1993 higher in untreated AP patients.

64. Fibronectin
• is a glycoprotein, promotes selective adhesion and
colonization of certain bacterial species, while
inhibiting others.
• Gibbons et al. (1986) Higher proteolytic activity
observed in saliva collected immediately after
awakening, and the levels of enzyme activity
correlated with the state of cleanliness.
• Lamberts et al. (1989) salivary fibronectin levels
(used as an index of GCF flow into the oral cavity),
did not differ significantly between individuals with
or without periodontal disease.

65. Defensins
• These are antimicrobial peptides which are induced in
epithelial tissues upon inflammation.
• These peptides are part of the innate immune system, have
broad spectrum antibacterial and antifungal activity.
Calprotectin
• Main source of salivary calprotectin are GCF and oral surface
epithelium.
• Salivary calprotectin levels are raised in patients with oral
candidiasis.

66. Epidermal growth factor (EGF)
• involved in oral wound healing and stimulates epithelial cells.
• Oxford(1998) found a transient increase in salivary EGF levels
in response to periodontal surgery.
Vascular endothelial growth factor (VEGF)
a multifunctional angiogenic cytokine important in inflammation
and wound healing.
Taichman 1998 Higher levels of VEGF were detected in whole
saliva from periodontitis patients

67. Epithelial keratins
• Epithelial cells from the lining of the oral cavity found in
saliva
• McLaughlin (1996) higher conc of keratin in GCF at sites
exhibiting gingivitis and periodontitis
• not observed in saliva.

68. Inflammatory cells
• leukocytes in saliva varies from person to person, and vary for
an individual during the course of the day.
• Klinkhammer (1968) standardized collection and counting of
leukocytes in saliva
• developed the orogranulocytic migratory rate (OMR). The
OMR was found to be correlated with gingival index
• Raeste(1978) the OMR reflects the presence of oral
inflammation

69. Salivary ions
Calcium(Ca)
• A high concentration of salivary Ca was correlated with good
dental health
• no relationship detected with periodontal bone loss as
measured from radiographs (Sewon & Makela 1990).
• Sewon et al. 1990 salivary Ca, and the saliva Ca to phosphate
ratio were higher in periodontitis- affected subjects

70. Volatiles
• Salivary volatiles suggested as possible diagnostic markers
and contributory factors in periodontal disease.
• primarily hydrogen sulfide and methylmercaptan(Rosenberg &
McCulloch 1992).
• pyridine and picolines were found only in subjects with
moderate to severe periodontitis (Kostelc et al. 1980).

71. Vitamins
thiamine, riboflavin, niacin, pyridoxine, pantothenic acid, biotin,
folic acid and vitamin C and B12, and vitamin K
Coagulation factors
coagulation factors VIII, IX, X, plasma thromboplastin
antecedent (PTA) and the Hageman factors
hasten blood coagulation and protect the wounds from bacterial
invasion.

72. Bacteria
• De Jong,1984 serve as a growth medium for oral Streptococus
species and A. viscous.
• Bowden ,1997 number of bacterial cells for a in unstimulated
saliva may show active growth in plaque.
Oral microbial rinse test (Oratest)
• described by Rosenberg,1989
• for estimating oral microbial levels.
• provides a reliable estimate of gingival inflammation

73. • Asikainen,1991 A.a when recovered from subgingival sites
was also found in 69.9% and 35.9% of the samples of
stimulated and unstimulated saliva.
• Umeda,1998 bacterial detection in whole saliva had a
sensitivity of 42.6% for A. a, 68.4% for T.f, 97.8% for P. g, and
88.7% for P. i. The specificity of bacterial detection for these
microorganisms in saliva was 88.5%, 71.2%, 77.9% and
77.1%.
• Von Troil-Linden,1995 salivary levels of the periodontal
pathogens reflected the periodontal status.
• Christoph A,2009 Elevated salivary MMP-8 and T. denticola
biofilm levels in periodontal disease

74. Systemic markers related to periodontal infection
C-reactive protein
• released during the acute phase of an inflammatory
response
• produced by liver and stimulated by circulating cytokines
such as TNFa,IL1
• may reach saliva via gcf or salivary glands.
• High levels associated with chronic and aggressive
periodontal diseases
• measurable from saliva using lab on chip method.

75. Markers of periodontal soft tissue inflammation
• During the initiation of an inflammatory response
PGE2,IL1β,IL6,TNFa are released from the cells of junctional
epithelium and from ct fibroblasts, macrophages and pmn
leckocytes.
• Subsequently enzymes such as MMP8,9,13 are produced by
pmn and osteoclasts leading to degradation of ct collagen and
alveolar bone.
• Bradon,2008 increase in TNF in periodontitis

76. Markers of alveolar bone loss
MMP
Host proteinases responsible for both tissue degradation and
remodeling.
MMP-8 most prevalent in diseased tissues and gcf
found in elevated level in saliva from patients with periodontal
disease
elevated in peri-implant sulcular fluid in peri-imlantitis lesions.
Integrated Microfluidic Platform for Oral Diagnostics
(IMPOD) mean MMP-8 concentration in the saliva of the
periodontally healthy individuals was 10-fold less than that of
the periodontally diseased patients Herr AE,2007

77. MMP-9,is produced by neutrophils and degrades collagen, intercellular
ground substance.
2 fold increase found in patients with progressive attachment loss.
Makela et al. (1994) higher concentration of MMP-9 in whole saliva
of periodontitis patients
MMP-13 has been implicated in peri-implantitis
found in elevated level corresponding to the vertical bone loss around
loosening dental implants.
Hayakawa et al. (1994) reported lower conc of TIMP-1 in whole
saliva of patients with periodontal disease with higher collagenase
activity
Increased TIMP-1 and decreased collagenase activity observed after
initial therapy

78. Uitto et al. (1990)
Collagenase originated from PMNs entering the oral cavity
through the gingival sulcus.
activity was in higher and active form in the periodontitis
patients.
Very little collagenase activity detected in saliva of edentulous
subjects.

79. Pyridinoline Cross-Linked Carboxyterminal Telopeptide
of Type I Collagen (ICTP)
• to detect bone resorption in periodontitis and
periimplantitis
• Palys et al. strongly correlated with whole subject levels
of several periodontal pathogens including T.f, P. g, P. i,
and T. d.
• Golub et al. 70% reduction in GCF ICTP levels after
treatment.

80. Osteonectin
polypeptide that binds strongly to hydroxyapatite and other
extracellular matrix proteins including collagens.
has been implicated in the early phases of tissue mineralization.
Osteopontin
• highly concentrated at sites where osteoclasts are attached to
the underlying mineral surface
• conc increased proportionally with the progression of disease

81. CONCLUSION
The knowledge of normal salivary composition, flow and
function is extremely important on a daily basis when treating
the patients.
Recognition should be given to saliva for the many contribution it
makes to the preservation and maintenance of oral and
systemic health
Saliva as a diagnostic specimen can give not only the same
information as serum testing but also additional or new
information that cannot be obtained from serum.

82. • The device performs rapid microfluidic chip-based
immunoassays (<3–10 min) with low sample volume
requirements (10 μL) and appreciable sensitivity (nM–
pM). Our microfluidic method facilitates hands-free
saliva analysis by integrating sample pretreatment
(filtering, enrichment, mixing) with electrophoretic
immunoassays to quickly measure analyte
concentrations in minimally pretreated saliva samples.
The microfluidic chip has been integrated with
miniaturized electronics, optical elements, such as
diode lasers, fluid-handling components, and data
acquisition software to develop a portable, self-
contained device.