The gingival tissue is constantly subjected to mechanical & bacterial aggressions. Resistance to these action is provided by: 1. Epithelium 2. gingival crevicular fluid 3. saliva


The role of gingival epithelium in defence by its degree of keratinization & turn over rate. Epithelium constitutes a continuous lining of stratified squamous epithelium. Principle cells of this epithelium is keratinocytes. Other cells are non keratinocytes which includes langerhan’s cells,merkel cells & melanocytes.

The main function of the epithelium is to protect the deep structure while allowing selective intechange with oral environment. This is achived by proliferation & differtiation of the keratinocytes. Differentiation of keratinocytes by keratinization process which leads to production of an orthokeratinized superficial horny layer which is made up of mainly keratin protein. This layer makes intact barrier between the oral environment & deep layers.

Other protein keratolinin & involucrin forms a chemically resistant structure (envelop) located below the cell membrane. The upper most cells of the stratum spinosum contains dense granules known as keratinosomes or odland bodies which are modified lysosomes. They contain large amount of acid phosphatase. Acid phosphatase activity is related to the degree of keratinization.

Langerhans cells located among keratinocytes at supra basal levels. They belong to the mononuclear phagocyte system as modified monoctes. They have an important role in immune reaction as antigen presenting cells for lymphocytes.

GCF is secreted by sulcular epithelium in gingival sulcus. The presence of crevicular fluid has been known since the 19th century. Its composition & possible role in oral defence mechanism were elucidated by WAERHANG,BRILL & KRASSE in 1950.

Studies of BRILL considered that GCF is a continuous transudate. LOE,HOLM- PEDESEN, WEINSTEIN, MANDEL ID & SALKIND demonstrated that GCF is a inflammatory exudate.

o These includes: 1. Use of absorbing paper strips 2.Twisteel threads 3.Micropipettes 4.Intra crevicular washing

1. Use of absorbing paper strips:  Paper strips are placed into the sulcus or pocket.  This method causes the irritation of the sulcular epithelium that can oozing of fluid. 2. Extra sulcular method:  Paper strips are placed at the entrance or over the sulcus or pocket.  The fluid seeping out is picked up by the strip.

3. Twisted threads:  Preweighed twisted threads are placed in the sulcus around the tooth & the amount of fluid collected is estimated by weighing the thread. 4. Micropipettes:  Micropipettes (capillary tubes) of standerized length & diameter are placed in the pocket & their content is centrifuged & analyzed.

4. Intra crevicular washing:  A acrylic plate appliance is used in this method.  Plate covering the maxilla with soft border & groove following the gingival margins.  This appliance is connected by 4 collection tubes ,2 on palatal sides & 2 on buccal side.  The washing is obtained by rinsing the crevicular area from 1 side to the other using a peristaltic pump.

o The amount of fluid collected on paper strip is evaluated by: 1. Staining 2. Electronic method

1. Staining: o Wetted paper strips is stain by ninhydrin. o It is than measured planimetrically on an enlarged photograph or a with help of magnifing glass or a microscope.

2.Electronic method: o Fluid collected on a ‘blotter’ (periopaper) employing an electronic transducer (periotron). o Wettners of strip affects flow of current & a digital read out. o Measurement performed by CIMASONI showed that a 1.5 mm wide strip paper inserted 1mm within the sulcus of inflammed gingiva absorbs about 0.1mg of fluid.

o CHALLACOMBE used an isotope dilution method to measure the amount of GCF present in particular space at any given time. o His calculation in human with mean gingival index of less than 1 showed that mean GCF volume in proximal spce of molar teeth ranged from 0.43 to 1.56 microlitre.

o It contains:

 Cellular elements
    Electrolytes Organic compounds Metabolic & bacterial products Enzymes & enzymes inhibitors

A. Cellular elements
1. Epithelial cells:
Oral sulcular epithelium & junctional epithelium are constantly renewing & shed cells will be found in GCF. Krekelar & ochs showed that fluid originated from area with more severe gingivitis contains a much higher proportion of these cells thus conferming the possible stimulating effect of inflammation upon the renewal of sulcular or junctional epithelium.

2. Leukocytes
The major site of entrance of leukocytes in oral cavity is the gingival sulcus. In sulcus the differential leukocytes count are present in following relative proportion. 95-97% neutrophils 1-2% lymphocytes 2-2% monocytes Among lymphocytes – 58% B lymphocytes 24% T lymphocytes

Number of leukocytes increase with the intensity of inflammatory process. Their main function is phagocytic & killing of bacteria therefore they constitute a major protective mechanism.

Bacteria cultured from GCF is similar those grown from adjacent dental plaque. Eg. Strepto sanguis Actinomyces viscosus Porphyromonas gingivalis Porphyromonas endodentalis Camphylobacter rectus Prevotella intermedia

B. Electrolytes
Na, k, Ca, F have been studied in GCF. 1. Na concentration: The investigation of GCF in inflammed gingiva by matsue (1967) show an average concentration of Na is 207- 222 meq Na \ litre. While normal gingival fluid contains 158 meq Na/litre.

2. K concentration: Matsue reported that K concentration is 69 meq K / litre in inflammed area. Normal GCF contains K conc. is 9.54 meq k / litre.

3.Na: k ratio: Study of krasse & egelberg (1962) showed that GCF of inflammed gingiva contain Na:k ratio is 3:9. While normal GCF shows Na:k ratio is 28:1.

4.concentration of other ions:

Ca, Mg, phosphate ion, chlorine ion have also been determined in known amount in GCF. Krasse & egelburg (1962) reported Na:Ca ratio average about 10 in normally healthy gingiva. In inflammed gingiva this ratio is 18. Weinstein et al (1967) reported that ca: protein ratio much higher in gingival fluid than serum this ratio tended to decrease in fluid from more inflammed areas.

C. Organic compounds
1. Carbohydrates
Mainly 3 substances reported in crevicular exudate. Glucose Hexasamine Hexuronic acid Exudate glucose content is higher in inflammed gingiva than normal gingiva. This is interpreted not only as a result of metabolic activity of tissues but also as a function of local microbial flora.

2.proteins 5 proteins alpha,beta,alpha 1,alpha 2 globulin & albumin were reported in GCF. Holmberg & killander confirmed that IgG,IgA & IgM immunoglobulin are present in GCF. These immunoglobulins might significantly contribute the oral defence mechanism.

3.Lipids: • Gingival fluid contains many classes of phospholipids as well as neutral lipids.

D. Metabolic & bacterial products
1. Lactic acid: Lactic acid present in gingival fluid was reported positively correlation to both the degree of inflammation & intensity of gingival fluid flow. Its origin considered mainly tissue origin.

2. Hydroxyproline: Hydroxyproline is a major break down products of collagen. Its presence in gingival fluid is on indicator of the rate of progression of periodontal disease.

3.Prostaglandins: It is a component of inflammatory reaction. Inflammed gingiva show more concentration of prostaglandins. It causes vasodilatation, bone deposition & inhibition of collagen synthesis.

4. Endotoxins: Endotoxins released from gram negative bacteria are highly toxic to gingival tissue & pathogenic factor in periodontal disease.

5.Cytotoxic substance: Cytotoxic substance like hydrogen sulphide which is toxic metabolite of bacteria origin also reported in gingival fluid & causes gingival inflammation.

6. PH of gingival fluid: Production of ammonia by microorganism causes elevation of PH. Elevated PH increases the severity of gingivitis & periodontitis. 7. Antibacterial factor: Antibacterial factor like leukocytes & flow of crevicular fluid is able to remove various kinds of bacteria from gingival pocket.

E. Enzyme & enzyme inhibitor:
1. Acid phosphatase: The main source of acid phosphatase in crevicular area are probably the PMNs & desqamating epithelial cells. In PMNs the enzyme is confined with in the azurophil granules. Acid phosphatase is bacteriocidal. It attacks teichoic acid which is 1 of the components of the bacterial all wall.

The enzyme is also found in bacteria including those of the gingival sulcular pocket. Eg. Actinobacillus Capnocytophaga Veilloonella

2. Alkaline phosphatase : • The concentration of this enzyme is significantly correlated with pocket depth. • This enzyme present in PMNs, exclusively in specific or secondary granules. • Some gram negative subgingival plaque bacteria also produces alkaline phosphatase activity.

3. Beta glucuronidase: Beta glucuronidase is 1 of the hydrolyses found in the azyrophilic or primary granules of PMNs. Beta glucuronidase is probably responsible for the final degradation of the oligosaccharides produced initially by the action of hyaluronidase. Beta glucuronidse also found in plaque bacteria.

4. LYSOZYME: Lysosome has bactericidal properties. Its ability to hydrolyze B-1, 4glycosidic bond of peptidoglycans of the bacterial cell wall. It is found in PMNs. The free enzyme may contribute to pocket formation by its detrimental effect upon epithelial cell stickness & lytic activity of connective tissue. It also accelerates the local release of intracellulr bacterial enzyme.

5. Hyaluronodase: Hyaluronidase splits B-1, 4-Nacetyl glucasaminide link in hyaluronic acid, condroitin 4 – sulphate & condroitin 6- sulphate which is components of bacterial cell wall.

6. Proteolytic enzyme: Proteinases might have major role in the destruction of tissue component during inflammation. Mammalian proteinase: (i) Cathepsin D: It is a carboxy endopeptidase 1 of the chief acid enzyme in lysosomes present at high concentration in inflammed tissues. It is abundant in mononuclear leukocytes.

(ii) Elastase: Elastase found in azurophilic granules of PMNs. These are analogus to lysosomes. (iii) Cathepsin G: It is the serine endopeptidase contained into the azurophilic granules of PMNs. It hydrolyze hemoglobin, fibrinogen, casein, collagen & proteoglycan.

(iv) Plasminogen activators: It is serine proteinase. It activates the components of complement which cause increased vascular permeability & accumulation of PMNs & monocytes. It also help in wound healing. (v) Collagenase: It is found in PMNs. (Specific granules) It causes degradation of collagen.

Bacterial proteinase: It includes serinr endopeptidase,fibrinolytic enzyme, bacterial collagenase etc. Serum proteinase inhibitor: These are mainly alpha-2 macroglobulin, alpha-1 anti tyypsin, alpha -1 anti chymotrypsin. These inhibits proteinase enzymes. Its concentration increased during inflammation.

Analysis of GCF has identified cellular & humoral response in both healthy individuals & these with periodontal disease. The cellular immune response includes the appearance of cytokines in GCF but there is no clear evidence of a relationship between them & disease. Interleukin-1 alpha & -1 beta are known to increase the binding of PMNs & monocytes to endothelial cells, stimulate the production of PGE2 & release of lysosomal enzyme & stimulate bone resorption.

Cellular & humoral activity in GCF

There is also preliminary evidence of the presence of y- interferon in GCF which may have protective role in periodontal disease because of its ability to inhibit the bone resorption activity of interleukin -1B. Presence of antibodies in GCF, its role in gingival defence mechanism is Hard to ascertain, there is a consensus indicating that : In a patient with periodontal disease a reduction in antibody response is deterimental. Antibody response play a protective role in periodontal disease.

Clinical significance of GCF:
Gingival fluid is an inflammatory exudate. Its presence in clinically normal sulcus can be explained by the fact that gingiva that appears clinically normal exhibits inflammation when examined microscopically.

A. General health & gingival fluid: (i) Circadian periodicity: There is a gradual increase in gingival fluid amount from 6.00 AM to 10.00 PM & decrease afterwards. (ii) Sex hormones: Female sex hormones increase the gingival fluid flow, probably they enhance vascular permeability. Clinical investigations have been shown an excerbation of gingivitis during pregnancy, menstrual cycle & at puberty.

B. Measurement of gingival inflammation: Increased GCF is a sign of inflammation.

C. Influence of mechanical stimuli: Chewing, vigrous gingival brushing, intrasulcular placement of paper strips increased the production of GCF.

D. Periodontal therapy: There is a increased in gingival fluid production during the healing period after periodontal therapy.

E. Smoking: Smoking causes marked increase in gingival fluid.

Drugs in GCF:
o Some antibiotics o Eg. Tetracyclin, metronidazole, are detected in GCF.

Salivary secretion are protective in nature because they maintain the oral tissue in a physiologic state. Saliva exerts major influences: o On plaque by mechanically cleansing the expose oral surfaces. o By buffering acids produced by bacteria. o By controlling bacterial activity.

• Saliva contains: (i) Antibacterial factor: Saliva contains lysozymes, myeloperoxidase, lactoperoxidase, glucoproteins, mucins & antibodies etc. • (a) Lysosomes: Lysosomes is a hydrolytic enzyme that clevage linkage beta-1, 4- glycosidic bond of peptidoglycans of bacterial cell wall. Eg. Veillnell species Actinobacillus actinomycetemcomitans

(b) Lacto- peroxidase –thiocyanate: Bactericidal is some strains of lactobacillus & streptococcus by preventing accumulation of lysine & glutamia acids both of which are essentials for bacterial growth. (c) Myeloperoxidase: It is bactericidal for actinobacillus.

It forms coating layer over tissue structures & provides lubrication & physical protection.

(d) Glycoproteins & mucin:

(e) Antibodies: Saliva contains IgG,IgM & IgA antibodies. IgG preponderant immunoglobulin found in saliva. Antibodies causes opsonization of bacteria.

(ii) Buffers & coagulation factor: Salivary buffer bicarbonate – carbonic acid system maintain the physiologic pH of oral cavity. Saliva contains coagulation factor- factor viii, ix, x, PTA & hagman factor that hasten blood coagulation & protect wound from bacterial invasion. (iii) Leukocytes: Leukocytes reach the oral cavity migrating through the gingival sulcus. PMNs leukocytes chiefly found in saliva that causes the phagocytosis.

o As we have seen that various component act in defence of gingiva o Eg. Sulcular fluid Saliva Gingival epithelium Leukocytes etc. o In which sulcular fluid is 1 of the most important component of defence mechanism. o These component through various mechanism & enzymes resist against the mechanical & bacterial aggressions & maintain the gingiva normal healthy state.


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