The aim of the present trial is to assess the gingival crevicular fluid level of caspase-3 and apoptosis inducing factor (AIF) in Generalized versus Molar-incisor grade C periodontitis. The present study will be carried out on patients selected from those attending on the outpatient clinics of Department of Oral Medicine, Periodontology, Oral Diagnosis and Dental Radiology, Faculty of Dental Medicine, Al-Azhar University, Assiut.
Periodontal diseases are inflammatory disorders that give rise to tissue damage and loss, as a result of complex interactions between pathogenic bacteria and host immune response. Gingivitis is generally regarded as a site-specific inflammatory condition initiated by dental biofilm accumulation and characterized by gingival redness and edema and the absence of periodontal attachment loss.(1,2) periodontitis, a disease characterized by gingival inflammation combined with connective tissue attachment and bone loss. Aggressive periodontitis is a form of periodontal disease characterized by rapid attachment loss, bone destruction, non-contributory medical history and family history of the cases Apoptosis is a fundamental and complex biological process that enables an organism to kill and remove unwanted cells during development, normal homeostasis and disease. Caspases are proteins implicated in the activation and execution of apoptosis was identified in humans as being crucial for this process. Caspases 3 appear to be activated in a protease cascade that leads to inappropriate activation or rapid disablement of key structural proteins and important signaling, homeostatic and repair enzymes and is essential for some of the characteristic changes in cell morphology and certain biochemical events associated with the execution and completion of apoptosis. However, the specific requirements of this caspase in apoptosis were until now largely unknown. Apoptosis-inducing factor (AIF) is a mitochondrion-localized flavoprotein with nicotinamide dinucleotide (NADH) oxidase activity is encoded by a nuclear gene. It has been shown to translocate from mitochondria to the cytosol as well as the nucleus when apoptosis is induced and mediate caspase-independent death because inhibition of caspase activation. Many authors view apoptosis as a process that is near-to-synonymous to massive caspase activation. The existence of a caspase-independent death effector thus does not fit the dominant scheme. During the last 2 years, several papers advocated the hypothesis that AIF is indeed a caspase-dependent death effector, meaning that the translocation of AIF from the mitochondrion (where it would be inert) to the nucleus (where it would induce apoptosis) would be caspase-dependent.
Study Type
Gingival crevicular fluid (GCF) samples will be collected to assess the level of caspase-3,8,9 and apoptosis inducing factor (Aif)
Faculty of dental medicine Al-Azhar University (Assiut branch)
Asyut, Asyut Governorate, Egypt
caspase 3 , picograms/millimeter (pg/ml) in gingival crevicular fluid
GCF from the mesio-buccal site of each tooth was collected using paper strips held in place for 30 seconds, and then removed. GCF volume (ml) was measured using a calibrated machine. After volume determination, strips were stored in sterile collection tubes that were labeled and then stored at - 80C. Enzyme-linked immunosorbent assays were used to quantify caspase 3 amount in picograms and its concentration (pg/ml) estimated according to the following formula= total caspase 3 amount (pg) / volume GCF (ml).
Time frame: through diagnosis completion, an average of 6 months
caspase 8 , picograms/millimeter (pg/ml) in gingival crevicular fluid
GCF from the mesio-buccal site of each tooth was collected using paper strips held in place for 30 seconds, and then removed. GCF volume (ml) was measured using a calibrated machine. After volume determination, strips were stored in sterile collection tubes that were labeled and then stored at - 80C. Enzyme-linked immunosorbent assays were used to quantify caspase 8 amount in picograms and its concentration (pg/ml) estimated according to the following formula= total caspase 8 amount (pg) / volume GCF (ml).
Time frame: through diagnosis completion, an average of 6 months
caspase 9 , picograms/millimeter (pg/ml) in gingival crevicular fluid
GCF from the mesio-buccal site of each tooth was collected using paper strips held in place for 30 seconds, and then removed. GCF volume (ml) was measured using a calibrated machine. After volume determination, strips were stored in sterile collection tubes that were labeled and then stored at - 80C. Enzyme-linked immunosorbent assays were used to quantify caspase 9 amount in picograms and its concentration (pg/ml) estimated according to the following formula= total caspase 9 amount (pg) / volume GCF (ml).
Time frame: through diagnosis completion, an average of 6 months
Apoptosis Inducing Factor(AIF), picograms/millimeter (pg/ml) in gingival crevicular fluid
GCF from the mesio-buccal site of each tooth was collected using paper strips held in place for 30 seconds, and then removed. GCF volume (ml) was measured using a calibrated machine. After volume determination, strips were stored in sterile collection tubes that were labeled and then stored at - 80C. Enzyme-linked immunosorbent assays were used to quantify AIF amount in picograms and its concentration (pg/ml) estimated according to the following formula= total AIF amount (pg) / volume GCF (ml).
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OBSERVATIONAL
Enrollment
50
Time frame: through diagnosis completion, an average of 6 months
Gingival Index
gingival index used to measure gingival inflammation. The degree of gingival inflammation was assessed as follow: 0 = Normal gingiva. 1. Mild inflammation, slight change in color, slight edema and no bleeding on probing. 2. Moderate inflammation, redness, edema and bleeding on probing. 3. Sever inflammation, marked redness, edema and tendency to spontaneous bleeding.
Time frame: through diagnosis completion, an average of 6 months
Plaque Index
Plaque index measured to assess plaque accumulation around gingival margin. The degree of plaque accumulation was assessed as follows: 0 = no plaque around the gingival margin. 1. = A thin film of plaque around the gingival margin. The plaque may be recognized only by running a probe across the tooth surface. 2. = moderate accumulation of soft deposits on the gingival margin and/or adjacent tooth surface, which can be seen by naked eye. 3. = Abundance plaque accumulation within the gingival pocket and/or on the gingival margin and adjacent tooth surface and hard deposits on the tooth surface are seen.
Time frame: through diagnosis completion, an average of 6 months
Attachment level
Attachment level by mm was measured using color -coded probe. The distance from the cemento-enamal junction to the free gingival margin and the distance from the free gingival margin to the base of the pocket where properly measured. The difference between the two measurements yields the attachment level in millimeter (mm).
Time frame: baseline, immediately after complete diagnosis and before any intervention or therapy
Bleeding on probing (BOP)
Bleeding on probing (BOP) at a force of 0.3N.with a manual pressure of sensitive probe, recorded on distal, facial, mesial, gingival surfaces. Calculation = (Number of bleeding surfaces / total number of tooth surface) \*100.Expressed in percentage (%) of gingival units bleeding on probing FMBS. In the rating system, 0 indicates absence of bleeding on probing, with 15%, 20% and \>20% indicating an increased percentage of inflammation/infection.
Time frame: through diagnosis completion, an average of 6 months
radiographic evaluation at molar-incisor area
Marginal bone loss around all central incisors and first molars was evaluated using periapical radiographs. The distance from cemento-enamel junction to the crest of alveolar bone will represent bone lost by mm
Time frame: through diagnosis completion, an average of 6 months