Identification of EMT Indicators in Healthy and Periodontally Diseased Gingival Samples

Localization of Indicators for Epithelial Mesenchymal Transformation in Gingiva From Chronic Periodontitis Patients as Compared to Healthy Individuals (Case Control Study)

The current study focuses on the localization and quantitative assessment of growth factors and cytokines related to the EMT process found in the human gingival tissue samples taken from periodontally diseased individuals compared to other samples taken from healthy individuals. Through this investigation the correlation between the severity of the disease and the amount of these factors will be studied aiming to alleviation of the high prevalence of periodontal diseases among the Egyptian population.

In a case control human study The existence and the amount of the indicators and cytokines for EMT in the gingival tissue samples taken from the periodontitis patients will be investigated and compared to those in healthy individuals using histopathology and real time PCR to reveal the role of these factors in periodontal disease incidence in humans.

Total of 36 participants will be recruited in this study The participants will be divided according to their periodontal status into four groups (healthy, mild, moderate and severe periodontitis) 9 patients in each group.

The periodontal screening of patients will be done using mouth mirror and graduated Williams's periodontal probe for the following:

• Gingival status will be assessed by using gingival index
• Periodontal pocket depth.
• Clinical attachment loss.
• Radiographic examination using panoramic technique. Screening criteria for chronic periodontitis was the presence of at least 5 sites with 4 mm horizontal alveolar bone loss on radiographs.

Gingival tissue samples will be harvested during periodontal surgery and extraction of the periodontally hopeless tooth for the periodontitis patients and during the premolar extraction (Orthodontic ttt) or third molar extraction for the healthy controls.

The dissected gingival samples will be washed in saline solution and fixed in 4% buffered formalin for 48 hours, dehydrated in ascending grades of ethyl alcohol, cleared in xylene and embedded in paraffin and will be subjected to the following:.

1. Sections of 4-5 µm in thickness will be obtained, deparaffinized, rehydrated and subjected to histological staining and histomorphometric analysis.
2. Real time- PCR for TGF-B1 ,Vimenten , E-Cadherin .

Inclusion Criteria:

• Inclusion criteria for healthy group:

  • Absence of any clinical sign of gingival inflammation
  • Probing depth ≤3 mm
  • No clinical attachment loss.

Inclusion criteria for periodontitis group:

• Generalized presence of clinical signs of gingival inflammation
• Generalized probing depth ≥4 mm
• Generalized clinical attachment loss of ≥2 mm.

Exclusion Criteria:

• • Patients with any systemic disease,

  • Smokers
  • Pregnant or lactating women,
  • Cervical/proximal/subgingival caries or restorations,
  • Periodontal or antimicrobial therapy within 3 months before sampling.