Periodontal Disease Clinical Trial
Official title:
Host DNA Methylation Patterns in Association With Periodontal Disease : MiRNA Changes in Obesity
The overall goal of this research is (1) to identify changes in gene expression and DNA methylation status in subjects who exhibit advanced chronic periodontal inflammation and (2) to identify microRNAs (miRNAs) and the interactive pathways associated with obesity as a modifier of periodontal infection pathogenesis.
Aim 1. To determine whether diseased periodontal tissues biopsied from patients with
periodontal disease will have altered DNA methylation patterns of selected genes as compared
to the DNA methylation status biopsies obtained from adjacent, non-diseased periodontal
sites, as well as periodontal biopsy samples obtained from non-diseased, healthy subjects.
Aim 2. To determine the role of tissue DNA Methylation on mRNA expression:
- 2a. To determine whether alterations in tissue mRNA expression (as determined by
quantitative PCR) are associated with an aberrant DNA methylation status of respective
gene promoter CpG islands in tissue biopsy samples obtained from diseased and
non-diseased tissues from patients with periodontal disease and from non-diseased
healthy subjects.
- 2b. By performing laser capture microdissection of biopsy samples from diseased
patients [(epithelium, connective tissue and inflammatory infiltrate)] we seek to
determine cellular patterns of mRNA expression and DNA methylation of targeted genes
comparing inflamed to non-inflamed sites within diseased patients.
- 2c. We will perform in vitro studies to analyze whether gingival fibroblasts from
patients with periodontal disease will show a differential methylation pattern and
whether these cells will display a differential inflammatory response when compared to
control fibroblasts isolated from healthy subjects.
Aim 3. The aim of this pilot investigation was to determine if miRNA expression differed in
the presence or absence of obesity, comparing gingival biopsies obtained from patients with
or without periodontal disease.
- 3a. To determine whether diseased periodontal tissues biopsied from patients with
periodontal disease will influence the level of miRNA expression as compared to the
biopsies obtained from non-diseased, healthy subjects.
- 3b. To determine whether BMI [nonobese subjects BMI <30kg/m2, obese subjects >30kg/m2]
will influence the level of miRNA expression in biopsies from diseased periodontal
tissues as compared to periodontal biopsy samples obtained from non-diseased, healthy
subjects.
Gingival samples, gingival crevicular fluid, saliva and plaque samples will be collected and
a periodontal examination will be performed. The periodontal assessment will record pocket
depth, clinical attachment level and percent bleeding on probing. Blood pressure, height and
weight (Body Mass Index assessment) will be obtained on participants as well.
Tissue gene expression by quantitative PCR and DNA methylation sequence analysis, proteomic
analyses of gingival crevicular fluid, Differential Methylation Hybridization and, microRNA
expression profile by Human miRNA Microarray and Real-time PCR quantification will all be
performed.
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Observational Model: Case Control, Time Perspective: Cross-Sectional
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