ABSTRACT
BACKGROUND: Little is known about the release of apoptotic proteins during periodontal breakdown. This pilot study investigates the presence of factors associated with apoptosis in serum, saliva, and gingival crevicular fluid (GCF) and their association with periodontal disease severity and activity. METHODS: GCF, whole saliva, and serum were obtained from 47 adult patients with chronic periodontitis (CP) and 10 healthy controls. Clinical measurements, including probing depth (PD), clinical attachment level (CAL), and radiographs, were used to classify patients into healthy, mild, and moderate/severe CP groups. Enzyme-linked immunosorbent assays were used to measure apoptosis or DNA fragmentation in GCF and active caspase-3, soluble Fas (sFas), and sFas ligand (sFasL) in saliva and serum. Western immunoblotting was used to detect Fas, FasL, sFasL, and caspase-3 expression in GCF. RESULTS: DNA fragmentation was positively correlated with PD and CAL regardless of patient disease status (P <0.001). sFas and sFasL were present in saliva and serum, but there were no differences between groups. In GCF, the greater odds of detecting Fas, sFasL, and caspase-3 increased with increasing PD and CAL (P <0.05). In addition, sites with inflammation and PD ≥5 mm had significantly greater odds of exhibiting Fas, sFasL, and caspase-3 expression compared with sites without inflammation and PD <5 mm (P <0.05). Caspase-3 was not detected in saliva or serum. At the patient level, only FasL and disease status were significantly correlated (P <0.05). CONCLUSION: Factors associated with apoptosis were detected in GCF in patients with CP.
Subject(s)
Apoptosis Regulatory Proteins/analysis , Chronic Periodontitis/metabolism , Periodontal Index , Adult , Aged , Alveolar Bone Loss/classification , Alveolar Bone Loss/metabolism , Apoptosis Regulatory Proteins/blood , Caspase 3/analysis , Caspase 3/blood , Chronic Periodontitis/classification , Cross-Sectional Studies , DNA Fragmentation , Fas Ligand Protein/analysis , Fas Ligand Protein/blood , Female , Gingival Crevicular Fluid/chemistry , Humans , Male , Middle Aged , Periodontal Attachment Loss/classification , Periodontal Attachment Loss/metabolism , Periodontal Pocket/classification , Periodontal Pocket/metabolism , Pilot Projects , Saliva/chemistry , Salivary Proteins and Peptides/analysis , fas Receptor/analysis , fas Receptor/bloodABSTRACT
INTRODUCTION: The aim of this study was to examine a potential link between apoptotic biomarkers in gingival crevicular fluid (GCF) and periodontal destruction in four cases of localized aggressive periodontitis (LAP), diagnostically enhanced by cone beam computed tomography. CASE SERIES: This study examined the GCF in four patients diagnosed with LAP (formerly localized juvenile periodontitis) at a routine periodontal examination. The LAP diseased sites had attachment loss ranging from 5-12 mm. Atotal of 62 samples of GCF were collected from diseased sites and from contralateral, matched healthy sites with minimal or no attachment loss. All samples were assayed for apoptotic markers, including Fas/FasL, DNAfragmentation, and nitric oxide. The GCF samples were analyzed utilizing enzyme-linked immunosorbent assays for DNA fragments and nitric oxide levels, whereas Western blotting was used for Fas/FasL analyses. Our results showed a significant increase in the apoptotic markers Fas/FasL and DNA fragmentation when comparing GCF from diseased versus non-diseased sites in patients with LAP. CONCLUSION: To our knowledge, this is the first report of apoptotic biomarkers associated with patients diagnosed with LAP. Finding significantly increased levels of these markers in localized areas may help us understand the pathophysiology associated with this specific form of periodontitis, and, furthermore, may provide a basis for a quantifiably prognostic test when attempting to treat this disease.
