Comparison of gingival crevicular fluid sampling methods in patients with severe chronic periodontitis

Abstract

Background: The analysis of samplings from periodontal pockets is important in the diagnosis and therapy of periodontitis. In this study, three different sampling techniques were compared to determine whether one method yielded samples suitable for the reproducible and simultaneous determination of bacterial load, cytokines, neutrophil elastase, and arginine-specific gingipains (Rgps). Rgps are an important virulence factor of Porphyromonas gingivalis, the exact concentration of which in gingival crevicular fluid (GCF) has not been quantified.

Methods: GCF was sampled from four sites per patient (one sample per quadrant using two samples per method) in 36 patients with chronic periodontitis. One week later, the procedure was repeated with alternative methods. Variables determined were loads of Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans) and P. gingivalis, levels of interleukin-6 and -8, activity of neutrophil elastase, and level of Rgps.

Results: The detected cytokine levels were higher using paper strips compared to paper points. Bacteria were found in similar loads from paper strips and paper points. Rgps were only detectable in high quantities by washing the periodontal pocket. The level of Rgps correlated with the load of P. gingivalis.

Conclusions: The use of paper strips was suitable for the simultaneous determination of microbial and immunologic parameters. Obtaining GCF by washing can be useful for special purposes. The gingipain concentration in periodontal pockets was directly determined to be ≤1.5 μM. This value indicated that most of the substrates of these proteases by in vitro assays identified until now can be easily degraded in P. gingivalis-infected sites.

Figures

Figure 1

Levels of Interleukin (IL)-6 and IL-8, activity of neutrophil elastase, bacterial loads of P. gingivalis and A. actinomycetemcomitans as well as levels of arginine specific cysteine proteases (R-gingipains derived from Porphyromonas gingivalis) in gingival crevicular fluid determined by using paper points and paper strips for sampling material

Figure 2

Levels of arginine specific cysteine protease (R-gingipains derived from P. gingivalis) in gingival crevicular fluid in relation to the load of P. gingivalis

Figure 3

Recovered levels of cytokines applied in vitro onto paper points and paper strips