Comparison of Gingival Crevicular Fluid Sampling Methods in Patients With Severe Chronic Periodontitis

Abstract

Background Analysis of samplings from periodontal pockets is important in diagnosis and therapy control of periodontitis. In this study, three different sampling techniques were compared to determine if one method can yield samples suitable for reproducible and simultaneous determination of bacterial load, cytokines, neutrophil elastase, and Arg-specific gingipains. R-gingipains are an important virulence factor of Porphyromonas gingivalis, the exact concentration of which in gingival crevicular fluid (GCF) has not yet been quantified. Methods GCF was sampled from four sites per patient (each two sites one method) in 36 chronic periodontitis patients. One week later, the procedure was repeated with alternative methods. The variables that had been determined were: loads of Aggregatibacter actinomycetemcomitans and P. gingivalis, levels of interleukin-6 and interleukin-8, activity of neutrophil elastase and level of R-gingipains. Results The detected cytokine levels were higher using paper strips compared to paper points. Bacteria were found in similar loads from the paper strips and paper points. R-gingipains were detectable in high quantities only by washing of the periodontal pocket. The level of R-gingipains correlated with the load of P. gingivalis. Conclusion The use of paper strips is suitable for simultaneous determination of microbial and immunological parameters. Obtaining GCF by washing can be useful for special purposes. Gingipain concentration in periodontal pockets was directly determined to be up to 1.5 μM. This value indicates that most of so far identified substrates of these proteases by in vitro assays can be easily degraded in P. gingivalis infected sites.

Cleavage of extracellular matrix in periodontitis: Gingipains differentially affect cell adhesion activities of fibronectin and tenascin-C

“…Although other virulence factors are present in P. gingivalis , the analysis of gingipain-defective strains has shown that these proteases are essential for survival, proliferation, and infectious potential of the pathogenic bacteria [3, 4]. It has been shown recently that the concentration of Rgps in the periodontal pockets of periodontitis patients can exceed 1 μM [5]. …”

The Effect of Systemic Doxycycline and Mechanical Therapy on GCF β-Glucuronidase Leves in Chronic Periodontitis Patients

“…However, certain limitations could not be overcome, such as the choice of collection device, length of collection time, number of sample repetitions. [26][27][28] It is a known fact that the amount of GCF collected varies from site to site, and hence the volume collected differs. In this study, we addressed this issue by standardizing the time of collection to 30 seconds, yet the volume of fluid collected differed.…”

Structural insights unravel the zymogenic mechanism of the virulence factor gingipain K from Porphyromonas gingivalis, a causative agent of gum disease from the human oral microbiome

“…Among the latter are the gingipains K (Kgp) and R (RgpA and RgpB), which specifically cleave substrates after lysines and arginines (18), respectively. They are soluble or outer membrane-anchored cysteine peptidases responsible for up to 85% of the total extracellular proteolytic activity of the bacterium (19 -21) and can be found at very high concentrations in gingival crevicular fluid from CP patients (22). Kgp accounts for most of this activity (23) and is essential for bacterial survival and progression of CP (18).…”