Protein recovery from several paper types used to collect gingival crevicular fluid

Abstract

The purpose of this study was to compare the efficiency of protein elution from several types of gingival crevicular fluid (GCF) collection papers when the volume of the inoculated protein and the elution methods were constant. Various concentrations of bovine serum albumin (BSA) and 14C-BSA were placed onto strips of Whatman #1 [W1], Whatman 3 MM chromatographic [W3], Periopaper (ProFlow) [P] and Periopaper (Harco) [H], and recovered proteins measured following a non-optimized centrifugal elution technique. There were significant differences in % recovery of BSA and 14C-BSA from the papers, which was dependent on both the type of paper and the concentration of the inoculated protein; that is, proteins at the lowest concentrations were less efficiently eluted from GCF collection papers than those at higher concentrations. Equations for regression lines of elution efficiency were quadratic. Thus, our data suggest significant differences in the efficiency for elution of BSA from absorbent papers when the volume of the inoculated fluid and the elution technique were constant. Previous variable or conflicting experimental data between research groups may have resulted from incomplete elution of proteins from GCF collection papers, possibly due to entrapment within, or binding of GCF proteins to the paper.

Drop in Transforming Growth Factor-α and Osteoprotegerin Level in Gingival Crevicular Fluid from Patients with Gingivitis

“…[21] It has been previously reported that protein recovery rates from Periopaper points is not consistent, and depends on nature of the proteins and their concentrations. [22] But, we confirmed the reproducible evidence that protein recovery rates are relatively high and constant (80-87%) from Periopaper using IL-1b, b2-MG, and albumin in a preliminary way. b2-MG was determined by a sandwich ELISA that consisted of solid-phase (polystyrene bead)-immobilized antibodies and antibodies labeled with b-D-galactosidase, as described previously.…”

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

“…This finding was supported by the in vitro analysis; the recovery rate of IL‐8 was much lower from endodontic paper points compared to paper strips. An earlier study 37 also reported an incomplete recovery of proteins from paper points supposedly because of binding of GCF proteins to the paper. The difference in elution of IL‐6 and IL‐8 was most likely due to difference in the structure, charge distribution, and hydrophobicity of these cytokines molecules.…”

The in vivo Expression of the Collagenolytic Matrix Metalloproteinases (MMP-2, -8, -13, and -14) and Matrilysin (MMP-7) in Adult and Localized Juvenile Periodontitis

“…Elevated MMP-8 levels and activation in AP GCF have been documented before by our and other groups Lee et al, 1995;Nomura et al, 1998;Chen et al, 2000), whereas Romanelli et al (1999) and Mancini et al (1999), analyzing mouthrinses or GCF diluted with tap-water , did not detect MMP-13 in GCF samples. It should be kept in mind that GCF sample collection methods and materials (Johnson et al, 1999) evidently affect the apparent levels and amounts of different MMPs (Ingman et al, 1996;Golub et al, 1997;Nomura et al, 1998;Mancini et al, 1999;Romanelli et al, 1999;Sorsa et al, 1999;Chen et al, 2000). The soluble pro-form of 58-kDa MT1-MMP or MMP-14 was demonstrated for the first time in human GCF.…”