A new ultrasensitive chemiluminescent assay for the site‐specific quantification of alkaline phosphatase in gingival crevicular fluid

Abstract

The search for markers of periodontal disease activity and progression has accelerated over the last decade, in an effort to replace existing subjective clinical measures of periodontal health status. Research is being aimed at establishing more objective and quantitative methodology, capable of rapid diagnosis prior to the appearance of clinical signs of destructive disease. Such tests need to be sensitive enough to evaluate individual periodontal sites in health as well as disease states. We report the development of a new chemiluminescent assay for the enzyme alkaline phosphatase, that is capable of quantifying the enzyme in sub-microliter volumes of gingival crevicular fluid and serum. The technique will measure alkaline phosphatase (ALP) whilst immobilised on paper strips, without the need for an elution stage. It is simple, versatile and amenable to chair-side use. We discuss in detail the assay procedure and have examined levels of ALP in 11 adult volunteers with clinically healthy periodontal tissues. The mean ALP concentration was 2135 IU/L for GCF and 183 IU/L for serum, a 12-fold difference. There also appeared to be an "oral pattern" of enzyme distribution in healthy periodontal sites, with levels being higher in the anterior region of the mouth and highest in the lower anterior region.

Association between interleukin-8 levels and chronic periodontal disease

“…Lamster et al [51] suggested that the total cytokine amount in GCF might be more representative of the disease status than the evaluation of protein concentration. [51] In addition, Chapple et al [52] emphasized that volumes are variable regardless of inflammatory status. Therefore, they proposed utilizing the total marker activity per 30 seconds in GCF samples, rather than the concentration of the marker, because it might provide a better correlation with health or disease status.…”

“…Therefore, they proposed utilizing the total marker activity per 30 seconds in GCF samples, rather than the concentration of the marker, because it might provide a better correlation with health or disease status. [52] To measure gingival fluid, the use of micropipettes, ninhydrin staining, and fluorescein is complicated and time-consuming, but an electric measuring device, the “Periotron,” is efficient and can digitally demonstrate the amount of gingival fluid. [53] Therefore, the use of a Periotron provides a confident measurement of GCF volume, which is required for calculating the protein concentration present in the GCF.…”

Assessment of the alkaline phosphatase level in gingival crevicular fluid, as a biomarker to evaluate the effect of scaling and root planing on chronic periodontitis: An in vivostudy

“…Conversely only extremely small volume of fluid is available from a single site, and so GCF needs highly sensitive techniques for quantitative analysis. [11131617] The composition and possible role of GCF in oral defense mechanisms were elucidated by the work of Waerhaug,[3] Brill and Krasse. [9] Gustafsson and Nilsson[15] found the presence of fibrinolytic activity in GCF.…”

Effect of periodontal therapy on crevicular fluid interleukin-1β and interleukin-10 levels in chronic periodontitis

“…It has been suggested that in GCF the total cytokine amount might be more representative of the disease status as compared to the concentration. 44 According to Chapple et al 45 GCF volumes are very variable irrespective of inflammatory status, while the large variation in GCF volume obtained from each periodontal site may result in a wide range of marker concentrations. Thus, it was proposed 45 that the total marker activity per 30 s GCF sample rather than the concentration of the marker might provide a better correlation with health or disease status.…”