In Vivo Crevicular Leucocyte Response in Humans to a Chemotactic Challenge: Effects of Periodontal Diseases

Abstract

An in vivo assay was recently developed to monitor the crevicular leucocyte response to chemotactic agents, e.g., casein and N‐formyl peptides. This method was used to monitor humans with little or no gingival disease (C group), gingivitis (G group), chronic periodontitis (CP group) and localized juvenile periodontitis (LJP group). Casein (0.2 μ1, 2 mg/ml) was placed into an isolated gingival crevice of each subject with a calibrated wire loop and the time recorded (t = 0). Leucocytes were counted in crevicular washes (10 μl) 15 minutes later and every 5 minutes thereafter up to t = 50 minutes. This protocol was repeated for the crevice of an adjacent tooth except that the crevicular fluid flow response to the chemotactic challenge was monitored. The C, G and CP subjects showed a similar pattern of response to the chemoattractant with a single “peak” of leucocytes at approximately t = 25 minutes. However, the peak cell count was much greater in the G and CP groups than in the C group. LJPs showed an abnormal pattern with two leucocyte peaks, one at approximately 25 minutes and the other at 45 minutes. Both peaks tended to be higher than the single peak seen in Cs but were significantly lower than that in Gs or CPs, even at similar levels of inflammation. In addition, the peak leucocyte response (to casein) in LJPs did not increase with increasing leucocyte counts in the unchallenged (resting) crevice, whereas a positive relationship was seen in the other groups of subjects. These data suggest that this new assay may provide important diagnostic information on in vivo neutrophil migration in the gingival crevice and on susceptibility to periodontal disease.

Neutrophil chemotaxis in refractory cases of periodontitis

“…Resting cell counts in refractory patients were often as high or higher than the peak values. This is similar to the relationship reported by Singh et al (1984) for counts of LJP patients.…”

“…All of the refractory cases examined in this study exhibited a multiple-peak response to an in vivo chemotactic agent. In a recent study by Singh et al (1984) using the in vivo chemotaxis assay, patients with chronic periodontitis exhibited a single-peak response while patients with LJP exhibited a double-peak response. In the current study, the patients with refractory disease showed the multiple-peak response, not the single.…”

1985 Kreshover lecture. Molecular Factors Influencing Neutrophil Defects in Periodontal Disease

“…In contrast, the UP patients showed an abnormal pattern with two peaks, one at 25 minutes and another at 45 minutes. The numbers of neutrophils migrating through the gingiva were lower than those in control subjects with comparable inflammation (Singh et al, 1984).…”

“…Gillett and Wilton (1981) also reported that neutrophils accumulate in lesions of juvenile periodontitis patients, and quantitative studies by Singh et al (1984) show a marked reduction in the numbers of neutrophils migrating into the gingival crevice in UP patients. Sandholm (1984), in a light and scanning electron microscopic study of gingival washings, found migratory forms of neutrophils in samples taken from adult periodontitis patients.…”

“…Furthermore, this functional abnormality in peripheral blood neutrophils is apparently expressed in vivo by a reduced rate of neutrophil influx into the gingival crevice in response to a chemotactic challenge (Singh et al, 1984). Normal subjects challenged at the gingival crevice with casein manifested a single "peak" of infiltrating leukocytes approximately 25 minutes after challenge.…”