L-Proline 4-methoxy-β-naphthylamide hydrochloride

Examining the relationships among indicators of the acute inflammatory response in gingival crevicular fluid (GCF) and specific bacterial species in subgingival plaque may provide indications of which bacterial species or groups of species may be associated with potentially destructive host-derived processes. Here we report on the relationship of the subgingival plaque flora to the activity of mammalian forms of the enzymes beta-glucuronidase (beta G), lactate dehydrogenase (LDH), and arylsulfatase (AS) in GCF from a total of 54 4-6 mm periodontal sites from 13 periodontitis patients. Sites were scored for probing depth (PD) and bleeding on probing, and GCF was collected using filter paper strips inserted into the sulcus for 30 s, eluted in buffer and assayed for enzyme activity. 1 week later, the patients were again evaluated for PD and bleeding, and subgingival plaque was removed with a curette oriented toward the pocket epithelium. Plaque samples were examined by darkfield microscopy and cultured anaerobically on selective and non-selective media. Various groups of bacteria, including species of black pigmenting Bacteroides (BPB), Fusobacterium sp., Capnocytophaga sp, Streptococcus sanguis, and total facultative organisms were enumerated. Relationships among the enzymes and bacterial groups expressed as colony-forming unit (CFU) counts or as a % of the total cultivable flora were assessed by Spearman correlation analysis. beta G levels were significantly correlated with populations of spirochetes, B. intermedius, B. gingivalis, and total lactose negative BPB's. Correlation between beta G and F. nucleatum sp. or Capnocytophaga sp. approached but did not reach statistically significant levels. In contrast, LDH activity showed a significant positive correlation with levels of B. gingivalis and total lactose negative BPB's. AS levels were significantly correlated only with B. gingivalis. beta G and LDH showed a significant negative correlation with levels of coccoid forms. Thus, beta G, an acid hydrolase which can serve as a marker for primary granule release from polymorphonuclear leukocytes, was most closely correlated with the micro-organisms found in other studies to be associated with chronic adult periodontitis.

This investigation analyzed, in a cross-sectional study, the possible relationship between gingival crevicular fluid (GCF) elastase-like protease (ELP) levels and the periodontal clinical parameters or the presence of specific bacteria in subgingival plaque. A total of 388 periodontal sites from 8 adult periodontitis patients were examined for plaque index (PII), gingival index (GI), pocket depth (PD) and alveolar bone loss (ABL). GCF ELP levels were determined as ELP alpha-1 protease inhibitor (ELP-alpha 1-PI) complex levels with a commercially available ELISA. Subgingival plaque samples were tested for the presence of Bacteroides gingivalis, B. intermedius and Actinobacillus actinomycetemcomitans by indirect immunofluorescence (IF) microscopy. GCF ELP-alpha 1-PI levels were then correlated with clinical periodontal indices and proportions of IF-positive bacteria per site. Statistically significant positive correlations were found between GCF ELP-alpha 1-PI concentrations and subgingival Bacteroides proportions. When the sites examined were analyzed depending on the level of each clinical parameter, the levels of these correlations changed. A. actinomycetemcomitans correlated highly (r = 0.716) with ABL for sites with low GI score. The correlations between GCF ELP-alpha 1-PI and B. gingivalis (r = 0.642) or B. intermedius (r = 0.774) were the highest for ABL less than or equal to 20% and PD less than or equal to 3 mm, respectively. The strong association between GCF ELP-alpha 1-PI concentrations and subgingival bacteria previously associated with advancing periodontitis indicates that measurement of GCF ELP-alpha 1-PI concentrations may be useful in the evaluation of periodontal sites, especially those with very little or no tissue destruction.

Two unique forms of periodontal disease, HIV-gingivitis and HIV-periodontitis, have been described in patients with Acquired Immunodeficiency Syndrome (AIDS). In order to determine the bacterial species associated with periodontitis in AIDS patients, the predominant cultivable microflora was examined in 21 subgingival plaque samples from 11 AIDS patients with periodontitis. The presence of putative periodontal pathogens including Actinobacillus actinomycetemcomitans, Bacteroides intermedius, Porphyromonas gingivalis (formerly B. gingivalis), and Wolinella recta was examined by immunofluorescence in 128 subgingival dental plaque samples from 50 AIDS patients including 32 patients with periodontitis. Of 666 bacterial strains isolated from the 21 subgingival plaque samples, Streptococcus sanguis II was the most frequently recovered species comprising 18.5% of the total number of isolates followed by Lactobacillus acidophilus (12.2%), Porphyromonas gingivalis (12%), Fusobacterium nucleatum (11.4%), Staphylococcus epidermidis (8.7%), Actinomyces naeslundii (7.5%), and Actinomyces viscosus (4.7%). Fusobacterium nucleatum was the most prevalent species and was found in 76% of the sites and 91% of the patients. Enteric species including Enterococcus avium and Enterococcus faecalis, Clostridium clostridiiforme and Clostridium difficle as well as Klebsiella pneumoniae also were recovered. Immunofluorescence assays detected similar carriage rates of A. actinomycetemcomitans, B. intermedius, and P. gingivalis in both gingivitis patients and periodontitis patients, while four times more periodontitis patients demonstrated W. recta. Subgingival yeast was a frequent finding in these AIDS patients, present in 62% of the subjects and 55% of the sites. This study indicates that subgingival plaque in AIDS patients with periodontitis can harbor high proportions of the same periodontal pathogens as are associated with periodontitis in non-HIV infected subjects as well as high proportions of opportunistic pathogens.