Association Between Porphyromonas gingivalis, Prevotella intermedia, Actinobacillus actinomycetemcomitans and Spirochaetes with Neutral Proteolytic Enzyme Activities in Deep Periodontal Pockets

Abstract

The aim was to study whether specific periodontopathogenic bacteria or inflammatory cells were associated with the neutral proteolytic enzyme activities in deep periodontal pockets. A total of 15 sites was sampled from six subjects. Samples were collected twice, before and after conventional periodontal treatment. First, a crevicular fluid (CF) sample was collected using a washing method, after which a subgingival plaque sample was taken with a curette. Bacteria were cultured on selective media and enumerated. Amounts of inflammatory cells were counted from the washing samples. The general neutral protease activity was measured using Azocoll and the trypsin-like activity was assessed using two substrates, carbobenzoxy-L-arginine-7-amino-4-methylcoumarin (CAAM) and 7V-benzoyl-DL-arginine-2-naphthylamide (BANA). The general protease activities of both plaque samples and CF samples were associated with the numbers of inflammatory cells in the pocket. BANA hydrolysis was most strongly associated with the amount of total bacteria. The proportions of spirochaetes were connected to BANA hydrolysis. CAAM activity was associated with the proportions of Porphyromonas gingivalis and spirochaetes. Of the studied bacteria, BANA seems to be the substrate of choice for studying the trypsin-like enzyme activity of spirochaetes, while CAAM best detected the trypsin-like activity of P. gingivalis.

Key Words:

• Periodontitis
• Proteolytic activity
• Porphyromonas gingivalis
• Prevotella intermedia
• Spirochaetes