Tetracycline inhibition of neutrophil-associated collagenolysis has been the focus of a number of investigations. Evidence has suggested that this inhibition results from the ability of this family of antimicrobial drugs to bind divalent cations such as Ca2+ and Zn2+, two cations that are required for full expression of activity of metalloproteinases such as collagenase and gelatinase. Data presented in this study demonstrate that tetracyclines can also inhibit neutrophil-mediated RBC lysis, superoxide anion synthesis, degranulation and migration. To some extent, tetracycline inhibition of neutrophil functions is mimicked by the Ca2+ binding agents, EDTA and TMB-8. However, Ca2+ enrichment restored full function to EDTA- and TMB-8-treated cells but not to tetracycline-treated neutrophils. This suggests that Ca2+ binding plays a role but is not the critical effect leading to tetracycline suppression of neutrophil functions. It has been suggested that tetracyclines can suppress leukocyte-associated tissue damage. Host tissues are protected from neutrophil-mediated damage by two mechanisms: 1. Neutrophil granule-associated enzymes are secreted in an inactive state; and, 2. tissues are protected from these enzymes by a potent inhibitor shield. Neutrophils can bypass these protective elements by activating enzymes and by destroying the shield through the synthesis of oxygen radicals. Therefore, tetracyclines may suppress neutrophil-mediated tissue damage by inhibiting their migration and degranulation and, potentially more importantly, by suppressing synthesis of oxygen radicals.(ABSTRACT TRUNCATED AT 250 WORDS)

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Human periodontal ligament-derived fibroblasts (HPLF) were grown to confluency in culture and were subjected to various combinations of neutrophils (PMNs), lipopolysaccharide (LPS) and the chemoattractant formylmethionyl-leucyl-phenylalanine (FMLP). After treatment, the cells were stained to distinguish between normal and damaged cells. The stain also allowed an estimation of PMN adherence to the HPLF monolayer. We report that FMLP, LPS or PMNs alone did not damage HPLF cells, nor did PMNs when combined with LPS or FMLP separately. However, PMNs subjected to combinations of LPS (10-1000 ng/ml) and FMLP (10(-9)-10(-6) M) caused significant PMN-mediated fibroblast damage. LPS concentrations greater than 1000 ng/ml inhibited the cytotoxic reaction. Furthermore, we found that FMLP alone did not significantly enhance PMN adherence to the HPLF monolayer but that LPS increased PMN adherence 3-fold and the combination of LPS and FMLP enhanced adherence 6-fold. We conclude that LPS promotes PMN adherence to fibroblasts and that such adherence appears to be a crucial, but insufficient stimulus, for the induction of PMN-mediated HPLF injury.

Material with endotoxin activity has been detected in extracts prepared from pooled, periodontally involved teeth, and it has been shown that root planing in vivo reduces the level of such material. However, questions concerning the concentration of endotoxin on the diseased surfaces of individual teeth and questions concerning how rapidly individual root planed tooth surfaces retoxify in vivo have not been addressed previously. Citric acid extracts were prepared from individual, periodontally diseased teeth that had been extracted either from the oral cavity without prior root planing or at varying times up to 12 weeks following root planing. Using a chromogenic Limulus Amebocyte Lysate (LAL) assay, we were able to quantitate the amount of endotoxin associated with diseased root surfaces of individual teeth. We concluded that the extracted material contained endotoxin since it activated LAL and since the LAL‐activation was heat‐stable, acid‐stable and neutralizeable by polymyxin B. The levels of endotoxin found on the root surfaces of these individual, periodontally involved teeth at varying times following in vivo root planing support the following conclusions: (1) the concentration of endotoxin present on diseased root surfaces is markedly reduced, but not eliminated, by in vivo root planing, (2) significant retoxification of root planed surfaces occurs within a relatively short time period after root planing and (3) biological responses to such toxification conceivably may lead to subsequent phases having reduced levels of endotoxin.

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