Smoking and periodontal disease: discrimination of antibody responses to pathogenic and commensal oral bacteria.

Smoking is an independent risk factor for the initiation, extent and severity of periodontal disease. This study examined the ability of the host immune system to discriminate commensal oral bacteria from pathogens at mucosal surfaces, i.e. oral cavity. Serum immunoglobulin (Ig)G antibody reactive with three pathogenic and five commensal oral bacteria in 301 current smokers (age range 21-66 years) were examined by enzyme-linked immunosorbent assay. Clinical features of periodontal health were used as measures of periodontitis. Antibody to the pathogens and salivary cotinine levels were related positively to disease severity; however, the antibody levels were best described by the clinical disease unrelated to the amount of smoking. The data showed a greater immune response to pathogens than commensals that was related specifically to disease extent, and most noted in black males. Significant correlations in individual patient responses to the pathogens and commensals were lost with an increasing extent of periodontitis and serum antibody to the pathogens. Antibody to Porphyromonas gingivalis was particularly distinct with respect to the discriminatory nature of the immune responses in recognizing the pathogens. Antibody responses to selected pathogenic and commensal oral microorganisms differed among racial groups and genders. The antibody response to the pathogens was related to disease severity. The level of antibody to the pathogens, and in particular P. gingivalis, was correlated with disease severity in black and male subsets of patients. The amount of smoking did not appear to impact directly serum antibody levels to these oral bacteria.

Dental treatment of Papillon-Lefèvre syndrome: 15-year follow-up.

A 9-year-old girl was initially treated for the periodontal component of Papillon-Lefèvre syndrome by extraction of all patient's erupted teeth, after unsuccessful clinical treatment with two different antibiotics. Follow-up dental records at age 24 showed the patient to have generalized gingivitis and poor oral hygiene; however, no additional teeth were lost or mobile. Radiographically, the alveolar crests, lamina dura, and periodontal ligament spaces appeared normal for a subject with missing teeth. Initially, the patient had depressed polymorphonuclear leukocyte (PMN) chemotaxis and adherence, as well as evidence of periodontal infection with Actinobacillus actinomycetemcomitans, (A.a.). The 6 and 15-year follow-ups showed normal PMN function and no detectable A.a. The improvement of the patient's PMN function was coincident with lack of detection of certain periodontopathic bacteria. If the PMN dysfunction of PLS is secondary to the infection, the reasons for the initiation of the disease still need to be clarified.

Colonization of retrieved polytetrafluoroethylene membranes: morphological and microbiological observations.

Polytetrafluoroethylene membranes (ePTFE) used in guided tissue regeneration (GTR) are accessible to colonization by oral bacteria. The bacterial composition of the adherent biomass is unknown. We examined a total of 6 membranes that were retrieved after 4 to 6 weeks from human periodontitis sites, using optical and transmission electron microscopy (TEM) as well anaerobic cultivation. Five of the 6 membranes provided the microbiological data and microscopic data. TEM revealed an organized microbial mass covering the surfaces and also within the interstices of the open microstructure and occlusive portions of the membranes. Numerous bacterial forms including cocci, rods, and filaments with an interbacterial matrix, frequently in microcolonies, were identified. Anaerobic cultivation yielded Streptococcus and Actinomyces species with a minor component of Gram-negative facultative rods comprised mainly of Haemophilus species. Candida species was recovered from one membrane. These data show that ePTFE is heavily colonized by oral bacteria during retention. The impact of bacterial colonization of ePTFE is not known but it seems reasonable to assume that colonization of membranes may affect connective tissue regeneration. Further studies will be needed to examine the effect of systemic antimicrobials on ePTFE colonization and in turn to examine the effect on GTR.

Prostaglandin E release from human monocytes treated with lipopolysaccharides isolated from Bacteroides intermedius and Salmonella typhimurium: potentiation by gamma interferon.

The purpose of this investigation was to examine gamma interferon potentiation of lipopolysaccharide (LPS) responses in human monocytes by using phenol-water-extracted (unfractionated) and highly purified LPS preparations isolated from Bacteroides intermedius and Salmonella typhimurium. Phenol-water-extracted LPS preparations from these bacteria were further purified by chromatography over Sepharose-CL-4B. LPS enrichment in pooled column fractions was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and quantitation of hydroxy-fatty acid and 2-keto-3-deoxyoctulosonic acid content, protein contamination, and anthrone-reactive material. Monocyte stimulation by LPS, measured as prostaglandin E (PGE) release, was assessed with and without gamma interferon treatment. Cells were either treated simultaneously with gamma interferon and LPS or pretreated with gamma interferon prior to LPS stimulation. PGE release from counterflow-isolated monocytes was quantitated during the 0- to 24-h and 24- to 48-h culture intervals. Contrary to previous results from this laboratory, phenol-water-extracted LPS preparations from B. intermedius and S. typhimurium were similar in their capacities to stimulate PGE release from monocytes. Molecular sieve chromatography was found to remove substantial amounts of high-molecular-weight polysaccharide contaminants only from the B. intermedius LPS but did not significantly alter the potency of either B. intermedius or S. typhimurium LPS. Gamma interferon cotreatment did not potentiate the release of PGE with any of the LPS preparations tested. However, 24-h pretreatment of monocytes with gamma interferon followed by a 24-h exposure to LPS resulted in significant potentiation of PGE release over LPS alone. In addition, B. intermedium preparations were approximately threefold more potent than similarly prepared LPS isolates from S. typhimurium following gamma interferon pretreatment. These results indicate that gamma interferon can selectively potentiate the effects of B. intermedius LPS in human monocyte isolates.

Use of adhesin-specific monoclonal antibodies to identify and localize an adhesin on the surface of Capnocytophaga gingivalis DR2001.

Monoclonal antibodies capable of inhibiting coaggregation between Capnocytophaga gingivalis DR2001 and Actinomyces israelii PK16 were used to identify the adhesin on C. gingivalis that mediates the interaction. The monoclonal antibodies were used to demonstrate that a 140-kilodalton polypeptide found in the outer membrane of C. gingivalis was the adhesin responsible for coaggregation. A coaggregation-defective mutant that was unable to coaggregate with A. israelii lacked this large polypeptide. The monoclonal antibodies were also used to estimate the number of binding sites on the surfaces of individual cells and show how the adhesin molecules were arranged on the outer membrane. Values of between 220 and 280 were obtained for the number of adhesin molecules per cell. Immunoelectron microscopy performed with the monoclonal antibodies revealed that the adhesin molecules were arranged nonuniformly on the bacterial surface and occurred singly, in pairs, and in small clusters.

Selective medium for the isolation of Haemophilus aphrophilus from the human periodontium and other oral sites and the low proportion of the organism in the oral flora.

We developed a medium for the selective recovery of Haemophilus aphrophilus. The medium, designated TSBVF, was composed of 4% tryptic soy agar, 10% heat-inactivated horse serum, 75 micrograms of bacitracin per ml, 5 micrograms of vancomycin per ml, and 50 micrograms of sodium fluoride per ml. TSBVF yielded a threefold higher recovery of oral H. aphrophilus than did chocolate agar with 75 micrograms of bacitracin per ml, which is a medium routinely used to diagnose human Haemophilus infections. H. aphrophilus and the few contaminating organisms on TSBVF were readily distinguished on the basis of colony morphology. The H. aphrophilus isolates exhibited variable fermentation of raffinose and dextrin but otherwise were biochemically similar. In a clinical study, H. aphrophilus was frequently recovered from supragingival plaque and saliva and occasionally from buccal mucosa and the tonsils. It was also isolated from 29 of 56 subgingival sites in 11 of 14 subjects. Its proportion of the subgingival microflora averaged 0.13% for healthy periodontal sites, 0.05% for adult periodontitis lesions, and 0.03% for localized juvenile periodontitis lesions. We concluded that H. aphrophilus is an indigenous bacterium of the human oral cavity. It occurs in low proportions in subgingival plaque and plays no apparent role in advanced periodontal disease in humans.