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Notes: Abstract: The aim of the in vitro study was to examine the accumulation of Streptococcus mutans on light-cured composite materials and amalgam. Bacteria cultures were grown in a brain heart infusion medium, and their growth rate was determined through turbidity measurements. The data, so obtained, were evaluated statistically by analysis of variance (ANOVA) followed by Scheffe test. Experiments on amalgam showed better results compared to those on composite materials. There were no statistically significant differences in plaque accumulation on different composite materials after finishing and polishing procedures, compared to plaque accumulation on composite materials against a Mylar strip.

Notes: Adults (35–45 yr old) and children (5 and 12 yr old) recently arrived in Israel from rural areas in Ethiopia were examined. Caries levels were low: e.g. DMFT = 0.31 among 12-yr-olds olds, and 1.27 amongst adults. The 5-yr-olds were 86.8% caries-free, while 12-yr-olds were 81.8% and adults 54% caries-Tree respectively. According to interview data, the diet in Ethiopia had been based on local agricultural products and was almost sugar-free. The mean total count of salivary bacteria, as determined on blood agar, was 3.4 × 108; mean count of Streptococcus viridans. on mitis salivarius, was 6.7 × 107; and mean count of S. mutans. as determined on mitis salivarius with bacitracin, was 1.7 × 107. These levels were all high and were not significantly different from a control group of 20 Israelis. The mean number of lactobacilli, on Rogosa agar, was 2.75 × 104, which was significantly higher than among the controls (3.6 × 103). Salivary pH levels were generally similar between the Ethiopian group and the controls. Salivary How was significantly higher for the Ethiopians (1.93 ml/min) than for controls (1.16 ml/min). Low levels of caries in this population can be attributed to an almost sugar-free diet and high salivary flow, but not to the composition of oral microflora.

Notes: This study was carried out to examine the release kinetics of chlorhexidine from a sustained release device (S. R. D.) prepared from ethyl cellulose (fast S. R. D.) or ethyl cellulose with polyethylene glycol (slow S. R. D.) and to examine the effects on the bacterial flora of pockets in patients with periodontal disease.It was shown that fast S. R. D.'s release up to 80% of the chlorhexidine within the first 3 days in insertion in periodontal pockets, whereas the slow S. R. D.'s release 50% of the chlorhexidine content after 6 days. The release kinetics of chlorhexidine from S. R. D.'s placed in pockets as expressed by the Higuchi system (Higuchi 1963) indicate that it is diffusion controlled. The rate of chlorhexidine release is dependent on the structure of S. R. D., the drag concentration within the device, and the effective surface area.The microbial flora of sixteen pockets from 6 patients were examined using darkfield microscopy at day 0, 3, 10, and 14 after treatment with S. R. D.'s containing, chlorhexidine or placebo S. R. D.'s. The pocket depths ranged from 5–8 mm. The chlorhexidine-treated group showed a marked decrease in the relative proportions of motile rods and spirochetes and a corresponding increase in non-motile organisms compared to the flora prior to chlorhexidine treatment or compared to the flora of the placebo treated pockets. These differences were significant up to 10 days post treatment (P 〈 0.0025).The study indicated the effectiveness of ethyl cellulose polymers as S. R. D.'s in vivo and their ability to reduce the relative proportions of the motile organisms of periodontal pockets to negligible amounts.

Notes: Guided tissue regeneration (GTR) techniques are increasingly used for the treatment of periodontal defects, or in conjunction with dental implant procedures. As adhesion of bacteria to barrier membranes used in these techniques may lead to failure, a prerequisite for treatment success is an infection-free healing process. The present study examined the adhesion of 3 periodontal pathogenic bacteria: Actinobacillus actinomycetemcomitans, Treponema denticolu and Porphyromonas gingivalis, to 3 barrier membranes: Collagen, (Biomend™) e-PTFE, (TefGen-FD™) and e-PTFE, (Gore-Tex®). The membranes were incubated with 3[H]-thymidine labeled bacteria, and the number of adherent bacteria was calculated using a scintillation counter. The effect of albumin coating on bacterial adherence to the membranes was also studied. Bacterial adherence to the membranes was further examined by scanning electron microscopy (SEM). The results show that the adherence of all bacterial strains to collagen membranes was significantly higher than to the other membranes tested. Precoating of the membranes with albumin did not change the bacterial adherence significantly. These findings are of importance in evaluating the ability of periodontal bacteria to colonize and infect different types of barrier membranes.

Notes: Abstract Previous studies have indicated that in certain types of chronic inflammatory periodontal diseases, polymorphonuclear leucocytes (PMN) functions are impaired. In view of the damage oxygen-free radicals may cause to the periodontal tissues, the present study focussed on superoxide (SO) formation and luminol-dependent chemiluminescence (LDCL) by peripheral PMN cells in rapidly progressive periodontitis patients (RPP). PMN cell preparations were stimulated by either opsonized bacteria or phorbol myristate acetate (PMA). The results indicate that PMN cells from RPP patients, stimulated by opsonized bacteria, have significantly enhanced SO formation and LDCL response as compared to healthy subjects. The hyperactivity was cell-associated. In the presence of PMA, no significant differences were detected between the groups. The results suggest that PMN cells from RPP patients are functionally activated, and produce elevated levels of oxygen radicals. These oxygen radicals may play a role in the pathogenesis of RPP.

Notes: Abstract: Bacterial infection in the vicinity of guided tissue regeneration barrier membranes was shown to have a negative effect on the clinical outcomes of this increasingly used technique. Several oral and specifically periodontal bacteria were shown to adhere to such membranes in vivo and in vitro with a higher affinity to membranes constructed from collagen. The present study examined the role of periodontal bacteria and their enzymes in the degradation of commercially used collagen membranes. Degradation of two collagen membranes [BiomendTM (Calcitek®, Colla-Tec Inc., Plainsboro, NJ) and Bio-GideTM (Geistlich Biomaterials, Wolhousen, Switzerland)] labeled by fluorescein isothiocyanate was examined by measuring soluble fluorescence. Porphyromonas gingivalis, Treponema denticola and Actinobacillus actinomycetemcomitans and their enzymes were evaluated. Collagenase from Clostridium hystolyticum was used as a positive control. While whole cells of P. gingivalis were able to degrade both types of membranes, T. denticola could degrade Bio-Gide membranes only and A. actinomycetemcomitans whole cells could degrade none of the membranes. Fractionation of P. gingivalis cells revealed that cell membrane associated proteases were responsible for the degradation of the two collagen membranes. In T. denticola, the purified major phenylalanine protease was found to be responsible for the degradation of Bio-Gide membranes. These results suggest that proteolytic bacterial enzymes may take part in the degradation of collagen barrier membranes used for guided tissue regeneration.

Notes: Abstract Previous studies have shown that a 3-day exposure of the pocket flora to the sustained release of chlorhexidine significantly reduced the relative numbers of spirochetes and motile rods in periodontal pockets to negligible amounts. By 14 days post-treatment, their numbers had returned to pre-treatment levels. The present study extended the exposure time of the pocket flora to the sustatined release of chlorhexidine in an attempt to prolong the suppression of the microbial flora for a clinically significant period of time. Clinical parameters were also studied. Sustained release devices (SRD) were inserted into 13 pockets from 8 patients. Pocket depth ranged between 5 and 8 mm. The SRD's were replaced every 3 days to give a total exposure of 9 days. Plaque index (P1I). bleeding on probing and pocket depth were measured, and bacterial samples taken for dark field microscopy and anaerobic culture.There was a marked decrease in the relative proportions of spirochetes and motile rods and the total anaerobic count post-treatment. Pocket depth was reduced in all 13 pockets. These results indicate that a prolonged exposure to chlorhexidine suppresses the pocket flora to negligible amounts and reduces pocket depth for up to 11 weeks post-treatment.

Notes: In this study, the adhesion of radioactively labeled Actinomyces viscosus (A.viscous), Actinobacillus actinomicetemcomitans (Aa) and Porphyomonas gingivalis (P gingivalis) to titanium (Ti) and Ti6Al4V alloy (Ti-alloy) coated with albumin or human saliva was investigated. All the tested bacteria displayed greater attachment to Ti-alloy than to Ti. P. gingivalis exhibited less adhesion to Ti and Ti-alloy than did the other bacterial strains. Adhesion of A. viscosus and Aa was greatly reduced when Ti or Ti-alloy were coated with albumin or saliva. P. gingivalis demonstrated a lesser reduction in adhesion to albumin or saliva-coated surfaces. The results show that oral bacteria have different adhesion ffinities for Ti and Ti-alloy and that both albumin and human saliva reduce bacterial adhesion.

Notes: The 160, 190 and 270 kDa outer sheath proteases of Treponema denticola ATCC 35404 were found to be multiple forms of the major 91 kDa phenylalanine protease (PAP) by immunoblotting using anti-91kDa specific antibodies. Multiple forms of the phenylalanine protease were also found in 2 other T. denticola strains studied, ATCC 33520 and the clinical isolate GM-1. Protein, proteolytic and Western blot analyses using antibodies against the PAP and the major outer sheath protein (MSP) indicated that the 190 and 270 kDa proteases were protein complexes formed by the MSP and the PAP. These complexes dissociated by storage in 0.3% or higher SDS concentrations. The purified PAP was found to completely degrade keratin, but was unable to degrade native actin either in its monomeric or polymerized form. The association of the MSP adhesin with a protease capable of degrading host native proteins may benefit the obtention of protein-based nutrients necessary to support the growth of these treponemes. These complexes may also play a role in the structural organization of T. denticola outer sheath.

Notes: Abstract A variety of cationic polyelectrolytes opsonized group A streptococci andCandida albicans to phagocytosis by human polymorphonuclear leukocytes and by mouse peritoneal macrophages. The most potent opsonins for streptococci were specific antibodies supplemented with complement, nuclear histone, polylysine, polyarginine, ribonuclease, leukocyte lysates, leukocyte cationic protein and, to a lesser extent, lysozyme and myeloperoxidase. Histone, RNAse, leukocyte extracts, and platelet extracts also functioned as opsonins for phagocytosis of streptococci in the peritoneal cavity, where phagocytic indices, higher than those obtained for the in vitro phagocytosis, were obtained. Fresh serum, polylysine, polyarginine, and nuclear histone acted as good opsonins forCandida, but none of the other factors tested were active. In order for the cationic proteins and leukocyte extracts to function as opsonins, they must be present on the particle surface. These agents were poor opsonins when applied on the macrophages. Nuclear histone, polylysine, polyarginine, and fresh human serum also functioned as good opsonins for the uptake ofCandida by mouse fibroblasts. On the other hand, none of the other substances which opsonized streptococci were effective withCandida. The phagocytic capabilities of fibroblast polykaryons were much higher than those of ordinary spindle-shaped mouse fibroblasts. Histone also functioned as a good opsonic agent for the uptake ofCandida by human fibroblasts, HeLa cells, epithelial cells, monkey kidney cells, and rat heart cells. On the other hand, neither leukocyte extracts nor ribonuclease LCP or MPO functioned as opsonins for these mammalian cells.Candida, taken up by fibroblasts, were present within tight phagosomes, but no fusion of lysosomes with the phagosome occurred. A small proportion of the internalized yeast cells underwent partial plasmolysis, but little damage to the rigid cell walls was observed within 24–48 h of internalization. Phagocytosis of streptococci andCandida by macrophages and the uptake ofCandida by fibroblasts were both strongly inhibited by liquoid (polyanethole sulfonic acid sodium salt). This anionic polyelectrolyte also markedly inhibited the release ofN-acetylglucosaminidase from macrophages without affecting cell viability (LDH release). Hyaluronic acid, DNA, and dextran sulfate markedly inhibited the uptake of histone-coated particles by macrophages. On the other hand, hyaluronic acid and DNA enhanced the uptake ofCandida by fibroblasts. The effect of these anionic polyelectrolytes on phagocytosis of serum-opsonized particles by macrophages was not consistent. While in some experiments it blocked phagocytosis, in others it either had no effect or even enhanced the uptake of the particles. Phagocytosis of microorganisms by “nonprofessional” phagocytes like fibroblasts and the paucity in these cells of hydrolases capable of breaking down microbial cell wall components may contribute to the persistence of non-biodegradable components of bacteria in tissues and to the perpetuation of chronic inflammatory sequellae. Cationic polyelectrolytes may also prove important as “helper” opsonins and as agents capable of enhancing the penetration into cells of both viable and nonviable particles, genetic material, and drugs.