The effects of endodontic irrigants and calcium hydroxide on lipopolysaccharide (LPS; endotoxin) were analyzed using the highly selective technique of mass spectrometry/gas chromatography with selected ion monitoring. An aqueous solution of LPS was mixed with one of a variety of endodontic irrigants for 30 min. Because it is a commonly used interappointment dressing, calcium hydroxide was also applied to LPS for 1, 2, or 5 days. LPS inactivation was measured by quantitation of free fatty acid release. Water, EDTA, ethanol, 0.12% chlorhexidine, chlorhexidine + sodium hypochlorite, and sodium hypochlorite alone showed little breakdown of LPS. Long-term calcium hydroxide--as well as 30-min exposure to an alkaline mixture of chlorhexidine, ethanol, and sodium hypochlorite--did detoxify LPS molecules by hydrolysis of ester bonds in the fatty acid chains of the lipid A moiety.

Bacteria from infected root canals can invade dentinal tubules, thus dentin disinfection is an important aspect of endodontic therapy. This study compares three endodontic irrigants for efficiency in killing bacteria established within human dentinal tubules. Root canals in extracted teeth were prepared and sterilized. Broth cultures of Enterococcus faecalis were allowed to grow within the canals to penetrate dentinal tubules. The infected canals were exposed individually to each of the irrigants for 1 min. Irrigants were 0.525% sodium hypochlorite, Tubulicid (0.2% EDTA), and 0.12% chlorhexidine (Peridex). Sterile water was the control. Viable bacteria were analyzed by drilling incrementally into dentin from the cementum toward the canal. Smaller diameter drills were used for each depth. Shavings were cultured at three depths, for each of three root levels: coronal, midroot, and apical. Although considerable variation occurred between roots, sodium hypochlorite seemed to be superior. Tubulicid and Peridex were better than water. More bacteria remained viable at greater distances from the pulp. These observations apparently apply to all levels in the canal.

The adherence of Streptococcus sanguis to hydroxylapatite beads has been analyzed by binding isotherms, Langmuir isotherms, and Scatchard plots. For saliva-coated beads, the Scatchard curves contained components with both positive and negative slopes. The results are interpreted as evidence for positive cooperativity in the binding process. Although all Scatchard curves were similar in shape, distinct differences were observed between saliva samples from different individuals. Salivary agglutinins against whole S. sanguis cells did not appear to influence the shapes of the curves or the extent of adherence. In addition, different strains of S. sanguis yielded similar Scatchard plots. When the binding of S. sanguis to buffer-coated hydroxylapatite beads was analyzed by Scatchard plots or binding isotherms, curves were generated which suggested that either direct ligand-ligand or nonspecific interactions were occurring. Hill plots of the adherence data yielded curves with slopes greater than unity for saliva-coated beads, providing additional support for the view that the interactions between S. sanguis and the pellicle involve cooperative phenomena. In contrast, a Hill plot for the binding data of S. sanguis to buffer-coated hydroxylapatite beads gave a curve with a slope of 0.91 ± 0.07, suggesting negative cooperativity or limited specificity. When adherence data were plotted by the Langmuir method, curves were obtained which could not discriminate between the binding of the bacteria to the hydroxylapatite beads coated with either saliva or buffer. It was also observed that several different proteins and whole saliva tended to inhibit adherence. Scatchard plots, however, describing the binding of S. sanguis to the proteincoated beads were unique and revealed possible specific and nonspecific interactions. Scatchard analyses of binding data may be useful in understanding the mechanism(s) of adherence of streptococci to smooth surfaces.Streptococcus sanguis can be observed in early dental plaque and comprises a significant portion of the oral microbiota found on the tooth surface (3,13,15). The mechanism(s) involved in the attachment of S. sanguis to the salivary pellicle which coats the tooth surface remains obscure. Little is known about the structural requirements of pellicle protein receptors for the bacterial ligands. In fact, several groups report that oral streptococci, including Streptococcus mutans as well as S. sanguis, can bind to hydroxylapatite in the absence of any pellicleforming salivary proteins (4,18).In addition to the difficulties in purifying and characterizing specific bacterial surface components and salivary proteins which interact to form stable complexes, there have been only a few efforts to describe the quantitative relationships in adherence phenomena. Clark et al. (4), Gibbons et al. (10), and Appelbaum et al. (1)have employed the Lapgmuir adsorption isotherm to characterize the quantitative aspects of the adherence of oral streptococci to smooth surfaces. Similarly, Wheeler et al. (27...

Adherence of Streptococcus mutans to smooth surfaces has been attributed to the production of sucrose-derived d -glucans. However, several studies indicate that the bacterium will adhere in the absence of sucrose. The present data confirmed that S. mutans adherence to saliva-coated hydroxyapatite beads in the absence of sucrose is described by the Langmuir equation. The nature of the sucrose-independent adherence was studied with the Persea americana agglutinin as a selective adherence inhibitor. Pretreatment of the bacterium with P. americana agglutinin caused a 10-fold reduction in adherence, and the inhibition was not reversed with the addition of sucrose. Pretreatment of S. mutans with proteases also reduced adherence, regardless of the sucrose content, whereas periodate oxidation and glucanohydrolase treatment of the bacteria reduced sucrose-mediated adherence to the levels found for sucrose-independent adherence. The P. americana agglutinin, glucanohydrolase, and pepsin pretreatment of the cells did not eliminate sucrose-induced agglutination. Scanning electron microscopy showed that short streptococcal chains were bound to saliva-coated hydroxyapatite crystals in the sucrose-independent system, whereas the presence of sucrose caused larger bacterial clumps to be found. A two-reaction model of S. mutans adherence was developed from these data. It is proposed that one reaction is attachment to the tooth pellicle which is mediated by cell-surface proteins rather than glucans or teichoic acids. The other reaction is cellular accumulation mediated by sucrose-derived d -glucans and cell surface lectins. A series of sequential adherence experiments with P. americana agglutinin as a selective inhibitor provided presumptive evidence for the validity of our model of S. mutans adherence.