Ageing-related effects of dye penetration in the apical 5 mm mesiobuccal and mesial roots of molars.

This study examined how the tubular permeability in the apical 5 mm of mesial roots was affected by age, direction, distance from root apex, canal shape and canal number. Twenty mesiobuccal/mesial roots of maxillary/mandibular molars from three age groups (<30, 30-60, >60 years) were prepared. Maximal dye penetration in buccal, lingual, mesial and distal directions was measured at 1-mm segment depth and analysed statistically. The <30 years group, when comparing to the >60 years group, had significantly deeper dye penetration in buccal and lingual directions. The permeability in proximal directions was not affected by either age or canal number. In the >60 years group, neither direction, segment depth nor canal number significantly impacted the permeability. However, these parameters significantly impacted samples younger than 60 years. Effective removal of infected root canal dentine would depend on age and its related preferred direction, distance from root apex and canal number.

An In Vitro Model to Study the Colonization and Tubular Invasion of Enterococcus faecalis.

INTRODUCTION: The purpose of this study was to examine the colonization and tubular invasion of Enterococcus faecalis in minimally altered canal walls. We hypothesized that age, axial directions (buccolingual or mesiodistal), regions (apical, midroot, or cervical), and disinfection treatment (sodium hypochlorite or autoclave) would impact bacterial colonization and invasion patterns. METHODS: Single roots of extracted teeth from 2 age groups (≤30 years old and ≥60 years old) were challenged with bacteria for 2 weeks. Colonization on the canal walls in 3 regions was evaluated with scanning electron microscopy. The prevalence of tubular invasion in the axial directions in 3 regions was examined with confocal laser scanning microscopy. The data were analyzed using the GLIMMIX procedure in SAS software (SAS Institute Inc, Cary, NC) with a Tukey adjustment for comparisons. RESULTS: Bacteria successfully colonized and invaded tubules in 2 weeks. The apical region in the ≥60-year age group was the least infected. A significantly higher invasion prevalence in a buccolingual (B/L) than a mesiodistal (M/D) direction was detected in both age groups and in all 3 regions. The ≤30-year age group had a significantly higher prevalence of tubular invasion than the ≥60-year age group in the B/L and M/D direction, respectively. Sodium hypochlorite treatment significantly impacted bacterial colonization and invasion in more calcified areas. CONCLUSIONS: Our data support a more conservative enlargement of the apical region of older teeth. An emphasis in a B/L direction rather than an M/D direction is recommended for debridement.

Mature dendritic cells in inflamed human pulps beneath deep caries.

OBJECTIVE: Dendritic cells (DCs) in pulps have been identified with markers for immature DCs and the relationship of DC maturity to pulpal health has not been carefully examined. We sought to test the hypothesis that the frequency of CD83+ mature DCs would correlate with caries invasion. STUDY DESIGN: Pulps were collected from extracted teeth exhibiting (I) no caries (n = 9), (II) shallow dentinal caries (n = 5), and (III) deep caries (n = 9). Immature DCs (CD209+), mature DCs (CD83+), and monocytes/macrophages (CD14+) in three groups were enumerated immunohistochemically. RESULTS: Mature DCs were frequently found beneath deep caries (21.3 mature DCs/3 grids versus <1 in groups I and II) with increasing numbers of CD209+ DCs and CD14+ cells. Co-localization of CD4+ T cells with mature DCs and macrophages was observed in deep caries. CONCLUSION: Mature DCs were frequently found only beneath deep caries and these DCs were co-localized with CD4+ T cells suggesting antigen presentation.

Update on the adaptive immune responses of the dental pulp.

Recent advances in immunology have disclosed the enormous complexity of the immune regulatory system. The dental pulp is equipped to mount adaptive immune responses to caries, which include at least antigen-presenting cells, lymphocytes, mast cells and their cytokines, and chemokines. The purpose of this review is to summarize our current understanding of the roles of these cellular and molecular components in the irreversibly inflamed pulp. The immunopathology of abscess formation and the mechanisms for painless pulpitis are also discussed.

Innate immune responses of the dental pulp to caries.

Various cells and inflammatory mediators are involved in the initial pulpal responses to caries. This review focuses on the cellular, neuronal, and vascular components of pulpal innate responses to caries. Discussion will include dentinal fluid, odontoblasts, neuropeptides, and neurogenic inflammation, which are not classic immune components but actively participate in the inflammatory response as the caries progress pulpally. Summaries of innate immune cells as well as their cytokines and chemokines in healthy and reversible pulpitis tissues are presented.

Relationships between caries bacteria, host responses, and clinical signs and symptoms of pulpitis.

Knowledge of caries bacteria and the inflammatory responses they elicit in the dental pulp is prerequisite to our understanding of the pathogenesis of pulpitis. Recent advances in immunology and neurophysiology can now explain some of the clinical manifestations of pulpitis. The purpose of this review is twofold. The first purpose is to review the literature of the caries microflora, the host immune responses they elicit, and how they do so. The relationship between both proinflammatory and anti-inflammatory cytokines and pulpitis is discussed. The proinflammatory properties of lipoteichoic acid, which is a common virulence factor among Gram-positive bacteria such as those found among the caries bacteria, are reviewed. The second purpose is to review how bacteria and their metabolites, as well as pulpal immune and inflammatory reactions to them, modify the pain sensation in pulpitis.

The antimicrobial effect of MTAD, sodium hypochlorite, doxycycline, and citric acid on Enterococcus faecalis.

This study compared the antimicrobial effect of MTAD, two of its components, doxycycline and citric acid, and sodium hypochlorite (NaOCl) in two in vitro models on Enterococcus faecalis. In the bovine tooth model, the lumens of 30 bovine dentin discs were infected with E. faecalis for 2 weeks before treating with either one of the experimental irrigants or saline. Bacteria in the shavings were collected with two sizes of burs and enumerated after overnight culturing. Zones of inhibition were recorded in the agar diffusion model for each irrigant. In the tooth model, NaOCl and doxycycline were more effective than control in killing E. faecalis at the shallow bur depth, but at the deeper bur depth only NaOCl was superior. In the agar diffusion model, NaOCl produced less inhibition than MTAD or doxycycline.

The pulpal origin of immunoglobulins in dentin beneath caries: an immunohistochemical study.

Immunoglobulins localized in uninfected dentin beneath caries are thought to be protective, but their origin remains controversial. We reasoned that the localization and dominance of serum IgG1 would support the pulpal origin of the immunoglobulins while a predominance of secretory component (SC) bearing IgA1 and IgA2 would support their salivary origin. The prevalence and staining intensity of IgG1, IgA1, IgA2, IgM, and SC in uninfected dentinal tubules beneath shallow, deep caries, and noncaries teeth were examined immunohistologically. SC was only localized in caries, and IgG1 was the predominant subclass in uninfected dentinal tubules beneath shallow and deep caries, followed by IgA1. In noncaries teeth, IgG1 was localized on the pulpal end. The intensity of IgG1 was significantly higher than either IgA1 or IgA2 in both shallow and deep caries. Our data support the serum origin of immunoglobulins in uninfected dentin beneath caries.

Endocarditis-associated oral streptococci promote rapid differentiation of monocytes into mature dendritic cells.

Endocarditis is frequently attributable to oral streptococci, but mechanisms of pathogenesis are not well understood, although monocytes appear to be important. High titers of interleukin-12 (IL-12) are produced by peripheral blood mononuclear cells (PBMC) after engaging Streptococcus mutans, but monocytes in developing endocardial vegetations tend to disappear rather than become macrophages. These data prompted the hypothesis that streptococcus-infected monocytes differentiate into short-lived IL-12-producing dendritic cells (DCs) rather than macrophages. PBMC from healthy subjects were stimulated with six isolates of oral streptococci, three nonstreptococcal oral bacteria, or IL-4 plus granulocyte-macrophage colony-stimulating factor, and the appearance of cells with markers typical of mature DCs (CD83(+), CD86(+), CD11c(+), and CD14(-)) was monitored. Supernatant fluids from the PBMC cultures were harvested and IL-12 p70 levels were determined. S. mutans-stimulated monocytes were analyzed for their ability to elicit allogeneic mixed-lymphocyte reactions. All streptococci examined, except one strain of Streptococcus oralis (35037), rapidly induced up-regulation of CD83 and CD86 and a loss of CD14 in the CD11c(+) monocyte population within 20 h. Induction of IL-12 was CD14 dependent and correlated with streptococcal isolates that promoted the DC phenotype. Major histocompatibility complex (MHC) class II expression was up-regulated by S. mutans, and these cells were short-lived and elicited potent allogeneic mixed-lymphocyte reactions typical of DCs. In summary, monocytes stimulated with endocarditis-associated oral streptococci rapidly exhibited the DC phenotype and functions. These data suggest that the initiation of bacterial endocarditis by oral streptococci may involve monocyte-to-DC differentiation, and this may help explain the low levels of macrophages in the site.

Comparison of type 1 and type 2 cytokine production by mononuclear cells cultured with streptococcus mutans and selected other caries bacteria.

A feature of pulpal immune responses is the predominance of type 1 cytokine mRNA under shallow caries and a mixed (type 1/type 2) profile under deep caries. These results prompted an examination of the cytokine profiles induced by bacteria in shallow caries (Streptococcus mutans and Actinomyces viscosus) and deep caries (Lactobacillus casei, Pseudoramibacter alactolyticus, and Prevotella intermedia). All isolates induced interferon-gamma and interleukin-10, whereas interleukin-4 and interleukin-2 titers were low to undetectable. S. mutans was the most potent and persistent interferon-gamma inducer. Differences in interleukin-10 were apparent at low doses but were less dramatic, with L. casei the dominant producer. S. mutans induced substantially more interferon-gamma than interleukin-10 over all doses and time points, suggesting strong type 1 polarization. P. alactolyticus induced significantly more interleukin-10 than interferon-gamma at higher concentrations, suggesting polarization toward type 2. A similar amount of interferon-gamma and interleukin-10 induced by L. casei, A. viscosus, and P. intermedia reflected a mixed profile. A better understanding of pulpal immune response to caries bacteria may enable us to develop an immune system-based pulp therapy in the future.