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PubMed2006

[Isolation, cultivation and initial identification of Nanobacteria from dental pulp stone].

OBJECTIVE: To isolate Nanobacteria from dental pulp stone and perform culturing and the identification of Nanobacteria. METHODS: Freshly collected 27 dental pulp stones were divided into nine samples. Each sample contained three dental pulp stones. All samples were used for the isolation and culture of Nanobacteria. The shape and the growth characteristics of the cultured bacteria were observed. Nanobacteria were identified by von Kossa staining, immunohistochemical staining and indirect immunofluorescence staining, double staining including Hoechst staining and von Kossa staining. RESULTS: The characteristics growth and morphology of the bacteria detected in seven samples were similar to Nanobacteria. von Kossa staining, immunohistochemical staining, indirect immunofluorescent staining were positive for Nanobacteria. In double staining method, Hoechst staining of the samples was negative for Nanobacteria, but von Kossa staining was positive. Hoechst staining of the dental pulp cells was positive. No Nanobacteria was found in the other two samples. CONCLUSIONS: The bacteria isolated from dental pulp stone in this study was similar to Nanobacteria in terms of growth rate, morphology and staining properties. These unusual properties of the bacteria may play an important role in the formation of pulp stone.

Bacteria↗
PubMed1991

Age-related changes of the dental pulp complex and their relationship to systemic aging.

The dental pulp and its associated structures, the dentin and the cementum, are discussed. Because many of the age-related pulpal changes have components considered in several of the current theories of aging, these theories are briefly reviewed. Part 2 describes the age-related changes of the dental pulp, the dentin, and the cementum (the dental pulp complex). An attempt is made to differentiate inherent aging changes from physiologic defensive changes and pathologic irritant-induced changes. Part 3 describes the relationship of age-induced changes in the dental pulp complex to components of the current aging theories together with a unified concept of the dental pulp complex aging. Part 4 considers whether dental pulp complex aging can be used as a biomarker for generalized aging. Whether age-related changes of the dental pulp complex can be altered by interventions is discussed.

Aging↗
PubMed2000

Defence reaction in dental pulp after pulp capping and partial pulpectomy in dogs.

The dental pulp was capped indirectly or directly, or partial vital pulpectomy was performed on the 12 functionally most important teeth of 24 beagle dogs. For pulp capping, calcium hydroxide was used, followed by zinc phosphate as a lining, and the preparation was restored with amalgam or composite material. Histological sections were prepared and examined for degree and type of pulp inflammation (hyperaemia, pulpitis, necrosis or gangrene). Degenerative changes of dental pulp (vacuolation, calcification, amyloid or hyaline changes) were also determined. For indirect pulp capping the width of predentine in crown and root dental pulp was measured, and for direct pulp capping and partial pulpectomy (vital pulpotomy) the width and quality of the dentinal bridge were graded. The radical method of partial vital pulpectomy of the coronal part of the dental pulp in dogs produced better quality and continuity of the dentinal bridge than the less radical method of direct pulp capping. This was particularly obvious in small single root teeth such as incisors.

Animals↗
PubMed2000

Identification of G-protein coupled receptor subunits in normal human dental pulp.

To respond appropriately to their environment, dental pulp cells must integrate informational input from multiple ligands, such as neuropeptides, growth factors, and vasoactive amines. These ligands act through multiple intracellular signaling pathways. G-protein coupled receptor subunits play a major role in this process, providing a mechanism for coordinated regulation of both messengers and effectors. Increasing number of neuropeptides have been found in pulpal tissue. However, there is no data about molecular identification of G-protein subunits in human dental pulp. To identify the postreceptor mechanism involved in dental pulp cell signal transduction, we performed a Western blot analysis of different G-protein subunits. Biopsy specimens of human dental pulp were prepared and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by analysis with appropriate antibodies. We detected G alpha q/alpha 11, short and long forms of G alpha s, beta common, Gio-3, and Gil-2 antigens with a molecular weight approximately 42 kDa, 42 and 45 kDa, 36 kDa, 40 kDa, and 40 kDa, respectively. These results indicate that human pulp cells possess the cellular machinery to respond to sensory neuropeptides when they are released from the peptidergic nerve fibers. On this basis, the relationships of postdevelopmental, age-dependent, and pathophysiological disorders of G-proteins subunits in dental pulp could be studied.

Dental Pulp↗
PubMed1996

Distribution and possible origin of galanin-like immunoreactive nerve fibers in the mammalian dental pulp.

Human, dog, cat and rat dental pulps were investigated for the presence and distribution of galanin-like immunoreactive (-IR) nerve fibers, and the possible origin of pulpal galanin-IR nerve fibers in the rat was examined. Galanin-IR nerve fibers were present in the dental pulps of all species examined. Two types of galanin-IR nerve fibers were distinguished with regard to morphology; thin varicose nerve fibers and thick smooth-surfaced nerve fibers. Thin varicose galanin-IR nerve fibers were seen to run along the blood vessel in the human, dog and cat root pulp. In the coronal pulp, galanin-IR nerve fibers ran toward the odontoblastic layer but they did not form the subodontoblastic nerve plexus. In rat molar pulp, few galanin-IR nerve fibers were observed; the distribution of these nerve fibers was similar to those in human, dog and cat pulp. In contrast, many thick smooth-surfaced galanin-IR nerve fibers were observed near the blood vessels in incisor pulp of the rat; occasionally a few varicose galanin-IR nerve fibers were also observed. Transection of the inferior alveolar nerve or mandibular nerve caused complete disappearance of galanin-IR nerve fibers in rat dental pulp, while surgical sympathectomy of the superior cervical ganglion did not affect their distribution. The present results indicate that galanin-IR nerve fibers are present in the mammalian dental pulp, and that the intrapulpal galanin-IR nerve fibers in the rat originate from the trigeminal ganglion and are primary afferents.

Animals↗
PubMedDate not supplied

[Biochemical study of the dental pulp in the calf].

A biochemical study of dental pulp of calves has been performed concerning: a) peroxydability b) A, E, C vitamins content c) glutation (GSH) content d) presence of paramagnetic compounds e) phosphorylation ratio The dental pulp from incisors of 5-months-old calves has been preserved. Immediately after decapitation the pulp was immersed in liquid nitrogen. Chromatographic (HPLC) and spectroscopic (NMR-ESR) techniques have been used. GSH in dental pulp are present and dosable (4.56 +/- 0.08 n moles/mg prot.) and GSSG (1.05 +/- 0.01 n moles/mg prot.). Because of blood traces in the extracted pulps, the AA. have determined the hemoglobin (Hb) dosage and GSH of erythrocytic derivation (Fig. 1). After deduction of GSH of erythrocytic derivation, the GSH really present in the pulp was 4.41 n moles/mg prot. and the GSSG was 0.90 n moles/mg prot. Peroxydability of the dental pulp has been evaluated with Lowry method with dental pulp homogenate and rat liver homogenate (see Table 1). The ESR spectre shows 4 resonances with the following values: g. 2.24-2.04-2.00-1.97; there are some free intermediary radicals (gr.-2.00) (Fig.2). The NMR spectre shows the presence of ATP (0.22 n moles/g) of inorganic phosphate (16.58 n moles g) (Fig.3). The pulp seems to have a lot of antioxidant factors. The next researches will be to study E, A and C vitamins concentrations. This high presence of GSH and GSSG may be an embryonic peculiarity.

Aged↗
PubMed2006

The existence of CD11c+ sentinel and F4/80+ interstitial dendritic cells in dental pulp and their dynamics and functional properties.

Dental caries and pulpitis are the most common bacterial infections in humans. However, the immune responses against bacterial stimulation in dental pulp that is bounded by special hard tissues are poorly understood. We examined the initial immune responses in mouse dental pulp after cusp trimming and acid treatment. Using fluorescence immunohistochemistry, two distinct cell populations were identified in the intact pulp; CD11c+F4/80- and CD11c-F4/80+ cells. CD11c+F4/80- cells were localized in the pulp-dentin (P-D) border of the central pulp beneath the dental fissure, whereas CD11c-F4/80+ cells with dendritic morphology were distributed in the perivascular region of the inner pulp and the sub-odontoblastic layer. CD11c+F4/80- cells, but not CD11c-F4/80+ cells, constitutively expressed toll-like receptors 2 and 4 and CD205, and migrated to the P-D border of the treated side within 2 h after the treatment. In parallel, some of the F4/80+ cells migrated to the inner pulp of the treated side, increased in size and enhanced CD86 expression. At 24 h, the CD86+ cells with high fluorescence intensity had disappeared entirely from the pulp. Concurrently, CD86high cells expressing intermediate levels of CD11c and high levels of MHC class II and F4/80, assessed by using flow cytometry, increased significantly in the regional lymph nodes, suggesting migration of these cells from the dental pulp. Our results are the first to demonstrate the existence of at least two types of dendritic cells (DCs) in dental pulp. The CD11c+ sentinel and F4/80+ interstitial DCs might have distinct territories and unique roles in responding to external stimuli via the dentinal tubules.

Animals↗
PubMed2000

A small-conductance Ca(2+)-activated K+ current and Cl- current in rat dental pulp cells.

We characterized a voltage-dependent ionic current in dental pulp cells on dental pulp slices using a nystatin perforated-patch recording configuration. The outward currents in dental pulp cells were inhibited by the following channel blockers: 1) Ca(2+)-free extracellular solution containing 10 mM Ba2+, 2) extracellular 400 nM apamin and 3) extracellular 300 nM 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS). On the other hand, 15 mM tetraethylammonium (TEA) did not inhibit the outward currents. The inhibitory effects of Ca(2+)-free extracellular solution, apamin and DIDS had voltage-dependency. These results indicated that dental pulp cells expressed a small-conductance Ca(2+)-activated K+ current (SK current) and a DIDS-sensitive Cl- current. The functional significance of these channels is discussed.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed2006

Capacity of dental pulp differentiation after tooth transplantation.

Under pathological conditions, dental pulp elaborates both bone and dentin matrix in which the contribution of periodontal tissue cannot be excluded. This study has aimed to clarify the capability of dental pulp to deposit bone matrix in an auto-graft experiment by using (1) immunohistochemistry for 5-bromo-2'-deoxyuridine (BrdU) and nestin and (2) histochemistry for tartrate-resistant acid phosphatase (TRAP). Following the extraction of the molars of 3-week-old mice, the roots and pulp floor were resected and immediately transplanted into the sublingual region. On Days 5-7, tubular dentin formation commenced next to the pre-existing dentin at the pulp horn in which nestin-positive odontoblast-like cells were arranged. Up until Day 14, bone-like tissue formation occurred in the pulp chamber in which intense TRAP-positive cells appeared. These results suggest that odontoblast- and osteoblast-lineage cells reside in the dental pulp. Overall, specific dental pulp regeneration should provide fundamental knowledge for the realization of human tooth regeneration in the near future.

Acid Phosphatase↗
PubMed2000

Detection of Coxiella burnetii DNA in dental pulp during experimental bacteremia.

Colonization of dental pulp by blood-borne bacteria in the absence of previous inflammation has been hypothetized but has never been convincingly demonstrated. In order to provide convincing support for this hypothesis we attempted to detect Coxiella burnetii DNA in the dental pulp of bacteremic, intraperitoneally inoculated guinea-pigs by PCR amplification and direct sequencing of two molecular targets. Coxiella burnetii DNA was recovered from 20-50% of the animals depending on the molecular target, from 15-20 days after experimental challenge. These results demonstrated, for the first time, that dental pulp is contaminated by blood-borne bacteria and can be detected by molecular tools.

Animals↗
PubMed1984

Identification of bacteria in immunopathologic mechanisms of human dental pulp.

Serial frozen sections from human dental pulp were used for the identification of oral bacteria in immunopathologic mechanisms. Sera raised against Actinomyces viscosus, A. naeslundii, Bacteroides gingivalis, and B. melaninogenicus ss. intermedius, commercial sera against human immunoglobulins, complement, and monoclonal antibodies against human T cells were used in a double-staining immunofluorescence technique. Sections of dental pulp from normal teeth showed no penetration of bacteria or bacterial antigens and no signs of inflammation. A unique aspect of the present study was the demonstration that penetrating bacteria and bacterial antigens in the pulp of involved teeth were always associated with antibodies and frequently also with complement. A. viscosus has been found most frequently in complement-fixing immune complexes followed by B. gingivalis. A. naeslundii and B. melaninogenicus ss. intermedius were found only in complexes with antibodies. The involvement of plasma cells and T cells was also demonstrated. In the dental pulps of diseased teeth, cytotoxic and Arthus type immunopathologic reactions occurred.

Actinomyces↗
PubMed2001

A voltage-dependent transient K(+) current in rat dental pulp cells.

We characterized a voltage-dependent transient K(+) current in dental pulp fibroblasts on dental pulp slice preparations by using a nystatin perforated-patch recording configuration. The mean resting membrane potential of dental pulp fibroblasts was -53 mV. Depolarizing voltage steps to +60 mV from a holding potential of -80 mV evoked transient outward currents that are activated rapidly and subsequently inactivated during pulses. The activation threshold of the transient outward current was -40 mV. The reversal potential of the current closely followed the K(+) equilibrium potential, indicating that the current was selective for K(+). The steady-state inactivation of the peak outward K(+) currents described by a Boltzmann function with half-inactivation occurred at -47 mV. The K(+) current exhibited rapid activation, and the time to peak amplitude of the current was dependent on the membrane potentials. The inactivation process of the current was well fitted with a single exponential function, and the current exhibited slow inactivating kinetics (the time constants of decay ranged from 353 ms at -20 mV to 217 ms at +60 mV). The K(+) current was sensitive to intracellular Cs(+) and to extracellular 4-aminopyridine in a concentration-dependent manner, but it was not sensitive to tetraethylammonium, mast cell degranulating peptide, and dendrotoxin-I. The blood depressing substance-I failed to block the K(+) current. These results indicated that dental pulp fibroblasts expressed a slow-inactivating transient K(+) current.

4-Aminopyridine↗
PubMed1983

The concentration, extractability and characterization of collagen in human dental pulp.

Premolar and third molar dental pulps were studied. The amount of collagen in the dried pulps was 25.7 per cent in premolars and 31.9 per cent in third molars. These percentages are much higher than those reported for pulps in other species. Significant differences were further found in the collagen content and cell distribution (DNA) of the coronal, middle and apical parts of the pulp. Collagen content was the lowest in the coronal part, while the cell content was the lowest in the middle part. The extractability of collagen in a neutral salt solution or 0.5 M acetic acid was found to be extremely low (less than 1 per cent). Pretreatment of the pulp with hyaluronidase in order to remove proteoglycans had no effect on the solubility. It is concluded that human pulp collagen is highly cross-linked and cannot be considered as immature. Characterization of collagen was performed by methods in which limited pepsin digestion or CNBr cleavage was used. The digests were analysed by means of quantitative electrophoresis which revealed an amount of 42.6 per cent type III of the total collagen. Because of the large differences between dental pulps from man and experimental animals, extreme caution should be exercised in drawing conclusions from data of other species to explain phenomena observed in human teeth.

Adolescent↗
PubMed1983

Fluoride-resistant acid phosphatase (FRAP) activity of nociceptive nerve terminals in the dental pulp.

Nerve fibers in the dental pulp of the lower molar teeth of the rat exert fluoride resistant acid phosphatase (FRAP) activity. FRAP-positive axons establish a three-dimensional nerve plexus within the pulp; the individual axons are very fine (calibre less than 1 micrometer) and only their varicosities measure 1...2 micrometer in diameter. Electron microscopically, FRAP-positive amyelinate axons containing lysosomes are partly embedded in Schwann cells. Removal of the cervical superior ganglion does not induce any alteration of FRAP-positive axons, while destruction of the Gasserian ganglion results in Wallerian degeneration. No FRAP-positive nerve fibers were found in rat incisors. Since, in the rat, only molar teeth are equipped with nociceptive terminals while continuously growing incisors lack pain fibers, it is concluded that FRAP-positive varicose axons in the dental pulp represent nerve endings of trigeminal primary nociceptive neurons.

Acid Phosphatase↗
PubMed2003

Effect of fluoride on human dental pulp cells in vitro.

Human dental pulp cells were cultured in fluoride containing medium of various concentrations (0, 1, 2, 5, 10, 20, 25, 30, 40, 50, 60 and 80 ppm) in order to study the biological effect on the cells' proliferation and alkaline phosphatase (ALP) activities. It was found that fluoride at 5 ppm concentration significantly stimulated cell proliferation and ALP activity between 24 and 48 hours after exposure whereas at higher concentrations (40 - 80 ppm), fluoride significantly inhibited cell growth and ALP activity after 48 hours (Student's t test). The maximum effect was around 80 ppm. These observations suggest that fluoride, if used at a low concentration, may be a useful therapeutic agent for the treatment of pulpal disease by means of stimulating the proliferation and differentiation of dental pulp cells. At higher concentrations, it will have negative effects on this kind of cell.

Adolescent↗