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PubMed2004

[Characterization of Notch gene involved in genetic regulatory networks in dental pulp stem cells].

OBJECTIVE: To investigate the characterization of Notch gene involved in genetic regulatory networks in dental pulp stem cells. METHODS: The pulp tissue was separated from mouse teeth and digested by collagenase type I. Single-cell suspensions of dental pulp were seeded into 6-well plates with alpha modification of Eagle's medium supplemented with ES cell qualified Fetal Bovine Serum. Colony-forming efficiency was assessed in 14ds culture. Transcripts for Notch were detected by reverse transcription-PCR by using total RNA isolated from cells. RESULTS: There were clonogenic cells in dental pulp cell and the incidence of colony-forming cells derived from mouse dental pulp cells was 1.6-2.5 colonies/10(4) plate. Mouse-specific Notch mRNA expressed in colony-forming cells. CONCLUSION: Notch mRNA expressing in colony-forming cells provided a more detailed understanding of mouse dental pulp stem cell biology.

Animals↗
PubMed1986

Calcitonin gene related peptide: a sensory transmitter in dental pulps?

Nerve fibers displaying calcitonin gene related peptide (CGRP) immunoreactivity occurred in dental pulps of several mammals, including man. The CGRP fibers were more numerous in the apical parts than in the coronal parts and were distributed around small blood vessels as well as in the pulpal stroma without any obvious relation to blood vessels. The trigeminal, spinal and jugular-nodose ganglia harbored a moderate supply of CGRP immunoreactive perikarya and nerve fibers. Immunocytochemic double staining revealed the coexistence of CGRP and SP in a population of perikarya in the sensory ganglia and suggested coexistence of the two peptides in perivascular nerve fibers in the cat dental pulp. The cervical sympathetic ganglia did not contain CGRP-immunoreactive perikarya. Cervical sympathectomy (studied in the guinea-pig and rat) did not affect the frequency or distribution of pulpal CGRP fibers. The distribution of CGRP fibers within the dental pulp and the presence of CGRP perikarya in sensory ganglia known to supply the dental pulps indicate that the pulpal CGRP fibers are sensory in nature and that CGRP together with SP may participate in the regulation of local blood flow and the response to local inflammation.

Animals↗
PubMed2005

Expression of macrophage inflammatory protein 3alpha in human inflamed dental pulp tissue.

Severe pulpitis resulting from dental caries is characterized by marked inflammatory infiltrate such as lymphocytes. Little is known about the recruitment of these cells into the dental pulp lesions of carious teeth. Macrophage inflammatory protein-3alpha (MIP-3alpha), a CC chemokine attracts CC chemokine receptor 6 (CCR6)-expressing T cells. We examined the distribution of MIP-3alpha-positive and/or CCR6-positive cells in human inflamed and normal dental pulp by immunohistochemistry. MIP-3alpha was observed in all inflamed pulp sections, and was mostly distributed in macrophages that had accumulated in the area adjacent to carious lesions. Furthermore, CCR6 expression was also observed in the infiltrating lymphocytes. In contrast, MIP-3alpha and CCR6 were rarely detected in normal pulp. These findings suggest that MIP-3alpha plays a role in the advancement of pulpal inflammation via the recruitment of CCR6-expressing lymphocytes.

Adult↗
PubMed1981

[Structure and vascularization of the dental pulp in horse incisors].

The dental pulp of 34 horses aged between 4 months and 20 years has been studied on India ink injected and on paraffin embedded specimens. The pulp dimensions of the first and especially of the second dentition decrease with age. This reduction is accompanied by pronounced changes of structure; the mucous connective tissue becomes a dense, irregular connective tissue. The vessels are numerous and make a network particularly rich at the margins of the pulp; they diminish with advancing age. The reduction of the pulp volume and the regression of its vascular network are particularly pronounced at the crown level whose pulp cavity is progressively filled with secondary dentin. The regression of the vascular network precedes the reduction of the pulp volume.

Aging↗
PubMed1990

An ultrastructural study of the process of hard tissue formation in amputated dental pulp dressed with alpha-tricalcium phosphate.

This study was undertaken to determine the reaction of amputated dental pulp to the capping agent, alpha-tricalcium phosphate (alpha TCP), and to reveal the exact nature of the processes involved in both the healing of, and new hard tissue formation in the pulp when dressed with alpha TCP. The dental pulps of the anterior teeth of Macaca fuscata were dressed with alpha TCP ceramic 4-10 microns in diameter. The teeth were fixed by perfusion, then observed with light and electron microscopes. At 10 and 14 days postoperatively, the phagocytosis of the alpha TCP layer by the many macrophages and multinucleated giant cells, both of which had emerged beneath it, was seen. The synthesis of collagenous matrix underlying the alpha TCP layer began at 14 or 21 days, the cells involved exhibiting morphologic characteristics similar to those seen in osteoblasts. By the 90th day, only a bone-like hard tissue remained. No inflammation was observed in the residual dental pulp at any time during the experimental period. At 14 days, needle-like crystals were seen to have formed on the surface of alpha TCP particles, and by the 21st day, these crystals had extended to the newly synthesized matrix, with the result that calcification of the matrix was initiated, a calcified layer being superficially produced in the matrix. In the region of the matrix adjacent to the superficial calcified layer, a number of matrix vesicles were also found to function as another mechanism of matrix mineralization. Thus, it was concluded that the bone-like hard tissue was induced in the pulp capped with alpha TCP, and its potential for clinical application to exposed dental pulp was confirmed.

Animals↗
PubMed1984

Immunohistochemical localization of fibronectin in structures of the human oral cavity: dental pulp and gingiva.

The distribution of fibronectin (FN) in the dental pulp and gingiva of the human adult was investigated by indirect immunofluorescence and indirect immunoperoxidase techniques. FN could not be demonstrated in the connective tissue of the dental pulp even in the blood vessels. On the contrary, a positive stain was obtained in the ground substance at the base of the odontoblast cell bodies and in various structures of a denticle which was accidentally found in the pulp of one of the examined teeth. The lack of FN in the dental pulp could mean that this tissue, sometimes considered to be of an immature and undifferentiated type, is instead a mature one. The subepithelial connective tissue and the epithelium of the gingiva were examined. The distribution of FN in the connective tissue of the gingiva was found to be similar to that described in other types of connective tissue; high concentrations were found both in the epithelial cells and around them. This finding is described here for the first time and remains to be verified in other types of stratified epithelia.

Dental Pulp↗
PubMed1989

[Interaction between bradykinin and enkephalins in rat dental pulp].

The content of bradykinin (BK)-like peptides in rat dental pulp was significantly increased 1, 6 and 24 h after cavity formation at the neck of incisor. We have reported that enkephalin (EK)-like peptides in rat dental pulp were increased by cavity formation or BK. In the present study, the mechanism of the production of EK enhanced by BK was investigated using benzoyl-L-arginine-2-naphthylamide (BANA), a synthetic substrate. BK and its products cleft by carboxypeptidase B, des-Arg9-BK and arginine (Arg), activated the degradation of BANA. It is suggested that these substances may enhance the processing of enkephalins from precursor proteins. The activating effects were inhibited by EGTA. The BANA-degrading enzymes in lysosomal fraction were activated by BK, des-Arg9-BK and Arg, but the enzymes in supernatant were activated by Arg only. On the other hand, morphine and met-EK inhibited the production of BK-like peptides by trypsin from plasma kininogen. It is suggested that BK is cleft by carboxypeptidase B in pulp cell to des-Arg9-BK and Arg, which activate the lysosomal or soluble EK processing enzymes, and then the produced EK inhibits the production of BK from plasma kininogens in the pulp.

Animals↗
PubMed2004

Neuropeptides stimulate production of interleukin-1 beta, interleukin-6, and tumor necrosis factor-alpha in human dental pulp cells.

OBJECTIVE: Orthodontic tooth movement causes inflammatory reactions in the periodontal membrane and dental pulp. It has been reported that substance P (SP) and calcitonin gene-related peptide (CGRP), both sensory neuropeptides, are manifested in the dental pulp of rats during experimental tooth movement, suggesting that they might be involved in the dental pulp inflammation during orthodontic tooth movement. However, the relationships between neuropeptides and pro-inflammatory cytokines have not been fully elucidated. MATERIALS AND METHODS: Human dental pulp (HDP) fibroblasts were prepared from 6 healthy young volunteers (3 males, 3 females; 15-25 years old) during the course of orthodontic treatment. HDP cells were incubated for 24 h in fresh medium containing 2% FCS in the presence of various concentrations of CGRP (10(-12) to 10(-4) M) and SP (10(-12) to 10(-4) M), and the levels of interleukin (IL)-1 beta, IL-6, and tumor necrosis factor (TNF)-alpha present in the media were determined using commercially available high-sensitivity enzyme-linked immunosorbent assay kit. RESULTS: We examined the effects of stimulation by these neuropeptides on the production of inflammatory cytokines in HDP fibroblasts, and found that the levels of IL-1 beta, IL-6, and TNF-alpha increased in a time- and concentration-dependent manner. However, the neuropeptides did not act synergistically to increase cytokine secretion in HDP cells or significantly modify LPS-induced cytokine production by HDP cells. CONCLUSIONS: Our results suggest that human pulp fibroblasts may be involved in the progress of inflammation in pulp tissue during orthodontic tooth movement, as they produced large amounts of IL-1 beta, IL-6, and TNF-alpha following stimulation with neuropeptides.

Calcitonin Gene-Related Peptide↗
PubMed1991

[Aging of the dental pulp].

During life, the modifications of the dental pulp are important mostly in the root portion. This area, more than the coronal part of the tooth is submitted to involution and histologic dedifferentiation. Fibrosis and calcification occur leading to severe difficulty in performing endodontic treatment.

Adult↗
PubMed1995

Histological study of the circulatory system of human dental pulp from individuals under local anesthesia and electro-acupuncture.

A transmission electron microscopic (TEM) study was conducted on dental pulp obtained from patients under acupuncture or infiltration local analgesia. It was difficult to differentiate lymphatic circulation in the dental pulp that received infiltration anesthesia, because the vessels were constricted, congested, and showed stasis and thrombosis. On the other hand, the dental pulp that received acupuncture showed normal arterioles, capillaries, and venules, as well as some lymph capillaries and small efferent lymphatic vessels that measured about 8 microns and 100 microns in diameter, respectively. The lymphatic endothelial walls had many intercellular gaps, an imperfect basal lamina, and a few discontinuous pericytes. Between the openings in the lymphatic vessels, there were bundles of junctional filaments extending towards the dental pulp connective tissue. Therefore, the lymphatic system, which contains mainly B-3-alpha capillaries, is a leaky tissue for regulating fluid in the dental pulp.

Acupuncture Analgesia↗
PubMed2006

[Expression of lipopolysaccharides signal receptors CD14 and TLR4 in human dental pulp tissue and fibroblasts].

OBJECTIVE: To investigate the expression of lipopolysaccharides (LPS) receptors CD14 and TLR4 in normal and inflamed human dental pulp tissue and fibroblasts and to determine the signal transduction pathway of LPS in pulpitis. METHODS: CD14 and TLR4 expression in healthy and inflammatory pulps was observed by immunohistochemistry. The rates of CD14 and TLR4 positive cells and the mean fluorescence intensity in in vitro cultured pulp fibroblasts before and after being stimulated by LPS. RESULTS: CD14 and TLR4 were not detected in healthy pulp tissues, but they were strongly positively stained in inflammatory pulps. Ratio of TLR4 positive cells and mean fluorescence intensity produced by the cells stimulated by LPS increased significantly compared with normal pulp cells. However, there were no CD14 expression in human dental pulp cells before and after LPS stimulation. CONCLUSIONS: This study revealed a significant increase in the expression of CD14 and TLR4 in inflamed human dental pulp tissue. Human dental pulp cells could be induced by LPS to express TLR4, which may play an important role in the process of pulpitis.

Cells, Cultured↗
PubMed2005

Histological analysis of rat dental pulp tissue capped with propolis.

The aim of the present study was to assess the response of rat dental pulp to direct pulp capping with propolis. Flavonoid and non-flavonoid materials were purified from an ethanol extract of propolis obtained from South Sulawesi, Indonesia. A Class I cavity was prepared on the occlusal surface of the right maxillary first molar in Sprague Dawley rats. The dental pulp was exposed and then capped with a zinc oxide-based filler as a control (group I), or with propolis flavonoids (group II) or non-flavonoids (group III). The animals were sacrificed at week 1, 2 or 4, biopsy samples were obtained, and these were stained and viewed by light microscopy. The results showed that pulp inflammation occurred in groups I and III as early as week 1. No dentin bridge formation was seen in these groups. In contrast, there was no evident inflammatory response in group II at week 1. Mild and moderate pulp inflammation in this group occurred at 2 and 4 weeks after treatment, respectively. Partial dentinal bridge formation was seen in group II at week 4. Therefore, the present results suggest that direct pulp capping with propolis flavonoids in rats may delay dental pulp inflammation and stimulate reparative dentin.

Animals↗
PubMed1998

Integrin expression in human dental pulp cells and their role in cell attachment on extracellular matrix proteins.

Integrins are a family of heterodimeric glycoproteins consisting of alpha and beta subunits that noncovalently interact to form cell surface adhesion receptors. The objective of this study was to identify integrins in human dental pulp cells and determine their role in human dental pulp cell attachment to the biological active molecules, laminin and fibronectin. Integrin expression was studied by immunoblot and immunoprecipitation using monoclonal integrin antibodies. The role of integrin in human dental pulp cell adhesion on laminin and fibronectin was determined by inhibition of cell adhesion with those antibodies. This study found human dental pulp cells expressed alpha 1, alpha 3, alpha 5, alpha 6, alpha v, and beta 1 integrin subunits. The adhesion of human dental pulp cells to laminin and fibronectin was not inhibited by monoclonal antibody to any subunit, except that anti-beta 1 antibody inhibited pulp cells adhesion on laminin. These data provide information for further studying the role of integrins in dental pulp cell biological function.

Cell Adhesion↗
PubMed1982

Biosynthesis of glycoproteins by rabbit dental pulp fibroblasts in culture.

Confluent dental pulp fibroblast cultures incorporated L-[3H]-fucose in a linear manner with time into non-diffusible macromolecules over 24 h. Ascorbate supplementation did not appear to alter the amount or type of macromolecules. Equilibrium CsCl-density-gradient centrifugation established that the [3H]-fucose-labelled macromolecules released into the medium were predominantly glycoproteins. Sodium dodecyl sulphate-polyacrylamide-gel electrophoresis showed that the major fucosylated glycoprotein had an apparent molecular weight of 230,000, but several other species were also seen. The major fucosylated glycoprotein was fibronectin by its affinity for gelatin and its immunoprecipitation with a specific anti-(cold-insoluble globulin).

Animals↗
PubMed1990

Properties of alkaline phosphatase of the human dental pulp.

Enzymatic and immunological properties of alkaline phosphatase [ALP; orthophosphoric monoester phosphohydrolase, alkaline optimum, EC 3.1.3.1.] in the human dental pulp were investigated. In inhibition and thermal inactivation studies, dental pulp ALP showed properties of universal-type ALP (kidney/bone/liver type). Dental pulp ALP cross-reacted with polyclonal and monoclonal antibodies against purified swine-kidney ALP, and with monoclonal antibody against ALP of human osteoblast-like cells in the same manner as ALPs of human bone and kidney. The sodium dodecyl sulfate-gel electrophoretic pattern showed a 140,000-Mr native protein band. These data suggest that dental pulp ALP can be classified as a universal-type ALP having antigenic determinants common to ALP of the kidney and bone.

Alkaline Phosphatase↗
PubMed1983

The chromatographic and electrophoretic micro-heterogeneity of alkaline phosphatase of rat dental pulp.

Alkaline phosphatase of rat dental pulp was separated into two forms by anion-exchange chromatography, gel-filtration and electrophoresis. The minor activity (AP I) was eluted at 0.1 M of NaCl from a DEAE-cellulose column, and the major activity (AP II) in the broad range of approx. 0.15-0.5 M. These two enzyme activities gave rise to distinct peaks on a TSK-GEL Toyopearl (Fractogel TSK) HW 55-S column and showed different electrophoretic behaviours on both sodium dodecyl sulphate (SDS) and non-SDS polyacrylamide gels. The molecular weights of AP I and AP II were estimated by gel-filtration to be 130,000-150,000 and above 500,000, respectively. On SDS gel electrophoresis, the molecular weight of AP II was changed to 130,000, while AP I had a molecular weight of 150,000. Judging from the sensitivities to heat and some inhibitors, AP I and AP II are biochemically indistinguishable from each other.

Alkaline Phosphatase↗