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PubMed1987

Immunocompetent cells in the normal dental pulp.

The existence and location of various immunocompetent cells in the human dental pulp were investigated. Pulp tissue for analysis was obtained both from clinically intact pre-molars and from third molars without restorations or caries. Frozen and acetone-fixed pulp tissue sections were subjected to indirect immunohistochemistry with monoclonal antibodies to the following cell types: all peripheral T cells, helper/inducer T cells, cytotoxic/suppressor T cells, macrophages, B cells, and Class II antigen-expressing cells. Dendritic cells expressing Class II antigens (HLA-DR, -DQ), indicating a capacity for presentation of antigen to T helper cells, were seen in the odontoblastic layer as well as in the central portions of the pulp tissue. T lymphocytes, divided into helper/inducer and cytotoxic/suppressor cells, were observed in all pulp specimens. B cells were not seen in any of the pulp samples examined. The data demonstrate that the human dental pulp is equipped with immunocompetent cells essential for the initiation of immunological responses.

Adolescent↗
PubMed1991

Flow cytometric analysis of human dental pulp tissue.

Existing knowledge regarding the cellular components of the dental pulp has been derived primarily from classical methods of histology and biochemistry. Since observations made from prepared tissue sections are static, it is not clear whether this accurately reflects the cellular dynamics of living pulp tissue. Therefore, we developed a method to analyze vital human pulpal tissue by flow cytometry. To test this method, two analyses of the prepared pulpal tissue were performed. First, the prepared tissue was stained with monoclonal antibodies to detect lymphocyte subpopulations. Second, the tissue was processed for DNA analysis of individual cells. Results demonstrated that lymphocytes bearing CD4 and CD8 antigens were clearly detected in pulpal tissue by this method. No B cells were found in any sample. DNA analysis revealed two distinct cell populations. Approximately 88% were small and 12% were large. According to DNA content, 90% of all cells were noncycling and 10% were cycling. These results demonstrate the feasibility of using flow cytometric analysis to examine, at a quantitative level, the cellular heterogeneity of the human dental pulp.

Antibodies, Monoclonal↗
PubMed1988

In vivo and in vitro effects of zinc oxide-eugenol (ZOE) on biosynthesis of cyclo-oxygenase products in rat dental pulp.

To determine the in vivo effects of a zinc oxide-eugenol mixture (ZOE) on the cyclo-oxygenase system in dental pulp, we used radioimmunoassay to measure the levels of prostaglandin E2 (PGE2), 13,14-dihydro-15-keto-PG (DHK-PG), thromboxane B2 (TXB2), and 6-keto-PGF1 alpha in the dental pulp of rats. When the dental pulp was irritated by a hole made in the dentin of the mandibular incisors without use of any coolants, the levels of these cyclo-oxygenase products in the pulp were increased to, respectively, 2.8, 1.7, 10.0, and 2.6 times those in the normal pulp at six hr after treatment. In contrast, these increases in cyclo-oxygenase products disappeared immediately when the artificial cavity in the dentin was filled with ZOE (P/L; 1 g/0.25 mL), but were not altered when the cavity was filled with zinc oxidewater (ZOW, 1 g/1.5 mL). Most of the eugenol portion of ZOE was released into the pulp within two hr after the cavity was filled with ZOE. The maximal eugenol content was 35 pmol per mg of pulp. Furthermore, when the cavity was filled either with ZOE or by the addition of 10 mumol/L eugenol to the pulp homogenate, biosynthesis of 14C-6-keto-PGF1 alpha, PGF2 alpha, and PGE2 from 14C-arachidonic acid in the homogenate was inhibited. These results suggest that eugenol released from ZOE in the cavity prepared in the dentin inhibited the biosynthesis of cyclo-oxygenase products during pulp irritation.

Animals↗
PubMed2004

Vascular endothelial growth factor, fibroblast growth factor 2, platelet derived growth factor and transforming growth factor beta released in human dental pulp following orthodontic force.

OBJECTIVE: The release of four diffusible angiogenic growth factors in human dental pulp following orthodontic force was investigated by using neutralising growth factor antibodies (NAs), individually and in four different combinations to block their effects. This study investigated if increasing the number of NAs (anti h vascular endothelial growth factor (VEGF), anti h fibroblast growth factor (FGF2), anti h platelet derived growth factor (PDGF) and anti Transforming growth factor beta (TGFbeta)) in combination resulted in a progressive reduction of the angiogenic response of the pulp. MATERIALS AND METHODS: The dental pulps from two groups of 40 premolar teeth, four teeth from each of 20 patients treated with fixed appliances for 2 weeks, were divided vertically, and sections from each half pulp co-cultured with sections of rat aorta in collagen. In one group, one of each of the four NAs, and in the other group, one of the four different NA combinations were added to the media of the co-cultures from one half of the pulp from each of the four teeth of each patient; the other half pulp co-cultures were controls. Cultures were examined daily by light microscopy for growth and number of microvessels. RESULTS: NAs significantly reduced microvessel numbers in the co-cultures when added individually (P<0.004), and in each of the four combinations (P<0.002), with a trend to progressively reduced microvessel numbers with increasing number of NAs in combination. CONCLUSIONS: Results indicated that all four angiogenic growth factors examined were released following orthodontic force application and play a role in the angiogenic response of the pulp, and that these factors may be more effective in combination.

Angiogenic Proteins↗
PubMed1992

Reaction of the dental pulp to hydroxyapatite.

The purpose of this study was to evaluate the action of hydroxyapatite (HA) (Osteogen HA Resorb, GBD Marketing Group Inc., Valley Stream, N.Y.) on the dental pulp of rats. Four upper molar pulps in 45 rats were exposed and capped with synthetic HA (Osteogen) with a stereoscopic microscope. Pulps capped with calcium hydroxide (Dycal, L.D. Caulk Co., Milford, Del.) served as controls. The cavities were filled with amalgam, and the molars on each side of the maxilla were protected by the placement of a pedodontic steel crown. Pulp inflammation and dentin repair were compared by histologic observations and computer image analysis after 7, 14, and 28 days. After 7 days a partial acute pulpitis were observed in specimens treated with Osteogen or Dycal. Reparative dentin formation along the pulp walls was also seen. After 14 days the pulpitis was more extensive in the Osteogen-treated teeth than in the control teeth. Dentin formation as measured by morphometric analysis was more pronounced in Osteogen-treated teeth. Neo-odontoblasts were observed after the use of both materials. After 28 days an acute inflammatory reaction was still evident in the Osteogen-treated group. A complete dentinal bridge was observed more frequently with Dycal than with Osteogen. Despite the putative abilities of HA to be osteoconductive, osteogenic, and dentinogenic, the results of this study indicate that it should not be used as a pulp-capping agent because of its tendency to cause scattered dystrophic calcification in the dental pulp, which could interfere with future endodontic treatment.

Animals↗
PubMed1997

Light-microscopical investigation of the distribution of extracellular matrix molecules and calcifications in human dental pulps of various ages.

The distribution of extracellular matrix molecules, especially collagen types I, III, V, and VI, in the extracellular matrix of the connective tissue of human dental pulp of various ages was studied by polarization and indirect immunofluorescence microscopy by using a conventional fluorescence microscope and a confocal laser scanning microscope. Polarization and immunofluorescence microscopy of paraffin sections showed thick fibers of collagen type I, which represented the main component of the connective tissue matrix of the dental pulp. By indirect immunofluorescence, thin fibers and small bundles of collagen type III were determined to be one of the main fibrillar elements present in the dental pulp matrix. Collagen type IV was detected by a clear intense staining of the basement membrane of blood vessels at all ages examined. Collagens type V and VI formed a dense meshwork of thin microfibrils throughout the stroma of the connective tissue of the dental pulp. These fibers were localized around blood vessels and appeared to be enriched in the subodontoblastic layer. Investigations by means of confocal laser scanning microscopy revealed fibers of collagen type VI spiralling between fully differentiated odontoblasts toward the predentin layer. With advancing age, the connective tissue matrix appeared to be condensed and aggregates of thick fiber bundles could be observed. Furthermore, the participation of various collagen types in the composition of pulp stones was shown. These calcifications and diffuse calcifications increased in frequency with advancing age in a statistically significant manner.

Adolescent↗
PubMed2002

Lymphatic networks in the periodontal tissue and dental pulp as revealed by histochemical study.

The structural organization and fine distribution of the lymphatic networks in the periodontal tissues (gingiva, periodontal membrane, and alveolar process) and dental pulp of animals and humans were reviewed with special reference to histochemical examination by light and electron microscopy. The distinction between lymphatics and blood vessels was made on cryostat sections of undecalcified and calcified teeth treated with EDTA solution and whole mount preparations of periodontal membranes using 5'-nucleotidase (5'-Nase)-alkaline phosphatase (ALPase) double staining. This staining procedure allowed lymphatic vessels in the periodontal tissue and dental pulp to be differentiated from blood vessels. The specificity and localization of the enzyme reactions were confirmed by comparative histochemical studies of the same specimen with light microscopy and scanning or transmission electron microscopy. Well-developed 5'-Nase-positive lymphatic networks were observed on the tissue sections and whole mount preparations of the gingiva, periodontium, and dental pulp. More lymphatic vessels were seen in the root area of the periodontium than in the cervical area. In the dental pulp, lymphatic vessels were more numerous in the central part than in the peripheral odontoblastic layer. These distributions of the lymphatic capillary networks are discussed in relation to their ability to supply lymph to the teeth.

5'-Nucleotidase↗
PubMed2002

Chelating resin-based extraction of DNA from dental pulp and sex determination from incinerated teeth with Y-chromosomal alphoid repeat and short tandem repeats.

A procedure utilizing Chelex 100, chelating resin, was adapted to extract DNA from dental pulp. The procedure was simple and rapid, involved no organic solvents, and did not require multiple tube transfers. The extraction of DNA from dental pulp using this method was as efficient, or more so, than using proteinase K and phenol-chloroform extraction. In this study, the Chelex method was used with amplification and typing at Y-chromosomal loci to determine the effects of temperature on the sex determination of the teeth. The extracted teeth were incinerated in a dental furnace for 2 minutes at 100 degrees C, 200 degrees C, 300 degrees C, 400 degrees C, and 500 degrees C. After the isolation of DNA from the dental pulp by the Chelex method, alphoid repeats, and short tandem repeats, the human Y chromosome (DYZ3), DYS19, SYS389, DYS390, and DYS393 could be amplified and typed in all samples incinerated at up to 300 degrees C for 2 minutes. The DYS389 locus in some samples could not be amplified at 300 degrees C for 2 minutes. An autopsy case is described in which genotypings of DYS19, DYS390, and DYS393 from dental pulp obtained from a burned body were needed. The data presented in this report suggest that Chelex 100-based DNA extraction, amplification, and typing are possible in burned teeth in forensic autopsy cases.

Chelating Agents↗
PubMed2000

Immunohistochemical localization of tenascin, fibronectin, and type III collagen in human dental pulp.

The distribution of tenascin (TN), fibronectin (FN), and type III collagen (col III) in the extracellular matrix of the connective tissue of normal, inflamed, and hyalinized human dental pulp was studied by immunohistochemical staining with monoclonal antibodies against these molecules. TN, FN, and col III were present in all normal tissues studied. In areas of hyalinization only col III was observed. None of the molecules studied were seen in areas of inflammatory exudate. Strong staining for TN and FN was found in the periphery of all specimens analyzed next to the odontoblastic layer. We therefore conclude that TN, FN, and col III are present in the extracellular matrix of normal human dental pulp. TN, FN, and col III distribution in inflammatory and degenerative processes is different from that observed in normal human dental pulp.

Antibodies, Monoclonal↗
PubMed2003

Substance P induces interleukin-8 secretion from human dental pulp cells.

OBJECTIVE: The role of neuropeptides in recruiting leukocytes in dental pulps is not known; therefore, we investigated whether interleukin-8 (IL-8) secretion from human pulp cells is increased after stimulation with substance P (SP) or calcitonin gene-related peptide (CGRP). METHODS: Primary pulp cells exhibiting a fibroblast-like phenotype and an endothelial cell line were stimulated with various doses of SP or CGRP, and IL-8 secretion was measured by enzyme-linked immunosorbent assays. RESULTS: IL-8 secretion from pulp cells increased significantly at 10(-8) to 10(-4) mol/L of SP stimulation (5- to 13.8-fold; P <.05); however, no significant IL-8 increase with CGRP (up to 10(-4) mol/L) stimulation was observed, nor was there synergistic induction of IL-8 with SP plus CGRP. The IL-8 increase reached its maximum at 8 hours after SP stimulation of the pulp cells. SP at a concentration of 10(-4) mol/L created minimal IL-8 induction in the endothelial cells and no synergistic induction by SP and CGRP. CONCLUSIONS: Pulp cells may up-regulate IL-8 secretion by SP stimulation, which suggests that SP released in dental pulp may play a role in the influx of leukocytes, attracted by IL-8, into the pulp tissue.

Adolescent↗
PubMed1991

Regulation of glycosaminoglycan synthesis by parathyroid hormone and prostaglandin E2 in cultured dental pulp cells.

The effects of parathyroid hormone (PTH) and prostaglandin E2 (PGE2) on glycosaminoglycan (GAG) synthesis in bovine dental pulp cells were studied. Dibutyryl cyclic adenosine 3',5'-monophosphate and isobutyl methylxanthine were used to assess whether their effects were mediated by intracellular cAMP. Glycosaminoglycan synthesis was assayed by measuring [35S]sulfate incorporation into the GAG fraction of dental pulp cells. Glycosaminoglycan synthesis was increased 1.3-fold by PTH (4 units per ml) alone, 1.6-fold by PTH in the presence of isobutyl methylxanthine, 1.2-fold by PGE2 (100 ng per ml) alone, and 1.5-fold by PGE2 in the presence of isobutyl methylxanthine. Dibutyryl cyclic adenosine 3',5'-monophosphate enhanced GAG synthesis in a concentration-dependent manner and mimicked the effects of PTH and PGE2. The effects of these hormones on pulp and gingival cells were compared; addition of PTH, PGE2, and dibutyryl cAMP had no effect on gingival cell GAG synthesis, whereas their addition induced significant increases of GAG in pulp cells. These results indicate that PTH and PGE2 are involved in the differentiation of dental pulp cells and that these effects are mediated by cAMP.

Animals↗
PubMed1998

Spherulitic brushite stone in the dental pulp of a cow.

Pulp stones of human teeth usually contain biological apatite. In the dental pulp of a cow, aged 12 years, we observed spherulitic pulp stones, up to approximately 200 microns in diameter, consisting of radially arranged crystals with several kinds of light microscopy and scanning electron microscopy including the secondary electron and the backscattered electron signals. The crystals extending fan-like towards the periphery showed a rectangular shape, and they had subunits of elongated crystallites and fine slits of NaOCl-dissolved substances, probably including organic matter and amorphous mineral. By using energy-dispersive X-ray microanalysis, large amounts of Ca and P, and trace elements of Na, Mg, Cl, and Fe were detected from the stones. The calcium phosphate crystals were identified as biological brushite by shape, Ca/ P molar ratio, and electron diffraction pattern. Core regions of the stones containing granular structures, probably erythrocytes, were always present in blood vessels, and rectangular crystals frequently penetrated the vessel walls. Under the decalcified sections, the nidi were completely or incompletely dissolved, but the peripheries showed a radial arrangement of erythrocytes. Thus, the nidi may be thrombus or necrotic blood cells including erythrocytes. Such calcospherulites in the dental pulp of a cow were similar to the spherulitic pulp stones in human deciduous teeth reported previously, however, their nidi were present in different regions.

Animals↗
PubMed2006

Human dental pulp cell culture and cell transplantation with an alginate scaffold.

Many studies on tissue stem cells have been conducted in the field of regenerative medicine, and some studies have indicated that cultured dental pulp mesenchymal cells secrete dentin matrix. In the present study we used alginate as a scaffold to transplant subcultured human dental pulp cells subcutaneously into the backs of nude mice. We found that when beta-glycerophosphate was added to the culture medium, dentin sialophosphoprotein mRNA coding dentin sialoprotein (DSP) was expressed. An increase in alkaline phosphatase, which is an early marker for odontoblast differentiation, was also demonstrated. At 6 weeks after implantation the subcutaneous formation of radio-opaque calcified bodies was observed in situ. Immunohistochemical and fine structure studies identified expression of type I collagen, type III collagen, and DSP in the mineralizing transplants. Isolated odontoblast-like cells initiated dentin-like hard tissue formation and scattered autolyzing apoptotic cells were also observed in the transplants. The study showed that subcultured dental pulp cells actively differentiate into odontoblast-like cells and induce calcification in an alginate scaffold.

Alginates↗
PubMed1986

Catecholamine contents of rabbit gingiva and dental pulp.

The contents of endogenous noradrenaline (NA) in rabbit gingiva and dental pulp were approximately 15-fold greater than either adrenaline (A) or dopamine (DA). Whereas catecholamine contents of maxillary incisor pulps were similar to those in mandibular incisor pulps, variations existed between gingiva excised from different regions. Both reserpine pretreatment and chronic sympathetic denervation greatly decreased the NA contents. These findings support the view that NA is the sympathetic neurotransmitter in dental pulp and gingiva.

Animals↗
PubMed2001

Streptococcus mutans lipoteichoic acid-induced apoptosis in cultured dental pulp cells from human deciduous teeth.

Herein, we suggest that Streptococcus mutans lipoteichoic acid-induced death of dental pulp cells on human deciduous teeth is caused by apoptosis. We provide evidence for the causal role of apoptosis in this process by demonstrating an increase in the proportion of fragmented DNA in such dental pulp cells, which results in a ladder pattern of DNA fragmentation. Additionally, Streptococcus mutans lipoteichoic acid-induced apoptotic cell death is suppressed by caspase inhibitor. Collectively, these findings suggest that Streptococcus mutans lipoteichoic acids may cause apoptosis in human dental pulp cells, and serve as an important factor in pulpitis.

Apoptosis↗
PubMed2003

Interleukin-10 receptor expression in human dental pulp cells in response to lipopolysaccharide from Prevotella intermedia.

Interleukin-10 receptor (IL-10R) expression in human, dental pulp, fibroblast cultures was investigated by using the reverse transcriptase-polymerase chain reaction (RT-PCR) technique. After exposure to lipopolysaccharide (LPS) from Prevotella intermedia, the IL-10R mRNA levels increased after 4 h, peaked at 7 h, and dropped back to the unstimulated level at 24 h. Maximal production of the IL-10R protein in dental pulp fibroblast cultures was detected by Western blot analysis after 12 h of LPS stimulation. In contrast, the human skin fibroblast (SF-MA) and human monocyte (U937) cell lines expressed IL-10R mRNA. Anti-CD14 antibodies inhibited P. intermedia LPS-induced IL-10R mRNA expression. These results indicate that P. intermedia LPS induces IL-10R gene expression in human, dental pulp fibroblasts in vitro.

Adolescent↗