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Dental pulp fibroblasts contain target cells for lysophosphatidic Acid.

Lysophosphatidic acid (LPA) is a locally produced bioactive phospholipid which is involved in tissue repair. The objective of this study was to determine whether dental pulp tissue also responds to the phospholipid. Effects of LPA on proliferation, differentiation, and mitogen-activated protein kinase (MAPK) signaling of dental pulp fibroblasts (DPF) were examined in vitro. We report that DPF express LPA receptors LPA1, LPA2, and LPA3 and respond to the ligand with increased mitogenic activity. Involvement of extracellular signal-regulated kinase, p38 MAPK, and c-Jun NH(2)-terminal kinase in LPA signaling could be demonstrated by use of specific inhibitors and detection of the phosphorylation status of the kinases. An increased mitogenic activity paralleled a decreased number of alkaline-phosphatase-positive cells and expression levels of dentin sialophosphoprotein and osteocalcin. Together, these results suggest that dental pulp fibroblasts can respond to LPA, a process that may play a role in pulp tissue repair.

Adult↗

The effect of collagen and indomethacin on inflamed dental pulp wounds of baboon teeth.

Following the induction of experimental pulpitis in baboon teeth, calf skin collagen, indomethacin, or zinc oxide-eugenol was used to dress exposed dental pulp wounds in selected teeth. The teeth were extracted after 7, 35, and 90 days fixed in formalin, and processed, for histopathologic examination. The inflammatory response to zinc oxide-eugenol remained moderate throughout the experiment, whereas the inflammatory response to indomethacin and collagen was present from the outset and became progressively more severe, especially with collagen. The amount of irritation dentin production, however, was greater in those dental pulps dressed with collagen and indomethacin.

Animals↗

[Incorporation of 3H-cortisol into the dental pulp and the induced changes].

The incorporation of labelled hydrocortisone (3H-cortisol) in the dental pulp of Wistar rats has been tested using autoradiography. Light microscopy showed that an hour after injection the hormone is incorporated in the nuclei of the odontoblasts, subodontoblastic and endothelial cells of the dental pulp. It was also demonstrated at the ultrastructural level that a 4 week-treatment with high rates of hydrocortisone (16 and 32 mg/kg) at the time of rat intensive growth induces degeneration of the odontoblasts, pulp connective tissue cells and capillaries. High rates of hydrocortisone thus induces a decrease in the protective and reparative responses of rat molar pulp.

Animals↗

Dentin regeneration by dental pulp stem cell therapy with recombinant human bone morphogenetic protein 2.

Regenerative medicine is based on stem cells, signals, and scaffolds. Dental pulp tissue has the potential to regenerate dentin in response to noxious stimuli, such as caries. The progenitor/stem cells are responsible for this regeneration. Thus, stem cell therapy has considerable promise in dentin regeneration. Culture of porcine pulp cells, as a three-dimensional pellet, promoted odontoblast differentiation compared with monolayers. The expression of dentin sialophosphoprotein (Dspp) and enamelysin/matrix metalloproteinase 20 (MMP20) mRNA confirmed the differentiation of pulp cells into odontoblasts and was stimulated by the morphogenetic signal, bone morphogenetic protein 2 (BMP2). Based on the in vitro experiments, an in vivo evaluation of pulp progenitor/stem cells in the dog was performed. The autogenous transplantation of the BMP2-treated pellet culture onto the amputated pulp stimulated reparative dentin formation. In conclusion, BMP2 can direct pulp progenitor/stem cell differentiation into odontoblasts and result in dentin formation.

Animals↗

Immunohistochemical observation on neuropeptides around the blood vessel in feline dental pulp.

Recent advances in immunohistochemistry have revealed the distribution of various neuropeptides in several mammalian dental pulps. These neuropeptides are substance P, neurokinin A, calcitonin gene-related peptide, vasoactive intestinal polypeptide, and neuropeptide Y. Nerve fibers showing immunoreactivity for these neuropeptides are mainly localized around the pulpal vessels, but some are apart from the blood vessel. The distribution of substance P-, neurokinin A-, and calcitonin gene-related peptide-containing nerve fibers is very similar; it appears that these three neuropeptides may be contained in the same nerve fibers. Denervation experiments indicate that substance P-, neurokinin A- and calcitonin gene-related peptide-containing nerve fibers originate from the trigeminal ganglion and that neuropeptide Y-containing nerve fibers come from the superior cervical ganglion. However, the exact origin of vasoactive intestinal polypeptide-containing fibers still remains unknown. These immunohistochemical data indicate that neuropeptides localized around the blood vessel may play a significant role in the regulation of the blood flow in the dental pulp.

Animals↗

Effects of deprivation of neonatal nerve growth factor on the expression of neurotrophin receptors and brain-derived neurotrophic factor by dental pulp afferents of the adult rat.

The dental pulp is richly innervated by peptidergic nociceptive neurones that are of special interest because of their central role in dental pain and because they have some features that are not typical of other somatic nociceptors. Here, (35)S-riboprobes were used to determine whether pulpal afferents of adult (2-month-old) rats express the nerve growth-factor (NGF) receptors, p75(NTR) and trkA, which are characteristic of peptidergic nociceptors, and additionally, whether these cells express receptors (trkB and trkC) for other members of the neurotrophin family. In order to begin characterizing the postnatal role of NGF in regulating these neurones, the susceptibility of pulpal afferents to antiserum-mediated early postnatal NGF depletion spanning the period of pulpal innervation development was also examined. In control animals, about 200 trigeminal ganglion cells were labelled after application of the retrograde tracer Fluoro-gold to the first maxillary molar. Among the labelled cells, 79% had positive hybridization signals for p75(NTR), 72% for trkA, 34% for trkB, 1% for trkC, and 77% for BDNF. Neonatal NGF depletion reduced the number of retrogradely labelled pulpal afferents by 33%, with numbers of smaller neurones being most strikingly subnormal. This reduction could be attributed to a partial depletion of the neurone population that expressed p75(NTR) and trkA. Consistent with reports that NGF-responsive neurones also express BDNF, NGF deprivation resulted in a reduction in the number of pulpal afferents that expressed BDNF to an extent similar to that seen for trkA. In contrast, anti-NGF exposure had little effect on the number of pulpal afferents that expressed trkB. These findings indicate that most pulpal afferents in the adult express the NGF receptors p75(NTR) and trkA, and thus have a continuing potential susceptibility to NGF-mediated regulation of functions such as neuropeptide and BDNF synthesis. However, only a subpopulation of this group of neurones requires NGF in order to develop connections to the pulp during the neonatal period. Few, if any, pulpal afferents express the high-affinity neurotrophin-3 (NT3) receptor trkC, although many have large cell bodies typical of NT3-responsive sensory neurones. A small subpopulation of pulpal afferents seems to express no neurotrophin receptors, yet it is unlikely that these cells belong to the class of small sensory cells known to bind isolectin IB4.

Age Factors↗

Differentiation ability of rat postnatal dental pulp cells in vitro.

The current rapid progression in stem cell research has enhanced our knowledge of dental tissue regeneration. In this study, rat dental pulp cells were isolated and their differentiation ability was evaluated. First, dental pulp cells were obtained from maxillary incisors of male Wistar rats. Immunochemistry by stem cell marker STRO-1 proved the existence of stem cells or progenitors in the isolated cell population. The dissociated cells were then cultured both on smooth surfaces and on three-dimensional (3-D) scaffold materials in medium supplemented with beta-glycerophosphate, dexamethasone, and L-ascorbic acid. Cultures were analyzed by light and scanning electron microscopy and, on proliferation, alkaline phosphatase activity and calcium content were determined and the polymerase chain reaction was performed for dentin sialophosphoprotein, osteocalcin, and collagen type I. These cells showed the ability to differentiate into odontoblast-like cells and produced calcified nodules, which had components similar to dentin. In addition, we found that the "odontogenic" properties of the isolated cells were supported by three-dimensional calcium phosphate and titanium scaffolds equally well.

Animals↗

A mineralizing rat dental pulp cell subline expressing collagen type I and dentin sialoprotein-phosphophoryn transcripts.

A mineralizing rat dental pulp cell subclone, termed M2H4, was selected from single-cell cloning of the rat dental pulp cell line RPC-C2A by screening confluent single-cell cultures for their ability to undergo mineralization. To induce mineralization, confluent single-cell cultures were treated for 8 d with ascorbic acid followed by the addition of inorganic phosphate to a final concentration of 4 mM for an additional 3 d. Confluent M2H4 subclones were shown by immunofluorescence and electron microscopy to form collagen type I fibrils. Furthermore, using reverse transcriptase-polymerase chain reaction, this subclone was found to be capable of expressing dentin sialoprotein-phosphophoryn (DSP-PP) transcripts, an odontoblast-specific marker. Thus, this newly identified mineralizing rat M2H4 subclone possesses odontoblast-like characteristics and can serve as an in vitro model for examining the role of DSP and PP in the formation of mineralized dentin.

Animals↗

Use of the dental pulp tester in veterinary dentistry.

Personal experience with a manually set electronic dental pulp tester is described. The small battery-operated hand-held unit is used in awake dogs and cats, and can rapidly indicate absence of vitality or increased or decreased responsiveness of the pulp tissue.

Animals↗

Stimulation of DNA synthesis by endothelin-1 in primary cultures of human dental pulp.

Endothelin (ET), besides being a powerful vasoactive agent, acts as a mitogen in some cell types. ET-like immunoreactivity has been recently detected by immunocytochemistry in the vascular endothelium of human tooth germ and dental pulp, thus providing evidence for local ET production in these tissues. The effects of ET-1 on DNA synthesis in primary cultures of human dental pulp were now investigated. DNA synthesis was evaluated by flow cytometric assay and by 5-bromo,2'deoxyuridine incorporation as detected by immunocytochemistry. Cultured cells were morphologically similar to dental pulp cells and displayed vimentin immunoreactivity. Incubation of cultures with ET-1 resulted in a dose-dependent increase in the number of S-phase-traversing cells over control (unstimulated) cultures. Control skin fibroblasts were also responsive to ET. This finding raises the possibility that the multifunctional peptide ET-1 might subserve growth-promoting activity in the human tooth. It is tentatively suggested that such as an activity might be important during tooth development and in pulp inflammation and healing.

Adolescent↗

Effect of IL-1ra on human dental pulp cells and pulpal inflammation.

AIM: This study was conducted to investigate the effect of interleukin-1 receptor antagonist (IL-1ra) on the LPS-induced interleukin-1beta (IL-1beta) synthesis in human dental pulp cells and to assess the role of IL-1ra in pulpal inflammation. METHODS: IL-1beta from human dental pulp cells (HDP) was measured by sandwich ELISA; IL-1ra expression in pulpal tissue was detected by immunohistochemical staining. RESULTS: Stimulation of HDP with increasing concentrations of FnLPS resulted in dose-dependent IL-1beta production. The addition of IL-1ra reduced FnLPS-induced IL-1beta synthesis in human dental pulp cells. Significant inhibition of the FnLPS-induced IL-1beta synthesis was observed when IL-1ra was added before treating with FnLPS for 60 min. Large numbers of IL-1ra positive neutrophils, plasmacytes, endothelial cells and lymphocytes were observed in inflamed pulp tissue. CONCLUSIONS: IL-1ra could reduce LPS-stimulated IL-1beta synthesis, suggesting that IL-1ra may play a role in pulpitis.

Anti-Inflammatory Agents↗

[Dental pulp diseases. Ultramicroscopic studies].

After the description of the anatomic and histologic aspects of the healthy dental pulp, stress is laid on pulpal pathologies and on the use of ultramicroscopic techniques to analyze the cytologic modifications in normal and pathologic dental pulp tissues. The results obtained are presented.

Dental Pulp↗

Sympathetic-sensory axon to axon contacts in the dental pulp.

The activity in stimulated intrapulpal sensory nerve fibres can be increased by simultaneous stimulation of the sympathetic system. The anatomical basis for this interaction is unknown. Unmyelinated nerve fibres in the dental pulp are characterized by incomplete isolation and ensheathment. Individual pockets of Schwann cells often contain several axons in close contact within a confined space. If a pocket contained both sensory and adrenergic axons it is possible that activity in one could affect the properties of the other. Four young adult ferrets were, under general anesthesia injected intraperitoneally with the false neurotransmitter 6-hydroxydopamine which accumulates as dark vesicles within adrenergic axons. The animals were, still under general anesthesia, perfused four hours later with a fixative mixture. The canine teeth were removed, decalcified and prepared for electron microscopy. Ultrathin sections of the dental pulp were examined in the electron microscope. Convincingly labelled axons were rare. Labelled sympathetic axons were found in common Schwann cell pockets with unlabelled, presumably sensory, axons. This confirms studies in which degeneration has been observed following either surgical removal of the superior cervical ganglion or chemical sympathectomy. While this "axonal mingling" probably occurs in other sites it may be more extensive in the dental pulp.

Adrenergic Fibers↗

Bacteria recovered from dental pulp induce apoptosis of lymph node cells.

Apoptosis is critical in the pathogenesis of several infectious diseases. The induction of apoptosis was assessed in mouse lymph node cells by four bacteria recovered from infected human dental pulp: Gemella morbillorum, Clostridium butyricum, Fusobacterium nucleatum and Bifidobacterium adolescentis. Smaller lymph nodes and smaller numbers of cells were observed after experimental dental pulp infection with C. butyricum, suggesting that this bacterium induces cell death. Apoptosis was evaluated by determination of cell ploidy and detection of DNA degradation in cells cultured with killed bacteria. Paraformaldehyde-killed C. butyricum and heat-killed G. morbillorum induced substantial cell death, while F. nucleatum and B. adolescentis induced cell death at lower levels. No bacterial preparations induced apoptosis in cells from mice genetically deficient for tumour necrosis factor receptor p55 (TNFRp55), implicating this receptor directly or indirectly as a mediator in the process. It was concluded that apoptosis may be induced during periapical lesions of pulpal origin.

Animals↗

Healing and revascularization of apical periodontium and dental pulps in apicoectomized and nonapicoectomized tooth replants in monkeys. A study of fully developed teeth.

This report primarily concerns an experimental study of the effect of apicoectomy on the healing and revascularization of the periodontal membrane and dental pulps in tooth replants. Five rhesus monkeys (Macaca mulatta) were subjected to surgical procedures in which twenty incisor teeth were extracted and immediately replanted. Ten of these incisors were subjected to apicoectomy. Healing and revascularization of periapical tissues and dental pulps were evaluated by histologic and India-ink-perfusion techniques. Apicoectomized replants were characterized by (1) loss of dental substance (tooth apex) and creation of large periapical spaces, (2) exposure of dental pulps and production of periapical inflammatory processes with destruction of alveolar bone, and (3) blocking of normal healing because of persistence of chronic periapical inflammatory processes. In contrast, nonapicoectomized replants depicted short-lived primary inflammatory responses and healing processes characterized by variable histologic responses.

Animals↗

[The effect of chlorhexidine on the pulp-dentin system (research on the canine dental pulp)].

The effect on the pulp-dentin system by chlorhexidine 0.2% in comparison to the physiological sodium chloride solution was investigated in animal experiments. The methods of tests were based on the requirements of FDI 1980. After 3, 21, 70 and 90 days the teeth were extracted and prepared for histological studies. The evaluation of the histological preparations including measurement on the residual dentin layer was achieved by light microscopy. Differences in the pulp reaction after treatment of the teeth with chlorhexidine or sodium chloride solution could not be observed. The changes of pulp tissue were slight to moderate and depended on the residual dentin layer. The cavity cleaning with chlorhexidine 0.2% did not cause any additional damage in the pulp.

Animals↗

Effects of antibacterial capping agents on dental pulps of monkeys mechanically exposed to oral microflora.

The effects of antibacterial drugs on bacterially contaminated dental pulps were investigated in monkeys. Class V buccal cavities with pulpal exposures were prepared and then left open to the oral environment for 24 h. The exposed pulps were capped with alpha-tricalcium phosphate (alpha-TCP) containing a mixture of antibacterial drugs. Either alpha-TCP or Ca(OH)2 was used as a control. Pulpal responses were histologically evaluated after 4 wk. Those teeth capped with alpha-TCP alone showed total pulp necrosis and bacterial growth within the pulp chamber. By contrast, the pulps capped with alpha-TCP containing mixed antibacterial drugs remained almost normal without any necrotic layer, but showed persistent absorbing response to capping materials and no signs of hard tissue barrier formation. In teeth capped with Ca(OH)2, a hard tissue barrier was formed below the exposure site, with a wide loss of pulp tissue. No inflammation was seen under the barrier. These results indicate that mixed antibacterial drugs added to alpha-TCP effectively disinfected pulpal lesions, without destroying any of the sound pulp tissue. However, hard tissue barrier formation was delayed by this mixture as compared with Ca(OH)2.

Animals↗