Recently, the RNA/DNA-binding protein FUS, Fused in sarcoma, was shown to play a role in growth, differentiation, and morphogenesis in vertebrates. Because little is known about Fus, we investigated its expression pattern in murine tooth development. In situ hybridization of mouse mandibles at specific developmental stages was performed with a DIG-labeled RNA probe. During early tooth development, Fus was detected in the dental epithelium and dental mesenchyme at 11 days postcoitum (dpc) and 12 dpc. From 14 dpc, Fus was strongly expressed in the dental papilla and the cervical loop of the dental epithelium. At postnatal day 4 (PN4), Fus expression was observed in the odontoblasts, ameloblasts, the proliferation zone of the pulp, and the cervical loop. At PN14, the expression pattern of Fus was found to be maintained in the odontoblasts and the proliferation zone of the pulp. Furthermore, Fus expression was especially strong in the Hertwig's epithelial root sheath (HERS). Therefore, this study suggests that Fus may play a role in the HERS during root development.
Temperature signaling can be initiated by members of transient receptor potential (thermo-TRP) channels. Hot and cold substances applied to teeth usually elicit pain sensation. Since odontoblasts constitute a well-defined layer between the pulp and the mineralized dentin, being first to encounter thermal stimulation from oral cavity, they may be involved in sensory transduction process, in addition to their primary function as formation of dentin. We investigated whether thermo-TRP channels are expressed in a odontoblast cell line, MDPC-23. The expressions of thermo-TRP channels were examined using reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry, fluorometric calcium imaging. Analysis of RT-PCR revealed mRNA expression of TRPV1, TRPV2, TRPV4 and TRPM8, but no TRPV3, TRPA1. Immunohistochemical approach failed to detect TRPV1 expression. Whereas the application of 4-phorbol-12,13-didecanoate($10\;{\mu}M$, a TRPV4 agonist), menthol(1 mM, a TRPM8 agonist) and icilin($10\;{\mu}M$, a TRPM8 agonist) produced the enhancement of intracellular calcium concentration, capsaicin($1\;{\mu}M$, a TRPV1 agonist) did not. Our results suggest that subfamily of thermo-TRP channels expressed in odontoblasts may serve as thermal or mechanical transducer in teeth.
Oh, Chang Young;Kim, Su-Gwan;Go, Dae-San;Yu, Sun-Kyoung;Kim, Tae-Hoon;Kim, Chun Sung;Park, Joo-Cheol;Kim, Do Kyung
International Journal of Oral Biology
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v.42
no.2
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pp.39-45
/
2017
Metformin (1,1-dimethylbiguanide hydrochloride), derived from French lilac (Galega officinalis), is a first-line anti-diabetic drug prescribed for patients with type 2 diabetes. However, the role of metformin in odontoblastic cell differentiation is still unclear. This study therefore undertook to examine the effect of metformin on regulating odontoblast differentiation in MDPC-23 mouse odontoblastic cells derived from mouse dental papilla cells. As compared to controls, metformin significantly accelerated the mineralization, significantly increased and accelerated the expressions of ALP and Col I mRNAs, and significantly increased the accelerated expressions of DSPP and DMP-1 mRNAs, during differentiation of MDPC-23 cells. There was no alteration in cell proliferation of MDPC-23 cells, on exposure to metformin. These results suggest that the effect of metformin on MDPC-23 mouse odontoblastic cells derived from mouse dental papilla cells, facilitates the odontoblast differentiation and mineralization, without altering the cell proliferation.
It is considered that self-curing resin will induce a little pulpal irritation, if mixing time is insufficient. So this study was designed to compare four different conditions between control group and three experimental groups. Canine teeth of 20 cats used, and class V cavities were prepared on maxillary canine of cats. Control group was filled with zone and other experimental groups were filled with zone after the application of the self-curing resin. Animals of the experimental and control group were sacrified at 3 days, 1, 2, 3, 4 weeks after the experiment. The finding lead to the following conclusions ; 1. Odontoblasts of first and third experimental groups were condensed irregularly and were changed at early stage, and reapir was delayed. 2. Odontoblasts of second experimental group were condensed slightly but, histologic repair was appeared at 4 weeks after the experiment 3. In all groups, the damaged odontoblastic cell layer were eventually disappeared at 4 weeks after the experiment.
The purpose of this study was to investigate the morphology and intercellular junctions of the odontoblast of dental pulp in the rat incisor by means of the freeze fracture electron microscopy. Twenty male Sprague-Dawley rats weighing $150{\sim}200g$ were used. After being anesthetized by an intraperitoneal injection of 0.5 ml sodium pentobarbital per kg in body weight(60 mg/ml) the animals were perfused with 2.5% glutaraldehyde-2% paraformaldehyde fixative in 0.1 M cacodylate buffer, pH 7.2 through the ascending aorta for one hour. The incisors were carefully extracted from the jaws and demineralized by suspending them in 0.1 M EDTA in 3% glutaraldehyde (pH 7.2) for two weeks. After demineralization, the specimens were obtained from the portion divided into five equal parts. For freeze-fracture replication, demineralized tissues were infiltrated for several hours with 10%, 25% glycerol in 0.1M cacodylate buffer as a cryoprotectant and then frozen in liquid Freon 22 and stored in liquid nitrogen. Fracturing and replication were done in Balzers BAF 400D high-vacuum freeze-fracture apparatus at $-120^{\circ}C$ under routine $5X10^{-7}$ Torr vacuum. The tissue was immediately replicated with platinum unidirectionally at $45^{\circ}$ angle and reinforced with carbon at $90^{\circ}$ angle unidirectionally or by using a rotary stage. The replication process was monitored by a quartz-crystal device. The replicas were immersed in 100% methanol overnight. The tissue was then digested from the replica by clorox (laundry bleach), placed into 5% EDTA, and washed repeatedly with distilled water. The replicas were picked up on 0.3% formvar-coated 75 mesh grids and examined in the JEOL 100B electron microscope. The results were as follows; 1. Both in thin sections and freeze-fracture replicas, three types of intercellular junctions were recognizable in the plasma membrane of odontoblast: gap junction, tight junction and desmosome-like junction. 2. The nuclear pores were evenly distributed over the nuclear envelope. The pore complex formed a ring about 70 nm in diameter. 3. Gap junctions were found between odontoblasts as well as odontoblasts and neighbouring pulp cells (fibroblast, subodontoblastic cell process, nerve-like fibre). Gap junctions, which were round, ellipsoid and pear-shaped and 600 nm in diameter, were observed in the odontoblast. 4. Numerous round and ellipsoid gap junctions could be frequently seen on the plasma membranes in cell body and apical part of the odontoblasts. On the P face, the junctions were recognized as a cluster of closely packed particles, measuring about 9 nm in diameter, and on the E face, the junctions were recognized as a shallow grooves.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.37
no.5
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pp.415-420
/
2011
Purpose: Calcium phosphate cement (CPC) is one of many useful materials for restoring tooth defects, periodontium and maxillofacial area. Chitosan is a biodegradable material that has been shown to promote the growth and differentiation of osteoblasts in culture. This study examined the interaction between odontoblasts and bio-calcium phosphate cement reinforced with chitosan. Materials and Methods: $5{\times}10^3$ odontoblastic cells were seeded into each well. Various concentrations of bio-calcium phosphate cement reinforced with chitosan (10, 20, 50, 100, 200, 500 ${\mu}g$/ml, 1, 2, 4 mg/ml) were diluted and added to the wells. The well was incubated for 24 h, 48 h and 72 h. After incubation, the number of cells was assessed to determine the cell viability. A cytokinesis-block micronucleus assay and chromosomal aberration test were carried out to estimate the extent of chromosomal abnormalities. Microscopic photographs and RT-PCR were performed to examine the adhesion potential of bio-calcium phosphate cement reinforced with chitosan. Results: Bio-CPC-reinforced chitosan did not show significant cytotoxicity. The number of damaged chromosomes in the cells treated with Bio-CPC-reinforced chitosan was similar to that in the control cells. There was no significant increase in the number of chromosomal aberrations in the Bio-CPC reinforced chitosan exposed cells. Microscopic photographs and RT-PCR confirmed the adhesive potential of bio-CPC reinforced chitosan to odontoblasts. Conclusion: Bio-CPC-reinforced chitosan did not affect the odontoblastic cell viability, and had no significant cytotoxic effect. Bio-CPC-reinforced chitosan showed adhesive potential to odontoblasts. These results are expected form the basis of future studies on the effectiveness of dental restorative materials in Bio-CPC reinforced with chitosan.
Dentin, a major component of teeth, is formed by odontoblasts which produce the dentin matrix beneath the dental epithelium and induce the mineralization of dentin. To date, the biochemical properties of dentin matrix proteins have been well characterized, but upstream regulators of these proteins are not yet well known. Recently in this regard, several transcription factors have been identified as potential regulators of matrix proteins. Most transcription factors are generally involved in diverse biological processes and it is essential to identify those that are odontoblast-specific transactivators to further understand the process of dentin formation. We thus analyzed the expression pattern of dentin matrix proteins and the activities of established transactivators containing a Cre-locus. Expression analyses using in situ hybridization showed that dentin matrix proteins are sequentially expressed in differentiating odontoblasts, including type-I collagen, Dmp-1 and Dspp. The activities of the transactivators were evaluated using ${\beta}$-galactosidase following the generation of double transgenic mice with each transactivator and the ROSA26R reporter line. The ${\beta}$-galactosidase activity of each transactivator paralled the expression of the matrix proteins. These results thus showed that these transactivators could be utilized for odontoblastspecific conditional gene targeting. In addition, time- and tissue-specific conditional gene targeting might also be achieved using a combination of these transactivators. Odontoblast-specific conditional gene targeting with these transactivators will likely also provide new insights into the molecular mechanisms underlying dentin formation.
After a vital pulpotomy in dogs' teeth, the responses of the remaining pulp tissue under hydroxides (calcium hydroxide, magnesium hydroxide, aluminium hydroxide and zinc hydroxide) were studied histologically. The class V cavities were prepared on the teeth and the pulp was amputated. Each hydroxide was placed over the amputated tissue and the cavity was sealed with zinc oxide eugenol cement. Animals were sacrificed after 3 days, 1, 2, and 3 weeks following the operation. The teeth were decalcified, sectioned and stained with hematoxylin and eosin. Microscopic examination reveals as follows; 1. Calcium hydroxide: Inflammatory change was seen in the superficial portion of the remaining pulp tissue at the 3rd day and 1st week. The incompleted calicified material began to be deposited from the canal wall at the 2nd week, and the advanced calcified material was seen at the 3rd week. 2. Magnesium hydroxide: Severe inflammatory change was seen in the superficial portion of the remaining pulp from the 3rd day and the 1st week samples. Inflammatory change was decreased at the 2nd week and the slight calcified material was deposited from the root canal at the 3rd week. 3. Aluminium hydroxide: Severe inflammatory changes were seen in the remaining pulp tissue, the blood vessel was dilated, and the odontoblasts were destroyed at the 3rd day and 1st week. The fibrous degeneration spread to the apex at the 2nd week. There was no evidence of newly formed odontoblasts or deposition of calcified material underneath aluminium hydroxide. 4. Zinc hydroxide: The micrscopic picture was destructive. A thick necrotic layer was found under the amputated surface at the 3rd day and 1st week. Granulation tissue formation as well as chronic inflammatory changes extended to the apical area in the pulp tissue. Also there were no sign of odontoblastic formation or calcified material at the 2nd and 3rd week.
Shin, Sang Hun;Kim, Jae-Sung;Kim, Su-Gwa;Go, Dae-San;Yu, Sun-Kyoung;Kim, Chun Sung;Park, Joo-Cheol;Kim, Do Kyung
International Journal of Oral Biology
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v.43
no.3
/
pp.133-140
/
2018
Resveratrol (3,4',5,-trihydroxystilbene), a phytoalexin present in grapes, exerts a variety of actions to reduce superoxides, prevents diabetes mellitus, and inhibits inflammation. Resveratrol acts as a chemo-preventive agent and induces apoptotic cell death in various cancer cells. However, the role of resveratrol in odontoblastic cell differentiation is unclear. In this study, the effect of resveratrol on regulating odontoblast differentiation was examined in MDPC-23 mouse odontoblastic cells derived from mouse dental papilla cells. Resveratrol significantly accelerated mineralization as compared with the control culture in differentiation of MDPC-23 cells. Resveratrol significantly increased expression of ALP mRNA as compared with the control in differentiation of MDPC-23 cells. Resveratrol significantly accelerated expression of Col I mRNA as compared with the control in differentiation of MDPC-23 cells. Resveratrol significantly increased expressions of DSPP and DMP-1 mRNAs as compared with the control in differentiation of MDPC-23 cells. Treatment of resveratrol did not significantly affect cell proliferation in MDPC-23 cells. Results suggest resveratrol facilitates odontoblast differentiation and mineralization in differentiation of MDPC-23 cells, and may have potential properties for development and clinical application of dentin regeneration materials.
The author observed the effect of x-ray irradiation on the tooth germ development of the rat fetuses. The lower right abdomen of the pregnant rats were exposed to x-ray irradiation (400 rads) on 9½th day of qestation. At 18½th day of qestation, the fetuses were removed from their mothers and histological sections of molar region were prepared. The results were as folows: 1. In the experimental fetuses, no significant changes appeared in the histological aspects of the enamel pulp, except the poor development of the innerenamel epithelium in the cusp region. 2. Pulp cells of cusp region in the irradiated fetuses were not differentiated to odontoblasts, The arrangement and population of pulp cells showed marked regional differences in the dental papilla. 3. Developmental features of dental follicle of irradiated fetuses were similar with controls.
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