This study summarizes the recent cutting-edge approaches for dentin regeneration that still do not offer adequate solutions. Tertiary dentin is formed when odontoblasts are directly affected by various stimuli. Recent preclinical studies have reported that stimulation of the Wnt/β-catenin signaling pathway could facilitate the formation of reparative dentin and thereby aid in the structural and functional development of the tertiary dentin. A range of signaling pathways, including the Wnt/β-catenin pathway, is activated when dental tissues are damaged and the pulp is exposed. The application of small molecules for dentin regeneration has been suggested as a drug repositioning approach. This study reviews the role of Wnt signaling in tooth formation, particularly dentin formation and dentin regeneration. In addition, the application of the drug repositioning strategy to facilitate the development of new drugs for dentin regeneration has been discussed in this study.

Dentin hypersensitivity (DH) is defined as short and sharp pain caused by external stimuli such as heat, vaporization, contact, osmotic pressure, and chemical stimulation in a normal tooth, rather than due to disease or tooth damage. Its solution is to block the flow of dentinal fluid by physically blocking the dentinal tubule. Of these treatments, fluoride and oxalate type for hypersensitivity can only have a temporary effect. Resins should be used with a suitable bonding system and they may cause hypersensitivity symptoms after treatment. Overcoming these limitations, there is a need for method that can effectively treat dental hypersensitivity lasting long without any side effects. For this reason, experiments with 200 plant extracts as candidates for dentin hypersensitivity, Buddleja officinalis was considered as a candidate for present study. The purpose of this study is to confirm whether the ethanol extract of Buddleja officinalis is effective to protect enamel and dentin by coating tooth surface and resistance to acid or alkali even after tooth coating.

Dentin hypersensitivity is an abrupt intense pain caused by innocuous stimuli to exposed dentinal tubules. Mechanosensitive ion channels have been assessed in dental primary afferent neurons and odontoblasts to explain dentin hypersensitivity. Dentinal fluid dynamics evoked by various stimuli to exposed dentin cause mechanical stress to the structures underlying dentin. This review briefly discusses three hypotheses regarding dentin hypersensitivity and introduces recent findings on mechanosensitive ion channels expressed in the dental sensory system and discusses how the activation of these ion channels is involved in dentin hypersensitivity.

The aim of this study was to evaluate the role of demineralized and particulate autogenous tooth, and interleukin-6 in bone regeneration. A demineralized and particulate autogenous tooth was prepared and human osteoblast-like cells (MG63) and human osteosarcoma cells were inoculated into the culture. The rate of cell adhesion, proliferation and mineralization were examined, and the appearance of cellular attachment was observed. An 8 mm critical size defect was created in the cranium of rabbits. Nine rabbits were divided into three groups including: An experimental group A (3 rabbits), in which a demineralised and particulate autogenous tooth was grafted; an experimental group B (3 rabbits), in which a demineralized, particulate autogenous tooth was grafted in addition to interleukin-6 (20 ng/mL); and a control group. The rabbits were sacrificed at 1, 2, 4 and 6 weeks for histopathological examination with H-E and Masson’s Trichrome, and immunohistochemistry with osteocalcin. The cell-based assay showed a higher rate of cell adhesion, mineralization and cellular attachment in the experimental group A compared with the control group. The animal study revealed an increased number of osteoclasts, newly formed and mature bones in the experimental group A compared with the control group. Eventually, a higher number of osteoclasts were observed in the experimental group B. However, the emergence of newly formed and mature bone was lower than in the experimental group A. The current results suggest that treatment with demineralized and particulate autogenous tooth and interleukin-6 is not effective in stimulating bone regeneration during the bone grafting procedure.

Background: Matrix metalloproteinases (MMPs) are proteolytic enzymes involved in degradation of the extracellular matrix. MMP-8 has been reported to be involved in the degradation of collagen in progression of dental caries. MMP-13 was found to be expressed in both normal and caries pulp, implying its involvement in the pathogenesis of dental caries. Methods: Four extracted teeth were used. They were categorized into four grades according to caries progression. Three sections were prepared from each separated crown and root. Immunofluorescence of the FITC of the MMP-8 and 13 in coronal and radicular dentin was analyzed by confocal microscopy. Results: Immunofluorescence signals that were indicative of MMP-8 were observed both in radicular and coronal dentin in the sound, C1 and C3 carious teeth. In C2 carious teeth, immunofluorescence signals that were indicative of MMP-8 were observed only in radicular dentin. Immunofluorescence signals that were indicative of MMP-13 were observed both in radicular and coronal dentin in the sound teeth. In C1, C2 and C3 carious teeth, immunofluorescence signals that were indicative of MMP-13 were not observed both in radicular and coronal dentin. Conclusion: Immunofluorescence signals revealed that MMP-8 and 13 were present in coronal and radicular dentin, and was differently expressed as caries progressed.

Matrix metalloproteinases (MMPs) are proteolytic enzymes involved in degradation of the extracellular matrix.In a previous study, MMP-13 was found to be expressed in pulp implying its involvement in the pathogenesis of dental caries. Two extracted teeth were used. A sound tooth and a tooth with wide range of dental caries were used. Two sections were obtained each from isolated crown and root. Immunofluorescence of the FITC of the MMP-13 in coronal and radicular dentin was analyzed by confocal microscopy. Immunofluorescence signals that were indicative of MMP-13 were observed in coronal dentin of sound teeth and in carious teeth with a wide range of caries. Marked immunofluorescent reaction was observed in the border line of caries infected and affected coronal dentin. MMP-13 expression was not detected in the root dentin. The expressions of MMP-13 in carious dentin imply the roles of MMP-13 in caries progression.

In this study, hydroxyapatite (HAp) was incorporated into toothpaste and its effect on the remineralization and restoration of dental enamel was evaluated. Different sets of toothpaste were incorporated with HAp levels of 0%, 5%, 10 %, and 15 %. The filler particles of the resulting toothpaste samples were observed via SEM and XRD and compared with compositions of several commercially available toothpastes, showing that the HAp was successfully incorporated into the toothpaste samples. Different sets of human enamel were inflicted with lesions and then treated with the different fabricated toothpaste samples for five minutes three times a day for seven days. During the treatment, the teeth were subjected to demineralization and remineralization cycles to simulate the effect of natural saliva. The surface of the enamel samples were observed using SEM before and after one week of treatment, showing the formation of HAp layers on the surfaces of the enamel samples. The effect of the toothpaste on the lesions was observed using an inverted light microscope and the lesion depth was found to decrease as the concentration of HAp in the toothpaste used increased. HAp was successfully incorporated in the toothpaste and its presence was found to lessen lesion depths and improve tooth remineralization.

We investigated the pulpal response to direct pulp capping in rat molar teeth using mineral trioxide aggregate (MTA) and calcium hydroxide (CH). A palatal cavity was prepared in rat maxillary molar teeth. Either MTA or CH was placed on the exposed pulp and all cavities were restored with composite. Rats were sacrificed for histological evaluation after 12 hours and at 2, 7, 14 and 21 days. In both the MTA and CH groups, reparative dentin formation was clearly observed on histology after 14 days. The MTA-capped pulps were found to be mostly free from inflammation, and hard tissue of a tubular consistent barrier was observed. In contrast, in CH-capped teeth, excessive formation of re¬parative dentin toward residual pulp was evident. The pulpal cell response beneath the reparative dentin layer was examined by immunofluorescence using antibodies against DSP. After 2 days, a few DSP immunopositive cells, most of which showed a cuboidal shape, appeared beneath the predentin layer. At 7 days, DSP-immunopositive cells with columnar odontoblast-like cells were seen beneath the newly formed hard tissues. At 14 and 21 days, DSP was more abundant in the vicinity of the odontoblastic process along the dentinal tubules than in the mineralized reparative dentin. The CH group showed strong expression patterns in terms of DSP immunoreactivity. Our results thus indicate that MTA may be a more effective pulp capping material as it induces the differentiation of odontoblast-like cells and the formation of reparative dentin without the loss of residual pulp functions.

The aim of this study was to evaluate surface character¬istics and biological properties of the dentin -derived hydroxyapatite (HA) coating on titanium substrate. Dentin-derived HA was obtained from extracted human teeth using a calcination method at 850℃. The commercially pure titanium (cp-Ti, ASTM Grade II) was used as a metallic substrate and a radio frequency magnetron sputtering method was employed as a coating method. Scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDX) were utilized to investigate the coating aspects and composition. Atomic forced microscopy (AFM) and a surface profiler were used to assess the surface morphology and roughness. Corrosion tests were performed in phosphate- buffered saline at a 36.5 ± 1℃ in order to determine the corrosion behavior of the uncoated and coated specimens. The biocompatibility of dentin-derived HA coated specimens with fetal rat calvarial cells and human gingival fibroblasts was assessed by SEM and cell prolif¬eration analysis. The results showed that the dentin-derived HA coatings appeared to cover thinly and homogeneously the surfaces without changing of the titanium substrate. The EDX analysis of this the coating surface indicated the presence of Ca and P elements. The mean surface roughness of cp-Ti and dentin-derived coating specimens was 0.27 µm and, 1.7 µm, respectively. Corrosion tests indicated a stable passive film of the dentin-derived HA coating specimens. SEM observations of fetal rat calvarial cells and human fibroblast cells on coated surfaces showed that the cells proliferated and developed a network of dense interconnections. The cells on all specimens proliferated actively within the culture period, showing good cell viability. At day 1 and 3, dentin-derived coating specimens showed 89% and 93% cell viability, respectively, when normalized to cp-Ti specimens. These results suggest that dentin-derived HA coating using the RF magnetron sputtering method has good surface characteristics and biocompatibility.

The polarizing images of hard tissues including bone and cementum show characteristic features of different birefringence fortheir microstructures. Nevertheless, the clear mechanism of the amplified birefringence under polarizing microscope has not been well understood. We hypothesized that the unique polarized light could be accumulated in the microtubules due to the decreased refractory angle by the inside lower-density matrix, and then the accumulated light in the microtubules could be dispersed brightly. In order to elucidate the polarizing effect on the microtubules, the dentinal tubules in different conditions were observed, and compared with each other to explain their birefringence phenomena. In the decalcified sections of normal tooth, the dentinal tubules located near the pulp chamber showed strong birefringence, while the sclerosed dentinal tubules near the dentino-enamel junction did not show the birefringence. The birefringence was more conspicuous in the longitudinal sectionsof dentinal tubules than in the cross sections. In the decalcified sections of complex odontoma, which produced abnormal and immature dentinal tubules, the birefringence was not observed in the shrunken dentinal tubules filled with dense materials, while the peritubular matrix showed clear birefringence. The birefringence was also observed in the collagen fibers in the connective tissue, and continuously strong in the immature cemental materials containing precollagen fibers. However, the highly mineralized osteodentine did not show the birefringence. Taken together, these data suggest that the microtubules composed ofless-dense matrix than the background tissue, i.e., dentinal tubules, Haversian canals, etc., produce the amplified birefringence by the polarizing light according to the hypothesis of microtubule refraction.

Cementum is a hard connective tissue, produced by cementoblasts during tooth root formation, which provides for the attachment of the periodontal ligament to the roots and surrounding alveolar bone. Establishment of this attachment is an important event in the regeneration of lost periodontal tissues. We examined whether or not odontoblast conditioned media(CM) have a regulatory influence on the differentiation and mineralization of cementoblasts(murine cementoblastic cell line, OCCM-30) in vitro. To identify the effect of odontoblast conditioned media and dentin non collagenous proteins (dNCPs) on cementoblast differentiation and mineralization, we treated CM and dNCPs to cementoblast then differentiated the cells for 14 days. To evaluate the formation of mineralized nodules alizarin-red S staining was performed at 0,4,7 and 14 days. Expression of cementum matrix genes was measured by RT-PCR. Mineralization of cementoblasts was accelerated with CM from odontoblastic MDPC-23 and OD-11. The expression of BSP, ALP, and OC mRNA in cementoblastic OCCM-30 cells was facilitated by the MDPC-23 and OD-11 cells. The extracted dNCPs had little influence on the proliferation, cell cycle modification, and chemotaxis of OCCM-30 cells. Although the dNCPs did not exhibit chemotactic activities for cementoblasts, the dNCPs promoted the differentiation and mineralization of cementoblasts. In conclusion, the dentin matrix protein, or the secreted products of odontoblast, facilitates cementoblast differentiation and mineralization. This represents a new approach and suggests another avenue for cementum regeneration.

Previous in vi tro studies demonstrated that H202 or carbamide peroxide cou ld penetrate i nto pul p chambers through enamel and dentin (Benetti et a l., 2004; G okay et a l. , 2004‘ Suli eman et al .. 2005) ‘ Recently. Lee et al.(2006) demonstrated that H20Z enhanced the diffe rentiation of odontoblast like cell line, whereas it inhibited osteogenic diffe rentiation in pre 。steobl astic cell line, as seen by its efl"ecLs on an early difï"erentiation marker. ALP activity. I-lowever. the effects of HZ02 have not been well elucidated in primary cultured human pulp cells ln th is study‘ we investigated whether HO- 1 is involved in H20 2-induced cytotoxicity and examined the production 0 1" dent in sia lophosphoprotein (DSPP) and other minera li zation markers, in human pulp cells H20Z dec1'eased cell viabili ty. but increased HO-l and DSPP expression in a concentra t ion and time dependent manner. Inhibitors of guanylate cyclase, PI3K. ERK, and p38 MAP kinase blocked J-!?,0 2- induced cytot oxicity and the expression of HO-1 and DSPP mRNAs in pulp cells. These data suggest that t he induction of HO-l by H202 in pu lp cells plays a protective role against the cytotoxic effects of H202 and stimulates DSPP expression. resulting in prematu re oclontoblast differentiation th rough pathways t hat involve cGMP. p38. ancl ERK

Al though the changes in tooth morphology and hardness by hydrogen peroxide(H20 z) have been r‘epor‘.ted .‘ the pαr。o야t뻐ec야tive role of heme oxygenase-l(HO-l) against the cytotoxic effects of H202 has not been clarifïed i n human pulp cells ln this st udy. we investigated whether HO-l is involved in Hz0 2-induced cytoLox icity a nd examined the production 0 1' dentin sia lophosphoprotein(DSPP) and other mineralization markers‘ in hllman pu lp cells H202 decreased cell viabi lity, but increased HO-l and DSPP expression in a concentra tion and time dependent manner . Inhibitors of gllanylate cyclase. PI3K, ERK. and p38 MAP kinase blocked H202-indllced cytotoxicity and the expression of HO-l and DSPP mRNAs in pulp cells. These data suggest that the induction of HO-l by H202 in plllp cells plays a protective role against the cytotoxic effects 0 1' HzOz and stimulates DSPP expression‘ reslllting in prematllre odontoblast dilTerentiation throllgh pathways that involve cGMP‘ p38. and ERK.

Hereditary dentin defects consist of dentin dysplasia(DD) and dentinogenesis imperfecta(DI). The DI associated with osteogenesis imperfecta has been classified as DI type I, whereas isolated inherited defects have been categorized as DI types II and III. However, whether DI type III should be considered a distinct phenotype or a variation of DI type II is debatable. Recent genetic findings have focused attention on the role of the dentin sialophosphoprotein(DSPP) gene in the etiology of inherited defects of tooth dentin. We have identified a novel mutation(c.727G → A, p.D243N) at the 243th codon of exon 4 of the DSPP gene in a Korean patient with DI type III. The radiographic and histologic features of the patient revealed the classic phenotype of shell teeth. These findings suggest that DI type II and III are not separate diseases but rather the phenotypic variation of a single disease.

System L amino acid transporter is a major route for providing living cells with neutral amino acids including several essential amino acids. To elucidate the expression pattern of L-type amino acid transporter 1 (LAT1) in the bone formation process, the expressions of LAT1 and its subunit 4F2 heavy chain (4F2hc) were investigated in the healing process after the implantation of bone graft materials in the calvarial osseous defected rats. Circular calvarial defects (1 cm in diameter) were made midparietally. The rats were divided into 4 groups of 1 control group and 3 experimental groups. In the control group, the defect was only covered with soft tissue flap. In the experimental groups, they were filled with human particulate dentin (particulate dentin group), with plaster of Paris (plaster of Paris group) and with the mixture of human particulate dentin and plaster of Paris with ratio of 2 : 1 by weight (mixture group). The rats were sacrificed at the 1, 2, 4 and 8 weeks after operation and the RT-PCR analysis and immunohistochemical analysis were performed. In the RT-PCR analysis, the mRNAs of LAT1 and 4F2hc were strongly detected in all 4 groups. In the immunohistochemical analysis, at 1 week after operation, the LAT1 protein and its subunit 4F2hc protein were mainly expressed in the osteoblasts, osteocytes and interstitial tissues of the around the defect and inner part of newly forming bone in all 4 groups. The expressions of LAT1 and 4F2hc proteins were decreased at 2 and 4 weeks after operation. The LAT1 and 4F2hc proteins were scarcely expressed at 8 weeks after operation in all 4 groups. These results suggest that the LAT1 and its subunit 4F2hc highly expressed at the early stage of new bone formation and may have an important role in providing cells with neutral amino acids including several essential amino acids at that stage.

Hereditary dentin defects consists of dentin dysplasia(DD) and denti nogenesis imperfecta(Dr) ‘ The Dl associated with osteogenesis imperfecta has been classified as DI type 1. whereas isolated inherited defects have been categori zed as DI types II and III , However‘ whether DI type III should be considered a distinct phenotype 01' a variation of DI type 1I is debatable , Recent genetic findings have focused attention on the role of the dentin sialo phosphoprotein(DSPP) gene in the etiology of inherited defects of tooth dentin, We have identified novel mlltation( c,727G - > A, p,D243N) at the 243th codon of exon 4 of the DSPP gene in a Korean patient with DI type III The radiographic and histologic features of the patient revealed the classic phenotype of shell teeth These findings sllggest that DI type II and III are not separate diseases bllt rather the phenotypic variation 01' a s ingle disease

The phylogeneticall y conserved nuclear factor 1 (NFI) gene fami ly encodes s ite-specific tra nscription factors essential for the development of a number of organ syst ems. There are four NFI genes in mamma ls (Nfi a , Nfib, Nfi c, and Nfix) and single NFI genes in Drosophila melanogaster, Caenorhabdi t is elegans, Anopheles spP. ‘ and other simpl e animals. It was reported that Nfia-defici ent mice exhi bit agenesis of the corpus call osum and other forebrain defects , wher eas Nfib-defi cient mice possess unique defects in lung ma turation and fo rebrain defect. Recently, it was also found that Nfic-defi cient mice exhibit agenesis of mo l ar서 roots and severe incisor defects. In the present study, we investigat ed the possible role of NFI-C in odon toblast diffe rent ia tion and root dentin formation using the innovative and invalua ble Nfic knockout mice model Nfi c-defi cient mice showed a berrant odontoblast differentiation and consequentl y abnormal dentin formation, while other t issues/organs in the body including ameloblasts of the enamel organ a ppeared to be unaffec ted and normal One of the most st r iking changes observed in these aberrant odontoblasts was t he absence of in tercellular junctions beLween them, r esulting in di ssociation of the cells and loss of th eir cellular polarity a nd organi zation. Surprisingly, these cells became trapped in dentin-like minerali zed t issue and thus their overa ll morphology r esembled osteoblasts and os t eocyt es. There was also an increased apoptotic activity in Nfic-deficient mice. These findings strongly s uggest ed that NFI -C plays a key role in odon tob last differentiation and survival in a cell type-specific manner.