Genetic Comparison of Stemness of Human Umbilical Cord and Dental Pulp.

This study focuses on gene expression patterns and functions in human umbilical cord (UC) and dental pulp (DP) containing mesenchymal stem cells (MSCs). DP tissues were collected from 25 permanent premolars. UC tissue samples were obtained from three newborns. Comparative gene profiles were obtained using cDNA microarray analysis and the expression of tooth development-associated and MSC-related genes was assessed by the quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Genes related to cell proliferation, angiogenesis, and immune responses were expressed at higher levels in UC, whereas genes related to growth factor and receptor activity and signal transduction were more highly expressed in DP. Although UC and DP tissues exhibited similar expression of surface markers for MSCs, UC showed higher expression of CD29, CD34, CD44, CD73, CD105, CD146, and CD166. qRT-PCR analysis showed that CD146, CD166, and MYC were expressed 18.3, 8.24, and 1.63 times more highly in UC, whereas the expression of CD34 was 2.15 times higher in DP. Immunohistochemical staining revealed significant differences in the expression of genes (DSPP, DMP1, and CALB1) related to odontogenesis and angiogenesis in DP. DP and UC tissue showed similar gene expression, with the usual MSC markers, while they clearly diverged in their differentiation capacity.

Microscopic analysis of molar--incisor malformation.

OBJECTIVE: Molar-incisor malformation (MIM) is a newly discovered type of dental anomaly that involves a characteristic root malformation of the permanent first molars. The aim of this study was to reveal the microstructure of MIM teeth in order to determine their origin. STUDY DESIGN: Four MIM teeth were extracted from a 9-year-old girl due to severe mobility. The detailed microstructure of the teeth was determined by examinations with micro-computed tomography (micro-CT), hematoxylin and eosin (H&E) staining, immunohistochemical staining, and scanning electron microscopy to reveal the detailed microstructure. RESULTS: Micro-CT and H&E staining revealed the pulpal floor comprising three layers: upper, middle, and lower. Amorphous hard tissues and hyperactive cells were observed in the middle layer of the pulpal floor, and the cells stained positively for dentin sialoprotein and osteocalcin, but not for collagen XII. CONCLUSION: The results of the present study imply that MIM-affected molars probably result from inappropriate differentiation of the apical pulp and dental follicle.

The Rect-spring: a new device for treating ectopically erupting permanent molars.

The Rect-spring, a new device that simply and effectively resolves ectopically erupting molars, may be applied in cases where the ectopically erupted molar is entrapped by more than two mm. Its design, scientifically logical for changing the direction of force, delivers a continuous distalizing and uprighting force. The Rect-spring can be easily manufactured at chairside and delivered on the same day without the need to take impressions or perform laboratory procedures. With proper activation, it can resolve a severely ectopically erupting molar within three months. It is relatively inexpensive and easy to deliver, causes little irritation, and is less time consuming for patients. It is worth utilizing before introducing Halterman or Humphrey appliances due to its simplicity and economical benefit. The purpose of this clinical report was to describe the design and the clinical application procedure for the Rect-spring and three successfully treated cases.

Distinctive genetic activity pattern of the human dental pulp between deciduous and permanent teeth.

Human deciduous and permanent teeth exhibit different developmental processes, morphologies, histological characteristics and life cycles. In addition, their pulp tissues react differently to external stimuli, such as the pulp sensitivity test, dental trauma and pulp therapy materials. These suggest differences in gene expression and regulation, and in this study we compared gene-expression profiles of the human dental pulp from deciduous and permanent teeth. Pulp tissues from permanent premolars and deciduous molars aged 11-14 years were extirpated and mRNA was isolated for cDNA microarray analysis, and quantitative real-time PCR (qPCR). Other teeth were used for immunohistochemical analysis (IHC). Microarray analysis identified 263 genes with a twofold or greater difference in expression level between the two types of pulp tissue, 43 and 220 of which were more abundant in deciduous and permanent pulp tissues, respectively. qPCR analysis was conducted for eight randomly selected genes, and the findings were consistent with the cDNA microarray results. IHC confirmed that insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) was broadly expressed in deciduous dental pulp tissue, but minimally expressed in permanent dental pulp tissue. Immunohistochemical analysis showed that calbindin 1 (CALB1), leucine-rich repeat-containing G-protein-coupled receptor 5 (LGR5), and gamma-aminobutyric acid A receptor beta 1 (GABRB1) were abundantly expressed in permanent predentin/odontoblasts, but only minimally expressed in deciduous dental pulp tissue. These results show that deciduous and permanent pulp tissues have different characteristics and gene expression, suggesting that they may have different functions and responses to therapies focused on pulp or dentin regeneration.

In vitro and in vivo characteristics of stem cells from human exfoliated deciduous teeth obtained by enzymatic disaggregation and outgrowth.

OBJECTIVE: Stem cells from human exfoliated deciduous teeth (SHED) are a good source of dental tissue for regeneration therapy, and can be obtained using different primary culture methods. The aim of this study was to determine the differences in the in vitro and in vivo characteristics between SHED isolated via enzymatic disaggregation (e-SHED) and outgrowth (o-SHED) primary culture methods. DESIGN: Dental pulp stem cells were isolated from 14 exfoliated deciduous teeth by enzymatic disaggregation (n=7) and outgrowth (n=7). Their proliferation potential and colony-forming ability were evaluated in vitro, as was their mesenchymal stem-cell-marker expression (using flow cytometry), and their differentiation was verified using quantitative real-time PCR (qPCR) and histochemical staining. In addition, the qualitative and quantitative characteristics of the hard tissue that was generated after in vivo transplantation were compared using haematoxylin and eosin staining, immunohistochemical staining, qPCR, and quantitative alkaline phosphatase analysis. RESULTS: The cell-proliferation potential, colony-forming ability, and Stro-1 and CD146 expression were higher in e-SHED than in o-SHED. While the in vitro adipogenic differentiation potential was greater in e-SHED than in o-SHED, the in vitro osteogenic differentiation did not differ significantly between the two cell types. Although in vivo hard tissue formation was greater following transplantation of o-SHED into mice, there was no difference in the quality of hard tissue generated by e-SHED and o-SHED. CONCLUSION: The findings of this study indicate that e-SHED exhibit stronger stemness characteristics, but that o-SHED are more suitable for hard-tissue regeneration therapy in teeth.

Viability of pulp stromal cells in cryopreserved deciduous teeth.

The cryopreservation of exfoliated deciduous teeth and harvesting of stem cells from them as required would reduce the costs and efforts associated with banking stem cells from primary teeth. The aim of this study was determine whether the viability of pulp stromal cells from deciduous teeth was influenced by the cryopreservation process itself or the period of cryopreservation. In total, 126 deciduous teeth were divided into three groups: (1) fresh, (2) cryopreserved for <3 months (cryo<3), and (3) cryopreserved for 3-9 months (cryo3-9). The viability of the pulp tissues was compared among the three groups by evaluating the outgrowth from pulp tissues and cell activity within those pulp tissues. In addition, the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay was performed to compare cell apoptosis within fresh pulp tissue and pulp tissue that had been cryopreserved for 4 months. The outgrowth from and cell activity within the pulp tissues did not differ significantly between the fresh and cryo<3 pulp tissues. However, these parameters were significantly reduced in the cryo3-9 pulp tissue. In TUNEL assay, 4-month cryopreserved pulp tissues has more apoptotic cells than fresh group. In conclusion, it is possible to acquire pulp stromal cells from cryopreserved deciduous teeth. However, as the period of cryopreservation becomes longer, it is difficult to get pulp cells due to reduced cell viability.

Multiple teeth fractures in dentinogenesis imperfecta: a case report.

Dentinogenesis imperfecta (DGI) is a hereditary defect consisting of opalescent teeth composed of irregularly formed and hypomineralized dentin. This paper presents the multiple fractures of DGI-affected teeth and suggests the reason of low fracture resistance by observing the dentin microstructures directly using scanning electron microscope (SEM) and by measuring its surface hardness using the Vickers hardness test. SEM revealed that while the enamel microstructure was similar in the DGI-affected and normal teeth, the microstructure of the DGI-affected dentin was poorly woven and more loosely packed than that of the normal dentin. The Vickers hardness of the DGI-affected dentin was 4.89 times softer than the normal dentin. The low fracture resistance of DGI-affected teeth can be attributed to the poorly woven microstructure of their dentin, which leads to a reduction in hardness.

Comparative gene-expression analysis of the dental follicle and periodontal ligament in humans.

The human dental follicle partially differentiates into the periodontal ligament (PDL), but their biological functions are different. The gene-expression profiles of the dental follicle and PDL were compared using the cDNA microarray technique. Microarray analysis identified 490 genes with a twofold or greater difference in expression, 365 and 125 of which were more abundant in the dental follicle and PDL, respectively. The most strongly expressed genes in the dental follicle were those related to bone development and remodeling (EGFL6, MMP8, FRZB, and NELL1), apoptosis and chemotaxis (Nox4, CXCL13, and CCL2), and tooth and embryo development (WNT2, PAX3, FGF7, AMBN, AMTN, and SLC4A4), while in the PDL it was the tumor-suppressor gene WIF1. Genes related to bone development and remodeling (STMN2, IBSP, BMP8A, BGLAP, ACP5, OPN, BMP3, and TM7SF4) and wound healing (IL1, IL8, MMP3, and MMP9) were also more strongly expressed in the PDL than in the dental follicle. In selected genes, a comparison among cDNA microarray, real-time reverse-transcription polymerase chain reaction, and immunohistochemical staining confirmed similar relative gene expressions. The gene-expression profiles presented here identify candidate genes that may enable differentiation between the dental follicle and PDL.

Activation of G Proteins by Aluminum Fluoride Enhances RANKL-Mediated Osteoclastogenesis.

Receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis is accompanied by intracellular Ca(2+) mobilization in a form of oscillations, which plays essential roles by activating sequentially Ca(2+)/calmodulin-dependent protein kinase, calcineurin and NFATc1, necessary in the osteoclast differentiation. However, it is not known whether Ca(2+) mobilization which is evoked in RANKL-independent way induces to differentiate into osteoclasts. In present study, we investigated Ca(2+) mobilization induced by aluminum fluoride (AlF4 (-)), a G-protein activator, with or without RANKL and the effects of AlF4 (-) on the osteoclastogenesis in primary cultured mouse bone marrow-derived macrophages (BMMs). We show here that AlF4 (-) induces intracellular Ca(2+) concentration ([Ca(2+)]i) oscillations, which is dependent on extracellular Ca(2+) influx. Notably, co-stimulation of AlF4 (-) with RANKL resulted in enhanced NFATc1 expression and formation of tartrate-resistant acid phosphatase (TRAP) positive multinucleated cells. Additionally, we confirmed that mitogen-activated protein kinase (MAPK) is also activated by AlF4 (-). Taken together, these results demonstrate that G-protein would be a novel modulator responsible for [Ca(2+)]i oscillations and MAPK activation which lead to enhancement of RANKL-mediated osteoclastogenesis.

Comparative gene expression analysis of the human periodontal ligament in deciduous and permanent teeth.

There are histological and functional differences between human deciduous and permanent periodontal ligament (PDL) tissues. The aim of this study was to determine the differences between these two types of tissue at the molecular level by comparing their gene expression patterns. PDL samples were obtained from permanent premolars (n = 38) and anterior deciduous teeth (n = 31) extracted from 40 healthy persons. Comparative cDNA microarray analysis revealed several differences in gene expression between the deciduous and permanent PDL tissues. These findings were verified by qRT-PCR (quantitative reverse-transcription-polymerase chain reaction) analysis, and the areas where genes are expressed were revealed by immunohistochemical staining. The expressions of 21 genes were up-regulated in deciduous relative to PDL tissues, and those of 30 genes were up-regulated in permanent relative to deciduous PDL tissues. The genes that were up-regulated in deciduous PDL tissues were those involved in the formation of the extracellular matrix (LAMC2, LAMB3, and COMP), tissue development (IGF2BP, MAB21L2, and PAX3), and inflammatory or immune reactions leading to tissue degradation (IL1A, CCL21, and CCL18). The up-regulated genes in permanent PDL tissues were related to tissue degradation (IL6 and ADAMTS18), myocontraction (PDE3B, CASQ2, and MYH10), and neurological responses (FOS, NCAM2, SYT1, SLC22A3, DOCK3, LRRTM1, LRRTM3, PRSS12, and ARPP21). The analysis of differential gene expressions between deciduous and permanent PDL tissues aids our understanding of histological and functional differences between them at the molecular level.

Continued root development of a surgically repositioned human incisor tooth germ.

Conventional orthodontic traction may not be the treatment of choice in cases of inverted impaction of a maxillary incisor, especially when located near the alveolar crest. Poor prognosis is associated with the limited space for proper root development, resulting in a root too short for normal function and/or a severely dilacerated root interrupting the force-induced positioning. The surgical repositioning of ectopic impacted toothgerm before the development of root could be a valuable alternative choice of treatment before the decision of extraction. In this case report, an impacted immature incisor toothgerm in complete inversion was surgically repositioned using a closed-flap technique in a boy who was 6 years 8 months old. Continued root formation and spontaneous eruption were observed after surgery over the 51-month follow-up period, without pulpal or periodontal complications.

Shh signaling is essential for rugae morphogenesis in mice.

Palatal ridges, or rugae palatinae, are corrugated structures observed in the hard palate region. They are found in most mammalian species, but their number and arrangement are species-specific. Nine palatal rugae are found in the mouse secondary palate. Previous studies have shown that epithelial Shh signaling in the palatal ridge plays an important role during rugae development. Moreover, Wnt family members, including LEF1, play a functional role in orofacial morphogenesis. To explore the function of Shh during rugae development, we utilized the maternal transfer of 5E1 (anti-Shh antibody) to mouse embryos. 5E1 induced abnormal rugae patterning characterized by a spotted shape of palatal ridge rather than a stripe. The expression patterns of Shh and Shh-related genes, Sostdc1, Lef1 and Ptch1, were disrupted following 5E1 injection. Moreover, rugae-specific cell proliferation and inter-rugae-specific apoptosis were affected by inhibition of Shh signaling. We hypothesize that the altered gene expression patterns and the change in molecular events caused by the inhibition of Shh signaling may have induced abnormal rugae patterning. Furthermore, we propose a reaction-diffusion model generated by Wnt, Shh and Sostdc1 signaling. In this study, we show that Sostdc1, a secreted inhibitor of the Wnt pathway, is a downstream target of Shh and hypothesize that the interaction of Wnt, Shh and Sostdc1 is a pivotal mechanism controlling the spatial patterning of palatal rugae.

Physical properties of resin-reinforced glass ionomer cement modified with micro and nano-hydroxyapatite.

Hydroxyapatite is a biologically compatible material and a major component of dental enamel and bone tissue. Because of its biocompatibility and structural similarity to human teeth and the skeletal system, a number of dental studies have evaluated its application as a bone substitute or dental restorative material. This study was to evaluate the differences in bonding strength and resistance to demineralization between micro-hydroxyapatite and nano-hydroxyapatite added to self-cured resin-reinforced/modified glass ionomer cement. RelyX was used as the base glass ionomer cement material and for the control group. 10% micro-hydroxyapatite added glass ionomer cement was named experimental group 1, and 10% nano-hydroxyapatite added glass ionomer cement was named experimental group 2. Physical tests for ISO9917-1:2007 in each group was acceptable, except the setting time of nano-hydroxyapatite added glass ionomer cement, which exceeded maximum setting time. Bonding strength was greatest in nano-hydroxyapatite glass ionomer cement, and cohesive failure was common in all specimens. When fractured surface was observed under SEM, spherical particles were observed in experimental groups containing hydroxyapatite particles, and they were more prevalent in nano-HA added glass ionomer cement group than in micro-hydroxyapatite added group. Both experimental groups exhibited greater resistance to demineralization compared to the control group, and there was no significant difference between the experimental groups. Under SEM, nano-hydroxyapatite added glass ionomer cement exhibited increased resistance to demineralization compared to micro-hydroxyapatite added glass ionomer cement.

Incidence and relationship of an additional root in the mandibular first permanent molar and primary molars.

OBJECTIVES: The mandibular first permanent and primary molars occasionally have an additional root located distolingually. This study aimed to determine the incidences of an additional root in these molars and their relationship. STUDY DESIGN: This study involved 4050 children for whom periapical radiographs of the mandibular molar area were available. The incidence of an additional root for each molar was calculated and the pattern of concurrent additional roots in different molars was also investigated. RESULTS: Additional roots were present in 33.1%, 27.8%, and 9.7% of the first permanent, second primary, and first primary molars, respectively. When an additional root was present in a primary molar, the probability of the posterior adjacent molar also having an additional root was greater than 94.3%. CONCLUSION: The presence of an additional root in a primary molar can be used to predict the presence of an additional root in molars posterior to it.

Oral manifestations of a child with chronic vomiting.

Perimolysis is a type of intrinsic erosion--an irreversible dental demineralization linked to chronic regurgitation--which causes teeth to be more susceptible to dental caries. The purpose of this case report was to study a 35-month-old chronic vomiting child who visited the Department of Pediatric Dentistry in Yonsei Dental Hospital, Seoul, South Korea, for an evaluation of and treatment for the loss of tooth structure of his primary teeth. To prevent further destruction of the teeth and maintain occlusal height, all the posterior teeth were restored with stainless steel crowns and all the anterior teeth were restored with resin veneer crowns after pulpal treatment under general anesthesia. Therefore, when a child suffering from chronic vomiting visits a pediatric dental clinic, it is prudent to: (1) perform all possible dental treatment to control vomiting's adverse influences on the oral structures; and (2) refer the patient to a pediatrician to determine the cause of vomiting.

Oral self-mutilation in the Lesch-Nyhan syndrome.

Lesch-Nyhan Syndrome is a disorder caused by congenital absence of a purine metabolic enzyme, hypoxanthine-guanine phosphoribosyl transferase (HPRT). This syndrome is characterized clinically by mental retardation, chorea, athetosis, hyperuricemia, uricosuria and self-mutilation. This report is of two children, who are cousins, both of whom have Lesch-Nyhan syndrome and presented with severe self-mutilation wounds on their lip(s). Vital pulpotomy and coronal resection was done as a more conservative approach than extracting all offending teeth. By maintaining the root portion of the teeth in the bone, it is expected that preservation of the alveolar bone will be achieved.