Seong Oh Kim

Ameloblast differentiation in the human developing tooth: effects of extracellular matrices.

Tooth enamel is formed by epithelially-derived cells called ameloblasts, while the pulp dentin complex is formed by the dental mesenchyme. These tissues differentiate with reciprocal signaling interactions to form a mature tooth. In this study we have characterized ameloblast differentiation in human developing incisors, and have further investigated the role of extracellular matrix proteins on ameloblast differentiation. Histological and immunohistochemical analyses showed that in the human tooth, the basement membrane separating the early developing dental epithelium and mesenchyme was lost shortly before dentin deposition was initiated, prior to enamel matrix secretion. Presecretary ameloblasts elongated as they came into contact with the dentin matrix, and then shortened to become secretory ameloblasts. In situ hybridization showed that the presecretory stage of odontoblasts started to express type I collagen mRNA, and also briefly expressed amelogenin mRNA. This was followed by upregulation of amelogenin mRNA expression in secretory ameloblasts. In vitro, amelogenin expression was upregulated in ameloblast lineage cells cultured in Matrigel, and was further up-regulated when these cells/Matrigel were co-cultured with dental pulp cells. Co-culture also up-regulated type I collagen expression by the dental pulp cells. Type I collagen coated culture dishes promoted a more elongated ameloblast lineage cell morphology and enhanced cell adhesion via integrin alpha2beta1. Taken together, these results suggest that the basement membrane proteins and signals from underlying mesenchymal cells coordinate to initiate differentiation of preameloblasts and regulate type I collagen expression by odontoblasts. Type I collagen in the dentin matrix then anchors the presecretary ameloblasts as they further differentiate to secretory cells. These studies show the critical roles of the extracellular matrix proteins in ameloblast differentiation.

The effect of epigallocatechin-3-gallate (EGCG) on human alveolar bone cells both in vitro and in vivo

OBJECTIVEnThe effects of epigallocatechin-3-gallate (EGCG), a major catechin in green tea, on human and mouse osteoblasts remain controversial. This study investigated the direct effects of EGCG on human alveolar bone-derived cells (hABCs) both in vitro and in vivo.nnnDESIGNnhABCs which were collected from eight children (aged 7-9 years, seven males and one female) were treated with EGCG at various concentrations (1, 5, 10, 25, and 50μM), and a proliferation assay, flow cytometric analysis for apoptosis evaluation, migration assay, and in vitro osteogenic differentiation were performed. hABCs that were pretreated with 10μM EGCG and mixed with calcium phosphate carrier combined with EGCG (0.1, 0.5, or 1.5mg) in vivo were transplanted into immunodeficient mouse. Histological staining, quantitative gene expressions, and alkaline phosphatase activity were evaluated in the retrieved transplants.nnnRESULTSnThe proliferation and migration were decreased when EGCG was present at over 25μM. The osteogenic differentiation increased slightly when EGCG was present at up to 10μM, and clearly decreased for higher concentrations of EGCG. In vivo, the potential for hard-tissue formation was slightly higher for the group with 0.1mg of EGCG than for the control group, and decreased sharply for higher concentrations of EGCG.nnnCONCLUSIONnThe present observations suggest that EGCG at a low concentration can slightly enhance the osteogenic effect in vivo, whereas at a higher concentration it can prevent the osteogenic differentiation of hABCs both in vitro and in vivo.

Comparative Study of Pulpal Responses to Pulpotomy with ProRoot MTA, RetroMTA, and TheraCal in Dogs' Teeth

INTRODUCTIONnThis study was conducted to evaluate and compare pulpal responses to ProRoot MTA (Dentsply Tulsa Dental, Tulsa, OK), RetroMTA (Meta Biomed Co, Ltd, Seoul, Korea), and TheraCal (Bisco Inc, Schamburg, IL) in dog partial pulpotomy models.nnnMETHODSnPartial pulpotomies were performed on 60 beagle teeth. The exposed pulp tissues were randomly capped with either ProRoot MTA (n = 15), RetroMTA (n = 15), TheraCal (n = 15), or interim restorative material as a negative control (n = 15). After 4 weeks, the teeth were extracted and processed for histologic and immunohistochemical examinations using osteocalcin and dentin sialoprotein. Calcific barrier formation, inflammatory reaction, and the odontoblastic layer were evaluated and scored in a blind manner. The areas of newly formed calcific barriers were measured for each group.nnnRESULTSnIn most of the ProRoot MTA and RetroMTA specimens, continuous calcific barriers were formed, and the pulps contained palisading patterns in the odontoblastic layer that were free of inflammation. However, the TheraCal specimens had lower quality calcific barrier formation, extensive inflammation, and less favorable odontoblastic layer formation. Overall, areas of newly formed calcific barrier were higher in the ProRoot MTA and RetroMTA specimens than in the TheraCal specimens. Also, immunohistochemistry revealed that osteocalcin and dentin sialoprotein were more clearly visible in the ProRoot MTA and RetroMTA specimens than in the TheraCal specimens.nnnCONCLUSIONSnRetroMTA could provide an alternative to ProRoot MTA. Both materials produced favorable pulpal responses that were similar in nature, whereas TheraCal produced less favorable pulpal responses.

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

OBJECTIVEnStem 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.nnnDESIGNnDental 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.nnnRESULTSnThe 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.nnnCONCLUSIONnThe 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 <3xa0months (cryo<3), and (3) cryopreserved for 3–9xa0months (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 4xa0months. 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.

Morphological changes in the crown of mandibular molars with an additional distolingual root

OBJECTIVEnThe mandibular molars typically have two roots placed mesiodistally, but they occasionally have an additional distolingual (DL) root. This study was to determine the morphological characteristics of the crown of such mandibular molars.nnnDESIGNnRadiographic records and study models were collected from a Korean population (n=86 patients). Each molar was assigned to either the experimental group (i.e. with a DL root) or the control group (i.e. without a DL root; n=41 patients) based on the radiographic findings. The intercuspal distances of the first permanent molars (total, n=100; control/experimental, n=50/50) and primary second molars (46, 23/23), and the largest buccolingual/mesiodistal widths of those molars and primary first molars (42, 21/21) were measured for molars with and without a DL root. In addition, the correlation between the existence of a sixth cusp and a DL root was examined.nnnRESULTSnThe crowns of first permanent and primary second molars with DL roots had significantly larger intercuspal distances between the distobuccal-distolingual cusp tips and a larger distal-area buccolingual width than those without the DL root (t-test; p<0.05). There was no significant correlation between the existence of a sixth cusp and the presence of a DL root.nnnCONCLUSIONSnThe existence of a DL root was associated with larger buccolingual dimensions, especially in the distal area.

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.

Biological efficacy of two mineral trioxide aggregate (MTA)-based materials in a canine model of pulpotomy

The aim of this study was to compare the biocompatibility of Endocem Zr® and ProRoot MTA® by histopathologic analysis in a canine model of pulpotomy. This study utilized 39 teeth of two beagle dogs. The exposed pulp tissues were treated by pulpotomy using ProRoot MTA (n=19) or Endocem Zr (n=20). After 8 weeks, the teeth were extracted and processed with hematoxylin-eosin staining for histologic evaluation. Most of the specimens in both groups developed a calcific barrier at the pulp amputation site and formed an odontoblast layer. However, some of the Endocem Zr specimens showed less calcific barrier formation with a greater inflammatory response and less odontoblast layer formation when compared with the ProRoot MTA specimens. ProRoot MTA and Endocem Zr specimens developed a calcific barrier; however, ProRoot MTA was more biocompatible than Endocem Zr.

Prevalence of delayed tooth development and its relation to tooth agenesis in Korean children

OBJECTIVEnThe aim of this study was to investigate the epidemiology of delayed tooth development (DTD) and the link between DTD and tooth agenesis (TA).nnnDESIGNnThe dental maturity of all of the developing permanent teeth of 4611 children (2417 males and 2194 females) was evaluated from panoramic radiographs. The prevalence of DTD and TA was analyzed, and gender difference for DTS and TA was investigated. The correlation of DTD and TA was investigated in intra-fields and inter-fields.nnnRESULTSnThe total prevalence of DTD among the 4611 children was 3.40%. The maxillary second premolar was the most frequently delayed tooth (1.02%), followed by the maxillary second molar (0.88%) and the mandibular second premolar (0.74%). DTD significantly correlated with TA in both intra-fields and inter-fields (p<0.05).nnnCONCLUSIONSnThe field of delayed development exhibited a significant correlation with that of TA.

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.