The Effect of MMP-13, MMP-12, and AMBN on Gingival Enlargement and Root Deformation In a New Type of Gingival Fibromatosis.

This case compared gene-expression between a new type of idiopathic gingival fibromatosis (IGF) and normal gingiva, to clarify the nature of the gingival overgrowth and dental anomaly. A 6-year-old girl with generalized gingival overgrowth and root deformations was diagnosed with IGF. Gene expression profiles were compared between normal gingiva (N=9) and one IGF gingiva using cDNA microarray. Genes related to regulation of cell proliferation and proteolytic degradation were expressed strongly in IGF. MMP-13 and MMP-12 expression were 120 times and 96 times lower in IGF, respectively, whereas AMBN expression was 79 times higher. RT-PCR and immunohistochemical staining supported the microarray results. Reduced proteolytic activity due to low MMP-13 and MMP-12 expression appears to be a potential mechanism for gingival overgrowth. Genetic investigations, such as expression levels of MMP-13, MMP-12, and AMBN, may enable classification of a new syndrome characterized by gingival enlargement with abnormal root development.

Changing the Angulation of the Tooth Germ in the Bony Crypt: A Case Report.

An ankylosed primary molar may cause rotation or ectopic impaction of succedaneous premolar. When conventional treatment modalities such as observation, surgical exposure with or without orthodontic traction, and autotransplantation are not possible, the simple surgical relocation method could be an alternative treatment option for a lingually rotated premolar during the tooth germ stage before opting to extraction. In the case reported herein, the lingually rotated permanent mandibular second premolar tooth germ was surgically relocated within its bony crypt. Continued root development and spontaneous eruption were observed without complications during the 3.5-year follow-up period.

Ectopic Hard Tissue Formation by Odonto/Osteogenically In Vitro Differentiated Human Deciduous Teeth Pulp Stem Cells.

There have been many attempts to use the pulp tissue from human deciduous teeth for dentin or bone regeneration. The objective of this study was to determine the effects of odonto/osteogenic in vitro differentiation of deciduous teeth pulp stem cells (DTSCs) on their in vivo hard tissue-forming potential. DTSCs were isolated from extracted deciduous teeth using the outgrowth method. These cells were exposed to odonto/osteogenic stimuli for 4 and 8 days (Day 4 and Day 8 groups, respectively), while cells in the control group were cultured in normal medium. The in vitro differentiated DTSCs and the control DTSCs were transplanted subcutaneously into immunocompromised mice with macroporous biphasic calcium phosphate and sacrificed at 8 weeks post-implantation. The effect of odonto/osteogenic in vitro differentiation was evaluated using alkaline phosphatase (ALP) staining and quantitative reverse transcription polymerase chain reaction (RT-PCR). The in vivo effect was evaluated by qualitative RT-PCR, assessment of ALP activity, histologic analysis, and immunohistochemical staining. The amount of hard tissue was greater in Day 4 group than Day 8 group (p = 0.014). However, Day 8 group generated lamellar bone-like structure, which was immunonegative to anti-human dentin sialoprotein with significantly low expression level of DSPP compared with the control group (p = 0.008). This study demonstrates that odonto/osteogenic in vitro differentiation of DTSCs enhances the formation of bone-like tissue, instead of dentin-like tissue, when transplanted subcutaneously using MBCP as a carrier. The odonto/osteogenic in vitro differentiation of DTSCs may be an effective modification that enhances in vivo bone formation by DTSCs.

Peptide-induced formation of a tethered lipid bilayer membrane on mesoporous silica.

Tethered bilayer lipid membranes (tBLMs) on solid supports have substantial advantages as models of artificial cell membranes for such biomedical applications as drug delivery and biosensing. Compared with untethered lipid membranes, tBLMs have more space between substrate and the bilayer and greater stability. The purpose of this work was to use these properties to fabricate and characterize a zwitterionic 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine lipid tBLM containing 2 mol% 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-maleimide(poly(ethylene glycol))-2000 (DSPE-PEG2000-NHS) lipid tethers on a 3-aminopropyltrimethoxysilane-modified mesoporous silica substrate. A quartz crystal microbalance with dissipation monitoring was used to monitor the process of vesicle adsorption and tBLM self-assembly, and atomic force microscopy was performed to characterize the structural properties of the tBLM obtained. Whereas tether-containing lipid vesicles ruptured neither spontaneously nor as a result of osmotic shock, introduction of an amphipathic α-helical (AH) peptide induced vesicle rupture and subsequent tBLM formation. Taken together, our findings suggest that the AH peptide is an efficient means of rupturing vesicles of both simple and complex composition, and is, therefore, useful for formation of tBLMs on solid and mesoporous materials for applications in biotechnology.

Characteristics of stem cells from human exfoliated deciduous teeth (SHED) from intact cryopreserved deciduous teeth.

The aim of this study is to compare the characteristics of stem cells derived from human exfoliated deciduous teeth (SHED) from cryopreserved intact deciduous teeth with those of fresh SHED. In total, 20 exfoliated deciduous teeth were randomly divided into a fresh group (f-SHED; n = 11) and cryopreserved group (c-SHED; n = 9; stored for 1-8 months). Following thawing and separation of the pulp, the SHED cells were cultured, and the characteristics as mesenchymal stem cells were investigated using proliferation assays, cell-cycle analysis, colony-forming unit-fibroblast (CFU-F) assays, and flow cytometry analyses. Furthermore, differentiation into adipogenic and osteogenic lineages was investigated in vitro as well as in vivo via transplantation in mice. We found no significant differences between the two groups in the proliferation analyses, in the expression of mesenchymal stem cell markers, or in the adipogenic and osteogenic differentiation in vitro (p < 0.05). Furthermore, the in vivo transplantation results showed no significant differences in the quantity of bone tissue that formed or in histochemistry performance (p < 0.05). In conclusion, cryopreservation of intact exfoliated deciduous teeth appears to be a useful method for preserving SHED.

Osteoconduction capacity of human deciduous and permanent teeth ash in a rat calvarial bone defect model.

The aim of this study was to confirm the osteoconduction capacities and determine the potential of permanent teeth ash (PTA), and deciduous teeth ash (DTA) as bone substitutes. Rats (n = 71) were divided randomly into four groups: sham, micro macroporous biphasic calcium phosphate (MBCP), PTA, and DTA. A sample of the each group was transplanted into preformed 8-mm calvarial defects (one per rat). The density of new bone was calculated and the crystallinities of the PTA and DTA were analyzed by X-ray diffraction. The degree of new bone formation was high in the MBCP and DTA groups but low in the PTA groups. The DTA was highly crystalline, whereas the PTA was not. The percentages of ß-tricalcium phosphate in the DTA and PTA were 10.7 and 3.7%, respectively. DTA has a high osteoconduction capacity, suggesting that it is a useful bone substitute.

Controlling lipid membrane architecture for tunable nanoplasmonic biosensing.

Tunable nanoplasmonic biosensing for lipid and protein applications is reported based on controlling lipid membrane architecture on surfaces. The interaction of a peptide with lipid membranes is highly sensitive to the membrane architecture on top of plasmonic nanodisks, and the measurement response varies in a manner which is consistent with the surrounding lipid environment.

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 stemness between periodontal ligament and umbilical cord tissues in humans.

Background/purpose: Due to their regenerative potential, periodontal ligament (PDL) and umbilical cord (UBC) tissues are an attractive potential mesenchymal stem cells (MSCs) source. This study compared the expression patterns of genes related to stemness between fresh PDL and UBC tissues. Materials and methods: PDL tissues were collected from 38 permanent premolars extracted for orthodontic purposes, and UBC tissues were obtained from three newborns. Each sample was immediately frozen to prevent RNA degradation. cDNA microarray analysis, quantitative real-time polymerase chain reaction (PCR), and immunohistochemical staining were performed. Gene expression patterns associated with dental stemness (DS) and induced pluripotent stemness (iPS) were compared between PDL and UBC tissues. Results: In the cDNA microarray analyses, the expressions of most iPS genes were greater in the PDL than in the UBC. Meanwhile, the expressions of most DS genes were greater in the UBC than in the PDL. Quantitative real-time PCR analyses showed that the expression levels of matrix metallopeptidase 13 (MMP13), ADAM metallopeptidase domain 22 (ADAM22), vascular cell adhesion protein 1 (VCAM1), and kruppel-like factor 4 (KLF4) genes were greater in the PDL than in the UBC, while the expressions of melanoma cell adhesion molecule (MCAM) and activated leukocyte cell adhesion molecule (ALCAM) were greater in the UBC than in the PDL. Conclusion: These results suggest that UBC and PDL tissues showed slightly different expression patterns of genes related to stemness, which warrants further investigation to use these tissues for future regeneration and implantation therapies.

Assessment of dental caries lesion activity status using quantitative parameters obtained from the quantitative light-induced fluorescence method and difference of microbial distribution in primary molars.

BACKGROUND: This study aimed to assess differences in quantitative measures obtained from the quantitative light-induced fluorescence method and microbial composition of carious dentin and saliva according to the activity status of caries lesions in primary molars. METHODS: A total of 34 teeth from 34 children were evaluated in this study. The activity status of carious lesions was classified using the International Caries Classification and Management System criteria (active or inactive). Images of the carious lesions were captured using a quantitative light-induced fluorescence device for quantitative analyses. Carious dentin and saliva were collected to detect and quantify selected bacterial species (S. mutans, S. sobrinus, Lactobacillus species, F. nucleatum, P. nigrescence, P. intermedia) and C. albicans by quantitative polymerase chain reaction. Mann-Whitney U tests were performed to evaluate differences in quantitative measures from quantitative light-induced fluorescence, the microbial composition of carious dentin, and saliva according to the activity status of carious lesions. RESULTS: Red fluorescence values (∆R, ∆Rmax) from the quantitative light-induced fluorescence method were significantly higher in active lesions (∆R, p = 0.009; ∆Rmax, p = 0.014). The quantitative mean levels of Lactobacillus species (p = 0.010) in carious dentin and S. sobrinus (p = 0.017) in saliva were significantly higher in the active-lesion group. CONCLUSIONS: Quantitative measures related to red fluorescence from the quantitative light-induced fluorescence method, levels of Lactobacillus species from carious dentin, and levels of S. sobrinus from saliva were associated with caries lesion activity.

General gene expression patterns and stemness of the gingiva and dental pulp.

BACKGROUND/PURPOSE: Due to the unique properties of healing processes and cellular differentiation, the gingiva and dental pulp have attracted attention as a potential source of mesenchymal stem cells (MSCs). The purpose of this study was to obtain molecular-level information on these tissues in terms of their function and differentiation processes and investigate stemness. MATERIALS AND METHODS: Healthy gingival tissues were collected from patients (n = 9; aged 7-12 years) who underwent simple surgical procedures, and normal dental pulp tissues were obtained from patients (n = 25; aged 11-25 years) undergoing tooth extraction for orthodontic reasons. Complementary DNA microarray, qRT-qPCR, and immunohistochemical staining were performed to assess general and MSC gene expression patterns. RESULTS: In the gingival tissue, genes related to keratinization, the formation of epithelial cells and ectoderm, and immune and/or inflammatory responses were highly expressed. Meanwhile, in the dental pulp tissue, genes related to ion transport, neuronal development and axon guidance, bone and enamel mineralization, extracellular matrix organization, and angiogenesis were highly expressed. When focusing on the expression of MSC genes, induced pluripotent stem (iPS) cell genes, such as Sox2, c-Myc, and KLF4, were expressed at higher levels in the gingival tissue, whereas dental stem cell genes, such as NT5E and VCAM1, were expressed in dental pulp tissue. CONCLUSION: We found different general and MSC gene expression patterns between the gingival and dental pulp tissue. These results have implications for future regenerative medicine, considering the application of gingival tissue as a potential source of iPS cells.

The Piston Elastic: A Novel Device for Treating Entrapped Ectopic Permanent Molars.

Ectopic eruption of the permanent molar may absorb the distal root of the primary second molar and may result in a decreased arch length or delayed eruption of the permanent tooth, requiring timely treatment. Therefore, we devised an effective and convenient method to unlock the entrapped tooth using a novel device called a "piston-elastic". This case report aims to explain the design and clinical application of this piston-elastic and to describe successful cases. Three patients (aged 6, 13, and 16 years) with ectopically erupted maxillary and mandibular molars, respectively, were treated with a piston-elastic. It was bound to the locked molar to improve the eruption path. After a certain time period, the repulsive force pushed the surface of the adjacent tooth, improving the eruption path of the entrapped tooth. The piston-elastic is a novel device that simply and effectively changes the direction of eruption of ectopically entrapped molars. As it can be manufactured and attached to the chair side, impression acquisition on a model cast and laboratory procedures are unnecessary. Compared to existing methods, the piston-elastic can be easily produced and delivered, causes little irritation, and is inexpensive.

An Analysis of the Temporomandibular Joint Range of Motion and Related Factors in Children and Adolescents.

This study was designed to establish safe guidelines for pediatric dental practice regarding temporomandibular joint (TMJ) range of motion (ROM) and mouth area (MA). A total of 438 children aged 3-15 years old of homogenous ethnicity participated in the study; the distribution of participants was approximately equal (sex; n = 15; age, n = 30). Maximum mouth opening (MMO), body height, weight, and age of each participant were recorded, and the TMJ ROM including anterior and lateral movements, MA, and mouth width were documented. Males showed higher mouth width, MMO, and MA values than females. MMO and MA increased with age, height, and weight in a statistically significant manner. MMO of 40 mm is reached by the age of 5.2 years, at a height of 105.9 cm and a weight of 18.6 kg. MMO showed a moderate correlation with age, height, weight, and mouth width, and MA moderately correlated with mouth width. Anterior and lateral movements did not show any close relation to these aforementioned factors. The findings of this study suggest that forcible mouth opening over 40 mm should be more cautiously considered, especially in children shorter than 105 cm, lighter than 18 kg and in children under 5 years old.

In Vivo Evaluation of Decellularized Human Tooth Scaffold for Dental Tissue Regeneration.

Conventional root canal treatment may result in loss of tooth vitality, which can lead to unfavorable treatment outcomes. Notably, a ceased tooth development of immature permanent teeth with open apices, regeneration of periodontal ligaments (PDL), and pulp is highly expected healing process. For regeneration, the scaffold is one of the critical components that carry biological benefits. Therefore, this study evaluated a decellularized human tooth as a scaffold for the PDL and pulp tissue regeneration. A tooth scaffold was fabricated using an effective decellularization method as reported in previous studies. PDL stem cells (PDLSCs) and dental pulp stem cells (DPSCs) obtained from human permanent teeth were inoculated onto decellularized scaffolds, then cultured to transplant into immunosuppressed mouse. After 9 weeks, PDLSCs and DPSCs that were inoculated onto decellularized tooth scaffolds and cultured in an in vivo demonstrated successful differentiation. In PDLSCs, a regeneration of the cementum/PDL complex could be expected. In DPSCs, the expression of genes related to revascularization and the hard tissue regeneration showed the possibility of pulp regeneration. This study suggested that the potential possible application of decellularized human tooth could be a scaffold in regeneration PDL and pulp tissue along with PDLSCs and DPSCs, respectively, as a novel treatment method.

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.

Cytokine Expression of Stem Cells Originating from the Apical Complex and Coronal Pulp of Immature Teeth.

INTRODUCTION: The aim of this study was to measure and compare the expression levels of cytokines from developing apical complex cells (DACCs) and dental pulp stem cells (DPSCs) of the immature tooth. METHODS: DPSC-conditioned medium (CM) and DACCs-CM were obtained from human young teeth, and 174 cytokines secreted from each CM were identified and compared. A cytokine membrane array and enzyme-linked immunosorbent assay were used to measure and compare the expression levels of the cytokines. Immunocytochemistry targeting insulin-like growth factor-1 and neurotrophin-3 was additionally performed. RESULTS: There were statistically significant differences in the expression levels of 25 cytokines: 22 and 3 were expressed more strongly in DPSCs-CM and DACCs-CM, respectively. Odontoblast differentiation-related cytokines were more strongly expressed in DPSCs-CM, while cell-proliferation-related cytokines were more strongly expressed in DACCs-CM. Proinflammatory and anti-inflammatory cytokines were predominantly expressed in DPSCs-CM and DACCs-CM, respectively. CONCLUSIONS: DPSCs may exert a stronger paracrine effect than DACCs on regeneration of the dentin-pulp complex, in terms of odontoblast differentiation.

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.

OBJECTIVE: The aim of this study was to investigate the epidemiology of delayed tooth development (DTD) and the link between DTD and tooth agenesis (TA). DESIGN: The 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. RESULTS: The 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). CONCLUSIONS: The field of delayed development exhibited a significant correlation with that of TA.