Kun Xuan

Comparative characterization of stem cells from human exfoliated deciduous teeth and dental pulp stem cells.

OBJECTIVE This study focused on the characterization of stem cells from human exfoliated deciduous teeth (SHED) in comparison with dental pulp stem cells (DPSCs) to certify SHED as a key element in tissue engineering. METHODS In the present study, SHED and DPSCs were assayed for their cell surface antigens and proliferation by measuring the cell cycles, growth rates, Ki67-positive efficiencies, and colony-forming units (CFUs). The evaluation of multi-differentiation was performed using alizarin red and oil red O and real-time PCR in vitro. The mineralization capability of the cells was examined in vivo by implanting with ceramic bovine bone (CBB) into subcutaneous of immunocompromised mice for 8weeks. A three-dimensional pellet cultivation system is proposed for SHED and DPSCs to recreate the biological microenvironment that is similar to that of a regenerative milieu. RESULTS SHED showed a higher proliferation rate and differentiation capability in comparison with DPSCs in vitro, and the results of the in vivo transplantation suggest that SHED have a higher capability of mineralization than the DPSCs. The mRNA expression levels of inflammatory cytokines, including matrix metalloproteinase-1 (MMP1), tissue inhibitors of metalloproteinase-1 (TIMP1), matrix metalloproteinase-2 (MMP2), tissue inhibitors of metalloproteinase-2 (TIMP2) and interleukin-6 (IL-6) were higher in SHED than that in DPSCs. In addition, the expression levels of Col I and proliferating cell nuclear antigen (PCNA) in SHED sheets were significantly higher than those in DPSCs sheets. CONCLUSIONS This study systematically demonstrated the differences in the growth and differentiation characteristics between SHED and DPSCs. Consequently, SHED may represent a suitable, accessible and potential alternative source for regenerative medicine and therapeutic applications.

In vitro study of the effects of fluoride-releasing dental materials on remineralization in an enamel erosion model.

OBJECTIVES This study was conducted to compare the remineralization effects of five regimens on the loss of fluorescence intensity, surface microhardness, roughness and microstructure of bovine enamel after remineralization. We hope that these results can provide some basis for the clinical application of these materials. METHODS One hundred bovine incisors were prepared and divided into the following five groups, which were treated with distinct dental materials: (1) Clinpro™ XT varnish (CV), (2) F-varnish (FV), (3) Tooth Mousse (TM), (4) Fuji III LC(®) light-cured glass ionomer pit and fissure sealant (FJ) and (5) Base Cement(®) glass polyalkenoate cement (BC). Subsequently, they were detected using four different methods: quantitative light-induced fluorescence, microhardness, surface 3D topography and scanning electron microscopy (SEM). RESULTS The loss of fluorescence intensity of CV, BC and FJ groups showed significant decreases after remineralization (p<0.05). The microhardness values of the BC group were significantly higher than those of the other groups (p<0.05) after 6 weeks of remineralization. The CV groups surface roughness was significantly lower than those of the other groups after 6 weeks of remineralization (p<0.05). Regarding microstructure values, the FV group showed many round particles deposited in the bovine enamel after remineralization. However, the other four groups mainly showed needle-like crystals. CONCLUSIONS Glass ionomer cement (GIC)-based dental materials can promote more remineralization of the artificial enamel lesions than can NaF-based dental materials. Resin-modified GIC materials (e.g., CV and FJ) have the potential for more controlled and sustained release of remineralized agents. The effect of TM requires further study.

Identification of a novel missense mutation of MSX1 gene in Chinese family with autosomal-dominant oligodontia

OBJECTIVES Oligodontia is defined as the congenital absence of 6 or more permanent teeth excluding the third molar. The occurrence of non-syndromic still remains poorly understood, but in recent years some cases have been reported where mutations or polymorphisms of PAX9 and MSX1 had been associated with non-syndromic oligodontia. The objective of the present work was to study the phenotype and genotype of three generations of a Han Chinese family affected by non-syndromic autosomal-dominant oligodontia. DESIGN We examined all individuals of the oligodontia family by clinical and radiographic examinations. Based on clinical manifestations, candidate genes MSX1 and PAX9 were picked up to analyse and screen mutations. RESULTS Dental evaluation showed that the most commonly missing teeth are the mandibular second premolars, followed by the maxillary second premolars and maxillary lateral incisors, and subsequently the maxillary first premolars. The probability of missing a particular type of tooth is not always bilaterally symmetrical, and differences exist between maxilla and mandible. PCR-SSCP analysis and DNA sequencing revealed a novel missense mutation c.662C>A in a highly conserved homeobox sequence of MSX1 and a known polymorphisms c.347C>G. CONCLUSION Our finding suggests the missense transversion (c.662C>A) and the polymorphisms (c.347C>G) may be responsible for oligodontia phenotype in this Chinese family.

Periodontal tissue engineering with stem cells from the periodontal ligament of human retained deciduous teeth

BACKGROUND AND OBJECTIVE Periodontal ligament stem cells from human permanent teeth (PePDLSCs) have been investigated extensively in periodontal tissue engineering and regeneration. However, little knowledge is available on the periodontal ligament stem cells from human retained deciduous teeth (DePDLSCs). This study evaluated the potential of DePDLSCs in periodontal tissue regeneration. MATERIAL AND METHODS DePDLSCs were isolated and purified by limited dilution. The characteristics of DePDLSCs were evaluated and compared with PePDLSCs both in vitro and in vivo. RESULTS DePDLSCs presented a higher proliferation rate and colony-forming capacity than PePDLSCs in vitro. During the osteogenic induction, alkaline phosphatase (ALP) activity, mineralized matrix formation and expression of mineralization-related genes, including runt-related transcription factor 2 (RUNX2), ALP, collagen type I (COLI) and osteocalcin (OCN) were significantly enhanced in DePDLSCs compared with PePDLSCs. Furthermore, DePDLSC cell sheets showed a stronger synthesis of collagen type I in the extracellular matrix than did PePDLSC cell sheets. After in vivo transplantation, DePDLSC cell sheets recombined with human dentin blocks were able to generate new cementum/periodontal ligament-like tissues. CONCLUSION Our findings suggest that DePDLSCs can be used as a promising candidate for periodontal tissue engineering.

Oral and Craniofacial Manifestations and Two Novel Missense Mutations of the NTRK1 Gene Identified in the Patient with Congenital Insensitivity to Pain with Anhidrosis

Congenital insensitivity to pain with anhidrosis (CIPA) is a rare inherited disorder of the peripheral nervous system resulting from mutations in neurotrophic tyrosine kinase receptor 1 gene (NTRK1), which encodes the high-affinity nerve growth factor receptor TRKA. Here, we investigated the oral and craniofacial manifestations of a Chinese patient affected by autosomal-recessive CIPA and identified compound heterozygosity in the NTRK1 gene. The affected boy has multisystemic disorder with lack of reaction to pain stimuli accompanied by self-mutilation behavior, the inability to sweat leading to defective thermoregulation, and mental retardation. Oral and craniofacial manifestations included a large number of missing teeth, nasal malformation, submucous cleft palate, severe soft tissue injuries, dental caries and malocclusion. Histopathological evaluation of the skin sample revealed severe peripheral nerve fiber loss as well as mild loss and absent innervation of sweat glands. Ultrastructural and morphometric studies of a shed tooth revealed dental abnormalities, including hypomineralization, dentin hypoplasia, cementogenesis defects and a dysplastic periodontal ligament. Genetic analysis revealed a compound heterozygosity- c.1561T>C and c.2057G>A in the NTRK1 gene. This report extends the spectrum of NTRK1 mutations observed in patients diagnosed with CIPA and provides additional insight for clinical and molecular diagnosis.

Exposure to a continuous low dose of tetrachlorodibenzo-p-dioxin impairs the development of the tooth root in lactational rats and alters the function of apical papilla-derived stem cells

OBJECTIVES Ubiquitous environmental pollutants such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) cause abnormalities in reproduction and development. TCDD inhibits the development of teeth, and its effects depend on its dose and the developmental stage of the tooth. Our aim here was to investigate the effect of lower doses of TCDD on the development of the tooth root in vivo and in vitro. DESIGN We observed tooth root development in lactational rats exposed to continuous low doses of TCDD starting on postnatal day 6 using Mico-CT analyses and histopathological examinations. And then the characteristics of stem cells derived from the apical papilla (SCAPs) were evaluated and compared with SCAPs induced by lower doses of TCDD both in vitro and in vivo. RESULTS The results of experiments showed that rat pups exposed to low dose TCDD at prenatal stage developed, dentine hypoplasia, and hypomineralization. Further, TCDD impaired the functions of SCAPs in vivo by inhibiting cell proliferation and osteogenic and odontogenic differentiation. The impairment of SCAPs after TCDD exposure was accompanied by increased expression of AHR, down-regulation of the expression of Runx2, and alkaline phosphatase, suggesting that the AHR pathway mediated the effects of TCDD. CONCLUSION These results provide the first insights into the toxicity of TCDD, which adversely affects the development of the tooth root through indirectly altering the function of SCAPs.

Clinical and genetic evaluation of a Chinese family with isolated oligodontia

OBJECTIVES Oligodontia is defined as the congenital absence of 6 or more permanent teeth excluding the third molar. Tooth agenesis may be classified as syndromic/non-syndromic and as familial/sporadic. To date, more than 300 genes have been found to be involved in tooth development, but only a few of these genes, such as MSX1, PAX9 and AXIN2, are related to the condition of non-syndromic oligodontia. The objective of the present work was to investigate the disease-causing gene of non-syndromic oligodontia in a Han Chinese family and analyse the pathogenesis of mutations that result in oligodontia. DESIGN We examined all individuals of the oligodontia family by clinical and radiographic examinations. Based on the clinical manifestations, the candidate genes MSX, PAX9 and AXIN2 were selected to analyse and screen for mutations. RESULTS The clinical evaluation suggested that the family might show non-syndromic oligodontia. DNA sequencing of the MSX1 gene revealed two mutations in the two patients with oligodontia: a heterozygotic silent mutation, c.348C>T (P.Gly116=), in exon 1 and a homozygotic deletion of 11 nucleotides (c.469+56delins GCCGGGTGGGG) in the intron. However, the silent mutation and the deletion mutation were thought to be known polymorphisms (rs34165410 and rs34341187) by bioinformatics analysis. We did not detect any mutations in the PAX9 and AXIN2 genes of oligodontia patients. CONCLUSION Our finding suggests that identified polymorphisms (c.348C>T and c.469+56delins GCCGGGTGGGG) may be responsible for the oligodontia phenotype in this Chinese family, but the association requires further study.

Comprehensive and sequential management of an impacted maxillary central incisor with severe crown–root dilacerations

Tooth dilaceration refers to a dental anomaly characterized by an abrupt deviation in the longitudinal axis of tooth. Crown-root dilaceration is diagnosed in teeth with sharp angles at the cement-enamel junction. The greater the bending degree is, the less chance there is for successful teeth preservation and relocation. In this report, a clinical case of an impacted maxillary central incisor with severe crown-root dilacerations was described by means of an operative evaluation using three-dimensional dental computed tomography and a multidisciplinary approach that included surgical, orthodontic, endodontic, prosthetic and periodontal therapy.