Homogeneity and heterogeneity of biological characteristics in mesenchymal stem cells from human umbilical cords and exfoliated deciduous teeth.

Mesenchymal stem cells (MSCs) have proven powerful potential for cell-based therapy both in regenerative medicine and disease treatment. Human umbilical cords and exfoliated deciduous teeth are the main sources of MSCs with no donor injury or ethical issues. The goal of this study was to investigate the differences in the biological characteristics of human umbilical cord mesenchymal stem cells (UCMSCs) and stem cells from human exfoliated deciduous teeth (SHEDs). UCMSCs and SHEDs were identified by flow cytometry. The proliferation, differentiation, migration, chemotaxis, paracrine, immunomodulatory, neurite growth-promoting capabilities, and acetaldehyde dehydrogenase (ALDH) activity were comparatively studied between these two MSCs in vitro. The results showed that both SHEDs and UCMSCs expressed cell surface markers characteristic of MSCs. Furthermore, SHEDs exhibited better capacity for proliferation, migration, promotion of neurite growth, and chondrogenic differentiation. Meanwhile, UCMSCs showed more outstanding adipogenic differentiation and chemotaxy. Additionally, there were no significant differences in osteogenic differentiation, immunomodulatory capacity, and the proportion of ALDHBright compartment. Our findings indicate that although both UCMSCs and SHEDs are mesenchymal stem cells and presented some similar biological characteristics, they also have differences in many aspects, which might be helpful for developing future clinical cellular therapies.

TGF-ß1 and FGF2 stimulate the epithelial-mesenchymal transition of HERS cells through a MEK-dependent mechanism.

Hertwig's epithelial root sheath (HERS) cells participate in cementum formation through epithelial-mesenchymal transition (EMT). Previous studies have shown that transforming growth factor beta 1 (TGF-ß1) and fibroblast growth factor 2 (FGF2) are involved in inducing EMT. However, their involvement in HERS cell transition remains elusive. In this study, we confirmed that HERS cells underwent EMT during the formation of acellular cementum. We found that both TGF-ß1 and FGF2 stimulated the EMT of HERS cells. The TGF-ß1 regulated the differentiation of HERS cells into periodontal ligament fibroblast-like cells, and FGF2 directed the differentiation of HERS cells into cementoblast-like cells. Treatment with TGF-ß1 or FGF2 inhibitor could effectively suppress HERS cells differential transition. Combined stimulation with both TGF-ß1 and FGF-2 did not synergistically accelerate the EMT of HERS. Moreover, TGF-ß1/FGF2-mediated EMT of HERS cells was reversed by the MEK1/2 inhibitor U0126. These results suggest that TGF-ß1 and FGF2 induce the EMT of HERS through a MAPK/ERK-dependent signaling pathway. They also exert their different tendency of cellular differentiation during tooth root formation. This study further expands our knowledge of tooth root morphogenesis and provides more evidence for the use of alternative cell sources in clinical treatment of periodontal diseases.

[Investigation on the prevalence of dental caries and family oral health behaviors in young children aged 1 to 3 years in Chengdu city].

PURPOSE: To investigate the prevalence of dental caries in young children aging 1 to 3 years in Chengdu city and the relationship with family oral hygiene habits. METHODS: One thousand children aging 1 to 3 years in 5 kindergartens in Chengdu city were selected as the study subjects, and examined for oral hygiene. They were divided into case group and control group according to the presence of dental caries. Questionnaires were sent out to two groups of children to find out their oral hygiene habits, and the results were compared and analyzed with SPSS 20.0 software package. RESULTS: Three hundred and thirty-one children suffered from dental caries, and the incidence of dental caries was 33.10%. There was no significant differences in sex, low birth weight, gestational age, medical history, parents' highest educational background and main caregivers between the case group and the control group (P>0.05); there were significant differences in age distribution, feeding methods and annual family income between two groups (P<0.05). In addition to the frequency of flossing and brushing, there were significant differences in other family oral hygiene behaviors between two groups (regular dental examination, frequency of sweets eaten by children, frequency of sweets eaten before sleep and frequency of sweets eaten by parents) (P<0.05). Multivariate analysis showed that age, annual family income, sleeping with bottles, regular examination of children's teeth, frequency of children eating sweets and frequency of children eating before sleep were all influencial factors of caries in young children (P<0.05). CONCLUSIONS: Prevalence of dental caries in young children in Chengdu city is high, and there is significant correlation between dental caries in young children and family oral health behavior, which is one of the influencing factors for dental caries in young children.

[Curative Efficacy of Rituximab for ITP Patients with Different Sensitivity to Hormone].

OBJECTIVE: To investigatc the curative efficacy of low dose rituximab for glucocorticoid ineffective on dependent ITP patients and its relation with sensitivity to glucocorticoid so as to provide reference basis for rational use of drugs in clinical treatmant. METHODS: Seventy-ninth ITP patients enrolled in this study included the glucocorticoid-ineffective patients (19 cases) and glucocorticoid-dependent patients (60 cases). All ITP patients were treated with regimen consisted of high dose dexamethasone plus low dose rituximab (dexal-methasone 40 mg/d for 4 days per os, ritaximab 100 mg by intravenous infusion at D7, 14, 21 and 28 respectively). The patients after treatment were followed-up for 12 month, and the relation of patients sensitivity to glucocorticoid with therapentic response of rituximab was analyzed. The changes of Treg cell ratio and BAFF, IL-2 and sCD40L levels before and after treatment were detected by flow cytometry and ELISA respectively. RESULTS: The overall response rate (ORR) of patients treated with above- mentioned regemen at 1, 3, 6 and 12 months after treatment was 79.7% (63/79), 69.6% (55/79), 63.3% (50/79) and 60.8% (48/79) respectivcly, out of which the ORR of glucocorticoid ineffective and glucocorticoid-dependent ITP patients treated with above-mentioned regimen at 1, 3, 6 and 12 months after treatment was 47.4% (9/19) vs 90.0% (54/60), 36.8% (7/19) vs 80.0% (48/60), 21.1% (4/19) vs 76.7% (46/60), 21.1% (4/19) vs 73.3% (44/60), and the difference between 2 groups was statistically significant. The detection of T reg cell showed that the T reg cell ratio in glucocorticoid- ineffective and dependent patients at 1, 3, 6 and 12 months after treatment was (1.70±0.43)% vs (3.47±0.72)%, (1.66±0.33)% vs (4.29±0.91)%, (1.71±0.37)% vs (4.44±0.97)%, (3.36±0.54)% vs (4.29±1.04)%, respectively. The detection of cytokines showed that the levels of BAFF, IL-2 and sCD40L in plasma of glucocorticoid-dependent patients at 1 month after treatment significanlly decreased (P＜0.05), the levels of BAFF, IL-2 and sCD40L in plasma of glucocorticoid-ineffective patients although decreased at 1 mouth after treatment, but there was no statistical difference as compared with glucocosticoid-depenment patients. CONCLUSION: The treatment of glucocorticoid-dependent ITP patients with rituximab is more effective. The regulatory effect of rituximab on the T-reg cells, BAFF, IL-2 and sCD40L may be one of its mechanisms.

Maternal diabetes modulates dental epithelial stem cells proliferation and self-renewal in offspring through apurinic/apyrimidinicendonuclease 1-mediated DNA methylation.

Maternal gestational diabetes mellitus (GDM) has many adverse effects on the development of offspring. Aberrant DNA methylation is a potential mechanism associated with these effects. However, the effects of GDM on tooth development and the underlying mechanisms have not been thoroughly investigated. In the present study, a GDM rat model was established and incisor labial cervical loop tissue and dental epithelial stem cells (DESCs) were harvested from neonates of diabetic and control dams. GDM significantly suppressed incisor enamel formation and DESCs proliferation and self-renewal in offspring. Gene expression profiles showed that Apex1 was significantly downregulated in the offspring of diabetic dams. In vitro, gain and loss of function analyses showed that APEX1 was critical for DESCs proliferation and self-renewal and Oct4 and Nanog regulation via promoter methylation. In vivo, we confirmed that GDM resulted in significant downregulation of Oct4 and Nanog and hypermethylation of their promoters. Moreover, we found that APEX1 modulated DNA methylation by regulating DNMT1 expression through ERK and JNK signalling. In summary, our data suggest that GDM-induced APEX1 downregulation increased DNMT1 expression, thereby inhibiting Oct4 and Nanog expression, through promoter hypermethylation, resulting in suppression of DESCs proliferation and self-renewal, as well as enamel formation.

Expression and roles of syndecan-4 in dental epithelial cell differentiation.

Syndecan-4 (SDC4), a transmembrane heparan sulfate proteoglycan, acts as a signal transducer. It affects the growth and differentiation of a number of tissues and organs. However, the specific mechanisms through which SDC4 regulates the differentiation of dental epithelial cells (amelogenesis) and tooth development remains largely unknown. In the present study, to identify the SDC4-regulated processes in dental epithelial cells, the SDC4 expression pattern was examined in mouse molar and postnatal incisor tooth germs during the late bell stage of development. Small interfering RNA (siRNA) was designed for this study and used to downregulate SDC4 expression in the rat dental epithelial cell line, HAT-7. The results revealed that SDC4 was mainly present in the oral epithelium, the dental epithelial cells of enamel organs in the molars and the cervical loops in the incisors. When the inner enamel epithelial cells gave rise to ameloblasts, however, the loss of SDC4 expression was evident. SDC4 was also expressed in stratum intermedium (SI) cells in the incisors and in dental mesenchymal cells adjacent to the cervical loops in molars (E18) and postnatal incisors. Fibroblast growth factor 10 (FGF10) promoted proliferation and slightly decreased cell differentiation. The knockdown of SDC4 using specific siRNA led to a decrease in cell proliferation and a highly significant increase in amelogenin, ameloblastin, kallikrein 4 and matrix metalloproteinase 20 expression, molecules that are known to participate in the formation of enamel. These effects were attenuated by FGF10, which upregulated SDC4 expression. Taken together, these results suggest that SDC4 participates in amelogenesis, and FGF10 may modulate dental epithelial cell behaviors through the regulation of SDC4 expression.

Hertwig's epithelial root sheath cells regulate osteogenic differentiation of dental follicle cells through the Wnt pathway.

The development of periodontal ligament-cementum complex (PLCC) originates from the interaction between epithelial cells of Hertwig's epithelial root sheath (HERS) and mesenchymal cells of the dental follicle. While previous studies have suggested that the Wnt pathway is involved in osteogenic differentiation of dental follicle cells (DFCs) during tooth root development, its involvement in the interaction between DFCs and HERS cells (HERSCs) in tooth root mineralization remains unclear. Here, we investigated the hypothesis that HERSCs control osteogenic differentiation of DFCs via the Wnt pathway. We found that during co-culture with HERSCs, DFCs exhibited a greater tendency to form mineralized nodules. Moreover, under these conditions, DFCs expressed high levels of cementoblast/osteoblast differentiation-related markers, such as bone sialoprotein (BSP) and osteocalcin (OCN), the periodontal ligament phenotype-related gene type I collagen (COL1), and ß-catenin (CTNNB1), a core player in the canonical Wnt pathway. In contrast, expression in DFCs of alkaline phosphatase (ALP) was greatly decreased in the presence of HERSCs. Expression of CTNNB1 in DFCs was stimulated by Wnt3a, a representative canonical member of the Wnt family of ligands, but suppressed by Dickkopf1 (DKK1), a Wnt/CTNNB1 signaling inhibitor. Furthermore, in the presence of treated dentin matrix (TDM), differentiation of DFCs was enhanced by Wnt3a when they were in direct contact with HERSCs, but was curtailed by DKK1. Taken together, these results indicate that during tooth root formation, HERSCs induce osteogenic differentiation of DFCs in a process involving the Wnt pathway and the dentin matrix. Our study not only contributes to our understanding of tooth root development and diseases of tooth root mineralization, but also proffers a novel potential strategy for controlling mineralization during tooth root regeneration.

[Influence of zirconia content on translucency of zirconia-toughened alumina glass-infiltrated ceramic].

OBJECTIVE: The objective of this study is to investigate the influence of zirconia content which is 0-30.0% weight percentage of matrix on translucency of zirconia-toughened alumina glass-infiltrated ceramics. METHODS: Seven groups were divided according to different weight percentage of zirconia (0, 2.5%, 5.0%, 7.5%, 10.0%, 20.0% and 30.0%). After sintering, infiltrating and polishing, spectral transmittance was determined with spectrophotometer under D65 standard source. Contrast ratio was also tested by whiteness colorimeter. RESULTS: With mass fraction of zirconia increasing from 0 to 30.0%, spectral transmittance reduced from 0.406% to 0.058%, while contrast ratio value increased from 0.849 +/- 0.005 to 1.015 +/- 0.006. When zirconia content was 10.0%, contrast ratio was 0.990 +/- 0.008. When it was more than 10.0%, transmission rate of the downward trend and contrast ratio of the rising trend became flat. CONCLUSION: Zirconia content has a direct impact on translucency of zirconia-toughened alumina glass-infiltrated ceramic, which is essentially opaque when zirconia content is 10.0%. When mass fraction of zirconia is more than 10.0%, the influence of zirconia content is reduced.