Human-induced pluripotent stem cells (hiPSCs) offer a promising source for generating dental epithelial (DE) cells. Whereas the existing differentiation protocols were time-consuming and relied heavily on growth factors, herein, we developed a three-step protocol to convert hiPSCs into DE cells in 8 days. In the first phase, hiPSCs were differentiated into non-neural ectoderm using SU5402 (an FGF signaling inhibitor). The second phase involved differentiating non-neural ectoderm into pan-placodal ectoderm and simultaneously inducing the formation of oral ectoderm (OE) using LDN193189 (a BMP signaling inhibitor) and purmorphamine (a SHH signaling activator). In the final phase, OE cells were differentiated into DE through the application of Purmorphamine, XAV939 (a WNT signaling inhibitor), and BMP4. qRT-PCR and immunostaining were performed to examine the expression of lineage-specific markers. ARS staining was performed to evaluate the formation of the mineralization nodule. The expression of PITX2, SP6, and AMBN, the emergence of mineralization nodules, and the enhanced expression of AMBN and AMELX in spheroid culture implied the generation of DE cells. This study delineates the developmental signaling pathways and uses small molecules to streamline the induction of hiPSCs into DE cells. Our findings present a simplified and quicker method for generating DE cells, contributing valuable insights for dental regeneration and dental disease research.
The current oral health status and possible dental risk factors among children in rural Shaanxi Province, western China are unreported. This study aimed to describe the oral health status and to analyze the possible risk factors for the oral health status in this population. A multi-stage cluster sampling method was used to survey 12- to 15-year-olds and 4- to 6-year-olds in villages in Shaanxi Province. The structured questionnaires were provided to the 12- to 15-year-olds and to the caregivers of the 4- to 6-year-olds to collect information on the subjects' oral health knowledge, attitudes and behavior. A clinical examination was performed to assess dental caries and gingival bleeding (only 12- to 15-year-olds). SPSS 17.0 statistical software was used to analyze the data. The decayed, missing, filled teeth (DMFT) index scores of 12- to 15-year-olds and 4-to 6-year-olds averaged 0.45 and 3.05, respectively. The caries prevalence was 23.9% in 12- to 15-year-olds and 67% in 4-to 6-year-olds. Additionally, 45.2% of the 12- to 15-year-olds had gingival bleeding and 62.8% had calculus. The oral health knowledge of the subjects was generally poor, whereas they held very positive attitudes toward oral health. A low number of participants reported that they brushed their teeth at least twice daily. Moreover, a statistically significant relationship was found between oral health knowledge scores, tooth brushing frequency and DMFT scores as well as gingival bleeding in the 12- to 15-year-olds. Frequency of sweets consumption was strongly related to dmft scores in the 4- to 6-year-olds. The oral health status, oral health knowledge and behaviors among village children in Shaanxi Province are poor. Oral health education to improve oral health knowledge and to increase the frequency of tooth brushing should be undertaken in the rural schools in western China.
Fenugreek (Trigonella foenum-graecum), a pharmacologically important herb, is widely known for its antidiabetic, hypolipidemic, and anticancer effects. The medicinal properties of this herb are accredited to the presence of bioactive steroidal saponins with one or more sugar moieties linked to the C-3 OH position of disogenin or its C25-epimer yamogenin. Despite intensive studies regarding pharmacology and phytochemical profiles of this plant, enzymes and/or genes involved in synthesizing the glycosidic part of fenugreek steroidal saponins are still missing so far. This study reports the molecular cloning and functional characterization of a key sterol-specific glucosyltransferase, designated as TfS3GT2 here, from fenugreek plant. The recombinant TfS3GT2 was purified via expression in Escherichia coli, and biochemical characterization of the recombinant enzyme suggested its role in transferring a glucose group onto the C-3 hydroxyl group of diosgenin or yamogenin. The functional role of TfS3GT2 in the steroidal saponin biosynthesis was also demonstrated by suppressing the gene in the transgenic fenugreek hairy roots via the RNA interference (RNAi) approach. Down-regulation of TfS3GT2 in fenugreek generally led to reduced levels of diosgenin or yamogenin-derived steroidal saponins. Thus, Tf3SGT2 was identified as a steroid-specific UDP-glucose 3-O-glucosyltransferase that appears to be involved in steroidal saponin biosynthesis in T. foenum-graecum.
Enamel fluorosis is characterized by hypomineralization, and forkhead box O1 ( Foxo1 ) is essential for mouse enamel biomineralization. This study investigated the effect of fluoride on Foxo1 expression and its implications for enamel fluorosis. Mandibular incisors were extracted from Sprague Dawley rats treated for 3 months with water containing 0, 50, or 100 p.p.m. F − . Immunohistochemistry was used to localize and quantify FOXO1 expression in dental epithelial layer cells of the incisors. The effect of fluoride on expression of Foxo1 , kallikrein‐4 ( Klk4 ), and amelotin ( Amtn ) mRNA s was analyzed by real‐time RT ‐ PCR , and western blotting was used to measure total and nuclear FOXO 1 protein levels in mature dental epithelial cells. The results revealed that nuclear FOXO 1 was mainly localized in the transition and the mature ameloblasts and exhibited weaker expression in the rats exposed to fluoride. In addition to the reduced levels of Foxo1 , Klk4 , and Amtn mRNA s, the protein levels of total and nuclear FOXO 1 were decreased in the mature dental epithelial cells exposed to fluoride. Thus, excessive fluoride may have an effect on the expression levels of Foxo1 in dental epithelial cells and thereby affect hypomineralization of the enamel during fluorosis.
Objective: To determine whether amoxicillin had an effect on the enamel mineralization of SD rats. Methods: Eighteen SD rats were randomly divided into three groups. The rats in the control group were given distilled water. The rats in two experimental groups were administered 50 or 100 mg/kg amoxicillin by intragastric administration from day 3 to day 17 after birth. The general condition, the structure of liver and kidney, the enamel surface changes of mandibular first molars and incisors were observed. The changes of Ca/P ratio on enamel surface were analyzed by X-ray energy dispersive spectrometer (EDS). The surface morphology after phosphoric acid treatment was examined by scanning electron microscopy (SEM). Histological changes in the ameloblasts of mandibular incisors were analyzed by hematoxylin and eosin (HE) staining. Results: Compared with the control group, the general conditions as well as liver and kidney structures of SD rats in 50 and 100 mg amoxicillin groups had no significant differences. There was no obvious chalky changes on the first mandibular molars of SD rats in each group. All the incisors in 50 and 100 mg groups showed different degrees of chalkiness in the labial incisal 1/3 enamel. X-ray EDS analysis showed that the Ca/P ratios of occlusal and incisal 1/3 enamel in 50 and 100 mg groups (occlusal 1/3 of mandibular first molars: 1.51±0.03 and 1.52±0.02, incisal 1/3 of mandibular incisors: 1.46±0.01 and 1.43±0.01) was significantly lower than that in the control group (occlusal 1/3 of mandibular first molars: 1.67±0.41, incisal1/3 of mandibular incisors: 1.73±0.07) (P 0.05). SEM observation showed that the enamel rods of the first molars and incisors in the 50 and 100 mg group varied in size and arranged disorderly. The spaces between the enamel rods were larger than that in the control group and some areas even appeared large pits. HE staining showed that the gaps between ameloblasts in 50 and 100 mg groups were significantly wider than that in the control group. Conclusions: Intake of amoxicillin during the period of enamel development of SD rats might affect enamel mineralization.
Objective To assess the oral health-related quality of life (OHRQoL) of adolescents in Northwest China, and to explore the relationship between sociodemographic characteristics, oral health-related behaviors and OHRQoL. Methods A cross-sectional survey of adolescents aged 12–15 years in Shaanxi, Gansu province and Ningxia Hui Autonomous Region of Northwest China was conducted by stratified cluster random sampling. Oral examinations were performed with World Health Organization (WHO) standards, and the condition of crowns and periodontal was recorded. Adolescents' oral health-related behaviors and OHRQoL were collected by questionnaire. Chi-square test and binary logistic regression analysis were used to examine the relationship. Results A total of 7,648 subjects were included. The prevalence of low OHRQoL which defined as a score higher than 0, was 83.8%. A low OHRQoL was most prevalent in Gansu Province and least prevalent in Shaanxi Province. Younger (OR = 0.73, CI = 0.60–0.87), female (OR = 1.27, CI = 1.11–1.44), rural (OR = 1.44, CI = 1.21–1.72), and mother with less education (OR = 0.69, CI = 0.60–0.81) all showed statistically significant influence on teenagers' OHRQoL. From a behavior perspective, teens' OHRQoL was correlated with their frequency of sugar consumption (OR = 1.72, CI = 1.41–2.10), dental appointment rate (OR = 1.29, CI = 1.064–1.57), self-assessment of oral health (OR = 3.09, CI = 2.29–4.19), DMFT index (OR = 1.19, CI = 1.04–1.37), number of teeth calculus (OR = 1.20, CI = 1.01–1.43), and dental trauma (OR = 0.47, CI = 0.39–0.57) over the previous year; however, brushing their teeth was not statistically significant. Conclusions Generally speaking, oral health-related behaviors—such as eating more sugary foods and scheduling dental appointments within the last year—have a significant negative impact on the quality of life of adolescents in Northwest China. Female, and adolescents from families with low educational backgrounds are more likely to be affected by such behaviors in their daily lives.
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