Inhibition of human embryonic palatal mesenchymal cell cycle by secalonic acid D: a probable mechanism of its cleft palate induction

OBJECTIVE: To assess the mechanism(s) of cleft palate induction by secalonic acid D (SAD) in human embryonic palatal mesenchymal (HEPM) cells and compare them with those evaluated in the murine embryonic palate. DESIGN: Effect of SAD on HEPM cell proliferation was studied by obtaining dose response curves for cell numbers, uptake of 3H-thymidine and the expression of proliferating cell nuclear antigen (PCNA). Effects of SAD on cell cycle were assessed by flowcytometry. Cell-labeling with 3H-glucosamine and immunoblot analysis were conducted to study SAD effects on the synthesis of glycosaminogycans (GAG) and the expression of fibronectin and tenascin, respectively. RESULTS: SAD induced a concentration-dependent decrease in HEPM cell number and 3H-thymidine uptake beginning at 0.1 microg of SAD/ml. Expression of PCNA and progression of cell cycle from G1 to S phase were inhibited following SAD exposure. Cell viability was significantly reduced only at 7.5 microg/ml of SAD or higher indicating that the reduction in cell numbers by SAD at lower concentrations is likely due to reduced proliferation and at higher concentrations due to both reduced proliferation and cell death. Synthesis of extra cellular matrix components (GAGs, fibronectin or tenascin) by HEPM cells, however, was not inhibited by SAD. CONCLUSION: The results of these studies confirmed those of our previous studies with mice and the MEPM cells that SAD may induce cleft palate by reducing numbers of palatal mesenchymal cells by inhibition of their proliferation thereby leading to a reduction in the size of the developing palate shelves.     

Immunochemical detection of CD14 on human gingival fibroblasts in vitro

The activation of monocytes and macrophages induced by lipopolysaccharide has been shown to contribute to the binding of lipopolysaccharide and lipopolysaccharide-binding protein complex to the cell surface CD14 molecule. To clarify the mechanism of the lipopolysaccharide-induced modulation of the function of gingival fibroblasts, we investigated the effect of anti-CD14 on interleukin 6 (IL-6) production on human gingival fibroblasts in vitro. Immunochemical staining revealed weak positivity for CD14 on fibroblasts from healthy gingiva, while strong positivity for CD14 was found on fibroblasts from inflamed gingiva. Western blot profiles of the fibroblasts and monocytes showed a CD14-positive reaction at 55 kDa. Fluorescein isothiocyanate-conjugated Escherichia coli lipopolysaccharide bound to fibroblasts more strongly in the presence of 10% fetal bovine serum than without serum. This binding, as well as IL-6 production, was blocked by anti-CD14 monoclonal antibody. The results showed that CD14 was present on human gingival fibroblasts, which suggests that lipopolysaccharide modulation of gingival fibroblast function depends on CD14.     

Fluoride distribution in human dental calculus obtained from different sites on the tooth surface

We examined the site specificity of fluoride (F) distribution in human dental calculus. Teeth with supra- and subgingival calculus were obtained from patients who resided in non-fluoridated areas in Japan and China. Sequential layers of the dental calculus (30 μm thick) were abraded by an abrasive micro-sampling technique and fluoride and phosphorus in the powdered samples were analyzed. Fluoride concentrations were highest in the outer, lowest in the middle and intermediate in the inner layers of dental calculus in general. In the outermost layers fluoride concentrations were highest in calculus found near the tooth cervix both in supra- and subgingival calculus. Fluoride concentrations decreased markedly toward the apical region in subgingival calculus. while it did not change toward the incisal or occlusal region in supragingival calculus. In the inner layers, fluoride concentrations in both supra- and subgingival calculus were not affected by position on the teeth. Fluoride concentrations in subgingival calculus near the apex were lower than in supragingival calculus near the incisal or occlusal region. It was concluded that the fluoride concentrations differ in different regions of dental calculus, probably due to their different mechanisms of formation.     

Immunoexpression of extracellular matrix proteins in human salivary gland development

Immunoexpression of the extracellular matrix (ECM) proteins laminin, fibronectin, tenascin and types I, III and IV collagen was analyzed in the major and minor salivary glands of seven human fetuses at different gestational ages. The results showed the presence and localization of laminin, collagen IV and fibronectin around glandular structures at all stages of development. Tenascin was only detectable around excretory ducts. In the earliest stages of development, type I and type III collagen were presented as fine fibers delineating the glandular structures and delimiting the extension of the future lobule. As glandular development proceeded, the lobule was gradually filled with collagens and glandular tissue.     

Calcium ion release from calcium hydroxide stimulated fibronectin gene expression in dental pulp cells and the differentiation of dental pulp cells to mineralized tissue forming cells by fibronectin

Aim The effect of calcium ions on dental pulp cells was examined and the mechanism of dentine bridge formation by calcium hydroxide was investigated. Methodology Human dental pulp cells were treated with high concentration of calcium or magnesium ions for 24 h and fibronectin gene expression was measured by the quantitative PCR method. Human dental pulp cells were then cultured on fibronecin‐coated dishes for 24 h, and osteocalcin and osteopontin gene expression, which are typical phenotypes of mineralized tissue forming cells, were measured by the quantitative PCR method. Results Fibronectin gene expression was stimulated by calcium ions dose‐dependently. On the other hand, magnesium ions did not influence fibronectin gene expression. Furthermore, pulp cells cultured on fibronectin‐coated dishes enhanced the expression of phenotypes of mineralized tissue forming cells. Conclusions Calcium ions released from calcium hydroxide stimulates fibronectin synthesis in dental pulp cells. Fibronectin might induce the differentiation of dental pulp cells to mineralized tissue forming cells that are the main cells to form dentine bridges, via contact with cells.     

Comparison of exposed dentinal surfaces resulting from abrasion and erosion

The aim of this study was to compare the shape of exposed dentinal surfaces caused by abrasion and erosion with a view to developing a diagnostic clinical test. The study material consisted of 80 natural teeth and 129 dental models obtained from Australian Aborigines known to display considerable dental abrasion due to their diet, and dental models of 37 Caucasians diagnosed with dental erosion through detailed history and dietary analysis. Polyvinyl siloxane impressions were obtained of all occlusal surfaces with dentinal scooping in both the 'abrasion' and 'erosion' groups. All impressions were sectioned buccolingually through the deepest point of the scooped dentine, and then the profiles were photocopied at X 2 magnification. The breadth and depth of dentinal profiles were measured to an accuracy of 0.1 mm, enabling ratios of depth:breadth to be determined, and the position of the deepest part of each scooped surface was recorded. The mean depth:breadth ratio of scooped dentine was significantly greater in the Aboriginal natural teeth (0. 19±60.06, mean±SE) than in the Aboriginal dental models (0.15±0.04). Both Aboriginal natural teeth and models with abrasion showed significantly smaller ratios (p<0.05) than the Caucasian models showing erosion (0.33±.07). Furthermore, in the abrasion samples, the deepest region of the scooped dentine tended to be lingually placed more often in maxillary teeth but buccally placed more often in mandibular teeth (p<0.05). These results indicate that scooped dentine on abraded occlusal surfaces of teeth displays significant differences in shape compared with that caused mainly by erosion.     

Effect of root curvature on post length in the restoration of endodontically treated premolars

Abstract Two hundred and eighteen bi-rooted maxillary premolars were examined radio graphically to determine the length from the apex at which root curvature occurred. The information may serve as a guide in determining post preparation length. The results of the study indicated that the lingual root was slightly straighter than the buccal root. The curvature of the root was on the average 6.47 mm from the apex on the buccal root and 5.18 mm from the apex on the lingual root.     

血小板源性生长因子对人牙周韧带细胞在壳聚糖-磷酸三钙支架材料上附着和增殖的影响

目的:研究血小板源性生长因子(platelet-derived growth factor BB,PDGF-BB)对人牙周韧带细胞(human periodontal ligament cells,hPDLC)在壳聚糖-磷酸三钙(chitosan/tricalcium phosphate,CTCP)支架材料上附着和增殖的影响。方法:将一定体积的CTCP支架材料置96孔培养板内,取第5代hPDLC接种在材料表面。实验组加入含PDGF-BB的DMEM培养液,使得PDGF-BB终浓度分别为1ng/mL、10ng/mL和100ng/mL;对照组仅加入DMEM培养液。孵育24h及72h后分别对材料上的细胞计数;72h后对标本行扫描电镜观察。结果:与对照组相比,各浓度PDGF-BB组培养24h后均可促进hPDLC在CTCP支架材料上的附着,呈浓度依赖关系,差异具有统计学意义(P<0.05),其中100ng/mL为最有效浓度;同时培养72h后各浓度PDGF-BB组均能促进hPDLC在CTCP支架材料上的增殖,差异具有统计学意义(P<0.05);扫描电镜观察发现PDGF-BB组较对照组hPDLC在材料上附着数量增加,伸展更充分。结论:hPDLC在CTCP支架材料上的附着和增殖可被PDGF-BB所增强。