Neurogenic differentiation of dental pulp stem cells to neuron-like cells in dopaminergic and motor neuronal inductive media.

BACKGROUND/PURPOSE: Dental pulp stem cells (DPSCs) have been proposed as a promising source of stem cells in nerve regeneration due to their close embryonic origin and ease of harvest. The aim of this study was to evaluate the efficacy of dopaminergic and motor neuronal inductive media on transdifferentiation of human DPSCs (hDPSCs) into neuron-like cells. METHODS: Isolation, cultivation, and identification of hDPSCs were performed with morphological analyses and flow cytometry. The proliferation potential of DPSCs was evaluated with an XTT [(2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide)] assay. Media for the induction of dopaminergic and spinal motor neuronal differentiation were prepared. The efficacy of neural induction was evaluated by detecting the expression of neuron cell-specific cell markers in DPSCs by immunocytochemistry and quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). RESULTS: In the XTT assay, there was a 2.6- or 2-fold decrease in DPSCs cultured in dopaminergic or motor neuronal inductive media, respectively. The proportions of ßIII-tubulin (ßIII-tub), glial fibrillary acidic protein (GFAP), and oligodendrocyte (O1)-positive cells were significantly higher in DPSCs cultured in both neuronal inductive media compared with those cultured in control media. Furthermore, hDPSC-derived dopaminergic and spinal motor neuron cells after induction expressed a higher density of neuron cell markers than those before induction. CONCLUSION: These findings suggest that in response to the neuronal inductive stimuli, a greater proportion of DPSCs stop proliferation and acquire a phenotype resembling mature neurons. Such neural crest-derived adult DPSCs may provide an alternative stem cell source for therapy-based treatments of neuronal disorders and injury.

Erosive potential of soft drinks on human enamel: an in vitro study.

BACKGROUND/PURPOSE: Most soft drinks are acidic in nature. Regular consumption of these drinks may result in dental erosion. The aim of this in vitro study was to evaluate the erosive potential of different soft drinks in Taiwan by a novel multiple erosive method. METHODS: Four commercially available soft drinks in Taiwan were selected for this study. The properties of each product were analyzed to measure their pH, titratable acidity, and ion contents. The erosive potential of the soft drinks was measured based on the amount of loss of human enamel surface following its exposure to the soft drinks tested for different periods (20 minutes, 60 minutes, and 180 minutes). The enamel loss was measured using a confocal laser scanning microscope. RESULTS: The pH values of the soft drinks were below the critical pH value (5.5) for enamel demineralization, and ranged from 2.42 to 3.46. The drink with ingredients of citric acid and ascorbic acid had the highest titratable acidity (33.96 mmol OH(-)/L to pH 5.5 and 71.9 mmol OH(-)/L to pH 7). Exposure to all the soft drinks resulted in loss of human enamel surface (7.28-34.07 µm for 180-minute exposure). The beverage with the highest calcium content had the lowest erosive potential. CONCLUSION: All tested soft drinks were found to be erosive. Soft drinks with high calcium contents have significantly lower erosive potential. Low pH value and high citrate content may cause more surface enamel loss. As the erosive time increased, the titratable acidity to pH 7 may be a predictor of the erosive potential for acidic soft drinks. The erosive potential of the soft drinks may be predicted based on the types of acid content, pH value, titratable acidity, and ion concentration.

Role of taurine on acid secretion in the rat stomach.

BACKGROUND: Taurine has chemical structure similar to an inhibitory neurotransmitter, γ-aminobutyric acid (GABA). Previous studies on GABA in the stomach suggest GABAergic neuron is involved in acid secretion, but the effects of taurine are poor understood. METHODS: The effects of taurine on acid secretion, signal transduction, and localization of taurinergic neurons were determined in the rat stomach using everted whole stomach, RIA kit and immunohistochemical methods. RESULTS: We used antibodies against taurine-synthesizing enzyme, cysteine sulfuric acid decarboxylase (CSAD), and taurine. CSAD- and taurine-positive cells were found in the muscle and mucosal layers. Distributions of CSAD- and taurine-positive cells in both mucosal and muscle layers were heterogeneous in the stomach. Taurine at 10-9~10-4 M induced acid secretion, and the maximum secretion was at 10-5 M, 1.6-fold higher than the spontaneous secretion. Taurine-induced acid secretion was completely inhibited by bicuculline and atropine but not by cimetidine, proglumide, or strychnine. Atropine and tetrodotoxin (TTX) completely inhibited the acid secretion induced by low concentrations of taurine and partially inhibited induced by high concentrations. Verapamil, a calcium blocker agent, inhibited acid output elicited by taurine. We assumed all Ca2+ channels involved in the response to these secretagogues were equally affected by verapamil. Intracellular cAMP (adenosine 3', 5'-monophosphate) in the stomach significantly increased with taurine treatment in a dose-dependent manner. High correlation (r=0.859, p < 0.001) of taurine concentrations with cAMP was observed. CONCLUSIONS: Our results demonstrated for the first time in taurine-induced acid secretion due to increase intracellular calcium may act through the A type of GABA receptors, which are mainly located on cholinergic neurons though cAMP pathway and partially on nonneuronal cells in the rat stomach.

Regeneration of Tooth with Allogenous, Autoclaved Treated Dentin Matrix with Dental Pulpal Stem Cells: An In Vivo Study.

INTRODUCTION: Biomaterials designed for tissue engineering should be nontoxic and nonimmunogenic and should achieve their intended functions. Treated dentin matrix (TDM), a bioactive extracellular matrix, is promising for tooth regeneration. However, the effect of sterilization on the surface properties of allogenous TDM in the animal model is unclear. METHODS: The biological characteristics and influences of dental pulp stem cells (DPSCs) with autoclaved TDM (a-TDM) were studied using scanning electron microscopy, immunofluorescence microscopy, immunohistochemistry, and reverse transcription polymerase chain reaction in vitro. In addition, a-TDM was implanted in a mouse model for 6 weeks and was a substrate with DPSCs for tooth reconstruction in a goat animal model in vivo. RESULTS: Allogenous a-TDM induced and supported DPSCs to develop new dentin pulp-like tissues, enamel dental pulp, and cementum periodontal complexes. Immunohistochemistry data showed that the markers dentin sialoprotein, ßⅢ-tubulin, dentin matrix protein 1, amelogenin, VIII factors, type I collagen, cementum-derived attachment protein, and scleraxis transcription factor were positive stained in toothlike tissue. CONCLUSIONS: Allogenous a-TDM served as an effective scaffold enabling DPSCs to proliferate and differentiate into a broad array of resident cells including odontoblasts, fibroblasts, vascular cells, and neural endings. Allogenous a-TDM with DPSCs can provide an ideal biomaterial for optimizing the regeneration of tooth material.

Effect of tricalcium aluminate on the physicochemical properties, bioactivity, and biocompatibility of partially stabilized cements.

BACKGROUND/PURPOSE: Mineral Trioxide Aggregate (MTA) was widely used as a root-end filling material and for vital pulp therapy. A significant disadvantage to MTA is the prolonged setting time has limited the application in endodontic treatments. This study examined the physicochemical properties and biological performance of novel partially stabilized cements (PSCs) prepared to address some of the drawbacks of MTA, without causing any change in biological properties. PSC has a great potential as the vital pulp therapy material in dentistry. METHODS: This study examined three experimental groups consisting of samples that were fabricated using sol-gel processes in C3S/C3A molar ratios of 9/1, 7/3, and 5/5 (denoted as PSC-91, PSC-73, and PSC-55, respectively). The comparison group consisted of MTA samples. The setting times, pH variation, compressive strength, morphology, and phase composition of hydration products and ex vivo bioactivity were evaluated. Moreover, biocompatibility was assessed by using lactate dehydrogenase to determine the cytotoxicity and a cell proliferation (WST-1) assay kit to determine cell viability. Mineralization was evaluated using Alizarin Red S staining. RESULTS: Crystalline phases, which were determined using X-ray diffraction analysis, confirmed that the C3A contents of the material powder differed. The initial setting times of PSC-73 and PSC-55 ranged between 15 and 25 min; these values are significantly (p<0.05, ANOVA and post-hoc test) lower than those obtained for MTA (165 min) and PSC-91 (80.5 min). All of the PSCs exhibited ex vivo bioactivity when immersed in simulated body fluid. The biocompatibility results for all of the tested cements were as favorable as those of the negative control, except for PSC-55, which exhibited mild cytotoxicity. CONCLUSION: PSC-91 is a favorable material for vital pulp therapy because it exhibits optimal compressive strength, a short setting time, and high biocompatibility and bioactivity.

Development of calcium phosphate/sulfate biphasic cement for vital pulp therapy.

OBJECTIVES: Bioactive calcium phosphate cement (CPC) has been used widely to repair bone defects because of its excellent biocompatibility and bioactivity. However, the poor handling properties, low initial mechanical strength, and long setting time of CPC limit its application in vital pulp therapy (VPT). The aim of this study was to synthesize biphasic calcium phosphate/sulfate cements and evaluate the feasibility of applying these cements in VPT. METHODS: The physical, chemical, and mechanical properties of CPC were improved by mixing the cement with various amounts of α-calcium sulfate hemihydrate (CSH). The hydration products and crystalline phases of the materials were characterized using scanning electron microscopy and X-ray diffraction analysis. In addition, the physical properties, such as the setting time, compressive strength, viscosity, and pH were determined. Water-soluble tetrazolium salt-1 and lactase dehydrogenase were used to evaluate cell viability and cytotoxicity. RESULTS: The developed CPC (CPC/CSH cement), which contains 50wt% CSH cement, exhibited no obvious temperature increase or pH change during setting when it was used as a paste. The initial setting time of the CPC/CSH biphasic cement was substantially shorter than that of CPC, and the initial mechanical strength was 23.7±5.6MPa. The CPC/CSH cement exhibited higher viscosity than CPC and, thus, featured acceptable handling properties. X-ray diffraction analysis revealed that the relative peak intensity for hydroxyapatite increased, and the intensity for calcium sulfate dehydrate decreased as the amount of CPC was increased. The cell viability and cytotoxicity test results indicated that the CPC/CSH cement did not harm dental pulp cells. SIGNIFICANCE: The developed CPC/CSH biphasic cement exhibits substantial potential for application in VPT.

Orientation-enhanced growth and optical properties of ZnO nanowires grown on porous silicon substrates.

ZnO nanowires have been synthesized on porous silicon substrates with different porosities via the vapour-liquid-solid method. The texture coefficient analysed from the XRD spectra indicates that the nanowires are more highly orientated on the appropriate porosity of porous silicon substrate than on the smooth surface of silicon. The Raman spectrum reveals the high quality of the ZnO nanowires. From the temperature-dependent photoluminescence spectra, we deduced the activation energies of free and bound excitons.