Concurrent expression of Oct4 and Nanog maintains mesenchymal stem-like property of human dental pulp cells.

Human dental pulp stem cells (DPSCs), unique mesenchymal stem cells (MSCs) type, exhibit the characteristics of self-renewal and multi-lineage differentiation capacity. Oct4 and Nanog are pluripotent genes. The aim of this study was to determine the physiological functions of Oct4 and Nanog expression in DPSCs. Herein, we determined the critical role of an Oct4/Nanog axis modulating MSCs properties of DPSCs by lentiviral-mediated co-overexpression or co-knockdown of Oct4/Nanog in DPSCs. MSCs properties including osteogenic/chondrogenic/adipogenic induction differentiation was assayed for expression of osteogenic/chondrogenic/adipogenic markers by quantitative real-time RT-PCR analysis. Initially, we observed that the expression profile of Oct4 and Nanog in dental pulp cells, which exerted properties of MSCs, was significantly up-regulated compared to that of STRO-1-CD146- dental pulp cells. Down-regulation of Oct4 and Nanog co-expression significantly reduced the cell proliferation, osteogenic differentiation capability, STRO-1, CD146, and Alkaline phosphatase (ALP) activity of DPSCs. In contrast, co-overexpression of Oct4 and Nanog enhanced the expression level of STRO-1 and CD146, proliferation rate and osteogenic/chondrogenic/adipogenic induction differentiation capability, and expression of osteogenic/chondrogenic/adipogenic induction differentiation markers. Our results suggest that Oct4-Nanog signaling is a regulatory switch to maintain properties in DPSCs.

Synergies of Human Umbilical Vein Endothelial Cell-Laden Calcium Silicate-Activated Gelatin Methacrylate for Accelerating 3D Human Dental Pulp Stem Cell Differentiation for Endodontic Regeneration.

According to the Centers for Disease Control and Prevention, tooth caries is a common problem affecting 9 out of every 10 adults worldwide. Dentin regeneration has since become one of the pressing issues in dentistry with tissue engineering emerging as a potential solution for enhancing dentin regeneration. In this study, we fabricated cell blocks with human dental pulp stem cells (hDPSCs)-laden alginate/fish gelatin hydrogels (Alg/FGel) at the center of the cell block and human umbilical vascular endothelial cells (HUVEC)-laden Si ion-infused fish gelatin methacrylate (FGelMa) at the periphery of the cell block. 1H NMR and FTIR results showed the successful fabrication of Alg/FGel and FGelMa. In addition, Si ions in the FGelMa were noted to be bonded via covalent bonds and the increased number of covalent bonds led to an increase in mechanical properties and improved degradation of FGelMa. The Si-containing FGelMa was able to release Si ions, which subsequently significantly not only enhanced the expressions of angiogenic-related protein, but also secreted some cytokines to regulate odontogenesis. Further immunofluorescence results indicated that the cell blocks allowed interactions between the HUVEC and hDPSCs, and taken together, were able to enhance odontogenic-related markers' expression, such as alkaline phosphatase (ALP), dentin matrix phosphoprotein-1 (DMP-1), and osteocalcin (OC). Subsequent Alizarin Red S stain confirmed the benefits of our cell block and demonstrated that such a novel combination and modification of biomaterials can serve as a platform for future clinical applications and use in dentin regeneration.

Rehardening and the Protective Effect of Gamma-Polyglutamic Acid/Nano-Hydroxyapatite Paste on Surface-Etched Enamel.

Many revolutionary approaches are on the way pertaining to the high occurrence of tooth decay, which is an enduring challenge in the field of preventive dentistry. However, an ideal dental care material has yet to be fully developed. With this aim, this research reports a dramatic enhancement in the rehardening potential of surface-etched enamels through a plausible synergistic effect of the novel combination of γ-polyglutamic acid (γ-PGA) and nano-hydroxyapatite (nano-HAp) paste, within the limitations of the study. The percentage of recovery of the surface microhardness (SMHR%) and the surface parameters for 9 wt% γ-PGA/nano-HAp paste on acid-etched enamel were investigated with a Vickers microhardness tester and an atomic force microscope, respectively. This in vitro study demonstrates that γ-PGA/nano-HAp treatment could increase the SMHR% of etched enamel to 39.59 ± 6.69% in 30 min. To test the hypothesis of the rehardening mechanism and the preventive effect of the γ-PGA/nano-HAp paste, the surface parameters of mean peak spacing (Rsm) and mean arithmetic surface roughness (Ra) were both measured and compared to the specimens subjected to demineralization and/or remineralization. After the treatment of γ-PGA/nano-HAp on the etched surface, the reduction in Rsm from 999 ± 120 nm to 700 ± 80 nm suggests the possible mechanism of void-filling within a short treatment time of 10 min. Furthermore, ΔRa-I, the roughness change due to etching before remineralization, was 23.15 ± 3.23 nm, while ΔRa-II, the roughness change after remineralization, was 11.99 ± 3.90 nm. This statistically significant reduction in roughness change (p < 0.05) implies a protective effect against the demineralization process. The as-developed novel γ-PGA/nano-HAp paste possesses a high efficacy towards tooth microhardness rehardening, and a protective effect against acid etching.

Corrosion resistance of different nickel-titanium archwires in acidic fluoride-containing artificial saliva.

OBJECTIVE: To test the hypothesis that different nickel-titanium (NiTi) archwires may have dissimilar corrosion resistance in a fluoride-containing oral environment. MATERIALS AND METHODS: Linear polarization test, a fast electrochemical technique, was used to evaluate the corrosion resistance, in terms of polarization resistance (R(p)), of four different commercial NiTi archwires in artificial saliva (pH 6.5) with various NaF concentrations (0%, 0.01%, 0.1%, 0.25%, and 0.5%). Two-way analysis of variance was used to analyze R(p) with the factors of archwire manufacturer and NaF concentration. Surface characterizations of archwires were analyzed using scanning electron microscopy, atomic force microscopy, and x-ray photoelectron spectroscopy. RESULTS: Both archwire manufacturer and NaF concentration had a significant influence on R(p) of NiTi archwires. Different surface topography was present on the test NiTi archwires that contained the similar surface chemical structure (TiO(2) and trace NiO). The surface topography did not correspond to the difference in corrosion resistance of the NiTi archwires. Increasing the NaF concentration in artificial saliva resulted in a decrease in R(p), or corrosion resistance, of all test NiTi archwires. The NiTi archwires severely corroded and showed similar corrosion resistance in 0.5% NaF-containing environment. CONCLUSIONS: Different NiTi archwires had dissimilar corrosion resistance in acidic fluoride-containing artificial saliva, which did not correspond to the variation in the surface topography of the archwires. The presence of fluoride in artificial saliva was detrimental to the corrosion resistance of the test NiTi archwires, especially at a 0.5% NaF concentration.

The In Vivo Toxicity and Antimicrobial Properties for Electrolyzed Oxidizing (EO) Water-Based Mouthwashes.

The objective of this study was to verify the feasibility of electrolyzed oxidizing (EO) water as a mouthwash through the evaluation of its in vivo toxicity by embryonic zebrafish and antimicrobial efficacy against Streptococcus mutans (S. mutans). METHODOLOGY: Each 1.5-3.0 g of sodium chloride (NaCl), sodium bromide (NaBr), or calcium chloride (CaCl2) were added into an electrolyzer with 300 mL of DD water to produce electrolyzed oxidizing (EO) water. A zebrafish embryo assay was used to evaluate acute toxicity of specimens. Antimicrobial property was conducted with 100 µL microbial count of 1 × 108 cfu/mL S. mutans to blend with each 10 mL specimen of chlorhexidine (CHX) gluconate or hypochlorous acid (HOCl) for various time points. The concentration of viable microorganisms was assessed according to individually standardized inoculum by a plate-count method. RESULTS: Among the EO water produced from NaCl, NaBr, and CaCl2, the EO water from NaCl showed a relatively low mortality rate of zebrafish embryos and was chosen for a detailed investigation. The mortality rates for the groups treated with EO water containing 0.0125% and 0.0250% HOCl were not statically different from those of a negative control, however the mortality rate was 66.7 ± 26.2% in 0.2% CHX gluconate for the same treatment time of 0.5 min. All of the HOCl or 2.0% CHX gluconate groups showed >99.9% antimicrobial effectiveness against S. mutans; while the 0.2% CHX gluconate group showed a bacterial reduction rate of 87.5% and 97.1% for treatment times of 0.5 min and 1.0 min, respectively. CONCLUSIONS: Except for the 0.2% CHX gluconate, all the HOCl specimens and 2.0% CHX gluconate revealed similar antimicrobial properties (>99.9%) against S. mutans. The EO water comprised of both 0.0125% and 0.0250% HOCl showed >99.9% antimicrobial efficacy but with little in vivo toxicity, illuminating the possibility as an alternative mouthwash for dental and oral care.

A unique hybrid-structured surface produced by rapid electrochemical anodization enhances bio-corrosion resistance and bone cell responses of ß-type Ti-24Nb-4Zr-8Sn alloy.

Ti-24Nb-4Zr-8Sn (Ti2448), a new ß-type Ti alloy, consists of nontoxic elements and exhibits a low uniaxial tensile elastic modulus of approximately 45 GPa for biomedical implant applications. Nevertheless, the bio-corrosion resistance and biocompatibility of Ti2448 alloys must be improved for long-term clinical use. In this study, a rapid electrochemical anodization treatment was used on Ti2448 alloys to enhance the bio-corrosion resistance and bone cell responses by altering the surface characteristics. The proposed anodization process produces a unique hybrid oxide layer (thickness 50-120 nm) comprising a mesoporous outer section and a dense inner section. Experiment results show that the dense inner section enhances the bio-corrosion resistance. Moreover, the mesoporous surface topography, which is on a similar scale as various biological species, improves the wettability, protein adsorption, focal adhesion complex formation and bone cell differentiation. Outside-in signals can be triggered through the interaction of integrins with the mesoporous topography to form the focal adhesion complex and to further induce osteogenic differentiation pathway. These results demonstrate that the proposed electrochemical anodization process for Ti2448 alloys with a low uniaxial tensile elastic modulus has the potential for biomedical implant applications.

Fracture resistance of Nd:YAG laser-welded cast titanium joints with various clinical thicknesses and welding pulse energies.

The purpose of this study was to evaluate the fracture resistance of Nd:YAG laser-welded cast titanium (Ti) joints with various clinical thicknesses and welding pulse energies. A four-point bending test was used to assess the effects of various specimen thicknesses (1-3 mm) and welding pulse energies (11-24 J) on the fracture resistance of Nd:YAG laser-welded Ti dental joints. Fracture resistance was evaluated in terms of the ratio of the number of fractured specimens to the number of tested specimens. As for the fracture frequencies, they were compared using the Cochran-Mantel-Haenszel test. Morphology of the fractured Ti joints was observed using a scanning electron microscope. Results showed that decreasing the specimen thickness and/or increasing the welding pulse energy, i.e., increasing the welded area percentage, resulted in an increase in the fracture resistance of the Ti joint. Where fracture occurred, the fracture site would be at the center of the weld metal.

Andrographolide impedes cancer stemness and enhances radio-sensitivity in oral carcinomas via miR-218 activation.

Current evidence suggests that oral cancer stem cells (OCSCs) possess high tumorigenic and metastatic properties as well as chemo- and radioresistance. In this study, we demonstrated that andrographolide, the main bioactive component in the medicinal plant Andrographis, significantly reduced oncogenicity and restored radio-sensitivity of ALDH1+CD44+ OCSCs. Mechanistic studies showed that andrographolide treatment increased the expression of microRNA-218 (miR-218), leading to the downregulation of Bmi1. We showed that knockdown of miR-218 in ALDH1-CD44- non-OCSCs enhanced cancer stemness, while silencing of Bmi1 significantly counteracted it. Furthermore, we found tumor growth was reduced in mice bearing xenograft tumors after andrographolide treatment via activation of miR-218/Bmi1 axis. Together, these data demonstrated that the inhibition of tumor aggressiveness in OCSCs by andrographolide was mediated through the upregulation of miR-218, thereby reducing Bmi1 expression. These findings suggest that andrographolide may be a valuable natural compound for anti-CSCs treatment of OSCC.

Surface analysis and corrosion resistance of different stainless steel orthodontic brackets in artificial saliva.

The purpose of this study was to investigate the variation in corrosion resistance of commercial stainless steel (SS) brackets with different brands and types for the same application, using the electrochemical technique. The linear polarization test was used to evaluate the corrosion resistance, in terms of polarization resistance (Rp), of as-received commercial SS brackets in acidic artificial saliva. A two-way analysis of variance was used to analyze the Rp with the factors of brand and type. A scanning electron microscope and an atomic force microscope were used to analyze the surface morphology and roughness, respectively. The X-ray photoelectron spectroscopy was used to identify the chemical composition of the passive film on SS brackets. Results showed that different brands of SS brackets had a statistically significant difference in Rp (P < .0001), whereas there was no statistical difference between the bracket types ("Roth" and standard) (P = .27). Different surface topography, including surface roughness and defect, was present among the tested SS brackets. The same passive film structure, containing Cr2O3/Fe2O3 with small amounts of NiO, was observed on all SS brackets. The surface topography of the commercial SS brackets with identical surface passive film structure did not correspond with the difference in corrosion resistance.

Electrochemical impedance spectroscopy study of Ti-6Al-4V alloy in artificial saliva with fluoride and/or bovine albumin.

OBJECTIVES: To investigate the electrochemical mechanism of Ti-6Al-4V alloy in acidic artificial saliva with NaF and/or bovine albumin (BA). METHODS: The electrochemical behavior of Ti-6Al-4V alloy was investigated using electrochemical impedance spectroscopy (EIS) measurements at the open circuit potentials. The electrolytes used included artificial saliva with 0-0.5% NaF, with 0.1% NaF+0.01-0.5% BA, and with 0.01-0.5% BA at pH 5 and 37 degrees C. One-way ANOVA was used to analyze the influence of NaF and BA concentration on the polarization resistance (Rp) obtained from the EIS data. Surface chemical analyses were performed with X-ray photoelectron spectroscopy. RESULTS: The impedance was mainly characterized by the resistance and capacitance of a protective TiO2-based passive film on Ti-6Al-4V alloy when NaF concentration was low (< or = 0.01%). When NaF concentration was higher than 0.1%, the impedance was characterized by two capacitive and one inductive effects, which were related to the metal's charge transfer reaction and the decayed TiO2-based oxide film (capacitive effects), and the relaxation of the adsorbed Ti-F complex compound (inductive effect). The addition of 0.01-0.5% BA in 0.1% NaF-containing artificial saliva significantly increased the impedance of Ti-6Al-4V alloy compared to that without BA addition (p < 0.001). The presence of BA in fluoride-free artificial saliva had no influence on the electrochemical impedance of Ti-6Al-4V alloy (p > 0.05). SIGNIFICANCE: The EIS measurement technique is suitable for the study of the electrochemical behavior of dental alloy. The electrochemical mechanism of Ti-6Al-4V alloy in artificial saliva is related to the fluoride and bovine albumin concentration.

Anti-hypertension effects of traditional Chinese medicine ju-ling-tang on renal hypertensive rats.

This research investigated the anti-hypertension effect of the traditional Chinese medicine (TCM) ju-ling-tang (JLT) on an animal model of hypertension induced by unilateral renal artery ligation. In the study of anti-hypertension effects, 60 minutes after oral administration with NG tube feeding of 240 mg/kg JLT, a significant decrease in blood pressure (p < 0.05) was observed and sustained till 120 minutes. In the group given 50 mg/kg alpha-methyldopa orally, the effect was obvious 90 minutes after medication (p < 0.01), and lasted until 240 minutes. In terms of organ pathology, a significant reduction in the extent of induced glomerular sclerosis was observed in rats given 240 mg/kg JLT compared with the control. From these results, we infer that JLT has a beneficial anti-hypertensive effect on renal hypertension.

Ion release from NiTi orthodontic wires in artificial saliva with various acidities.

NiTi orthodontic wire products from different manufacturers would have different corrosion resistance. We assayed the corrosion resistance, in terms of ion release, of different NiTi orthodontic wires in artificial saliva with various acidities. Four types of as-received commercial NiTi orthodontic wires were immersed in artificial saliva (37 degrees C) at pH 2.5-6.25 for different periods (1-28d). The amount of Ni and Ti ions released from NiTi wires was determined using an atomic adsorption spectrophotometer. Surface morphology and roughness of wires were related to the corrosion resistance. Results showed that the manufacturer, pH value, and immersion period, respectively, had a significantly statistical influence on the release amount of Ni and Ti ions. The amount of Ni ions released in all test solutions was well below the critical value necessary to induce allergy and below daily dietary intake level. The amount of Ti ions released in pH>/=3.75 solution was mostly not detectable, representing that the TiO(2) film on NiTi wires exhibited a good protection against corrosion. Pre-existed surface defects on NiTi wires might be the preferred locations for corrosion. The NiTi wire with the highest release amount of metal ions had the maximal increase in surface roughness after immersion test, while a rougher surface did not correspond to a higher metal ion release.

Effect of surface roughness of ground titanium on initial cell adhesion.

The effect of surface roughness of ground Ti on the initial adhesion of osteoblast-like U-2 OS cells was investigated in this study. Different numbers (#120, #600, and #1500) of SiC sandpaper and two Al2O3 polishing powder (0.3 and 1 microm) were used to prepare the metal specimens with varying degrees of surface roughness. Surface roughness (Ra) was measured by profilometry. Surface topography was observed using an atomic force microscope. MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay was used to measure the optical density (OD) of specimens after 2 h of cell incubation. The OD value was analyzed by one-way ANOVA for analyzing the factor of surface roughness. Crystal violet staining technique was used to characterize the cell spreading. Results showed that the specimen of #1500 Ti (Ra: 0.15 microm) had the highest OD value. The specimens polished with 0.3 and 1 microm Al2O3 powder (Ra: 0.05 and 0.07 microm) exhibited the worst cell adhesion behavior. Contact guidance of cells could be observed on the rougher #600 and #120 specimens (Ra: 0.33 and 1.20 microm). This study concludes that the surface roughness (Ra: 0.05-1.20 microm) of ground Ti has a highly significant influence on the initial adhesion of osteoblast-like U-2 OS cells. The ground Ti with an Ra of 0.15 microm shows the optimal cell adhesion behavior with respect to either the rougher or smoother specimens.