This study was to explore a better three-dimensional (3-D) culture method of chondrocyte. The interpenetrating network (IPN) gel beads were developed through a photo-cross linking reaction with mixed barium ions and calcium ions at the ratio of 5:5 with the methacrylic alginate (MA), which was a chemically conjugated alginate with methacrylic groups. The second generation of primary cartilage cells was encapsulated in the MA gel beads for three weeks. In the designated timing, HE stain, Alamar blue method and Scanning electron microscopic were used to determine the cartilage cells growth, proliferation and the cell distribution in the scaffolds, respectively. The expression of type II collagen was investigated by an immunohistochemistry assay and the glycosaminoglycan content was quantitatively evaluated with the spectrophotometry of 1, 9 dimethylene blue assay. Compared to the alginate control group, the deposition of glycosaminoglycan was significantly upregulated in IPN-MA gel beads with higher cell proliferation. The secretion of extracellular matrix and proliferation of chondrocyte in methacrylic alginate gel beads were higher than that in Alginate beads. Cells were able to attach, to grow well on the scaffolds under scanning electron microscopy. The result of immunohistochemistry staining of collagen type II was positive, confirming the maintenance of chondrocyte phenotype in methacrylic alginate gel beads. This study shows a great potential for three-dimensional culture of cartilage.
Alginates ; chemistry ; Barium ; chemistry ; Calcium ; chemistry ; Cartilage ; cytology ; Cations ; Cell Culture Techniques ; instrumentation ; Cells, Cultured ; Chondrocytes ; cytology ; Collagen Type II ; chemistry ; Glucuronic Acid ; chemistry ; Glycosaminoglycans ; chemistry ; Hexuronic Acids ; chemistry ; Metals ; chemistry ; Microscopy, Electron, Scanning

The aim of this study was to evaluate the effect of controlled intraoral grinding and polishing on the roughness of full-contour zirconia compared to classical veneered zirconia. Thirty bar-shaped zirconia specimens were fabricated and divided into two groups (n=15). Fifteen specimens (group 1) were glazed and 15 specimens (group 2) were veneered with feldspathic ceramic and then glazed. Prior to grinding, maximum roughness depth (Rmax) values were measured using a profilometer, 5 times per specimen. Simulated clinical grinding and polishing were performed on the specimens under water coolant for 15 s and 2 N pressure. For grinding, NTI diamonds burs with grain sizes of 20 µm, 10 µm, and 7.5 µm were used sequentially. The ground surfaces were polished using NTI kits with coarse, medium and fine polishers. After each step, Rmax values were determined. Differences between groups were examined using one-way analysis of variance (ANOVA). The roughness of group 1 was significantly lower than that of group 2. The roughness increased significantly after coarse grinding in both groups. The results after glazing were similar to those obtained after fine grinding for non-veneered zirconia. However, fine-ground veneered zirconia had significantly higher roughness than venerred, glazed zirconia. No significant difference was found between fine-polished and glazed zirconia, but after the fine polishing of veneered zirconia, the roughness was significantly higher than after glazing. It can be concluded that for full-contour zirconia, fewer defects and lower roughness values resulted after grinding and polishing compared to veneered zirconia. After polishing zirconia, lower roughness values were achieved compared to glazing; more interesting was that the grinding of glazed zirconia using the NTI three-step system could deliver smooth surfaces comparable to untreated glazed zirconia surfaces.
Aluminum Silicates ; chemistry ; Ceramics ; chemistry ; Crowns ; Dental Materials ; chemistry ; Dental Polishing ; instrumentation ; methods ; Dental Prosthesis Design ; Dental Veneers ; Diamond ; chemistry ; Humans ; Materials Testing ; Microscopy, Electron, Scanning ; Particle Size ; Potassium Compounds ; chemistry ; Pressure ; Surface Properties ; Time Factors ; Water ; chemistry ; Yttrium ; chemistry ; Zirconium ; chemistry

The time domain entombment of bacteria by intratubular mineralization following orthograde canal obturation with mineral trioxide aggregate (MTA) was studied by scanning electron microscopy (SEM). Single-rooted human premolars (n=60) were instrumented to an apical size #50/0.06 using ProFile and treated as follows: Group 1 (n=10) was filled with phosphate buffered saline (PBS); Group 2 (n=10) was incubated with Enterococcus faecalis for 3 weeks, and then filled with PBS; Group 3 (n=20) was obturated orthograde with a paste of OrthoMTA (BioMTA, Seoul, Korea) and PBS; and Group 4 (n=20) was incubated with E. faecalis for 3 weeks and then obturated with OrthoMTA-PBS paste. Following their treatments, the coronal openings were sealed with PBS-soaked cotton and intermediate restorative material (IRM), and the roots were then stored in PBS for 1, 2, 4, 8 or 16 weeks. After each incubation period, the roots were split and their dentin/MTA interfaces examined in both longitudinal and horizontal directions by SEM. There appeared to be an increase in intratubular mineralization over time in the OrthoMTA-filled roots (Groups 3 and 4). Furthermore, there was a gradual entombment of bacteria within the dentinal tubules in the E. faecalis inoculated MTA-filled roots (Group 4). Therefore, the orthograde obturation of root canals with OrthoMTA mixed with PBS may create a favorable environment for bacterial entombment by intratubular mineralization.
Aluminum Compounds ; therapeutic use ; Calcification, Physiologic ; physiology ; Calcium Compounds ; therapeutic use ; Crystallization ; Dental Pulp Cavity ; microbiology ; Dentin ; microbiology ; Drug Combinations ; Enterococcus faecalis ; ultrastructure ; Humans ; Methylmethacrylates ; therapeutic use ; Microscopy, Electron, Scanning ; Oxides ; therapeutic use ; Root Canal Filling Materials ; therapeutic use ; Root Canal Obturation ; methods ; Root Canal Preparation ; instrumentation ; Silicates ; therapeutic use ; Time Factors ; Zinc Oxide-Eugenol Cement ; therapeutic use

In this study, we evaluate the influence of post surface pre-treatments on the bond strength of four different cements to glass fiber posts. Eighty extracted human maxillary central incisors and canines were endodontically treated and standardized post spaces were prepared. Four post pre-treatments were tested: (i) no pre-treatment (NS, control), (ii) sandblasting (SA), (iii) silanization (SI) and (iv) sandblasting followed by silanization (SS). Per pre-treatment, four dual-cure resin cements were used for luting posts: DMG LUXACORE Smartmix Dual, Multilink Automix, RelyX Unicem and Panavia F2.0. All the specimens were subjected to micro push-out test. Two-way analysis of variance and Tukey post hoc tests were performed (α=0.05) to analyze the data. Bond strength was significantly affected by the type of resin cement, and bond strengths of RelyX Unicem and Panavia F2.0 to the fiber posts were significantly higher than the other cement groups. Sandblasting significantly increased the bond strength of DMG group to the fiber posts.
Aluminum Oxide ; chemistry ; Composite Resins ; chemistry ; Curing Lights, Dental ; classification ; Cuspid ; pathology ; Dental Bonding ; Dental Etching ; methods ; Dental Materials ; chemistry ; Dental Stress Analysis ; instrumentation ; Glass ; chemistry ; Humans ; Incisor ; pathology ; Materials Testing ; Microscopy, Electron, Scanning ; Polymerization ; Post and Core Technique ; instrumentation ; Resin Cements ; chemistry ; Root Canal Preparation ; methods ; Self-Curing of Dental Resins ; methods ; Silanes ; chemistry ; Stress, Mechanical ; Surface Properties ; Tooth, Nonvital ; therapy

The present investigation assessed the effect of acid etching on marginal adaptation of white- and gray-colored mineral trioxide aggregate (MTA) to apical dentin using microcomputed tomography (micro-CT) and scanning electron microscopy (SEM). Sixty-four extracted single-rooted human maxillary teeth were used. Following root-end resection and apical preparation, the teeth were equally divided into four groups according to the following root end filling materials: (i) white-colored MTA (WMTA), (ii) etched WMTA (EWMTA), (iii) gray-colored MTA (GMTA) and (iv) etched GMTA (EGMTA). After 48 h, the interface between root-end filling materials and the dentinal walls was assessed using micro-CT and SEM. Data were statistically analyzed using the Kruskal-Wallis and Dunn tests. Micro-CT analysis revealed gap volumes between the apical cavity dentin walls and EGMTA, GMTA, EWMTA and WMTA of (0.007 1±0.004) mm(3), (0.053±0.002) mm(3), (0.003 6±0.001) mm(3) and (0.005 9±0.002) mm(3) respectively. SEM analysis revealed gap sizes for EGMTA, WMTA, EWMTA and GMTA to be (492.3±13.8) µm, (594.5±17.12) µm, (543.1±15.33) µm and (910.7±26.2) µm respectively. A significant difference in gap size between root end preparations filled with GMTA and EGMTA was found (P<0.05). No significance difference in gap size between WMTA and EWMTA were found in either SEM or micro-CT analysis. In conclusion, pre-etching of apical dentin can provide a better seal for GMTA but not for WMTA.
Acid Etching, Dental ; methods ; Aluminum Compounds ; chemistry ; Apicoectomy ; methods ; Calcium Compounds ; chemistry ; Dental Bonding ; Dental Marginal Adaptation ; Dental Pulp Cavity ; ultrastructure ; Dentin ; ultrastructure ; Drug Combinations ; Humans ; Materials Testing ; Microscopy, Electron, Scanning ; Oxides ; chemistry ; Retrograde Obturation ; methods ; Root Canal Filling Materials ; chemistry ; Root Canal Preparation ; instrumentation ; methods ; Silicates ; chemistry ; Surface Properties ; Time Factors ; Tooth Apex ; ultrastructure ; X-Ray Microtomography

This in vitro study aims to evaluate the crystal and surface microstructure of dental enamel after cold-light bleaching treatment. Twelve sound human premolars were cross-split into four specimens, namely, mesio-buccal (Group LP), disto-buccal (Group P), mesio-lingual (Group NP) and disto-lingual (Group L) specimens. These four groups were treated using the standard cold-light bleaching procedure, a bleaching agent, a peroxide-free bleaching agent and cold-light, respectively. Before and after treatment, all specimens were analyzed by high-resolution, micro-area X-ray diffraction and scanning electron microscopy. Using a spectrometer, tooth color of all specimens was measured before and after treatment. The phase of the enamel crystals was identified as hydroxyapatite and carbonated hydroxyapatite. After treatment, specimens in Groups LP and P showed significantly weaker X-ray diffraction peaks, significant reduction in crystal size and crystallinity, significant increase in L* but decrease in a* and b*, and obvious alterations in the surface morphology. However, specimens in Groups NP and L did not show any significant changes. The cold-light bleaching treatment leads to demineralization in the enamel surface. The acidic peroxide-containing bleaching agent was the major cause of demineralization, whereas cold-light did not exhibit significant increase or decrease effect on this demineralization.
Color ; Crystallography ; Dental Enamel ; drug effects ; radiation effects ; ultrastructure ; Durapatite ; radiation effects ; Humans ; Hydrogen Peroxide ; pharmacology ; Hydrogen-Ion Concentration ; Lighting ; instrumentation ; Materials Testing ; Microscopy, Electron, Scanning ; Silicon Dioxide ; pharmacology ; Spectrum Analysis ; Tooth Bleaching ; methods ; Tooth Bleaching Agents ; classification ; pharmacology ; Tooth Demineralization ; pathology ; X-Ray Diffraction

OBJECTIVETo evaluate the microleakage of fiber post and core systems after high-speed handpiece preparation at different time points.

METHODSThe crowns of forty-five extracted human premolar were removed and the roots were endodontically treated. The samples were devided into five groups. Root canal preparation was performed on each premolar followed by fiber post cementation and core build up. Tooth preparation was applied at 5 min in group 1, at 15 min in group 2 and at 30 min in group 3 after post cementation. Five teeth with only 5 mm apical sealing were served as a positive control group, and ten with fiber post and core build-up but no coronal preparation were taken as a negative control group. Microleakage was evaluated using a fluid filtration system. The bonding interface was observed by scanning electronic microscope (SEM).

RESULTSThe microleakage was significantly increased after coronal preparation with high-speed handpiece. The negative control group has less leakage [(1.50 × 10(-6) ± 0.37 × 10(-6)) µl×min(-1)×Pa(-1)] than the groups with coronal preparation (P < 0.05); Group 1 leaked significantly more [(6.02 × 10(-5) ± 1.02 × 10(-5)) µl×min(-1)×Pa(-1)] than group 2 [(1.50 × 10(-5) ± 0.26 × 10(-5)) µl·min(-1)×Pa(-1)] and group 3 [(1.50 × 10(-5) ± 0.39 × 10(-5)) µl×min(-1)×Pa(-1)] did (P < 0.05). Corresponding to microleakage, the micro gaps between the resin cement and dentine in group 1 were wider than those in the other groups. The coronal section was wider than the apical part.

CONCLUSIONSHigh-speed handpiece had negative effects on microleakage of fiber post and core systems. Coronal preparation should be performed 15 min or more after post cementation.

Cementation ; Dental Bonding ; Dental Leakage ; Dentin-Bonding Agents ; Humans ; Microscopy, Electron, Scanning ; Post and Core Technique ; instrumentation ; Resin Cements ; Root Canal Preparation ; Time Factors

OBJECTIVEIt aims to investigate the changes in composition and structure of bacterial communities developing on biological activated carbon (BAC) particles, and the bacterial functions.

METHODA pilot plant had been in service for 180 days, aiming to develop bacterial communities on activated carbon naturally. After 180 days of operation, the bacterial communities were determined by denaturing gradient gel electrophoresis (DGGE) analyses of PCR-amplified 16S rRNA genes. The study on community composition and the phylogenetic relationships of the organisms was complemented by a sequence analysis of cloned PCR products from 16S rRNA genes. Gas chromatography-mass (GC-MS) measurement was used to determine organic chemical composition of inflow and outflow water on the 300th day. TOC and NH(4)(+)-N were also tested in this experiment.

RESULTSIt showed that the stable bacterial structure did not develop on BAC particles until the 9th month during running time of the BAC filter. The communities were finally dominated by Pseudomonas sp., Ba-cillus sp., Nitrospira sp., and an uncultured bacterium. Stable bacterial communities played an important role in removal of NH(4)(+)-N and total organic carbon (TOC). Results from gas chromatography-mass (GC-MS) showed that 36 kinds of chemicals in feed water were eliminated, and concentrations of 5 kinds of chemicals decreased. These chemicals served as nutrients for the dominant bacteria.

CONCLUSIONThe findings from the study suggested that the stability of microbial structure was beneficial for improving NH(4)(+)-N and TOC removal efficiencies. The dominant bacteria had the advantage of biode-grading a wide range of organic chemicals and NH(4)(+)-N.

Ammonia ; chemistry ; Bacteria ; classification ; genetics ; Biofilms ; Carbon ; chemistry ; Filtration ; instrumentation ; methods ; Microscopy, Electron, Scanning ; Pilot Projects ; Waste Disposal, Fluid ; instrumentation ; methods ; Water Microbiology ; Water Purification ; instrumentation ; methods ; Water Supply ; standards

AIMTissue engineering is a promising area with a broad range of applications in the fields of regenerative medicine and human health. The emergence of periodontal tissue engineering for clinical treatment of periodontal disease has opened a new therapeutic avenue. The choice of scaffold is crucial. This study was conducted to prepare zein scaffold and explore the suitability of zein and Shuanghuangbu for periodontal tissue engineering.

METHODOLOGYA zein scaffold was made using the solvent casting/particulate leaching method with sodium chloride (NaCl) particles as the porogen. The physical properties of the zein scaffold were evaluated by observing its shape and determining its pore structure and porosity. Cytotoxicity testing of the scaffold was carried out via in vitro cell culture experiments, including a liquid extraction experiment and the direct contact assay. Also, the Chinese medicine Shuanghuangbu, as a growth factor, was diluted by scaffold extract into different concentrations. This Shuanghuangbu-scaffold extract was then added to periodontal ligament cells (PDLCs) in order to determine its effect on cell proliferation.

RESULTSThe zein scaffold displayed a sponge-like structure with a high porosity and sufficient thickness. The porosity and pore size of the zein scaffold can be controlled by changing the porogen particles dosage and size. The porosity was up to 64.1%-78.0%. The pores were well-distributed, interconnected, and porous. The toxicity of the zein scaffold was graded as 0-1. Furthermore, PDLCs displayed full stretching and vigorous growth under scanning electronic microscope (SEM). Shuanghuangbu-scaffold extract could reinforce proliferation activity of PDLCs compared to the control group, especially at 100 microg x mL(-1) (P < 0.01).

CONCLUSIONA zein scaffold with high porosity, open pore wall structure, and good biocompatibility is conducive to the growth of PDLCs. Zein could be used as scaffold to repair periodontal tissue defects. Also, Shuanghuangbu-scaffold extract can enhance the proliferation activity of PDLCs. Altogether, these findings provide the basis for in vivo testing on animals.

Biocompatible Materials ; chemistry ; toxicity ; Cell Adhesion ; drug effects ; Cell Culture Techniques ; Cell Proliferation ; drug effects ; Cell Shape ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Chemical Phenomena ; Drugs, Chinese Herbal ; chemistry ; toxicity ; Humans ; Materials Testing ; Microscopy, Electron, Scanning ; Particle Size ; Periodontal Ligament ; cytology ; drug effects ; Plant Extracts ; chemistry ; pharmacology ; Polypodiaceae ; chemistry ; Porosity ; Scutellaria baicalensis ; chemistry ; Sodium Chloride ; chemistry ; Time Factors ; Tissue Engineering ; instrumentation ; methods ; Tissue Scaffolds ; chemistry ; Zein ; chemistry ; toxicity

OBJECTIVETo investigate the stress shielding rate, ultrastructural and biomechanical property of the healing bone fixed by Swan-like Shape Memory Connector (SMC) during experimental fracture healing in rabbits.

METHODSUnilateral osteotomy of left humeral diaphysis was performed in 140 adult New Zealand rabbits. The humeral fracture was fixed with SMC in study group and 4-hole dynamic compression plate (DCP) in control group. The animals were sacrificed at the 2nd, 4th, 8th, 12th, and 16th weeks postoperatively. The humeral shafts were harvested for three-point bending tests and stress shielding rate examination. Samples from the fracture gaps were observed with scanning electron microscope.

RESULTSWithin an observation period of 12 weeks after operation, the stress shielding rates were significantly lower in study group than those in control group, and the bending strength and the bending rigidity were statistically higher. It was also found that collagen fibers in callus was arranged more regulate and early mineral deposition in study group.

CONCLUSIONThese results suggest that SMC contributes to the proliferation and differentiation of osteoblasts, promoting the formation and calcification of bone matrix and enhancing biomechanical property of the healing bone, which might be closely correlated with a less stress shielding effect produced by SMC.

Animals ; Bony Callus ; physiopathology ; Fracture Fixation, Internal ; instrumentation ; Fracture Healing ; Humeral Fractures ; pathology ; physiopathology ; surgery ; Male ; Microscopy, Electron, Scanning ; Nickel ; Rabbits ; Stress, Mechanical ; Titanium