Cui Huang

Natural killer cells suppress full cycle HCV infection of human hepatocytes

Summary.  The role of natural killer (NK) cells in controlling hepatitis C virus (HCV) infection and replication has not been fully delineated. We examined NK cell‐mediated noncytolytic effect on full cycle HCV infection of human hepatocytes. Human hepatocytes (Huh7.5.1 cells) co‐cultured with NK cells or treated with supernatants (SN) from NK cells cultures had significantly lower levels of HCV RNA and protein than control cells. This NK cell‐mediated anti‐HCV activity could be largely abolished by antibody to interferon‐gamma (IFN‐γ). The investigation of the mechanisms for NK cell‐mediated anti‐HCV activity showed that NK SN‐treated hepatocytes expressed higher levels of IFN‐α/β than the control cells. NK SN also enhanced IFN regulatory factor‐3 and 7 expression in the hepatocytes. In addition, NK SN enhanced the expression of signal transducer and activator of transcription 1 and 2, the nuclear factors that are essential for the activation of IFN‐mediated antiviral pathways. These data provide direct evidence at cellular and molecular levels that NK cells have a key role in suppressing HCV infection of and replication in human hepatocytes.

SIRT6 Regulates Osteogenic Differentiation of Rat Bone Marrow Mesenchymal Stem Cells Partially via Suppressing the Nuclear Factor‐κB Signaling Pathway

Sirtuin 6 (SIRT6) is a NAD‐dependent deacetylase involved in lifespan regulation. To evaluate the effect of SIRT6 on osteogenesis, rat bone marrow mesenchymal stem cells (rBMSCs) with enhanced or reduced SIRT6 function were developed. We observed that SIRT6 knockdown significantly reduced the mRNA levels of several key osteogenic markers in vitro, including alkaline phosphatase (ALP), Runt‐related transcription factor 2 (RUNX2), and osteocalcin, while overexpression of SIRT6 enhanced their expression. Additionally, SIRT6 knockdown activated nuclear factor‐κB (NF‐κB) transcriptional activity and upregulated the expression of acetyl‐NF‐κB p65 (Lys310). The decreased osteogenic differentiation ability of rBMSCs could be partially rescued by the addition of NF‐κB inhibitor BAY 11–7082. Furthermore, SIRT6 overexpression in rBMSCs combined with the use of collagen/chitosan/hydroxyapatite scaffold could significantly boost new bone formation in rat cranial critical‐sized defects, as determined by microcomputed tomography and histological examination. These data confirm that SIRT6 is mainly located in the nuclei of rBMSCs and plays an essential role in their normal osteogenic differentiation, partly by suppressing NF‐κB signaling. Stem Cells 2014;32:1943–1955

Epigallocatechin-3-gallate (EGCG) enhances the therapeutic activity of a dental adhesive

OBJECTIVES The purpose of this study was to evaluate the antibacterial potential and physicochemical properties of a dental adhesive incorporated with epigallocatechin-3-gallate (EGCG) in different concentration over time. METHODS EGCG was incorporated at a ratio of 100, 200, and 300 μg/ml into a dental adhesive. The effects of the cured adhesives on the growth of Streptococcus mutans were determined by direct contact test immediately or one month later and by scanning electron microscopy (SEM), respectively. Microtensile bond strength (μTBS) test was used to test the mechanical property of the adhesives immediately or six months later. The degree of conversion (DC) of the adhesives was evaluated by Fourier transform infrared spectroscopy (FTIR). RESULTS Compared with negative control, the 200 μg/ml and 300 μg/ml EGCG-incorporated dental adhesive were found to exhibit inhibitory effect on the growth of S. mutans. The μTBS of the EGCG-incorporated dental adhesive was higher than the control. The DC of the adhesive system was not affected by the addition of EGCG. CONCLUSIONS 200 μg/ml EGCG incorporated dental adhesives could accomplish therapeutic goals that play in antimicrobial function whilst keeping the durability of resin-dentine bond.

Synthesis and characterization of triethylene glycol dimethacrylate nanocapsules used in a self-healing bonding resin

OBJECTIVES To date, the production of highly durable dentine bonding is still a challenge. Self-healing bonding resins may provide a new direction for the improvement of the bonding durability. The objective of the current study was to synthesize polyurethane nanocapsules encapsulated with the core material triethylene glycol dimethacrylate (TEGDMA) for use as a major component in a self-healing bonding resin. METHODS TEGDMA nanocapsules were synthesized via interfacial polycondensation in a miniemulsion, and the TEGDMA nanocapsules were then characterized via Fourier-transform infrared (FTIR) spectrometer, field emission scanning electron microscopy (FESEM), and high-performance liquid chromatography (HPLC) to investigate the morphology, the average TEGDMA loading (DL%), and encapsulation efficiency (EE%). The mechanical property of dental adhesive with different concentrations (0, 3, 6, 9, and 12 wt%) of the TEGDMA nanocapsules were also measured, and the cytotoxicity was investigated using an MTT assay. RESULTS FTIR confirmed that the TEGDMA nanocapsules were successfully synthesized. These nanocapsules showed a high drug load. The bond strength of the dental adhesive incorporated with 9 wt% TEGDMA nanocapsules was significantly higher compared with those of the other groups (P<0.001). Moreover, the biocompatibility of the dental adhesive was not affected by the incorporation of the TEGDMA nanocapsules. CONCLUSIONS The current study demonstrated the successful synthesis of TEGDMA nanocapsules, and the overall properties of the dental adhesive were not compromised.

Antibacterial and physical properties of EGCG-containing glass ionomer cements.

OBJECTIVES To evaluate the effect of the addition of epigallocatechin-3-gallate (EGCG) on the antibacterial and physical properties of glass ionomer cement (GIC). METHODS A conventional GIC, Fuji IX, was used as a control. EGCG was incorporated into GIC at 0.1% (w/w) and used as the experimental group. Chlorhexidine (CHX) was added into GIC at 1% (w/w) as a positive control. The anti-biofilm effect of the materials was assessed by a colorimetric technique (MTT assay) and scanning electron microscopy (SEM). The leaching antibacterial activity of the materials on Streptococcus mutans was evaluated by an agar-diffusion test. The flexural strength of the materials was evaluated using a universal testing machine and the surface microhardness was measured using a microhardness tester. The fluoride-releasing property of the materials was tested by ion chromatography. RESULTS The optical density (OD) values of the GIC-EGCG group were significantly decreased at 4h compared with the GIC group, but only a slightly decreased tendency was observed at 24h (P>0.05). No inhibition zones were detected in the GIC group during the study period. Significant differences were found between each group (P<0.05). Compared with the control group, there was a significant increase in the flexural strength and surface microhardness for the GIC-EGCG group (P<0.05). The fluoride ion release was not influenced by EGCG-incorporation (P>0.05). CONCLUSIONS These findings suggested that GIC-containing 0.1% (w/w) EGCG is a promising restorative material with improved mechanical properties and a tendency towards preferable antibacterial properties. CLINICAL SIGNIFICANCE Modification of the glass ionomer cements with EGCG to improve the antibacterial and physical properties showed some encouraging results. This suggested that the modification of GIC with EGCG might be an effective strategy to be used in the dental clinic. However, this was only an in vitro study and clinical trials would need to verify true outcomes.

Effects of different artificial ageing methods on the degradation of adhesive-dentine interfaces.

OBJECTIVES To compare the effects of four commonly used artificial ageing methods (water storage, thermocycling, NaOCl storage and pH cycling) on the degradation of adhesive-dentine interfaces. METHODS Fifty molars were sectioned parallel to the occlusal plane, polished and randomly divided into two adhesive groups: An etch-and-rinse adhesive Adper SingleBond 2 and a self-etch adhesive G-Bond. After the composite built up, the specimens from each adhesive group were sectioned into beams, which were then assigned to one of the following groups: Group 1 (control), 24h of water storage; Group 2, 6 months of water storage; Group 3, 10,000 runs of thermocycling; Group 4, 1h of 10% NaOCl storage; and Group 5, 15 runs of pH cycling. The microtensile bond strengths were then tested. The failure modes were classified with a stereomicroscope and representative interface was analyzed with a field-emission scanning electron microscopy (FESEM). Nanoleakage expression was evaluated through FESEM in the backscattered mode. RESULTS The four artificial ageing methods decreased the bonding strength to nearly 50% and increased the nanoleakage expression of both adhesive systems compared with the control treatment. Adhesive failures were the predominant fracture modes in all groups. However, differences in detailed morphology were observed among the different groups. CONCLUSIONS Water storage, thermocycling, NaOCl storage and pH cycling could obtain similar degradation effectiveness through appropriate parameter selection. Each in vitro artificial ageing method had its own mechanisms, characteristics and application scope for degrading the adhesive-dentin interfaces. CLINICAL SIGNIFICANCE Water storage is simple, low-cost but time-consuming; thermocycling lacks of a standard agreement; NaOCl storage is time-saving but mainly degrades the organic phase; pH cycling can resemble cariogenic condition but needs further studies. Researchers focusing on bonding durability studies should be deliberate in selecting an appropriate ageing model based on the differences of test material, purpose and time.

Effect of pretreatment with calcium- containing desensitizer on the dentine bonding of mild self-etch adhesives

Desensitizing agents are frequently applied to sensitive teeth and may affect subsequent resin bonding. The current study aimed to evaluate the bonding performance of two self-etch adhesives containing functional monomers to dentine pretreated with three new calcium-containing desensitizers. No desensitizer was applied in the control group. Groups 1, 2, and 3 were treated with an arginine-calcium carbonate-containing polishing paste, a casein phosphopeptide-amorphous calcium phosphate (CPP-ACP)-containing paste, and an experimental hydroxyapatite paste, respectively. G-Bond and Clearfil S(3) Bond were used for bonding after desensitizer treatments. The microtensile bond strength (μTBS) was tested (n = 20 beams per group) and failure mode distribution was analyzed. Scanning electron microscopy was used to observe the occlusion of dentinal tubules. The mean (±SD) μTBS values, expressed in MPa, of groups 1, 2, and 3 and the control group were, respectively, 30.81 (7.79), 44.41 (8.02), 31.49 (6.13), and 41.40 (8.67) for G-Bond and 39.63 (9.59), 32.55 (7.86), 37.50 (8.60), 27.90 (6.52) for S3 Bond. Most failures were recorded as adhesive failure (69.375%), instead of cohesive failure or mixed failure. The dentinal tubules were seldom plugged in group 2, but were mostly occluded in groups 1 and 3. Two-way anova indicated that desensitizer application in association with a compatible adhesive system should be used when endeavoring to control hypersensitivity without adverse interference in bonding.

Biomimetic Intrafibrillar Mineralization of Type I Collagen with Intermediate Precursors-loaded Mesoporous Carriers

Limited continuous replenishment of the mineralization medium is a restriction for in-situ solution-based remineralization of hypomineralized body tissues. Here, we report a process that generated amine-functionalized mesoporous silica nanoparticles for sustained release of biomimetic analog-stabilized amorphous calcium phosphate precursors. Both two-dimensional and three-dimensional collagen models can be intrafibrillarly mineralized with these released fluidic intermediate precursors. This represents an important advance in the translation of biomineralization concepts into regimes for in-situ remineralization of bone and teeth.

Translation of a solution-based biomineralization concept into a carrier-based delivery system via the use of expanded-pore mesoporous silica

UNLABELLED Mineralization of collagen fibrils using solution-based systems containing biomimetic analogs of matrix proteins to stabilize supersaturated calcium phosphate solutions have been predictably achieved in vitro. Solution-based systems have limitations when used for in-situ remineralization of human hypomineralized tissues because periodic replenishment of the mineralizing solution is infeasible. A carrier-based platform designed for delivering mineral precursors would be highly desirable. In the present work, mesoporous silica nanoparticles with expanded pores (eMSN; 14.8nm) were synthesized. Polyacrylic acid-stabilized amorphous calcium phosphate (PA-ACP) was generated from a supersaturated calcium and phosphate ion-containing solution, and chosen as the model mineralizing phase. After amine functionalization (AF) of the eMSN through a post-grafting method, the positively-charged AF-eMSN enabled loading of PA-ACP by electrostatic interaction. In-vitro cytotoxicity testing indicated that PA-ACP@AF-eMSN was highly biocompatible. The release kinetics of mineralization precursors from PA-ACP@AF-eMSN was characterized by an initial period of rapid calcium and phosphate release that reached a plateau after 120h. Intrafibrillar mineralization was examined using a 2-D fibrillar collagen model; successful mineralization was confirmed using transmission electron microscopy. To date, this is the first endeavor that employs expanded-pore mesoporous silica to deliver polymer-stabilized intermediate precursors of calcium phosphate for intrafibrillar mineralization of collagen. The carrier-based delivery system bridges the gap between contemporary solution-based biomineralization concepts and clinical practice, and is useful for in-situ remineralization of bone and teeth. STATEMENT OF SIGNIFICANCE Concepts of collagen biomineralization have been reasonably well established in the past few years and intrafibrillar mineralization of collagen fibrils can be predictably achieved with analogs of matrix proteins using solution-based systems. However, solution-based systems have their limitations in clinical applications that require direct application of mineralization precursors in-situ because periodic replenishment of the mineralizing solution is impossible. The present work presents for the first time, the use of amine-functionalized mesoporous silica with expanded pores for loading and release of polyacid-stabilized amorphous calcium phosphate mineralization precursors, and for intrafibrillar mineralization of type I collagen fibrils. This strategy represents an important step in the translational application of contemporary biomineralization concepts for in-situ remineralization of bone and teeth.

Effect of intracanal dentine wettability on human dental pulp cell attachment

AIM To evaluate the effect of intracanal dentine wettability on human dental pulp cell (HDPC) attachment after application of endodontic irrigants. METHODOLOGY Twenty extracted human premolar teeth with straight canals were sectioned at the apical and coronal thirds of each root, and the remaining mid-root portions were bisected longitudinally. After polishing, the specimens were divided into two groups for wettability and cell attachment measurements, respectively. The intracanal surface wettability was assessed by atomic force microscopy after a rinse of H(2) O (control), 5.25% sodium hypochlorite (NaOCl), 17% ethylenediaminetetraacetic acid (EDTA) and MTAD (n = 5). Additionally, HDPCs were seeded onto the irrigated root canal dentine and the cell attachment was evaluated by calcein-CAM and propidium iodide (PI) dying under fluorescence microscope (n = 5). Data were analysed by means of one-way anova and LSD test. RESULTS Compared with the control group, 5.25% NaOCl, 17% EDTA and MTAD treatment significantly increased surface wettability (P < 0.001). The rank order of adhesion force was MTAD >17% EDTA >5.25% NaOCl > H(2) O. The highest average numbers of viable HDPCs were observed on the root canals irrigated with MTAD, followed by the 17% EDTA group (P < 0.001), and the lowest average numbers of viable cells were detected in the 5.25% NaOCl and H(2) O groups. CONCLUSIONS Surface wettability was affected by the irrigants. Increasing in wettability produced higher levels of HDPC attachment to irrigated dentine.