Development of an Antibacterial Dentin Adhesive.

Nisin is a peptide that possesses potent antibacterial properties. This study evaluated the antibacterial activity of a nisin-doped adhesive against Streptococcus mutans, as well as its degree of conversion and microtensile bond strength (µTBS) to dentin. Nisin was added to the adhesive Adper Single Bond 2 (3M ESPE), resulting in four groups: Control Group (Single Bond 2); Group 1% (1 wt% nisin-incorporated), Group 3% (3 wt% nisin-incorporated) and Group 5% (5 wt% nisin-incorporated). Antibacterial activity against S. mutans was evaluated using colony-forming unit counts (CFU). The degree of conversion was tested using FTIR. Forty human teeth were restored for µTBS evaluation. Data were statistically analyzed with ANOVA and Tukey tests at α = 0.05. The nisin-doped adhesives, for all concentrations, exhibited a significant inhibition of the growth of S. mutans (p < 0.05); Incorporation of 5% and 3% nisin decreased the degree of conversion of the adhesive (p < 0.05). The µTBS (in MPa): Control Group­38.3 ± 2.3A, Group 1%­35.6 ± 2.1A, Group 3%­27.1 ± 1.6B and Group 5%­22.3 ± 1.0C. Nisin-doped adhesives exerted a bactericidal effect on S. mutans. The µTBS and degree of conversion of adhesive were not affected after incorporation of 1% nisin.

Effects of calcium-phosphate, laser and adhesive on dentin permeability and bond strength.

OBJECTIVES: The present study examined a technique for reducing dentin permeability through the application of a calcium phosphate (CaP)-based desensitiser with a laser-assisted process and evaluated adhesive-dentin bond strength. Methods: Thirty dentin discs were divided into two groups according to whether the selected desensitiser (TeethMate; Kuraray Noritake) was used prior to dentin bonding. Each group was subdivided into three subgroups (n = 5): A- Adhesive (Single Bond Universal, 3M ESPE), AL- Adhesive + Laser (Nd:YAG 60 mJ) and LAL- Laser + Adhesive + Laser. Dentin permeability values (%) were recorded before and after desensitiser application. Resin composites were placed over the bonded specimens; the latter were aged prior to microtensile bond strength evaluation. Gelatinolytic activity within the hybrid layers was examined with in-situ zymography using confocal laser scanning microscopy. Data were analysed with ANOVA and Tukey test (α = 0.05). RESULTS: Significant differences in dentin permeability were identified for all groups (p = 0.00). Both laser treatment (p = 0.182) and desensitiser application (p = 0.687) did not significantly improve dentin bond strength. Ultrastructure of the resin-dentin interface identified presence of calcium phosphate within dentinal tubules. Laser treatment did not affect hybrid layer ultrastructure. Both treatment modalities (intratubular CaP occlusion and laser) had no influence on gelatinolytic activity within hybrid layers. CONCLUSION: Although intratubular CaP occlusion and laser treatment were effective in reducing dentin permeability, they did not affect bond strength, interfacial ultrastructure and gelatinolytic activity within hybrid layers. CLINICAL RELEVANCE: Treatment of etched dentin with Nd:YAG Laser at 60 mJ does not adversely affect collagen ultrastructure and gelatinolytic activity within the hybrid layer. The application of a calcium phosphate-based desensitiser to etch dentin does not affect dentin bond strength.

Anesthesia upstream of the alcoholic lesion point alleviates the pain of alcohol neurolysis for intercostal neuralgia: a prospective randomized clinical trial.

OBJECTIVES: Alcohol for intercostal neuralgia may induce severe injection pain. Although nerve block provided partial pain relief, alcohol might be diluted, and the curative effect decreased when the local anesthetic and alcohol were given at the same point. Therefore, we observed the modified method for intercostal neuralgia, a Two-point method, in which the local anesthetic and alcohol were given at different sites. METHOD: Thirty patients diagnosed with intercostal neuralgia were divided into 2 groups: Single-point group and Two-point group. In the Single-point group, alcohol and local anesthetic were injected at the same point, named the "lesion point", which was the lower edge of ribs and 5 cm away from the midline of the spinous process. In the Two-point group, alcohol was injected at the lesion point, whereas the local anesthetic was administered at the "anesthesia point", which was 3 cm away from the midline of spinous process. RESULTS: After alcohol injection, visual analog scale (VAS) in the Two-point group was lower than the Single-point group, and the satisfaction ratio of patients in the Two-point group was higher (p<0.05). The degree of numbness in the Two-point group was greater than the Single-point group at 1 month and 3 months after operation (p<0.05). However, the long-term effects did not differ. CONCLUSIONS: Local anesthetic was given upstream of the point where alcohol was administered, was a feasible and safe method to relieve pain during the operation, and improved the satisfaction of the patients and curative effect.

Anesthesia upstream of the alcoholic lesion point alleviates the pain of alcohol neurolysis for intercostal neuralgia: a prospective randomized clinical trial

OBJECTIVES: Alcohol for intercostal neuralgia may induce severe injection pain. Although nerve block provided partial pain relief, alcohol might be diluted, and the curative effect decreased when the local anesthetic and alcohol were given at the same point. Therefore, we observed the modified method for intercostal neuralgia, a Two-point method, in which the local anesthetic and alcohol were given at different sites. METHOD: Thirty patients diagnosed with intercostal neuralgia were divided into 2 groups: Single-point group and Two-point group. In the Single-point group, alcohol and local anesthetic were injected at the same point, named the "lesion point", which was the lower edge of ribs and 5 cm away from the midline of the spinous process. In the Two-point group, alcohol was injected at the lesion point, whereas the local anesthetic was administered at the "anesthesia point", which was 3 cm away from the midline of spinous process. RESULTS: After alcohol injection, visual analog scale (VAS) in the Two-point group was lower than the Single-point group, and the satisfaction ratio of patients in the Two-point group was higher (p<0.05). The degree of numbness in the Two-point group was greater than the Single-point group at 1 month and 3 months after operation (p<0.05). However, the long-term effects did not differ. CONCLUSIONS: Local anesthetic was given upstream of the point where alcohol was administered, was a feasible and safe method to relieve pain during the operation, and improved the satisfaction of the patients and curative effect.

Social-Emotional Functioning Explains the Effects of Physical Activity on Academic Performance among Chinese Primary School Students: A Mediation Analysis.

OBJECTIVE: To examine the mediating effects of socioemotional and behavioral functioning in the association between physical activity and academic achievement in Chinese children. STUDY DESIGN: Chinese children (n = 17 318; aged 6-11 years) from the Shanghai Children's Health, Education and Lifestyle Evaluation study in 2014 were the source of data. Children reported the time spent in moderate-to-vigorous physical activities (MVPA) during weekdays and weekends. Parents rated their children's socioemotional and behavioral difficulties. Head teachers rated the academic performance of each student. A mediation analysis with multiple mediators was performed to test the mediation effects of 5 socioemotional and behavioral variables (emotional problems, conduct problems, hyperactivity and inattention, peer relationship, and prosocial behaviors) in the association between MVPA and academic achievement, adjusting for demographic and health-related variables. RESULTS: Results revealed that MVPA was positively associated with academic performance (b = 0.78; SE = .01; P < .001). Conduct problems, hyperactivity and inattention, and peer problems showed significant mediating effects in the association between MVPA and academic achievement, with hyperactivity and inattention being the strongest mediator (b = 0.38; SE = 0.04; P < .001) accounting for 49% of the total effect. CONCLUSIONS: This study provides evidence that MVPA is associated with fewer socioemotional difficulties, which, in turn, is associated with better academic achievement. Promoting physical activity among school-aged children may benefit their overall development.

Water-associated attributes in the contemporary dentin bonding milieu.

OBJECTIVES: The water-associated attributes of resin-dentin interfaces created by contemporary adhesives are important determinants of bond integrity and stability. In the present work, these attributes were estimated from the perspectives of causality, to examine the behavior of the first and most-recently launched versions of universal adhesives when applied in either the etch-and-rinse mode or the self-etch mode. METHODS: The immediate cause of interfacial permeability and the time-dependent cause of water sorption were investigated in conjunction with the intermediate effect of interface degradation and the more long-term effect of loss of mechanical strength, before and after thermomechanical cycling. The results were compared with control etch-and-rinse and self-etch adhesives. RESULTS: Although the introduction of this new class of universal adhesives has brought forth significant changes to the dental adhesion arena, including more application options, reduced bonding armamentarium and increased user friendliness, the water-associated attributes that are critical for making resin-dentin bonds more durable to environmental challenges and less susceptible to degradation have remained unchanged at large, when compared with benchmarks established by former classes of adhesives. CONCLUSION: It appears that the current trend of adhesive development has brought forth significant changes but lacks the vigor that demarcates progress and technological sublimity. CLINICAL SIGNIFICANCE: The advent of the user friendly universal adhesives has brought forth significant changes to the dental adhesion arena. However, the elements that are critical for making resin-dentin bonds more durable to environmental challenges and less susceptible to degradation have remained unchanged at large.

Intrafibrillar silicified collagen scaffold promotes in-situ bone regeneration by activating the monocyte p38 signaling pathway.

Intrafibrillar silicified collagen scaffold (SCS) is a promising biomaterial for bone regeneration because it promotes cell homing and angiogenesis in bone defects via monocyte modulation. In the present study, a rat femoral defect model was used to examine the contribution of monocyte signaling pathways to SCS modulation. Activation of the monocyte p38 signaling pathway by SCS resulted in monocyte differentiation into TRAP-positive mononuclear cells. These cells demonstrated increased secretion of SDF-1α, VEGFa and PDGF-BB, which, in turn, promoted homing of bone marrow stromal cells (BMSCs) and endothelial progenitor cells (EPCs), as well as local vascularization. Monocyte differentiation and secretion were blocked after inhibition of the p38 pathway, which resulted in reduction in cell homing and angiogenesis. Taken together, these novel findings indicate that the p38 signaling pathway is crucial in SCS-modulated monocyte differentiation and secretion, which has a direct impact on SCS-induced bone regeneration. STATEMENT OF SIGNIFICANCE: Intrafibrillar silicified collagen scaffold (SCS) is a promising biomaterial for bone regeneration. The present work demonstrates that SCS possesses favorable bone regeneration potential in a rat femoral defect model. The degrading scaffold modulates monocyte differentiation and release of certain cytokines to recruit MSCs and EPCs, as well as enhances local vascularization by activating the p38 MAPK signaling pathway. These findings indicate that SCS contributes to bone defect regeneration by stimulating host cell homing and promoting local angiogenesis and osteogenesis without the need for loading cytokines or xenogenous stem cells.

Susceptibility of contemporary single-bottle self-etch dentine adhesives to intrinsic water permeation.

OBJECTIVES: To evaluate the effect of intrinsic water permeation on the microtensile bond strengths of different adhesive systems to dentine and the quality of resin-dentine interfaces. METHODS: Ninety-six non-carious human third molars were divided into 4 groups: Clearfil S3 Bond Plus (CSBP; Kuraray); Clearfil S3 Bond (C3S; Kuraray); iBond Self-Etch (IB; Heraeus-Kulzer) and Prime&Bond NT (PB, control etch-and-rinse adhesive, Dentply-Sirona). For each adhesive, specimens from one subgroup (N=10) were bonded using zero pulpal pressure, while specimens from the other subgroup (N=10) were bonded using 15cm water pressure (PP). Each bonded tooth was sectioned into 1×1mm sticks and stressed to failure. Data were analysed using two-way ANOVA and Holm-Sidak pairwise comparisons to examine the effects of "adhesive", "pulpal pressure" and their interaction on bond strength (α=0.05). Representative fractured sticks were examined by SEM. The remaining tooth slabs in each subgroup were used for TEM and CLSM. RESULTS: Microtensile bond strengths (mean±SD; in MPa) were: 33.4±6.9 (CSBP), 33.2±4.7 (CSBP-PP), 35.0±8.6 (C3S), 25.5±7.3 (C3S-PP), 18.4±4.0 (IB), 16.5±6.9 (IB-PP), 28.2±5.5 (PB), 20.5±7.2 (PB-PP). "Adhesive-type" (P<0.001), "pulpal-pressure" (P<0.001) and their interactions (P<0.001) significantly affected bond strength results. No difference between no-PP and PP subgroups was found for CSBP and IB (P>0.05). Water droplets were identified along the resin-dentine interface for IB, IB-PP and C3S-PP. CONCLUSION: IB exhibits water sensitivity when bonding is performed with/without pulpal pressure. C3S exhibits water sensitivity when bonding is performed with pulpal pressure. CSBP does not exhibit water sensitivity when bonding is performed with/without pulpal pressure. CLINICAL SIGNIFICANCE: Intrinsic water permeation during bonding procedures significantly affects bond strength results and the resin-dentine interface of contemporary single-bottle self-etch dentine adhesive systems.

Can long-term dentine bonding created in real life be forecasted by parameters established in the laboratory?

Tooth-coloured plastic dental fillings secured by adhesives to tooth structures are widely used to fix decayed teeth. Whereas laboratory tests demonstrate rapid deterioration of the ability of the adhesives to stick to dentine, clinical studies show that these fillings are relatively durable. This discrepancy suggests that the parameters used for simulating bond degradation in the laboratory setting do not correlate well with clinical outcomes. The present study examined the long-term tensile bond strength of resin composite fillings performed in real life and under different laboratory-simulated bonding conditions to identify parameters that may be used to forecast the durability of adhesive bonds created in dentine. Fillings placed in vivo were subjected to different periods of intraoral function. In vitro specimens were bonded based on whether simulated pulpal pressure (SPP) or thermomechanical cycling was implemented, and how long the completed fillings were stored in water. Thermomechanical cycling used in combination with long-term water ageing are useful in forecasting the decline in strength of resin-dentine bonds created in vivo. These parameters should be adopted for future evaluations. Conversely, the use of SPP does not appear to be a significant parameter in the simulation of long-term clinical deterioration of bond integrity.

Cytotoxicity and osteogenic potential of silicate calcium cements as potential protective materials for pulpal revascularization.

OBJECTIVES: In pulpal revascularization, a protective material is placed coronal to the blood clot to prevent recontamination and to facilitate osteogenic differentiation of mesenchymal stem cells to produce new dental tissues. Although mineral trioxide aggregate (MTA) has been the material of choice for clot protection, it is easily displaced into the clot during condensation. The present study evaluated the effects of recently introduced calcium silicate cements (Biodentine and TheraCal LC) on the viability and osteogenic differentiation of human dental pulp stem cells (hDPSCs) by comparing with MTA Angelus. METHODS: Cell viability was assessed using XTT assay and flow cytometry. The osteogenic potential of hDPSCs exposed to calcium silicate cements was examined using qRT-PCR for osteogenic gene expressions, alkaline phosphatase enzyme activity, Alizarin red S staining and transmission electron microscopy of extracellular calcium deposits. Parametric statistical methods were employed for analyses of significant difference among groups, with α=0.05. RESULTS: The cytotoxic effects of Biodentine and TheraCal LC on hDPSCs were time- and concentration-dependent. Osteogenic differentiation of hDPSCs was enhanced after exposure to Biodentine that was depleted of its cytotoxic components. This effect was less readily observed in hDPSCs exposed to TheraCal LC, although both cements supported extracellular mineralization better than the positive control (zinc oxide-eugenol-based cement). SIGNIFICANCE: A favorable tissue response is anticipated to occur with the use of Biodentine as a blood clot-protecting material for pulpal revascularization. Further investigations with the use of in vivo animal models are required to validate the potential adverse biological effects of TheraCal LC on hDPSCs.

Efficient Expression and Purification of Recombinant Human Enteropeptidase Light Chain in Esherichia coli

Human enterokinase (synonym: enteropeptidase, EC 3.4.21.9) light chain (hEKL) gene was designed and artificially synthesized with built-in codon blas towards Escherichia coli codon preference. The synthetic hEKL gene was cloned into prokaryotic expression vector pMAL-s and transferred into the expression strain E. coli BL21 (DE3). Recombinant hEKL protein with a maltose binding protein (MBP) tag was expressed at high levels in soluble form, which yielded about 42% of the total cellular protein. The target protein was then purified to the homogeneity (> 95%) by affinity chromatography. The peptide substrate GST-Melittin with enterokinase recognition site was completely cleaved by the purified MBP-hEKL at the molar ratio of 1:5000 (enzyme:substrate). Tricine SDS-PAGE analysis showed that the activity of MBP-hEKL was approximately seven times that of bovine enterokinase catalytic subunit (EKMaxTM, Invitrogen). From 1 L flask culture, 206 mg pure active MBP-hEKL was with specific activity of 1.4×104 U/mg.