Silica-based silver nanocomposite 80S/Ag as Aggregatibacter actinomycetemcomitans inhibitor and its in vitro bioactivity.

Background/purpose: As a commonly-found pathogen in periodontal disease, Aggregatibacter actinomycetemcomitans has been reported with several antibiotic resistance. Thus, to develop an alternative and protective therapy for A. actinomycetemcomitans infections is urgently needed in dentistry. In this study, we sought to synthesize a silica-based material to deliver silver nanoparticles for antibacterial purposes. Also, the bioactivities were examined via analyzing the formation of hydroxyapatite. Materials and methods: The 80S/Ag powders were prepared by the evaporation-induced self-assembly method, with Si, Ca, P, and Ag composition ratios of 80, 15, 5, and 1/5/10 (mole percentage), respectively. The nitrogen adsorption-desorption isotherms, transmission electron microscope, selected area electron diffraction, and Fourier transform infrared spectroscopy were conducted for textural analyses. The disk diffusion test was carried out against A. actinomycetemcomitans strain ATCC 29523. In vitro bioactivity assessment involved soaking 80S/Ag membrane powders in acellular simulated body fluid. Results: We successfully developed a material consisting of Si, Ca, P, and Ag, namely the 80S/Ag. In the antibacterial testing, the 80S/Ag demonstrated antibacterial activity against the commonly-found oral pathogen, A. actinomycetemcomitans, with a long-lasting effect for 168h. The formation of hydroxyapatite in simulated body fluid highlighted the characteristic of dentine remineralization for the 80S/Ag. The increased pH values after immersion in simulated body fluid would help neutralize the acidic oral environment. Conclusion: Our results indicate that 80S/Ag possesses remarkable antibacterial properties, hydroxyapatite formation, and increased pH values after immersion in simulated body fluid, supporting the potential therapeutic application of 80S/Ag for treating periodontal disease.

Synthesis, Characterization, and Bioactivity of Mesoporous Bioactive Glass Codoped with Zinc and Silver.

Due to the overconsumption of antimicrobials, antibiotic-resistant bacteria have become a critical health issue worldwide, especially methicillin-resistant S. aureus (MRSA) and vancomycin-resistant E. faecalis (VRE). Recently, many efforts have been made to load metals into bioactive glasses to enhance the multifunctionality of materials, such as antibacterial and osteoinductive functions. Zinc has been documented to stimulate the gene expression of various regulatory factors in bone cells. Meanwhile, previous studies have reported that silver and zinc could be a promising antibacterial combination with synergistic antimicrobial effects. Here, we sought to develop a biomaterial coreleasing zinc and silver, designated 80S-ZnAg, and to evaluate its antibacterial activity and biocompatibility. The textural analyses demonstrated different coreleasing patterns of zinc and silver for the materials. The chemical characterization revealed that the zinc in 80S-ZnAg could be the network modifier when its molar ratio was high, releasing more zinc; zinc could also be the network former when its molar ratio was low, showing an extremely low rate of release. However, the ICP results for 80S-Zn3Ag2 demonstrated up to 7.5 ppm of zinc and 67.6 ppm of silver. Among all the 80S-ZnAg materials, 80S-Zn3Ag2 demonstrated more marked antibacterial activity against MRSA and VRE than the others, with inhibition zones of 11.5 and 13.4 mm, respectively. The cytotoxicity assay exhibited nearly 90% cell viability at 20 mg/mL of 80-Zn3Ag2. Further clinical study is needed to develop an innovative biomaterial to address the issue of antibiotic resistance.

Synergistic effect of drug/antibiotic-impregnated micro/nanohybrid mesoporous bioactive glass/calcium phosphate composite bone cement on antibacterial and osteoconductive activities.

Calcium phosphate bone cements (CPC) can be used in minimally invasive surgery because of their injectability, and they can also be used to repair small and irregular bone defects. This study aimed to release the antibiotic gentamicin sulfate (Genta) to reduce tissue inflammation and prevent infection in the early stages of bone recovery. Subsequently, the sustained release of the bone-promoting drug ferulic acid (FA) mimicked the response of osteoprogenitor D1 cells interaction, thereby accelerating the healing process of the overall bone repair. Accordingly, the different particle properties of micro-nano hybrid mesoporous bioactive glass (MBG), namely, micro-sized MBG (mMBG) and nano-sized MBG (nMBG), were explored separately to generate different dose releases in MBG/CPC composite bone cement. Results show that nMBG had better sustained-release ability than mMBG when impregnated with the same dose. When 10 wt% of mMBG hybrid nMBG and composite CPC were used, the amount of MBG slightly shortened the working/setting time and lowered the strength but did not hinder the biocompatibility, injectability, anti-disintegration, and phase transformation of the composite bone cement. Furthermore, compared with 2.5wt%Genta@mMBG/7.5 wt% FA@nMBG/CPC, 5wt.%Genta@mMBG/5wt.%FA@nMBG/CPC exhibited better antibacterial activity, better compressive strength, stronger mineralization of osteoprogenitor cell, and similar 14-day slow-release trend of FA. The MBG/CPC composite bone cement developed can be used in clinical surgery to achieve the synergistic sustained release of antibacterial and osteoconductive activities.

In Vitro Characterizations of Post-Crosslinked Gelatin-Based Microspheres Modified by Phosphatidylcholine or Diammonium Phosphate as Antibiotic Delivery Vehicles.

Hydrogel-based microspheres prepared by emulsification have been widely used as drug carriers, but biocompatibility remains a challenging issue. In this study, gelatin was used as the water phase, paraffin oil was used as the oil phase, and Span 80 was used as the surfactant. Microspheres were prepared using a water-in-oil (W/O) emulsification. Diammonium phosphate (DAP) or phosphatidylcholine (PC) were further used to improve the biocompatibility of post-crosslinked gelatin microspheres. The biocompatibility of DAP-modified microspheres (0.5-10 wt.%) was better than that of PC (5 wt.%). The microspheres soaked in phosphate-buffered saline (PBS) lasted up to 26 days before fully degrading. Based on microscopic observation, the microspheres were all spherical and hollow inside. The particle size distribution ranged from 19 µm to 22 µm in diameter. The drug release analysis showed that the antibiotic gentamicin loaded on the microspheres was released in a large amount within 2 h of soaking in PBS. It was stabilized until the amount of microspheres integrated was significantly reduced after soaking for 16 days and then released again to form a two-stage drug release curve. In vitro experiments showed that DAP-modified microspheres at concentrations less than 5 wt.% had no cytotoxicity. Antibiotic-impregnated and DAP-modified microspheres had good antibacterial effects against Staphylococcus aureus and Escherichia coli, but these drug-impregnated groups hinder the biocompatibility of hydrogel microspheres. The developed drug carrier can be combined with other biomaterial matrices to form a composite for delivering drugs directly to the affected area in the future to achieve local therapeutic effects and improve the bioavailability of drugs.

In Vitro Bioactivity and Antibacterial Effects of a Silver-Containing Mesoporous Bioactive Glass Film on the Surface of Titanium Implants.

Peri-implantitis is defined as a bacterial infection-induced inflammation and suppuration of soft and hard tissues surrounding a dental implant. If bacteria further invade the alveolar bone, they can easily cause bone loss and even lead to the early failure of a dental implant surgery. In the present study, an 80SiO2-15CaO-5P2O5 mesoporous bioactive glass film system containing 1, 5, and 10 mol% of silver was prepared on titanium implant discs (MBG-Ag-coated Ti) using sol-gel and spin coating methods. The wettability and adhesion strength of the films were evaluated using contact angle measurements and adhesion strength tests, respectively. The phase composition, chemical bonding, morphology, and oxidation states of the films were analyzed via X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), and X-ray photoelectron spectroscopy (XPS). In vitro bioactivity analysis of the films was performed by immersion in a simulated body fluid (SBF) for 24 h. Disk diffusion tests were performed on the early colonizing bacteria Aggregatibacter actinomycetemcomitans and Streptococcus mutans to evaluate the antibacterial ability of the films. A silver-containing mesoporous bioactive glass film with excellent biocompatibility and antibacterial properties was successfully prepared.

Morphological Changes, Antibacterial Activity, and Cytotoxicity Characterization of Hydrothermally Synthesized Metal Ions-Incorporated Nanoapatites for Biomedical Application.

The objective of this study was to prepare hydroxyapatite (HA) with potential antibacterial activity against gram-negative and gram-positive bacteria by incorporating different atomic ratios of Cu2+ (0.1-1.0%), Mg2+ (1.0-7.0%), and Zn2+ (1.0-7.0%) to theoretically replace Ca2+ ions during the hydrothermal synthesis of grown precipitated HA nanorods. This study highlights the role of comparing different metal ions on synthetic nanoapatite in regulating the antibacterial properties and toxicity. The comparisons between infrared spectra and between diffractograms have confirmed that metal ions do not affect the formation of HA phases. The results show that after doped Cu2+, Mg2+, and Zn2+ ions replace Ca2+, the ionic radius is almost the same, but significantly smaller than that of the original Ca2+ ions, and the substitution effect causes the lattice distance to change, resulting in crystal structure distortion and reducing crystallinity. The reduction in the length of the nanopatites after the incorporation of Cu2+, Mg2+, and Zn2+ ions confirmed that the metal ions were mainly substituted during the growth of the rod-shape nanoapatite Ca2+ distributed along the longitudinal site. The antibacterial results show that nanoapatite containing Cu2+ (0.1%), Mg2+ (3%), and Zn2+ (5-7%) has obvious and higher antibacterial activity against gram-positive bacteria Staphylococcus aureus within 2 days. The antibacterial effect against the gram-negative bacillus Escherichia coli is not as pronounced as against Staphylococcus aureus. The antibacterial effect of Cu2+ substituted Ca2+ with an atomic ratio of 0.1~1.0% is even better than that of Mg2+- and Zn2+- doped with 1~7% groups. In terms of cytotoxicity, nanoapatites with Cu2+ (~0.2%) exhibit cytotoxicity, whereas Mg2+- (1-5%) and Zn2+- (~1%) doped nanoapatites are biocompatible at low concentrations but become cytotoxic as ionic concentration increases. The results show that the hydrothermally synthesized nanoapatite combined with Cu2+ (0.2%), Mg2+ (3%), and Zn2+ (3%) exhibits low toxicity and high antibacterial activity, which provides a good prospect for bypassing antibiotics for future biomedical applications.

Evaluation of Antibacterial Effects of Matrix-Induced Silver Ions against Antibiotic-Resistant ESKAPE Pathogens.

Recently, drug-resistant bacterial infections, especially ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.), have become a critical health issue worldwide, highlighting the emerging need for novel antibacterial agents. In this study, silver nanoparticles were extracted from silver-containing mesoporous bioactive glass (MBG-Ag) using four different matrixes, including water, phosphate buffer saline (PBS), tryptic soy broth (TSB), and taurine (Tau). The inductively coupled plasma-mass spectrometer (ICP-MS) results demonstrated that the silver concentration of Tau-Ag was the highest among the four matrixes. The Tau-Ag was also observed to have 87.35% silver ions in its X-ray photoelectron spectrometer (XPS) spectra. The micrograph of transmission electron microscope (TEM) displayed a uniform distribution of silver nanoparticles, which was confined in a smaller size compared to that in TSB-Ag. Moreover, the peak shifts observed in the Fourier-transform infrared spectrometer (FTIR) spectrum implied that the -SO32- and -NH groups in taurine may interact with silver. A low cytotoxicity was noted for Tau-Ag, with approximately 70% of cells surviving at 0.63 mg/mL. Compared to the other three matrix-induced silver agents, Tau-Ag represented a better antibacterial effect against methicillin-resistant Staphylococcus aureus, with a minimum inhibitory concentration (MIC) value of 0.63 mg/mL and a postponed growth of 0.31 mg/mL observed. Further antibacterial examinations illustrated the presence of remarkable antibacterial activities against vancomycin-resistant Enterococcus feacium, carbapenem-resistant Klebsiella pneumoniae, carbapenem-resistant Acinetobacter baumannii, and carbapenem-resistant Pseudomonas aeruginosa. Given our observations and multiple bioactive functions of taurine (prevent patients from inflammation and oxidative-stress injuries), we anticipate that taurine matrix-induced silver ions would be a biomedical material with a high potential for combatting drug-resistant ESKAPE pathogens.

The Antibacterial and Remineralization Effect of Silver-Containing Mesoporous Bioactive Glass Sealing and Er-YAG Laser on Dentinal Tubules Treated in a Streptococcus mutans Cultivated Environment.

The aim of this study was to evaluate the remineralization and antibacterial effect of silver-containing mesoporous bioactive glass (MBG-Ag) sealing combined with Er:YAG laser irradiation on human demineralized dentin specimens in a Streptococcus mutans cultivated environment. A total of 48 human dentin specimens were randomly divided into four groups. The characteristics of MBG-Ag and the occlusion efficiency of the dentinal tubules were analyzed using X-ray diffraction patterns, Fourier-transform infrared spectroscopy, scanning electron microscope images and energy dispersive X-ray spectroscopy. Moreover, the antibacterial activity against Streptococcus mutans was evaluated by colony formation assay. The results showed that the dentin specimens with Er:YAG laser irradiation can form a melted occlusion with a size of 3-4 µm. MBG-Ag promoted the deposition of numerous crystal particles on the dentinal surface, reaching the deepest penetration depth of 70 µm. The results suggested that both MBG-Ag and laser have the ability to enhance the remineralization and precipitation of hydroxyapatite crystals. While the results showed that MBG-Ag sealing combined with the thermomechanical subablation mode of Er:YAG laser irradiation-induced dense crystalline deposition, reaching a penetration depth of more than 300 µm, silver nanoparticles without good absorption of the Er:YAG laser resulted in a heterogeneous radiated surface. Er:YAG laser irradiation with a low energy and pulse rate cannot completely inhibit the growth of S. mutans, but MBG-Ag sealing reached the bactericidal concentration. It was concluded that the simultaneous application of MBG-Ag sealing and Er:YAG laser treatment can prevent the drawbacks of their independent uses, resulting in a superior form of treatment for dentin hypersensitivity.

Strength and Biocompatibility of Heparin-Based Calcium Phosphate Cement Grafted with Ferulic Acid.

The biomimetic synthesis of carbonated apatites by biomolecule-based templates is a promising way for broadening apatite applications in bone tissue regeneration. In this work, heparin was used as an organic template to prepare uniform carbonate-based apatite nanorods (CHA) and graft ferulic acid (F-CHA) for enhanced bone mineralization. Next, by combining calcium phosphate cement (CPC) with different F-CHA/CPC ratios, a new type of injectable bone cement combined with F-CHA bioactive apatite was developed (CPC + F-CHA). The physicochemical properties, biocompatibility, and mineralization potential of the CPC + F-CHA composites were determined in vitro. The experimental results confirmed the preparation of highly biocompatible CHA and the compatibility of F-CHA with CPC. Although CPC + F-CHA composites with F-CHA (2.5 wt%, 5 wt%, and 10 wt%) showed a significant reduction in compressive strength (CS), compositing CPC with 10 wt% F-CHA yielded a CS suitable for orthopedic repair (CS still larger than 30 MPa). Spectroscopic and phase analyses revealed that the phase of the hydrothermally synthesized CHA product was not modified by the heparin template. Injection and disintegration tests indicated that the CPC + F-CHA composites have good biocompatibility even at 10 wt% F-CHA. D1 osteoprogenitor cells were cultured with the composites for 7 days in vitro, and the CPC + 10%F-CHA group demonstrated significantly promoted cell mineralization compared with other groups. Given these results, the use of over 10% F-CHA in CPC composites should be avoided if the latter is to be applied to load-bearing areas. A stress-shielding device may also be recommended to stabilize these areas. These newly developed biocompatible CPC + F-CHA have great potential as osteoconductive bone fillers for bone tissue engineering.

Evaluation of the Grafting Efficacy of Active Biomolecules of Phosphatidylcholine and Type I Collagen on Polyether Ether Ketone: In Vitro and In Vivo.

Biomolecule grafting on polyether ether ketone (PEEK) was used to improve cell affinity caused by surface inertness. This study demonstrated the sequence-polished (P) and sulfonated (SA) PEEK modification to make a 3D structure, active biomolecule graftings through PEEK silylation (SA/SI) and then processed with phosphatidylcholine (with silylation of SA/SI/PC; without SA/PC) and type I collagen (COL I, with silylation of SA/SI/C; without SA/C). Different modified PEEKs were implanted for 4, 8, and 12 weeks for histology. Sulfonated PEEK of SA showed the surface roughness was significantly increased; after the silylation of SA/SI, the hydrophilic nature was remarkably improved. The biomolecules were effectively grafted through silylation, and the cells showed improved attachment after 1 h. Furthermore, the SA/SI/PC group showed good in vitro mineralization. The new bone tissues were integrated into the 3D porous structures of SA/SI/PC and SA/SI/C in vivo making PEEK a potential alternative to metals in orthopedic implants.

Synergistic Antibacterial Effect of Casein-AgNPs Combined with Tigecycline against Acinetobacter baumannii.

Acinetobacter baumannii (A. baumannii) is a common and challenging pathogen of nosocomial infections, due to its ability to survive on inanimate objects, desiccation tolerance, and resistance to disinfectants. In this study, we investigated an antibacterial strategy to combat A. baumannii via the combination of antibiotics and silver protein. This strategy used a functional platform consisting of silver nanoparticles (AgNPs) resurrected from silver-based calcium thiophosphate (SSCP) through casein and arginine. Then, the silver protein was combined with tigecycline, the first drug in glycylcycline antibiotic, to synergistically inhibit the viability of A. baumannii. The synergistic antibacterial activity was confirmed by the 96-well checkerboard method to determine their minimum inhibitory concentrations (MIC) and calculated for the combination index (CI). The MIC of the combination of silver protein and tigecycline (0.31 mg/mL, 0.16 µg/mL) was significantly lower than that of the individual MIC, and the CI was 0.59, which indicates a synergistic effect. Consequently, we integrated the detailed synergistic antibacterial properties when silver protein was combined with tigecycline. The result could make for a promising approach for the treatment of A. baumannii.

In vitro and in vivo assessments of inspired Ag/80S bioactive nanocomposites against carbapenem-resistant Klebsiella pneumoniae.

In 2017 the World Health Organization listed carbapenem-resistant K. pneumoniae as a critical priority for developing a novel antimicrobial agent. Here we report on our investigation of the antibacterial efficacy of silver nanoparticles (AgNPs), confined to a mesostructured material and designated as an Ag/80S bioactive nanocomposite, against carbapenem-resistant K. pneumoniae. Results from a textural analysis indicate a 7.5 nm mesopore size and 307.6 m2/g surface area for Ag/80S. UV-Vis spectrum and transmission electron microscope images of Ag/80S revealed a uniform AgNP size distribution with an approximately 3.5 nm average. ICP-MS analysis demonstrated a significantly higher silver content in TSB (a protein-rich environment) compared to ultrapure water, suggesting a controllable release of Ag/80S and thus designated as the inspired Ag/80S. Minimum inhibitory concentration (MIC) values against 16 K. pneumoniae isolates ranged from 0.25 to 0.5% (2.5 to 5.0 mg/ml). NIH 3T3 fibroblast viability at 0.25% exceeded 80% and at 0.5% just under 70%, suggesting low cytotoxicity. Mechanistic study results indicate that the inspired Ag/80S attached to and deformed bacterial cells and induced a time-dependent accumulation of reactive oxygen species, leading to bacterial death. Further, inspired Ag/80S significantly extended median survival time in a Caenorhabditis elegans animal model infected with carbapenem-resistant K. pneumoniae ATCC BAA-1705. Combined, we found a novel Ag/80S which could prevent aggregation of AgNP and control its release via a specific environment for medical use against carbapenem-resistant K. pneumoniae.

Antibacterial silver-containing mesoporous bioglass as a dentin remineralization agent in a microorganism-challenged environment.

OBJECTIVES: To provide a suitable material capable of treating dentin hypersensitivity with simultaneous active antibacterial activity. METHODS: We developed silver-containing mesoporous bioglass (MBG-Ag) using the sol-gel technique, which loaded silver nanoparticles as promising bacteriostatic agents. The MBG-Ag with a powder-to-liquid ratio of 0.5 g: 0.01 mL were uniformly mixed with 20 %, 30 %, and 40 % phosphoric acid for 5, 10 and 20 min, respectively. Furthermore, we evaluated the occlusion efficiency, depth of penetration, and antibacterial activity of dentin specimens by simulating a Streptococcus mutans (S. mutans) infection on dentin. Scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR) were used to characterize the powders and assess tubule occlusion. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the MBG-Ag against S. mutans were determined via time-killing curves and colony formation assays. RESULTS: The MIC ranged from 2.5 to 5 mg/mL, and the MBC ranged from 5 to 10 mg/mL. The highest dentinal tubule occlusion efficiency was over 90 %. The colony formation assay confirmed that 5 mg/mL MBG-Ag mixed with phosphoric acid reached the bactericidal concentration. CONCLUSION: The MBG-Ag 40PA achieved a good occlusion efficiency and deep apatite precipitation in a short time, implying its superiority in clinical applications. CLINICAL RELEVANCE: The MBG-Ag formed in this study is a promising candidate for the treatment of demineralized dentin and confers antibacterial effects on the remineralized dentin surface against S. mutans.

Antibacterial Activity of Silver Nanoparticles (AgNP) Confined to Mesostructured, Silica-Based Calcium Phosphate Against Methicillin-Resistant Staphylococcus Aureus (MRSA).

Staphylococcus aureus, which is commonly found in hospitals, has become a major problem in infection control. In this study, Ag/80S bioactive ceramics used for enhanced antibacterial applications have been developed. An in vitro bioactivity test of the Ag/80S bioactive ceramic powders was performed in a phosphate-buffered saline (PBS). To explore the antibacterial activity of the Ag/80S bioactive ceramic powders, the Kirby-Bauer susceptibility test, the kinetics of microbial growth analysis and the colony-forming capacity assay were used to determine their minimum inhibitory concentration (MIC) against methicillin-resistant Staphylococcus aureus (MRSA). The results confirmed that the Ag/80S bioactive ceramic powders have antibacterial activity against MRSA (ATCC 33592) and MRSA (ATCC 49476).

Synthesis of hierarchically mesoporous silica with encapsulated avobenzone as a UV protection filter.

In this study, hierarchically mesoporous silica (HMS) with properties such as high specific surface area, high photostability, and no cellular toxicity was synthesized. The synthesized silica can be considered as an excellent carrier candidate material. Through the use of nitrogen adsorption and desorption analysis, the shape of the hysteresis loop implied the presence of mesoporous structures in the HMS powder. In addition, the encapsulation efficiency was more than 90%. These results showed that avobenzone could be encapsulated into the HMS powder because of its high specific surface area and pore volume. Additionally, X-ray diffractometry (XRD), Fourier transform infrared spectroscopy (FTIR), thermal gravimetric analysis (TGA), and UV-visible (Vis) spectrophotometry were used to prove that the hierarchically mesoporous silica was able to effectively encapsulate avobenzone. In addition, the new synthetic sunscreen kept its excellent UVA absorption properties after being encapsulated.

Microwave assisted synthesis of negative-charge carbon dots with potential antibacterial activity against multi-drug resistant bacteria.

In this research, negative-charge carbon dots (CDs) were synthesized in one-step using a microwave and found to have potential antibacterial ability against multi-drug resistant bacteria. The CDs were synthesized by using citric acid and urea as precursors, and characterized by FT-IR, TEM and fluorescence spectrophotometry. The average size of CDs was about 2.5 nm, and the ζ potential was -11.06 mV. In the following antibacterial activity test, time-killing curve experiments and colony-forming assay were carried out to determine the minimum bactericidal concentration (MBC) and minimum inhibitory concentration (MIC) of the CDs against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-intermediate Staphylococcus aureus (VISA). The data showed the MBC of the CDs against MRSA is 2.5 mg mL-1, and the MIC of the CDs against MRSA is 0.63 mg mL-1; the MBC of the CDs against VISA is 1.25 mg mL-1, and the MIC of the CDs against VISA is 0.63 mg mL-1. The results demonstrated that the negative-charge CDs have potential against multi-drug resistant Staphylococcus aureus (S. aureus), and may serve as alternatives for therapy in the future.

Determination of phenylenediamines in hair colors derivatizated with 5-(4, 6-dichlorotriazinyl)aminofluorescein via micellar electrokinetic chromatography.

Phenylenediamines (PDs), which are reported to cause allergic dermatitis and possess genotoxicity and carcinogenicity, are the ingredients used in permanent hair dyes. The fluorescent derivatization strategy coupled with micellar electrokinetic chromatography (MEKC) were established to analyze four PDs, including o-phenylenediamine (OPD), m-phenylenediamine (MPD), p-phenylenediamine (PPD) and toluene-2,5-diamine (PTD). Additionally, 5-(4, 6-dichlorotriazinyl) aminofluorescein (DTAF) was used as a fluorescent reagent derived at amino groups of PDs and underwent nucleophilic substitution reaction to improve the detection sensitivity. The derivatization condition reacted at 90 °C for 10 min in alkaline conditions. The optimized separation conditions were 20 mM borate (pH 8.0) containing 10 mM Brij 35 and 35% (v/v) methanol. The limits of detection (S/N = 3) for MPD, PTD, PPD and OPD were 25, 25, 50 and 100 nM, respectively. Compared to MEKC-UV, the sensitivity enhancements were 30- to 81-fold when PDs were derived with DTAF. The high-sensitivity MEKC-LIF method was successfully established and applied to determine PDs in commercial hair colors for quality control and in real hair samples for evaluating the location of PDs in dyed hair samples, as well as in percutaneous absorption samples for evaluating the ability of PDs to penetrate skin.

Control of Ag nanoparticle distribution influencing bioactive and antibacterial properties of Ag-doped mesoporous bioactive glass particles prepared by spray pyrolysis.

Mesoporous bioactive glasses (MBGs) have become important bone implant materials because of their high specific surface area resulting in high bioactivity. Doping MBGs with Ag removes one of the remaining challenges to their applications, namely their lack of intrinsic antibacterial properties. In present work we demonstrate that Ag-doped MBGs can be prepared in one-step spray pyrolysis (SP) process. The SP preparation method offers the advantages of short processing times and continuous production over the sol-gel method previously used to prepare MBGs. Using scanning electron microscopy, transmission electron microscopy, and selected area electron diffraction we demonstrate that the synthesized MBG particles have amorphous structure with nanocrystalline Ag inclusions. The scanning transmission electron microscopy-X-ray energy dispersive spectrometry of cross-sectional samples shows that the distribution of the Ag dopant nanoparticles within MBGs can be controlled by using the appropriate formulation of the precursors. The distribution of the Ag dopant nanoparticles within the MBG particles was found to affect their surface areas, bioactivities and antibacterial properties. Based on the observations, we propose a mechanism describing MBG particle formation and controlling dopant distribution.

Thermal cycling effects on adhesion of resin-bovine enamel junction among different composite resins.

Thermal cycling is used to mimic the changes in oral cavity temperature experienced by composite resins when used clinically. The purpose of this study is to assess the thermal cycling effects of in-house produced composite resin on bonding strength. The dicalcium phosphate anhydrous filler surfaces are modified using nanocrystals and silanization (w/NP/Si). The resin is compared with commercially available composite resins Filtek Z250, Z350, and glass ionomer restorative material GIC Fuji-II LC (control). Different composite resins were filled into the dental enamel of bovine teeth. The bond force and resin-enamel junction graphical structures of the samples were determined after thermal cycling between 5 and 55°C in deionized water for 600 cycles. After thermal cycling, the w/NP/Si 30wt%, 50wt% and Filtek Z250, Z350 groups showed higher shear forces than glass ionomer GIC, and w/NP/Si 50wt% had the highest shear force. Through SEM observations, more of the fillings with w/NP/Si 30wt% and w/NP/Si 50wt% groups flowed into the enamel tubule, forming closed tubules with the composite resins. The push-out force is proportional to the resin flow depth and uniformity. The push-out tubule pore and resin shear pattern is the most uniform and consistent in the w/NP/Si 50wt% group. Accordingly, this developed composite resin maintains great mechanical properties after thermal cycling. Thus, it has the potential to be used in a clinical setting when restoring non-carious cervical lesions.

Genotyping of intron 22 inversion of factor VIII gene for diagnosis of hemophilia A by inverse-shifting polymerase chain reaction and capillary electrophoresis.

This is the first capillary electrophoresis (CE) analysis for diagnosis of hemophilia A (HA). The intron 22 inversion of factor VIII gene (F8) causes 40-50 % of severe bleeding disorder of HA in all human populations. Consequently, identification of the disease-causing mutations is becoming increasingly important for accurate genetic counseling and prenatal diagnosis. In this study, the key steps of inverse-shifting polymerase chain reaction (IS-PCR) and of short-end injection capillary electrophoresis were used for more specific and rapid genotyping of intron 22 inversion of F8. In IS-PCR, three specific primers were used to amplify 512-bp amplicon for wild type and 584-bp amplicon for patients with intron 22 inversion. The capillary gel electrophoresis (CGE) system was performed using 1× Tris-borate-EDTA (TBE) buffer containing 0.3 % (w/v) polyethylene oxide (PEO). The PCR amplicons were electrokinetically injected at 10 kV for 10 s at a temperature of 25 °C. The optimal short-end injection CGE was applied to detect the F8 gene of HA patients and carriers within 5 min. Intron 22 inversion was indeed found on some HA patients (13/35, 37.1 %). All genotyping results showed good agreement with DNA sequencing method and long-distance polymerase chain reaction (LD-PCR). The IS-PCR combined with short-end injection CGE method was feasible and efficient for intron 22 inversion screening of F8 in the HA populations.