Enhanced biomedical applicability of ZrO2-SiO2 ceramic composites in 3D printed bone scaffolds.

Zirconia (ZrO2) has been widely used in clinical applications, such as bone and dental implantation, because of its favorable mechanical properties and resistance to fracture. However, the poor cell affinity of ZrO2 for bone regeneration and tissue binding, as well as its shrinkage due to crystal phase transformation during heat treatment, limits its clinical use and processing plasticity. This study aims to investigate an appropriate ZrO2-SiO2 composite recipe for ceramic 3D printing processes that can strike a balance between the mechanical properties and cell affinity needed in clinical applications. Specimens with different ZrO2-SiO2 composite recipes were fabricated by a selective laser gelling method and sintered at temperatures ranging from 900 to 1500 °C. The S5Z5 composite, which consists of 50 wt% ZrO2, 35 wt% SiO2 and 15 wt% SiO2 sol, showed an appropriate compressive strength and bending strength of 82.56 MPa and 55.98 MPa, respectively, at a sintering temperature of 1300 °C. The shrinkage rate of the S5Z5 composite was approximately 5% when the sintering temperature was increased from 900 to 1500 °C. All composites exhibited no cytotoxicity after 144 h of MG63 cell incubation, and the S5Z5 composite exhibited the most obvious cell affinity among the composite recipes. From these results, compared with other composites, the S5Z5 composite was shown to possess mechanical properties and a cell affinity more comparable to those of natural human bone.

Enhanced mechanical and biological performances of CaO-MgO-SiO2 glass-ceramics via the modulation of glass and ceramic phases.

This work reports a new CaO-MgO-SiO2 (CMS) bioactive glass-ceramic, using ZrO2 as a nucleus to modulate the ratios of glass and ceramic phases as a function of sintering temperature. Mg-rich bioactive CMS glass-ceramics exhibit advantages regarding mechanical strength (flexural strength ~190 MPa and compressive strength ~555 MPa), in-vitro and in-vivo biocompatibilities, and bone ingrowth. The high mechanical strengths could be attributed to the CaMgSi2O6 glass-ceramic and lower porosity. X-ray absorption spectra indicate an increased SiO covalent bond via the development of CaMgSi2O6 glass-ceramics. From the in-vitro cytotoxicity and BMSC differentiation assays, the CMS samples sintered above 800 °C exhibited better cell attachment and differentiation, possibly due to structural stability, appropriate pore, and ion release to boost osteogenesis. Compared to hydroxyapatite (HA) ceramics, the CMS glass-ceramics display higher mechanical strengths, biocompatibility, and osteoconductivity. An in-vivo experiment demonstrated a fine bone-ingrowth profile around the CMS implant. This study may further the application of CMS glass-ceramics in bone implants.

Facilitating GL13K Peptide Grafting on Polyetheretherketone via 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide: Surface Properties and Antibacterial Activity.

In the present work, the antimicrobial peptide (AMP) of GL13K was successfully coated onto a polyetheretherketone (PEEK) substrate to investigate its antibacterial activities against Staphylococcus aureus (S. aureus) bacteria. To improve the coating efficiency, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) was mixed with a GL13K solution and coated on the PEEK surface for comparison. Both energy-dispersive X-ray spectroscopy (EDX) and X-ray photoelectron spectroscopy (XPS) data confirmed 30% greater peptide coating on PEEK/GL13K-EDC than PEEK without EDC treatment. The GL13K graft levels are depicted in the micrograms per square centimeter range. The PEEK/GL13K-EDC sample showed a smoother and lower roughness (Rq of 0.530 µm) than the PEEK/GL13K (0.634 µm) and PEEK (0.697 µm) samples. The surface of the PEEK/GL13K-EDC was more hydrophilic (with a water contact angle of 24°) than the PEEK/GL13K (40°) and pure PEEK (89°) samples. The pure PEEK disc did not exhibit any inhibition zone against S. aureus. After peptide coating, the samples demonstrated significant zones of inhibition: 28 mm and 25 mm for the PEEK/GL13K-EDC and PEEK/GL13K samples, respectively. The bacteria-challenged PEEK sample showed numerous bacteria clusters, whereas PEEK/GL13K contained a little bacteria and PEEK/GL13K-EDC had no bacterial attachment. The results confirm that the GL13K peptide coating was able to induce antibacterial and biofilm-inhibitory effects. To the best of our knowledge, this is the first report of successful GL13K peptide grafting on a PEEK substrate via EDC coupling. The present work illustrates a facile and promising coating technique for a polymeric surface to provide bactericidal activity and biofilm resistance to medical implantable devices.

Size-Dependent Antibacterial Activity of Silver Nanoparticle-Loaded Graphene Oxide Nanosheets.

A series of graphene oxide (GO) suspensions with different particle sizes (<100 nm, ~100 nm, ~1 µm and >1 µm) were successfully fabricated after 0, 30, 60 and 120 min of sonication, respectively. The antibacterial properties of GO suspensions showed that >1 µm GO size resulted in a loss of nearly 50% of bacterial viability, which was higher than treatment by ~100 nm GO size (25%) towards Escherichia coli (E. coli). Complete entrapment of bacteria by the larger GO was observed in transmission electron microscopy (TEM). Silver nanoparticles (Ag NPs) were doped onto GO samples with different lateral sizes to form GO-Ag NP composites. Resulting larger GO-Ag NPs showed higher antibacterial activity than smaller GO-Ag NPs. As observed by Fourier transform infrared spectroscopy (FTIR), the interaction between E. coli and GO occurred mainly at the outer membrane, where membrane amino acids interact with hydroxyl and epoxy groups. The reactive oxygen species (ROS) and the considerable penetration of released Ag+ into the inner bacterial cell membrane result in loss of membrane integrity and damaged morphology. The present work improves the combined action of GO size effect with constant Ag loadings for potential antibacterial activity.

Nanoarchitectured structure and population dynamics of anaerobic ammonium oxidizing (anammox) bacteria in a wastewater treatment plant.

Studies on microbial community and population dynamics are essential for the successful development, monitoring and operation of biological wastewater treatment systems. Especially for novel or sustainable systems such as the anaerobic ammonium oxidizing (anammox) process that are not yet well explored. Here we collected granular microbial sludge samples and investigated a community of anammox bacteria over a period of four years, divided into eight stages in a full scale simultaneous partial nitrification, anammox and denitrification (SNAD) process for treating landfill leachate. Specific qPCR primers were designed to target and quantify the two most abundant anammox species, Candidatus Kuenenia stuttgartiensis (KS) and Candidatus Brocadia anammoxidans (BA). The two species were monitored and could explain the dynamic shift of the anammox community corresponding to the operating conditions. Using the newly designed KS-specific primer (KSqF3/KSqR3) and BA-specific primer (BAqF/BAqR), we estimated the amounts of KS and BA to be in the range of 6.2 × 106 to 5.9 × 108 and 1.1 × 105 to 4.1 × 107 copies µg-1 DNA, respectively. KS was found to be the dominant species in all anammox granules studied and played an important role in the formation of granules. The KS/BA ratio was positively correlated to the size of granules in the reactor and ammonia nitrogen removal efficiency of the treatment plant.

Toward Antibiofouling PVDF Membranes.

Membranes for biologically and biomedically related applications must be bioinert, that is, resist biofouling by proteins, human cells, bacteria, algae, etc. Hydrophobic materials such as polysulfone, polypropylene, or poly(vinylidene fluoride) (PVDF) are often chosen as matrix materials but their hydrophobicity make them prone to biofouling, which in turn limits their application in biological/biomedical fields. Here, we designed PVDF-based membranes by precipitation from the vapor phase and zwitterionized them in situ to reduce their propensity to biofouling. To achieve this goal, we used a copolymer containing phosphorylcholine groups. An in-depth physicochemical characterization revealed not only the controlled presence of the copolymer in the membrane but also that bicontinuous membranes could be formed. Membrane hydrophilicity was greatly improved, resulting in the mitigation of a variety of biofoulants: the attachment of Stenotrophomonas maltophilia, Streptococcus mutans, and platelets was reduced by 99.9, 99.9, and 98.9%, respectively. Besides, despite incubation in a plasma platelet-poor medium, rich in plasma proteins, a flux recovery ratio of 75% could be measured while it was only 40% with a hydrophilic commercial membrane of similar structure and physical properties. Similarly, the zwitterionic membrane severely mitigated biofouling by microalgae during their harvesting. All in all, the material/process combination presented in this work leads to antibiofouling porous membranes with a large span of potential biomedically and biologically related applications.

Anti-ST2 monoclonal antibody inhibits eosinophil infiltration in Angiostrongylus cantonensis-infected mice.

BACKGROUND/PURPOSE: Interleukin-33 (IL-33) could play an important role in the pathogenesis of angiostrongylosis. However, the role of IL-33/ST2 pathway in this parasitic infection is uncertain. METHODS: C57BL/six mice were each infected with 35 Angiostrongylus cantonensis larvae. One group of mice received an intraperitoneal injection of anti-ST2 monoclonal antibody (mAb; 50 µg) 3 days postinfection and subsequent booster shots of the same dose at 5-day intervals. Blood samples from each group were collected every week for assays. RESULTS: The level of IL-5 significantly decreased in the mAb-treated group, and the infiltration of eosinophils in the meninges was also significantly reduced. CONCLUSION: The IL-33/ST2 axis may play a crucial role in the pathogenesis of angiostrongylosis and the results of this study could be useful for the development of strategies to reduce the neurological damage caused by this parasitic infection.

Gene polA as a Suitable Reference for Studying Antibacterial Effect of Hydroxyapatite.

Hydroxyapatite (HAP: Ca10(PO4)6(OH)2) is a widely used biocompatible material; however, information on the reaction mechanism between HAP and microorganisms is insufficient. This study aimed to identify a stable reference gene for studying the antibacterial activity of HAP. The half maximal inhibitory concentration (C50) and gene expression of a human symbiotic Escherichia coli strain TOP10, was investigated at various concentrations of HAP. Our uniformly sized HAP nanoparticles (HANPs) had a high surface area and an estimated IC50 of 75 mg/mL. The expressions of genes, including those of DNA polymerase I (poIA), DNA polymerase II (poIB), cytochrome d complex (cyd), glucan biosynthesis protein G (mdoG), D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and 16S ribosomal RNA (16S rRNA), were analyzed by performing quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and using reported and newly designed primers. The changes in the copy numbers compared to non-HANPs-treated conditions for polB, cyd, mdoG, GAPDH, and 16S rRNA were 50%-381%, 0.7%-66.3%, 1.1%-7.8%, 1.6%-86.3%, and 0.3%-8.1%, respectively. The expressions of polA remained stable under all conditions (62.8%-88.4%). Therefore, we identified polA as a suitable reference gene. Moreover, the expressions of cyd and mdoG were inhibited, indicating that the antibacterial activity of HANPs is related to cell membrane or cell wall proteins. Our findings provide a thorough understanding of HAP-microorganism interactions for potential biomedical applications.

Interleukin-27 Protects Cardiomyocyte-Like H9c2 Cells against Metabolic Syndrome: Role of STAT3 Signaling.

The present results demonstrated that high glucose (G), salt (S), and cholesterol C (either alone or in combination), as mimicking extracellular changes in metabolic syndrome, damage cardiomyocyte-like H9c2 cells and reduce their viability in a time-dependent manner. However, the effects were greatest when cells were exposed to all three agents (GSC). The mRNA of glycoprotein (gp) 130 and WSX-1, both components of the interleukin (IL)-27 receptor, were present in H9c2 cells. Although mRNA expression was not affected by exogenous treatment with IL-27, the expression of gp130 mRNA (but not that of WSX-1 mRNA) was attenuated by GSC. Treatment of IL-27 to H9c2 cells increased activation of signal transducer and activator of transcription 3 (STAT3) and protected cells from GSC-induced cytochrome c release and cell damage. The protective effects of IL-27 were abrogated by the STAT3 inhibitor, stattic. The results of the present study clearly demonstrate that the STAT3 pathway triggered by anti-inflammatory IL-27 plays a role in protecting cardiomyocytes against GSC-mediated damage.

Interleukin-27, a novel cytokine induced by ischemia-reperfusion injury in rat hearts, mediates cardioprotective effects via the gp130/STAT3 pathway.

Patients with coronary artery disease show high serum levels of interleukin (IL)-27, a novel member of the IL-6 family. However, the function of IL-27 in hearts suffering ischemia/reperfusion (IR) injury is unclear. Here, we showed increased expression of mRNA for the IL-27 subunits, EBI3 and p28, in rat hearts after 40 min of coronary ligation and release for 7 days. This increase was associated with a peak in the release of the cardiac enzyme, creatine kinase-MB, on day 2 post-release. Moreover, levels of IL-27 receptor subunit gp130 mRNA, but not those of subunit WSX-1 mRNA, decreased in post-ischemic hearts. These results suggest that increased IL-27 production may compensate for receptor downregulation during myocardial recovery. Lactate dehydrogenase release and crystal violet staining revealed that IL-27 or IL-6 significantly attenuated severe hypoxia (SH, 2 % O2)-induced cell damage in H9c2 cardiomyoblasts and primary rat neonatal cardiomyocytes. Incubating cardiomyocytes with IL-27 or IL-6 resulted in time-dependent activation of signal transducers and activators of transcription 3 (STAT3). Interestingly, IL-27-induced STAT3 activation was attenuated by pre-treatment with a gp130-neutralizing antibody. Blocking gp130 also reduced the cytoprotective effects of IL-27 or IL-6. Moreover, IL-27-mediated protection against SH was blocked by stattic, a small-molecule inhibitor of STAT3. IL-27 markedly improved post-ischemic recovery and reduced tissue damage in isolated perfused hearts when administered 5 min before reperfusion. These results indicate that IL-27 protects the myocardium against IR injury and facilitates the recovery of damaged cardiomyocytes via the gp130/STAT3 pathway.

Start-up of simultaneous partial nitrification, anammox and denitrification (SNAD) process in sequencing batch biofilm reactor using novel biomass carriers.

Simultaneous partial nitrification, anammox and denitrification (SNAD) process was started-up in a 2.5L sequencing batch biofilm reactor (SBBR) using novel biomass carriers. The SNAD process took only 51d for start-up at nitrogen loading rate (NLR) and organic loading rate (OLR) of 120 and 60g/m(3)-d, respectively. Long-term stable operation of SNAD process was observed at NLR and OLR of 360 and 180g/m(3)-d with average total nitrogen and COD removal efficiencies of >88% and >90%, respectively. The values of conversion ratio [Formula: see text] remained below 0.11 after the start-up period, which further confirmed the long-term stability of SNAD process. Results of polymerase chain reaction (PCR), qualitative PCR, and scanning electron microscopic (SEM) analysis of sludge samples confirmed the co-existence and enrichment of AOB, anammox bacteria and denitrifying bacteria in the reactor and biofilm formation on to the carriers.

Functionalized Fe3O4@silica core-shell nanoparticles as microalgae harvester and catalyst for biodiesel production.

Core-shell Fe3O4@silica magnetic nanoparticles functionalized with a strong base, triazabicyclodecene (TBD), were successfully synthesized for harvesting microalgae and for one-pot microalgae-to-fatty acid methyl ester (FAME, or so-called biodiesel) conversion. Three types of algae oil sources (i.e., dried algae, algae oil, and algae concentrate) were used and the reaction conditions were optimized to achieve the maximum biodiesel yield. The results obtained in this study show that our TBD-functionalized Fe3O4@silica nanoparticles could effectively convert algae oil to biodiesel with a maximum yield of 97.1 %. Additionally, TBD-Fe3O4@silica nanoparticles act as an efficient algae harvester because of their adsorption and magnetic properties. The method presented in this study demonstrates the wide scope for the use of covalently functionalized core-shell nanoparticles for the production of liquid transportation fuels from algal biomass.

Evaluating the antibacterial property of gold-coated hydroxyapatite: a molecular biological approach.

Hydroxyapatite nanoparticles (HAP NPs) are one of the widely used biocompatible materials. However, information about the reaction between HAP NPs and microorganisms is insufficient. This paper aims to understand the antibacterial property of a new nanocomposite consisting of gold-coated HAP and alginate polymer (namely, Au-HAP@Alg NPs). To the best of our knowledge, we reported the first information regarding to MIC (25mg/mL), DIZ (no visible zone), and IC50 (0.5mg/mL) of Au-HAP@Alg NPs toward the microorganism Escherichia coli TOP10. The real-time gene expression levels of polA, polB, cyd, mdoG, GAPDH, and 16S rRNA were maintained at stable levels up until conditions of 2.5mg/mL Au-HAP@Alg NPs. The results showed that 16S rRNA can be a good reference under these conditions. The expressions of GAPDH, cyd, and mdoG were inhibited obviously under condition of 10mg/mL of Au-HAP@Alg NPs. Our results indicated that the possible antibacterial mechanism of Au-HAP@Alg was through the interaction with these carbohydrate and cell wall-related genes. This novel biocompatible and antibacterial material can potentially be applied in medical and environmental fields.

A comparison of two 16S rRNA gene-based PCR primer sets in unraveling anammox bacteria from different environmental samples.

Two 16S rRNA gene-based PCR primer sets (Brod541F/Amx820R and A438f/A684r) for detecting anammox bacteria were compared using sediments from Mai Po wetlands (MP), the South China Sea (SCS), a freshwater reservoir (R2), and sludge granules from a wastewater treatment plant (A2). By comparing their ability in profiling anammox bacteria, the recovered diversity, community structure, and abundance of anammox bacteria among all these diverse samples indicated that A438f/A684r performed better than Brod541F/Amx820R in retrieving anammox bacteria from these different environmental samples. Five Scalindua subclusters (zhenghei-I, SCS-I, SCS-III, arabica, and brodae) dominated in SCS whereas two Scalindua subclusters (zhenghei-II and wagneri) and one cluster of Kuenenia dominated in MP. R2 showed a higher diversity of anammox bacteria with two new retrieved clusters (R2-New-1 and R2-New-2), which deserves further detailed study. The dominance of Brocadia in sample A2 was supported by both of the primer sets used. Results collectively indicate strongly niche-specific community structures of anammox bacteria in different environments, and A438f/A684r is highly recommended for screening anammox bacteria from various environments when dealing with a collection of samples with diverse physiochemical characteristics.

Microbial community and population dynamics of single-stage autotrophic nitrogen removal for dilute wastewater at the benchmark oxygen rate supply.

Microbial communities and their kinetic performance in a single-stage autotrophic nitrogen-removal filter at an optimal oxygen supply were examined to determine the presence and activity of denitrifiers, anaerobic ammonia-oxidizing (anammox), ammonia-oxidizing, and nitrite-oxidizing bacteria. To this end, different molecular biology techniques such as real-time quantitative polymerase chain reaction (qPCR) and biomarkers such as 16S rRNA revealed a diverse microbial community along the filter. It was important to survey the specific species of anammox bacteria using a newly designed Candidatus Brocadiafulgida (BF) specific primer, as well as Candidatus Brocadia anammoxidans (BA) and Candidatus Kuenenia stuttgartiensis (KS) specific primers. An unexpected finding was that the predominant anammox species switched from KS in concentrated wastewater to BA in dilute wastewaters. The Eckenfelder model of the NH3-N transformation along the filter was Se=S0 exp(-0.192D/L(2.3217)). These results provide a foundational understanding of the microbial structure and reaction kinetics in such systems.

Preparation of ordered mesoporous alumina-doped titania films with high thermal stability and their application to high-speed passive-matrix electrochromic displays.

Ordered mesoporous alumina-doped titania thin films with anatase crystalline structure were prepared by using triblock copolymer Pluronic P123 as structure-directing agent. Uniform Al doping was realized by using aluminum isopropoxide as a dopant source which can be hydrolyzed together with titanium tetraisopropoxide. Aluminum doping into the titania framework can prevent rapid crystallization to the anatase phase, thereby drastically increasing thermal stability. With increasing Al content, the crystallization temperatures tend to increase gradually. Even when the Al content doped into the framework was increased to 15 mol %, a well-ordered mesoporous structure was obtained, and the mesostructural ordering was still maintained after calcination at 550 °C. During the calcination process, large uniaxial shrinkage occurred along the direction perpendicular to the substrate with retention of the horizontal mesoscale periodicity, whereby vertically oriented nanopillars were formed in the film. The resulting vertical porosity was successfully exploited to fabricate a high-speed and high-quality passive-matrix electrochromic display by using a leuco dye. The vertical nanospace in the films can effectively prevent drifting of the leuco dye.

Engineering of the growth environment of microalgae with high biomass and lipid productivity.

Pure cultures of Botryococcus sp. microalgae have great potential for generating huge amounts of algae lipid that can be further converted into biodiesel. Lipids with nanometer in size can be applied to medicine and pharmacy recently. In this study, the effects of light intensity and CO2 concentration on the biomass productivity, lipid content, and lipid productivity of Botryococcus braunii were examined in 21-day intervals. The optimum cultivating conditions for biomass accumulation were 6,000 lux with 0.04% CO2 and 21 days of culturing; this provided the highest biomass productivity of 140.46 mg L(-1) d(-1). The highest lipid productivity of 44.46 mg L(-1) d(-1) occurred at 6,000 lux with 5% CO2 and 21 days of culturing. The maximum specific growth rate (micro(max)) was similar among different concentrations of CO2 (0.682 d(-1) under 12,000 lux at 10% CO2; 0.585 d(-1) under 6,000 lux at 5% CO2). Culturing at 5% or 10% CO2 has been shown to enhance the accumulation of lipids, introducing the possibility of using flue gas as a carbon source. The nanotechnology in this study will be helpful towards research in green science and engineering such as bio-fixation of CO2 and drug delivery systems.

Partial nitrification and anammox process: a method for high strength optoelectronic industrial wastewater treatment.

Completely autotrophic nitrogen removal over nitrite (CANON) process was employed in an 18 L sequencing batch reactor (SBR) for treatment of optoelectronic industrial wastewater containing high strength ammonium nitrogen (3712 ± 120 mg NH4(+) - N L(-1)). About 89% of total nitrogen and 98% of NH4(+) - N removal efficiencies were observed at the loading rate of 909 g N m(-3) d(-1) and the HRT of 4 d. A profound variation in the performance of CANON process was experienced at high DO exposure (above 1 mg L(-1)) and high nitrite concentration (above 100 mg L(-1)). Inhibition due to high DO exposure was found to be reversible phenomenon whereas the synergistic inhibition of nitrite, free ammonia and free nitrous acid was irreversible. The fluctuation of reactor temperature between 17 and 37 °C did not affect the performance of CANON system. The CANON process was stably controlled at high nitrogen loading rate for more than one month. The co-existence of aerobic and anaerobic ammonium oxidizing bacteria in the reactor was detected by The PCR analysis. About 5 fold increase in amount of anammox bacteria over a period of 258 days was confirmed from the results of qPCR on day 487.

Size distribution of airborne fungi in vehicles under various driving conditions.

Circulation or air conditioning (AC) system was proven to improve the air quality inside the vehicles; however, the quantified study was limited. In this study, fungal concentration under various driving mode inside the vehicle was proposed. The driving conditions were classified into 4 states: (1) window closed without AC and circulation, (2) window open without AC and circulation, (3) window closed with only circulation on, and (4) window closed with only AC on. Results show that at state 4, the mean respirable fraction was 83.3%, with a number median diameter of the fungi being 1.73 µm. More attention should be paid for these smaller fungi easily penetrating into the alveoli and probably lead to allergic alveolitis. Turning on AC for reducing the normalized concentration for each size range of fungi was suggested; however, the respirable fraction increased. Those who are prone to allergies or asthma are suggested to switch between AC and the circulation mode while driving a long time.

Efficient single-stage autotrophic nitrogen removal with dilute wastewater through oxygen supply control.

Autotrophic nitrogen removal via ammonia oxidizing (AOB) and anaerobic ammonium oxidizing (anammox) bacteria was evaluated for treatment of a dilute 50mg/L ammonia-containing solution in a single-stage nitrogen-removal filter at 25°C. Important was an external oxygenation system that permitted close control and measurement of oxygen supply, a difficulty with the generally used diffused air systems. Hydraulic retention time (HRT) was reduced in steps from 15 to 1h. At 1h HRT, total nitrogen (TN) removals varied between 73% and 94%, the maximum being obtained with a benchmark oxygenation ratio of 0.75mol O(2)/mol ammonia fed. At higher ratios, nitrate was formed causing TN removal efficiency to decrease. With lower ratios, TN and ammonia removals decreased in proportion to the decrease in BOR. When operating at or below the BOR, nitrate formation equaled no more than 2% of the ammonia removed, a value much less than has previously been reported.