Effect of Geopolymerization Reaction on the Flexural Strength of Kaolin-Based Systems.

Geopolymers exhibit broad application prospects, including construction and radiation shielding, which require excellent mechanical performances. However, investigations on the nature of geopolymerization reactions and their consequential impact on mechanical performance are still vague. In this study, the effect of the major factors of Si/Al ratio and curing time on the geopolymerization reaction and flexural strength were studied based on the microstructure evolution and chemical bonding formation analyzed using the SEM, FTIR, peak deconvolution, and XRD methods. The microstructure of geopolymers was transferred from initially layered smooth particles of kaolinite to a 3D network porous structure, corresponding to sodalite. A spectrum exclusive to the geopolymer structure occurred at 973 cm-1, corresponding to the sodium aluminum silicate hydrate (N-A-S-H) links, the integral area of which represents the degree of geopolymerization reaction. Furthermore, a controllable reaction degree was achieved by adjusting the Si/Al ratio and curing time, where the maximum reaction degree of 55% was achieved at a Si/Al ratio of 1.94 when cured for 7 d. The correlation between the flexural strength and reaction degree was found to follow a proportional relationship, achieving a flexural strength of 21.11 MPa with a degree of 45%. This study provides insight into the development of mechanical strength through controlling the reaction process.

3D PEEK Objects Fabricated by Fused Filament Fabrication (FFF).

PEEK (poly ether ether ketone) materials printed using FFF 3D printing have been actively studied on applying electronic devices in satellites owing to their excellent light weight and thermal resistance. However, the PEEK FFF process generated cavities inside due to large shrinkage has degraded both mechanical integrity and printing reliability. Here, we have investigated the correlations between nozzle temperatures and PEEK printing behaviors such as the reliability of printed line width and surface roughness. As the temperature increased from 360 to 380 °C, the width of the printed line showed a tendency to decrease. However, the width of PEEK printed lines re-increased from 350 to 426 µm at the nozzle temperatures between 380 and 400 °C, associated with solid to liquid-like phase transition and printed out distorted and disconnected lines. The surface roughness of PEEK objects increased from 49 to 55 µm as the nozzle temperature increased from 380 to 400 °C, where PEEK is melted down and quickly solidified based on more energy and additional heating time at higher printing temperatures at 400 °C. Based on these printing trends, a reliability analysis of the printed line was performed. The printed line formed the most uniform width at 380 °C and had a highest Weibull coefficient of 28.6 using the reliability analysis technique called Weibull modulus.

Development of ceramic additive manufacturing: process and materials technology.

Ceramic additive manufacturing (C-AM) is highlighted as a technology that can overcome the inherent limitations of ceramics such as processability and formability. This process creates a structure by slicing a 3D model and stacking ceramic materials layer-by-layer without mold or machining. C-AM is a technology suitable for the era of multiple low-volume because it is more flexible than conventional methods for shape complexity and design modification. However, many barriers to practical use remain due to process speed, defects, and lack of knowledge. This review focuses on studies to overcome the limitations of C-AM in terms of process and materials. The C-AM process has been advanced through various studies such as model/equation-based parameter control and high-speed sintering using external energy. Besides, by improving and fusing existing technologies, high-precision high-speed printing technology has been improved. A variety of material studies have been made of manufacturing ceramic structures with superior properties using preceramic polymers and composite materials. Through these studies, C-AM has been applied to various fields such as medicine, energy, environment, machinery, and architecture. These continued growths and diverse results demonstrate the importance and potential of C-AM based ceramic manufacturing technology.

Overexpression of N141I PS2 increases γ-secretase activity through up-regulation of Presenilin and Pen-2 in brain mitochondria of NSE/hPS2m transgenic mice.

Alzheimer's disease (AD) is known to induce alterations of mitochondrial function such as elevation of oxidative stress and activation of apopotosis. The aim of this study was to investigate the effects of human Presenilin 2 mutant (hPS2m) overexpression on the γ-secretase complex in the mitochondrial fraction. To achieve this, alterations of γ-secretase complex expression and activity were detected in the mitochondrial fraction derived from brains of NSE/hPS2m Tg mice and Non-Tg mice. Herein, the following were observed: i) overexpression of the hPS2m gene significantly up-regulated the deposition of Aß-42 peptides in the hippocampus and cortex of brain, ii) overexpression of hPS2m protein induced alterations of γ-secretase components such as main component protein and activator protein but not stabilization-related proteins, iii) changes in γ-secretase components induced by overexpression of hPS2m protein up-regulated γ-secretase activity in the mitochondrial fraction, and iv) elevation of γ-secretase activity induced production of Aß-42 peptides in the mitochondrial fraction. Based on these observations, these results indicate that alteration of γ-secretase activity in cells upon overexpression of hPS2m is tightly linked to mitochondrial dysfunction under the specific physiological and pathological conditions of AD.

3,4,5-trihydroxycinnamic acid inhibits lipopolysaccharide (LPS)-induced inflammation by Nrf2 activation in vitro and improves survival of mice in LPS-induced endotoxemia model in vivo.

NF-E2-related factor 2 (Nrf2) has been demonstrated to be a key transcription factor regulating the anti-inflammatory genes including heme oxygenase-1 (HO-1) in experimental sepsis models. Based on the fact that 3,4,5-trihydorxycinnamic acid (THC) has been reported to possess anti-inflammatory properties in BV2 microglial cells, the possible effects of THC and its underlying mechanism was examined against lipopolysaccharide (LPS)-induced RAW 264.7 cell culture and septic mouse models. Pretreatment of RAW 264.7 cells with THC significantly attenuated LPS-induced NO, PGE2 production, and expression of iNOS and COX-2. THC also significantly suppressed LPS-induced release of pro-inflammatory cytokines and degradation of IκB-α. Increased phosphorylation of Nrf2 and nuclear translocation of Nrf2 were observed with THC treatment with consequent expression of HO-1. The data demonstrated that multiple signaling pathways including Akt, p38, and PKC are involved in the THC-induced activation of Nrf2/HO-1 pathway. Treatment of THC resulted in significantly increased survival of LPS-induced septic mice. THC also significantly ameliorated LPS-induced septic features such as hypothermia and increased vascular leakage. In accordance with the data from cell culture model, THC exhibited increased expression of HO-1 in kidney and decreased serum level of pro-inflammatory mediators such as TNF-α, IL-1ß, and NO. Taken together, the present study for the first time demonstrates that THC inhibits inflammation in LPS-induced RAW264.7 cells by Nrf2 activation and improves survival of mice in LPS-induced endotoxemia model.

Monitoring of antibiotic resistance in bacteria isolated from laboratory animals.

The drug resistance of microorganisms isolated from laboratory animals never treated with antibiotics is being reported consistently, while the number of laboratory animals used in medicine, pharmacy, veterinary medicine, agriculture, nutrition, and environmental and health science has increased rapidly in Korea. Therefore, this study examined the development of antimicrobial resistance in bacteria isolated from laboratory animals bred in Korea. A total of 443 isolates (7 species) containing 5 Sphingomonas paucimobilis, 206 Escherichia coli, 60 Staphylococcus aureus, 15 Staphylococcus epidermidis, 77 Enterococcus faecalis, 27 Citrobacter freundii, 35 Acinetobacter baumannii were collected from the nose, intestine, bronchus and reproductive organs of ICR mice and SD rats. Of these species, Acinetobacter baumannii and Enterococcus faecalis showed significant antimicrobial resistance according to the minimum inhibition concentration (MIC) in E-test. In case of Acinetobacter baumannii, several isolates showed MIC values 16-128 µg/mL for cefazolin and cefoxitin, and higher resistance (128-512 µg/mL) to nitrofurantoin than that of standard type. Resistance to cefazolin, cefoxitin and nitrofurantoin was detected in 17.14, 20.00, and 8.57% of the Acinetobacter baumannii isolates, respectively. In addition, 44.1% of the Enterococcus faecalis isolates collected from the laboratory animals were resistant to oxacillin concentration of 16-32 µg/mL range, while MIC value of standard type was below oxacillin concentration of 6 µg/mL. These results suggest that in rodent species of laboratory animals, Acinetobacter baumannii are resistance to cefazolin, cefoxitin and nitrofurantoin, whereas those of Enterococcus faecalis were resistance to oxacillin.

3,4,5-Trihydroxycinnamic Acid Inhibits LPS-Induced iNOS Expression by Suppressing NF-κB Activation in BV2 Microglial Cells.

Although various derivatives of caffeic acid have been reported to possess a wide variety of biological activities such as neuronal protection against excitotoxicity and anti-inflammatory property, the biological activity of 3,4,5-trihydroxycinnamic acid (THC), a derivative of hydroxycinnamic acids, has not been clearly examined. The objective of the present study is to evaluate the anti-inflammatory effects of THC on lipopolysaccharide (LPS)-stimulated BV2 microglial cells. THC significantly suppressed LPS-induced excessive production of nitric oxide (NO) and expression of iNOS, which is responsible for the production of iNOS. THC also suppressed LPS-induced overproduction of pro-inflammatory cytokines such as IL-1ß and TNF-α in BV2 microgilal cells. Furthermore, THC significantly suppressed LPS-induced degradation of IκB, which retains NF-κB in the cytoplasm. Therefore, THC attenuated nuclear translocation of NF-κB, a major pro-inflammatory transcription factor. Taken together, the present study for the first time demonstrates that THC exhibits anti-inflammatory activity through the suppression of NF-κB transcriptional activation in LPS-stimulated BV2 microglial cells.

Pen-2 overexpression induces Aß-42 production, memory defect, motor activity enhancement and feeding behavior dysfunction in NSE/Pen-2 transgenic mice.

Pen-2 is a key regulator of the γ-secretase complex, which is involved in the production of the amyloid ß (Aß)-42 peptides, which ultimately lead to Alzheimer's disease (AD). While Pen-2 has been studied in vitro, Pen-2 function in vivo in the brains of transgenic (Tg) mice overexpressing human Pen-2 (hPen-2) protein has not been studied. This study aimed to determine whether Pen-2 overexpression could regulate the AD-like phenotypes in Tg mice. NSE/hPen-2 Tg mice were produced by the microinjection of the NSE/hPen-2 gene into the pronucleus of fertilized eggs. The expression of the hPen-2 gene under the control of the NSE promoter was successfully detected only in the brain and kidney tissue of NSE/hPen-2 Tg mice. Also, 12-month-old NSE/hPen-2 Tg mice displayed behavioral dysfunction in the water maze test, motor activity and feeding behavior dysfunction in food intake/water intake/motor activity monitoring system. In addition, tissue samples displayed dense staining with antibody to the Aß-42 peptide. Furthermore, NSE/hPen-2 Tg mice exhibiting feeding behavior dysfunction were significantly more apt to display symptoms related to diabetes and obesity. These results suggest that Pen-2 overexpression in NSE/hPen-2 Tg mice may induce all the AD-like phenotypes, including behavioral deficits, motor activity and feeding behavior dysfunction, Aß-42 peptide deposition and chronic disease induction.

GATA binding protein 3 overexpression and suppression significantly contribute to the regulation of allergic skin inflammation.

GATA binding protein 3 (GATA3) is a key molecule regulating the balance in the ratio of type 1 helper T (Th1) cells to type 2 helper T (Th2) cells, which is thought to be indicative of the pathogenesis of allergic diseases such as asthma and atopic dermatitis. The aim of this study was to investigate the role of GATA3 in allergic skin inflammation. Transgenic (Tg) mice overexpressing human GATA3 (hGATA3) were produced by the microinjection of pCMV/hGATA3 constructs into fertilized mouse eggs. The hGATA3 gene was successfully expressed at the protein level in the lymph node and thymus of CMV/hGATA3 transfected cells and Tg mice. CMV/hGATA3 Tg mice showed a significant increase in the allergic skin inflammation response such as ear thickness, draining auricular lymph node (aLN) weight, epidermal thickness, inflammatory cell number and Th2 immunoglobulin (Ig) concentration compared to wild-type (WT) mice after phthalic anhydride (PA) treatment. Furthermore, the secretion of Th2 type cytokines was increased by PA treatment in CMV/hGATA3 Tg mice, while the secretion of Th1 type cytokine was suppressed under the same conditions. However, the increased levels of Th2 type cytokines in CMV/hGATA3 Tg mice were almost recovered by the down-regulation of GATA3 expression with D-pinitol treatment. Therefore, these findings suggest that GATA3 could be considered as a potential target regulating the mechanism responsible for the differences in allergic skin inflammation.

Aß-42 deposition significantly increases the insolubility of synaptophysin in the brains of hAPPsw/hPS2m double transgenic mice.

Synaptophysin is a synaptic vesicle glycoprotein involved in the regulation process for neurotransmitter release, which is distributed throughout neuroendocrine cells and all neurons in the brain and spinal cord. In an effort to determine whether amyloid ß (Aß)-42 peptides could influence the quantity and biochemical properties of synaptophysin, alterations in the levels of the synaptophysin protein in various soluble fractions were detected in the brains of four genotypes of transgenic mice (Tg) including Non-Tg, neuron-specific enolase (NSE)-hPS2m, NSE-hAPPsw and hAPPsw/hPS2m double Tg mice. Among the four genotypes of Tg mice, the highest levels of Aß-42 peptides were noted in hAPPsw/hPS2m, followed by NSE-hAPPsw, NSE-hPS2m and Non-Tg mice. In the brains of these mice displaying different levels of Aß-42 peptides, the levels of soluble synaptophysin were reduced significantly only in the hAPPsw/hPS2m double Tg mice compared to the Non-Tg mice. However, immunohistochemical analysis revealed no differences in the levels of total synaptophysin protein between the neocortex and hippocampus of the four different genotypes of mice. Western blot analysis using four-step fractions with differing solubility revealed a marked decrease in synaptophysin levels in the Tris-buffer saline fraction of hAPPsw/hPS2m double Tg mice and a significant increase in the formic acid fraction, relative to the Non-Tg mice. The results obtained from our in vivo experiments in mice are identical to the results observed in SK-N-MC cells treated with 100 nM Aß-42 peptides. Therefore, our experiments collectively suggest that Aß-42 peptides may alter the solubility without changing the total amount of synaptophysin.

Differential regulation of the biosynthesis of glucose transporters by the PI3-K and MAPK pathways of insulin signaling by treatment with novel compounds from Liriope platyphylla.

The insulin signaling pathway, involving protein kinase B (PKB) and mitogen-activated protein kinase (MAPK), mediates the biological response to insulin and several growth factors and cytokines. To investigate the correlation between glucose transporter (Glut) biosynthesis and the insulin signaling pathway activated by novel compounds of Liriope platyphylla (LP9M80-H), alterations in Glut and key protein expression in the insulin signaling pathway were analyzed in the liver and brain of ICR mice treated with LP9M80-H. An in vitro assay showed that the highest level of insulin concentration was observed in the LP9M80-H-treated group, followed by the LP-H, LP-M, LP-E, and LP9M80-C-treated groups. Therefore, LP9M80-H was selected for use in studying the detailed mechanism of the insulin signaling pathway in animal systems. In an in vivo experiment, LP9M80-H induced a significant increase in glucose levels and a decrease of insulin concentration in the blood of mice, while their body weight remained constant over 5 days. The expression level of Glut-3 was down-regulated in the liver, or maintained at the same level in the brain of LP9MH80-H-treated mice. These changes corresponded to the phosphorylation of the p38 protein rather than to ERK and JNK in the MAPK signaling pathway. In addition, the expression level of Glut-1 increased significantly after LP9MH80-H treatment of both insulin target tissues in mice. Western blot analysis showed that Akt in the PI3-K pathway mainly participated in Glut-1 biosynthesis. Thus, these results suggest the possibility that the LP9M80-H-induced regulation of Glut-1 and Glut-3 biosynthesis may be mediated by the Akt and p38 MAPK signaling of the insulin signaling pathway in the liver and brain of mice.

Identification of origin of unknown derivative chromosomes by array-based comparative genomic hybridization using pre- and postnatal clinical samples.

Microarray-based comparative genomic hybridization (array CGH) is a high-resolution and comprehensive method for detecting both genome-wide and chromosome-specific copy-number imbalance. We have developed an array CGH analysis system (consisting of an array CGH chip plus its exclusive analysis software) for constitutional genetic diagnosis and have evaluated the suitability of our system for molecular diagnosis using pre- and postnatal clinical samples. In a blind study, each of the 264 sample karyotypes identified by array CGH analysis was consistent with that identified by traditional karyotype analysis--with one exception, case (47, XXX)--and we were able to identify origins, such as small supernumerary marker chromosomes, which cannot be determined by conventional cytogenetics. We also acquired very accurate, fast and reliable results using a diminutive amount of clinical samples. Taken together, the array CGH platform developed in this study is a rapid, powerful and sensitive technology for pre- and postnatal diagnosis using a very small amount of clinical sample.

Fabrication of hydroxyapatite-poly(epsilon-caprolactone) scaffolds by a combination of the extrusion and bi-axial lamination processes.

Hydroxyapatite (HA)/poly(epsilon-caprolactone) (PCL) composite scaffolds were fabricated using a combination of the extrusion and bi-axial lamination processes. Firstly, HA/PCL composites with various HA contents (0, 50, 60, 70 wt%) were prepared by mixing the HA powders and the molten PCL at 100 degrees C and then extruded through an orifice with dimensions of 600 x 600 microm to produce HA/PCL composite fibers. Isobutyl methacrylate (IBMA) polymer fiber was also prepared in a similar manner for use as a fugitive material. The 3-D scaffold was then produced by the bi-axial lamination of the HA/PCL and IBMA fibers, followed by solvent leaching to remove the IBMA. It was observed that the HA/PCL composites had a superior elastic modulus and biological properties, as compared to the pure PCL. The fabricated HA/PCL scaffold showed a controlled pore structure (porosity of approximately 49% and pore size of approximately 512 microm) and excellent welding between the HA/PCL fibers, as well as a high compressive strength of approximately 7.8 MPa.

Macrochanneled poly (epsilon-caprolactone)/ hydroxyapatite scaffold by combination of bi-axial machining and lamination.

A combination of bi-axial machining and lamination was used to fabricate macrochanneled poly (epsilon-caprolactone) (PCL)/hydroxyapatite (HA) scaffolds. Thermoplastic PCL/HA sheets with a thickness of 1 mm, consisting of a 40 wt% PCL polymer and 60 wt% HA particles, were bi-axially machined. The thermoplastic PCL/HA exhibited an excellent surface finish with negligible tearing of the PCL polymer and pull-out of the HA particles. The bi-axially machined sheets were laminated with a solvent to give permanent bonding between the lamina. This novel process produced three-directionally connected macrochannels in the dense PCL/HA body. The macrochanneled PCL/HA scaffold exhibited excellent ductility and reasonably high strength. In addition, good cellular responses were observed due to the osteoconductive HA particles.

Hydroxyapatite (HA) bone scaffolds with controlled macrochannel pores.

Hydroxyapatite (HA) macrochanneled porous scaffolds, with a controlled pore structure, were fabricated via a combination of the extrusion and lamination processes. The scaffold was architectured by aligning and laminating the extruded HA and carbon filaments. The macrochannel pores were formed by removing the carbon filaments after thermal treatments (binder removal and sintering). The porosity of the scaffolds was varied between 48 and 73% with a controlled pore size of approximately 450 microm, by adjusting the fractions of HA and carbon filaments. As the porosity was increased from 48 to 73%, the compressive strength decreased from 11.5 to 3.2 MPa. However, the osteoblast-like cell responses on the scaffold, such as the proliferation rate and alkaline phosphatase (ALP) activity, were significantly enhanced as the porosity was increased.

Improvement in biocompatibility of ZrO2-Al2O3 nano-composite by addition of HA.

The biocompatibility of zirconia-alumina (ZA) nano-composites in load-bearing applications such as dental/orthopedic implants was significantly enhanced by the addition of bioactive HA. The ZA matrix was composed of nano-composite powder obtained from the Pechini process and had higher flexural strength than conventionally mixed zirconia-alumina composite. Because the ZA nano-composite powder effectively decreased the contact area between HA and zirconia for their reaction during the sintering process, the HA-added ZA nano-composites contained biphasic calcium phosphates (BCP) of HA/TCP and had higher flexural strength than conventionally mixed ZA-HA composite. From the in vitro test with osteoblastic cell-lines, the proliferation and the differentiation (as expressed by the alkaline phosphatase activity) of the cellular response on the HA-added ZA nano-composites gradually increased as the amount of HA added increased. From the mechanical and biological evaluations of the HA-added ZA nano-composites, 30HA (30 vol% HA + 70 vol% ZA) was found to be the optimal composition for load-bearing biological applications.

Stability and cellular responses to fluorapatite-collagen composites.

Fluorapatite (FA)-collagen composites were synthesized via a biomimetic coprecipitation method in order to improve the structural stability and cellular responses. Different amounts of ammonium fluoride (NH4F), acting as a fluorine source for FA, were added to the precipitation of the composites. The precipitated composites were freeze-dried and isostatically pressed in a dense body. The added fluorine was incorporated nearly fully into the apatite structure (fluoridation), and a near stoichiometric FA-collagen composite was obtained with complete fluoridation. The freeze-dried composites had a typical biomimetic network, consisting of collagen fibers and precipitates of nano-sized apatite crystals. The human osteoblast-like cells on the FA-collagen composites exhibited significantly higher proliferation and differentiation (according to alkaline phosphatase activity) than those on the hydroxyapatite-collagen composite. These enhanced osteoblastic cell responses were attributed to the fluorine release and the reduced dissolution rate.

Sol-gel derived fluor-hydroxyapatite biocoatings on zirconia substrate.

Fluor-hydroxyapatite (FHA) film was coated on a zirconia (ZrO(2)) substrate by a sol-gel method. An appropriate amount of F ions was incorporated into the hydroxyapatite (HA) during the preparation of the sols. The apatite phase began to crystallize after heat treatment at 400 degrees C, and increased in intensity above 500 degrees C. No decomposition was detected by X-ray diffraction analyses up to 800 degrees C, which illustrates the high thermal stability of the FHA films. The films showed a uniform and dense morphology with a thickness of approximately 1 microm after a precisely controlled heat treatment process. These FHA films adhered firmly to the zirconia substrate, representing notable adhesion strengths of approximately 70 MPa after heat treatment above 500 degrees C. The dissolution rate of the FHA coating layer varied according to the heat treatment temperature, which was closely related to the film crystallinity. The dissolution rate of the FHA film was lower than that of the HA film, suggesting the possibility of a functional gradient coating of HA and FHA. The MG63 cells seeded onto the FHA films proliferated in a similar manner to those seeded onto pure HA ceramic and a plastic control.

Porous ZrO2 bone scaffold coated with hydroxyapatite with fluorapatite intermediate layer.

Highly porous zirconia (ZrO(2)) bone scaffolds, fabricated by a replication technique using polymeric sponge, were coated with hydroxyapatite (HA). To prevent the chemical reactions between ZrO(2) and HA, an intermediate fluorapatite (FA) layer was introduced. The strength of the porous ZrO(2) was higher than that of pure HA by a factor of 7, suggesting the feasibility of ZrO(2) porous scaffolds as load-bearing part applications. The coated HA/FA layer, with a thickness of about 30 microm, was firmly adhered to the ZrO(2) body with a bonding strength of 22MPa. The osteoblast-like cells were attached and spread well on the coating layer throughout the porous scaffolds. The alkaline phosphatase activity of the proliferated cells on the HA/FA coated ZrO(2) was comparable to that on pure HA and higher than that on pure ZrO(2).