Improved Biocompatibility of Intra-Arterial Poly-L-Lactic Acid Stent by Tantalum Ion Implantation : 3-Month Results in a Swine Model.

OBJECTIVE: Biodegradable poly-L-lactic acid (PLLA) with a highly biocompatible surface via tantalum (Ta) ion implantation can be an innovative solution for the problems associated with current biodegradable stents. The purpose of this study is to develop a Ta-implanted PLLA stent for clinical use and to investigate its biological performance capabilities. METHODS: A series of in vitro and in vivo tests were used to assess the biological performance of bare and Ta-implanted PLLA stents. The re-endothelialization ability and thrombogenicity were examined through in vitro endothelial cell and platelet adhesion tests. An in vivo swine model was used to evaluate the effects of Ta ion implantation on subacute restenosis and thrombosis. Angiographic and histologic evaluations were conducted at one, two and three months post-treatment. RESULTS: The Ta-implanted PLLA stent was successfully fabricated, exhibiting a smooth surface morphology and modified layer integration. After Ta ion implantation, the surface properties were more favorable for rapid endothelialization and for less platelet attachment compared to the bare PLLA stent. In an in vivo animal test, follow-up angiography showed no evidence of in-stent stenosis in either group. In a microscopic histologic examination, luminal thrombus formation was significantly suppressed in the Ta-implanted PLLA stent group according to the 2-month follow-up assessment (21.2% vs. 63.9%, p=0.005). Cells positive for CD 68, a marker for the monocyte lineage, were less frequently identified around the Ta-implanted PLLA stent in the 1-month follow-up assessments. CONCLUSION: The use of a Ta-implanted PLLA stent appears to promote re-endothelialization and anti-thrombogenicity.

Graphene Oxide-Supported Microwell Grids for Preparing Cryo-EM Samples with Controlled Ice Thickness.

Cryogenic-electron microscopy (cryo-EM) is the preferred method to determine 3D structures of proteins and to study diverse material systems that intrinsically have radiation or air sensitivity. Current cryo-EM sample preparation methods provide limited control over the sample quality, which limits the efficiency and high throughput of 3D structure analysis. This is partly because it is difficult to control the thickness of the vitreous ice that embeds specimens, in the range of nanoscale, depending on the size and type of materials of interest. Thus, there is a need for fine regulation of the thickness of vitreous ice to deliver consistent high signal-to-noise ratios for low-contrast biological specimens. Herein, an advanced silicon-chip-based device is developed which has a regular array of micropatterned holes with a graphene oxide (GO) window on freestanding silicon nitride (Six Ny ). Accurately regulated depths of micropatterned holes enable precise control of vitreous ice thickness. Furthermore, GO window with affinity for biomolecules can facilitate concentration of the sample molecules at a higher level. Incorporation of micropatterned chips with a GO window enhances cryo-EM imaging for various nanoscale biological samples including human immunodeficiency viral particles, groEL tetradecamers, apoferritin octahedral, aldolase homotetramer complexes, and tau filaments, as well as inorganic materials.

The impact of surface treatment in 3-dimensional printed implants for early osseointegration: a comparison study of three different surfaces.

3D printing technology has been gradually applied to various areas. In the present study, 3D-printed implants were fabricated with direct metal laser sintering technique for a dental single root with titanium. The 3D implants were allocated into following groups: not treated (3D-None), sandblasted with a large grit and acid-etched (3D-SLA), and target-ion-induced plasma-sputtered surface (3D-TIPS). Two holes were drilled in each tibia of rabbit, and the three groups of implants were randomly placed with a mallet. Rabbits were sacrificed at two, four, and twelve weeks after the surgery. Histologic and histomorphometric analyses were performed for the evaluation of mineralized bone-to-implant contact (mBIC), osteoid-to-implant contact (OIC), total bone-to-implant contact (tBIC), mineralized bone area fraction occupancy (mBAFO), osteoid area fraction occupancy (OAFO), and total bone area fraction occupancy (tBAFO) in the inner and outer areas of lattice structure. At two weeks, 3D-TIPS showed significantly higher inner and outer tBIC and inner tBAFO compared with other groups. At four weeks, 3D-TIPS showed significantly higher outer OIC than 3D-SLA, but there were no significant differences in other variables. At twelve weeks, there were no significant differences. The surface treatment with TIPS in 3D-printed implants could enhance the osseointegration process in the rabbit tibia model, meaning that earlier osseointegration could be achieved.

Biodegradable magnesium alloy (WE43) in bone-fixation plate and screw.

The purpose of the present study was to evaluate the mechanical strength and the absorption rate of WE43 material and to develop an absorbable metallic plate and screw for craniofacial application. The extruded WE43 plate and screw were evaluated using a LeFort I osteotomy canine model of 10 beagle dogs. Animals were divided into two groups: five dogs in the experimental group and five dogs in the control group. µCT was acquired at 4, 12, and 24 weeks. At 24 weeks after the operation, all animals were sacrificed, and histologic evaluation was performed. Swelling and gas formation were observed in three dogs in the experimental groups at 8 weeks. From 12 weeks, infraorbital fistula and inflammation were observed in three dogs in the experimental group, which gradually decreased and disappeared at 24 weeks. Other two dogs showed less gas formation at 12 weeks. The plates were completely absorbed, and gas formation was not observed at 24 weeks in these two dogs. New bone was well formed around the plates and screws in both groups. Histologic examination showed no specific differences between two groups. The mechanical strength of extruded WE43 was sufficient for mid-facial application. Plates and screws made with appropriately treated WE43 have the potential to be useful clinically.

Comparison of the Pain-relieving Effects of Human Milk, Sucrose, and Distilled Water during Examinations for Retinopathy of Prematurity: A Randomized Controlled Trial.

PURPOSE: This study compared the pain-relieving effects of human milk, sucrose, and distilled water during examinations for retinopathy of prematurity. METHODS: Forty-five preterm infants were randomly assigned to receive a pacifier dipped in one of three solutions: human milk (n=14), 24% sucrose (n=15), or distilled water (n=16), 2 minutes before an eye examination. Their pain score, pulse rate, and oxygen saturation were measured at three time points: 5 minutes before the examination, 30 seconds after speculum introduction, and 2 minutes after the examination. RESULTS: The infants' mean gestational age and weight at birth were 33.1±2.1 weeks and 1,842±470 g, respectively. There were no between-group differences in pain relief during the eye examination. The pain score significantly increased both during (p<.001) and after the examinations (p=.003). Oxygen saturation decreased during the examinations (p<.001); however, the infants in the 24% sucrose group showed higher oxygen saturation (p=.047) during the examinations than the infants in the other groups. CONCLUSION: Sucking on a pacifier dipped in human milk or 24% sucrose did not reduce the pain associated with eye examinations in preterm infants. Pacifiers dipped in sucrose can be used to maintain better oxygen saturation during these examinations.

Hydroxyapatite Microspheres as an Additive to Enhance Radiopacity, Biocompatibility, and Osteoconductivity of Poly(methyl methacrylate) Bone Cement.

This study demonstrates the utility of hydroxyapatite (HA) microspheres as an additive to enhance the radiopaque properties, biocompatibility, and osteoconductivity of poly(methyl methacrylate) (PMMA)-based bone cements. HA microspheres were synthesized using spray drying. They had well-defined spherical shapes, thus allowing for the production of PMMA/HA composites with a very high HA content (20 vol % and 40 vol %). The uniform distribution of these HA microspheres in the PMMA matrix resulted in a remarkable increase in compressive modulus (p < 0.05), while preserving a reasonably high compressive strength. The PMMA/HA bone cements showed much higher radiopacity than PMMA containing BaSO4 as the additive. This was attributed to the high HA content up to 40 vol %. In addition, the biocompatibility and osteoconductivity of PMMA/HA bone cements were significantly enhanced compared to those of PMMA bone cements containing BaSO4, which were assessed using in vitro tests and in vivo animal experiments.

Clinical significance of overexpression of NRG1 and its receptors, HER3 and HER4, in gastric cancer patients.

BACKGROUND: Neuregulin 1 (NRG1), a ligand for human epidermal growth factor (HER) 3 and HER4, can activates cell signaling pathways to promote carcinogenesis and metastasis. METHODS: To investigate the clinicopathologic significance of NRG1 and its receptors, immunohistochemistry was performed for NRG1, HER3, and HER4 in 502 consecutive gastric cancers (GCs). Furthermore, HER2, microsatellite instability (MSI), and Epstein-Barr virus (EBV) status were investigated. NRG1 gene copy number (GCN) was determined by dual-color fluorescence in situ hybridization (FISH) in 388 available GCs. RESULTS: NRG1 overexpression was observed in 141 (28.1%) GCs and closely correlated with HER3 (P = 0.034) and HER4 (P < 0.001) expression. NRG1 overexpression was significantly associated with aggressive features, including infiltrative tumor growth, lymphovascular, and neural invasion, high pathologic stage, and poor prognosis (all P < 0.05), but not associated with EBV, MSI, or HER2 status. Multivariate analysis identified NRG1 overexpression as an independent prognostic factor for survival (P = 0.040). HER3 and HER4 expressions were observed in 157 (31.3%) and 277 (55.2%), respectively. In contrast to NRG1, expression of these proteins was not associated with survival. NRG1 GCN gain (GCN ≥ 2.5) was detected in 14.7% patients, including two cases of amplification, and was moderately correlated with NRG1 overexpression (κ, 0.459; P < 0.001). CONCLUSIONS: Although our results indicate a lack of prognostic significance of HER3 and HER4 overexpression in GC, overexpression of their ligand, NRG1, was associated with aggressive clinical features and represented an independent unfavorable prognostic factor. Therefore, NRG1 is a potential prognostic and therapeutic biomarker in GC patients.

Biocompatibility and Biocorrosion of Hydroxyapatite-Coated Magnesium Plate: Animal Experiment.

Magnesium (Mg) has the advantage of being resorbed in vivo, but its resorption rate is difficult to control. With uncontrolled resorption, Magnesium as a bone fixation material has minimal clinical value. During resorption not only is the strength rapidly weakened, but rapid formation of metabolite also occurs. In order to overcome these disadvantages, hydroxyapatite (HA) surface coating of pure magnesium plate was attempted in this study. Magnesium plates were inserted above the frontal bone of Sprague-Dawley rats in both the control group (Bare-Mg group) and the experimental group (HA-Mg group). The presence of inflammation, infection, hydrogen gas formation, wound dehiscence, and/or plate exposure was observed, blood tests were performed, and the resorption rate and tensile strength of the retrieved metal plates were measured. The HA-Mg group showed no gas formation or plate exposure until week 12. However, the Bare-Mg group showed consistent gas formation and plate exposure beginning in week 2. WBC (White Blood Cell), BUN (Blood Urea Nitrogen), Creatinine, and serum magnesium concentration levels were within normal range in both groups. AST (Aspartate Aminotransferase) and ALT (Alanine Aminotransferase) values, however, were above normal range in some animals of both groups. The HA-Mg group showed statistically significant advantage in resistance to degradation compared to the Bare-Mg group in weeks 2, 4, 6, 8, and 12. Degradation of HA-Mg plates proceeded after week 12. Coating magnesium plates with hydroxyapatite may be a viable method to maintain their strength long enough to allow bony healing and to control the resorption rate during the initial period.

Innovative micro-textured hydroxyapatite and poly(l-lactic)-acid polymer composite film as a flexible, corrosion resistant, biocompatible, and bioactive coating for Mg implants.

The utility of a novel ceramic/polymer-composite coating with a micro-textured microstructure that would significantly enhance the functions of biodegradable Mg implants is demonstrated here. To accomplish this, bioactive hydroxyapatite (HA) micro-dots can be created by immersing a Mg implant with a micro-patterned photoresist surface in an aqueous solution containing calcium and phosphate ions. The HA micro-dots can then be surrounded by a flexible poly(l-lactic)-acid (PLLA) polymer using spin coating to form a HA/PLLA micro-textured coating layer. The HA/PLLA micro-textured coating layer showed an excellent corrosion resistance when it was immersed in a simulated body fluid (SBF) solution and good biocompatibility, which was assessed by in vitro cell tests. In addition, the HA/PLLA micro-textured coating layer had high deformation ability, where no apparent changes in the coating layer were observed even after a 5% elongation, which would be unobtainable using HA and PLLA coating layers; furthermore, this allowed the mechanically-strained Mg implant with the HA/PLLA micro-textured coating layer to preserve its excellent corrosion resistance and biocompatibility in vitro.

The Bioresorption and Guided Bone Regeneration of Absorbable Hydroxyapatite-Coated Magnesium Mesh.

INTRODUCTION: Nonabsorbable metallic membrane for guided bone regeneration is remained permanently even though after complete healing. There would be metallic exposure followed by the risk of infection; the membrane should be removed for the additional procedure such as implant installation. Since absorbable nonmetallic mesh is absorbed within 3 to 6 months, it is unnecessary to be removed. However, the absorbable membrane shows lower retention, lower mechanical strength, and difficulty of manipulation than the nonabsorbable ones.The purpose of this study is to evaluate the ability of absorbable metallic mesh (hydroxyapatite-coated magnesium mesh) with acceptable mechanical properties and satisfying biocompatibility. METHODS: The bioresorption and fate of magnesium were evaluated in Sprague Dawley rat (SD rat) with critical defect of calvarium. The critical defect with a diameter of 8 mm was made on calvarium using trephine bur in 18 SD rats. The defected models were divided into 2 groups: the control group (9 SD rat) without mesh and the experimental group (9 SD rat) with the insertion of prototype HA-coated magnesium mesh. The 3 SD rats were sacrificed at 6, 12, and 18 weeks. The histopathological and radiographic examinations were performed afterward. RESULTS: In the control group, there was no specific symptom. The experimental group also showed no specific symptom including swelling and dehiscence related to hydrogen gas formation. From 6 to 18 weeks, the experimental group showed the progressive absorption and fracture of magnesium mesh. However, there was no specific effectiveness of guided bone regeneration in both groups. There was no significant difference in bone volume, bone surface, and bone volume fraction between the negative control group and the group with magnesium mesh (P >0.05). CONCLUSION: Hydroxyapatite-coated magnesium mesh showed reasonable process of bioresorption and bony reaction; however, the effectiveness of guided bone regeneration and management of the bioresorption rate should be reconsidered.

Radiological, histological, and hematological evaluation of hydroxyapatite-coated resorbable magnesium alloy screws placed in rabbit tibia.

Titanium (Ti) screw has excellent mechanical property, and osseointegration capacity. However, they require surgery for removal. In contrast, polymer screws are resorbable, but they have poor mechanical properties. In this research, magnesium alloy screws (WE43: Mg-Y-Nd-Zr) that have advantages of titanium and polymer were manufactured. In addition, to increase biocompatibility and control degradation rate, the Mg alloy was coated with hydroxyapatite (HA). Torsion test and corrosion test were performed in vitro. For clinical, radiological and histological evaluation, on the eight rabbits, two HA-coated screws were installed in left tibia, and two noncoated screws were installed in right tibia. Each four rabbits were sacrificed 6 and 12 weeks postoperatively. For hematological evaluation, the same type of screws were installed on both legs. Complete blood count (CBC), Mg2+ concentrate were sampled from the ear central artery on the operation day for a control point, and at 1, 2, 4, 6, 8, and 12 weeks. Mg alloy screws have no differences of biocompatibility according to the HA coating. However, resorption of screw was slower in case of the HA coating. The hematological problem related releasing of Mg was not found. The results suggest that Mg alloy screws have feasibility for clinical application. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1636-1644, 2017.

Hydroxyapatite-coated magnesium implants with improved in vitro and in vivo biocorrosion, biocompatibility, and bone response.

Magnesium and its alloys are candidate materials for biodegradable implants; however, excessively rapid corrosion behavior restricts their practical uses in biological systems. For such applications, surface modification is essential, and the use of anticorrosion coatings is considered as a promising avenue. In this study, we coated Mg with hydroxyapatite (HA) in an aqueous solution containing calcium and phosphate sources to improve its in vitro and in vivo biocorrosion resistance, biocompatibility and bone response. A layer of needle-shaped HA crystals was created uniformly on the Mg substrate even when the Mg sample had a complex shape of a screw. In addition, a dense HA-stratum between this layer and the Mg substrate was formed. This HA-coating layer remarkably reduced the corrosion rate of the Mg tested in a simulated body fluid. Moreover, the biological response, including cell attachment, proliferation and differentiation, of the HA-coated samples was enhanced considerably compared to samples without a coating layer. The preliminary in vivo experiments also showed that the biocorrosion of the Mg implant was significantly retarded by HA coating, which resulted in good mechanical stability. In addition, in the case of the HA-coated implants, biodegradation was mitigated, particularly over the first 6 weeks of implantation. This considerably promoted bone growth at the interface between the implant and bone. These results confirmed that HA-coated Mg is a promising material for biomedical implant applications.

Toxicity generated through inhibition of pyruvate carboxylase and carnitine palmitoyl transferase-1 is similar to high glucose/palmitate-induced glucolipotoxicity in INS-1 beta cells.

This work was initiated to determine whether toxicity generated through inhibition of mitochondrial fuel metabolism is similar to high glucose/palmitate (HG/PA)-induced glucolipotoxicity. Influx of glucose and free fatty acids into the tricarboxylic acid (TCA) cycle was inhibited by treatment with the pyruvate carboxylase (PC) inhibitor phenylacetic acid (PAA) and carnitine palmitoyl transferase-1 (CPT-1) inhibitor etomoxir (Eto), or knockdown of PC and CPT-1. Treatment of PAA/Eto or knockdown of PC/CPT-1 induced apoptotic death in INS-1 beta cells. Similar to HG/PA treatment, PAA/Eto increased endoplasmic reticulum stress responses but decreased the Akt signal. JNK inhibitor or chemical chaperone was protective against both PAA/Eto- and HG/PA-induced cell death. All attempts to reduce [Ca²âº](i), stimulate lipid metabolism, and increase the TCA cycle intermediate pool protected PAA/Eto-induced death as well as HG/PA-induced death. These data suggest that signals induced from impaired mitochondrial fuel metabolism play a critical role in HG/PA-induced glucolipotoxicity.

Use of a poly(ether imide) coating to improve corrosion resistance and biocompatibility of magnesium (Mg) implant for orthopedic applications.

This study investigated the utility of poly(ether imide) (PEI) coating for improving the corrosion resistance and biocompatibility of magnesium (Mg) implants for orthopedic application. In particular, the microstructure of the PEI coating layers was controlled by the adjustment of the temperature used to dry the spin-coated wet PEI films. When a wet PEI film was dried at 4°C, a relatively thick and porous coating layer was achieved as a result of an extensive exchange of the solvent with water in a moist environment. In contrast, when a wet PEI film was dried at 70°C, a relatively thin and dense layer was created due to the faster evaporation of the solvent with a negligible exchange of the solvent with water. The porous PEI coating layer showed higher stability than did the dense one when immersed in a simulated body fluid (SBF), which was presumably attributed to the formation of chemical bonding between the PEI and the Mg substrate. Both the porous and the dense PEI coated Mg specimens showed significantly improved in vitro biocompatibility, which were assessed in terms of cell attachment, proliferation and differentiation. However, interestingly, the dense PEI coating layer showed greater cell proliferation and differentiation than did the porous layer. .

Involvement of the TLR4 (Toll-like receptor4) signaling pathway in palmitate-induced INS-1 beta cell death.

Fatty acid-induced cytotoxicity is believed to recapitulate lipotoxicity seen in obese type-2 diabetes, and, thus, contribute to beta cell loss in the disease. These studies were initiated to determine whether the Toll-like receptor (TLR) signaling pathway was involved in palmitate-induced beta cell death. Treatment of INS-1 beta cells with palmitate enhanced interaction between TLR and myeloid differentiation factor88 (MyD88). Concomitant with TLR/MyD88 interaction, the level of phospho-C-Jun N-terminal kinase (phospho-JNK) showed an increase; however, the level of inhibitory factor kappa B alpha (IκBα) showed a decrease. Gene knockdown of TLR4 prevented palmitate-induced INS-1 cell death, while knockdown of TLR2 did not. In addition, gene knockdown of TLR4 prevented palmitate-induced increase of phospho-JNK and decrease of IκBα. JNK inhibitor SP60125 significantly protected against palmitate-induced INS-1 cell death, while IκB kinase (IKK) inhibitor acetylsalicylate did not. These data suggest involvement of JNK activation through the TLR4 signaling pathway in palmitate-induced INS-1 beta cell death.

Protective role of autophagy in palmitate-induced INS-1 beta-cell death.

Autophagy, a vacuolar degradative pathway, constitutes a stress adaptation that avoids cell death or elicits the alternative cell-death pathway. This study was undertaken to determine whether autophagy is activated in palmitate (PA)-treated beta-cells and, if activated, what the role of autophagy is in the PA-induced beta-cell death. The enhanced formation of autophagosomes and autolysosomes was observed by exposure of INS-1 beta-cells to 400 microm PA in the presence of 25 mm glucose for 12 h. The formation of green fluorescent protein-LC3-labeled structures (green fluorescent protein-LC3 dots), with the conversion from LC3-I to LC3-II, was also distinct in the PA-treated cells. The phospho-mammalian target of rapamycin level, a typical signal pathway that inhibits activation of autophagy, was gradually decreased by PA treatment. Blockage of the mammalian target of rapamycin signaling pathway by treatment with rapamycin augmented the formation of autophagosomes but reduced PA-induced INS-1 cell death. In contrast, reduction of autophagosome formation by knocking down the ATG5, inhibition of fusion between autophagosome and lysosome by treatment with bafilomycin A1, or inhibition of proteolytic degradation by treatment with E64d/pepstatin A, significantly augmented PA-induced INS-1 cell death. These findings showed that the autophagy system could be activated in PA-treated INS-1 beta-cells, and suggested that the induction of autophagy might play an adaptive and protective role in PA-induced cell death.