Gene expression in human adipose-derived stem cells: comparison of 2D films, 3D electrospun meshes or co-cultured scaffolds with two-way paracrine effects.

Finding the appropriate cues to trigger the desired differentiation is a challenge in tissue engineering when stem cells are involved. In this regard, three-dimensional environments are often compared to cells' two-dimensional culture behaviour (plastic culture dish). Here, we compared the gene expression pattern of human adipose-derived stem cells (ASC) seeded in a three-dimensional (3D) electrospun mesh and on a two-dimensional (2D) film - both of exactly the same material. Additionally, we conducted experiments with a scaffold floating above a film to investigate two-way paracrine effects (co-system). Electrospun meshes (3D scaffolds) and films (2D), consisting either of pristine poly-lactic-co-glycolic acid (PLGA) or of PLGA-containing dispersed amorphous calcium phosphate nanoparticles (PLGA/aCaP), were seeded with ASCs and cultured either in Dulbecco Minimum Essential Medium (DMEM) or in osteogenic medium. After two weeks, minimum stem cell criteria markers as well as typical markers for osteogenesis, endothelial cell differentiation, adipogenesis and chondrogenesis were analysed by quantitative real-time PCR. Interestingly, mostly osteogenic genes of cells seeded on 3D meshes were upregulated compared to those on 2D films, while stem cell markers seemed to be only slightly affected. Runx2 and osteocalcin showed an especially strong upregulation under all conditions, while most other factors analysed for 2D/3D changes were highly dependent on the material composition, the culture medium and on paracrine signalling effects. The beneficial 3D environment for stem cells found in many studies has therefore not to be attributed to the third dimension alone and should carefully be compared to 2D films fabricated of the same material. Furthermore, paracrine interactions triggering differentiation are not negligible.

Industrial applications of nanoparticles.

Research efforts in the past two decades have resulted in thousands of potential application areas for nanoparticles - which materials have become industrially relevant? Where are sustainable applications of nanoparticles replacing traditional processing and materials? This tutorial review starts with a brief analysis on what makes nanoparticles attractive to chemical product design. The article highlights established industrial applications of nanoparticles and then moves to rapidly emerging applications in the chemical industry and discusses future research directions. Contributions from large companies, academia and high-tech start-ups are used to elucidate where academic nanoparticle research has revolutionized industry practice. A nanomaterial-focused analysis discusses new trends, such as particles with an identity, and the influence of modern instrument advances in the development of novel industrial products.

Insight into the beneficial immunomodulatory mechanism of the sevoflurane metabolite hexafluoro-2-propanol in a rat model of endotoxaemia.

Volatile anaesthetics such as sevoflurane attenuate inflammatory processes, thereby impacting patient outcome significantly. Their inhalative administration is, however, strictly limited to controlled environments such as operating theatres, and thus an intravenously injectable immunomodulatory drug would offer distinct advantages. As protective effects of volatile anaesthetics have been associated with the presence of trifluorinated carbon groups in their basic structure, in this study we investigated the water-soluble sevoflurane metabolite hexafluoro-2-propanol (HFIP) as a potential immunomodulatory drug in a rat model of endotoxic shock. Male Wistar rats were subjected to intravenous lipopolysaccharide (LPS) and thereafter were treated with HFIP. Plasma and tissue inflammatory mediators, neutrophil invasion, tissue damage and haemodynamic stability were the dedicated end-points. In an endotoxin-induced endothelial cell injury model, underlying mechanisms were elucidated using gene expression and gene reporter analyses. HFIP reduced the systemic inflammatory response significantly and decreased endotoxin-induced tissue damage. Additionally, the LPS-provoked drop in blood pressure of animals was resolved by HFIP treatment. Pathway analysis revealed that the observed attenuation of the inflammatory process was associated with reduced nuclear factor kappa B (NF-κΒ) activation and suppression of its dependent transcripts. Taken together, intravenous administration of HFIP exerts promising immunomodulatory effects in endotoxaemic rats. The possibility of intravenous administration would overcome limitations of volatile anaesthetics, and thus HFIP might therefore represent an interesting future drug candidate for states of severe inflammation.

Magnetic separation-based blood purification: a promising new approach for the removal of disease-causing compounds?

Recent studies report promising results regarding extracorporeal magnetic separation-based blood purification for the rapid and selective removal of disease-causing compounds from whole blood. High molecular weight compounds, bacteria and cells can be eliminated from blood within minutes, hence offering novel treatment strategies for the management of intoxications and blood stream infections. However, risks associated with incomplete particle separation and the biological consequences of particles entering circulation remain largely unclear. This article discusses the promising future of magnetic separation-based purification while keeping important safety considerations in mind.

ß-D-glucosidase assisted gold dissolution as non-optical and quantifiable detection technique for immunoassays.

Immunoassays are used for detecting protein targets for various applications. Here, a modification of immunoassays to allow a purely electrical detection of the target protein concentration is shown. The modification comprises a ß-D-glucosidase as reporter enzyme and a cyanogenic glycoside as substrate. The enzymatic reaction produces cyanide in small quantities. For electrical detection of the cyanide, a novel sensor is developed, based on a gold micro wire. The cyanide dissolves the gold wire and changes the electrical resistance of the wire. Monitoring the resistance change allows a quantitative measurement of the target human C-reactive protein (an inflammatory marker) in blood plasma in the physiological relevant concentration range.

One-step large scale gas phase synthesis of Mn(2 + ) doped ZnS nanoparticles in reducing flames.

Metal sulfide nanoparticles have attracted considerable interest because of their unique semiconducting and electronic properties. In order to prepare these fascinating materials at an industrial scale, however, solvent-free, dry processes would be most advantageous. In the present work, we demonstrate how traditional oxide nanoparticle synthesis in flames can be extended to sulfides if we apply a careful control on flame gas composition and sulfur content. The ultra-fast (<1 ms) gas phase kinetics at elevated temperatures allow direct sulfidization of metals in flames ([Formula: see text]). As a representative example, we prepared air-stable Mn(2 + ) doped zinc sulfide nanoparticles. Post-sintering of the initially polycrystalline nanopowder resulted in a material of high crystallinity and improved photoluminescence. An analysis of the thermodynamics, gas composition, and kinetics in these reducing flames indicates that the here-presented extension of flame synthesis provides access to a broad range of metal sulfide nanoparticles and offers an alternative to non-oxide phosphor preparation.

Radio-opaque nanosized bioactive glass for potential root canal application: evaluation of radiopacity, bioactivity and alkaline capacity.

AIM: To produce novel nanosized bioactive glass particles with radio-opaque properties and high alkaline capacity and to evaluate their performance as a potential bioactive root canal dressing or filling material. METHODOLOGY: Flame spray-derived bioactive glass particles in the nanometre range were produced including bismuth oxide as a radiopacifier. Calcium hydroxide, barium sulphate and bismuth oxide served as controls. Corresponding materials were compressed to obtain dense specimens with increased alkaline capacity. Radiopacity was evaluated, and in vitro bioactivity was monitored using Raman spectroscopy and scanning electron microscopy. Leaching of bismuth was controlled using atomic absorption spectroscopy. RESULTS: Bioactive glass particles with up to 50 wt% bismuth oxide revealed radiopacity with an equivalent of 4.94-mm aluminium. The introduction of bismuth into the bioactive glass altered the alkaline capacity and the in vitro bioactivity only for high bismuth oxide quantities. Bismuth oxide leaching out of the glass matrix was hardly detectable. CONCLUSION: Bioactive glass can be modified with bismuth oxide to become radio-opaque.

Composites made of flame-sprayed bioactive glass 45S5 and polymers: bioactivity and immediate sealing properties.

AIM: To engineer systems using polyisoprene (PI) or polycaprolactone (PCL) and nanometric bioactive glass 45S5 (BG) that could create a hydroxyapatite interface and thus ultimately make the use of an endodontic sealer unnecessary. METHODOLOGY: Different composites using PI or PCL as matrix material were prepared with BG contents of up to 30 wt%. Unfilled PI and PCL, commercially available filled PI (Obtura gutta-percha) and PCL pellets (Resilon) served as control materials. Bioactivity (in vitro precipitate formation in simulated body fluid) was investigated using scanning electron microscopy and X-ray diffraction analysis. To test immediate sealing ability, simulated root canals were filled with heated materials, and dye leakage was assessed. Leakage was statistically compared between groups using Kruskal-Wallis analysis of variance followed by Mann-Whitney U tests and Bonferroni correction. The alpha-type error was set at 0.05. RESULTS: Both composite systems revealed hydroxyapatite formation on their surface. This was not observed on control materials. Incorporating 30 wt% BG into PI and PCL significantly (P < 0.05) improved their immediate sealing ability compared to that of unfilled polymers, so that dye leakage in simulated root canals was prevented completely. CONCLUSION: Polyisoprene and PCL composites with BG showed promising results as single root canal filling materials. Incorporation of BG fillers into the polymers under investigation made the resulting composite materials bioactive and improved their immediate sealing ability.

Magnetic switching of optical reflectivity in nanomagnet/micromirror suspensions: colloid displays as a potential alternative to liquid crystal displays.

Two-particle colloids containing nanomagnets and microscale mirrors can be prepared from iron oxide nanoparticles, microscale metal flakes and high-density liquids stabilizing the mirror suspension against sedimentation by matching the constituent's density. The free Brownian rotation of the micromirrors can be magnetically controlled through an anisotropic change in impulse transport arising from impacts of the magnetic nanoparticles onto the anisotropic flakes. The resulting rapid mirror orientation allows large changes in light transmission and switchable optical reflectivity. The preparation of a passive display was conceptually demonstrated through colloid confinement in a planar cavity over an array of individually addressable solenoids and resulted in 4 x 4 digit displays with a reaction time of less than 100 ms.

Do bioactive glasses convey a disinfecting mechanism beyond a mere increase in pH?

AIM: To test whether bioactive glasses kill microbiota via mineralization or the release of ions other than sodium. METHODOLOGY: Flame-spray synthesis was applied to produce nanometric glasses of different sodium content and constant Ca/P ratio: 28S5, 45S5 and 77S. Calcium hydroxide and nanometric tricalcium phosphate (TCP) were used as controls. Apatite induction was monitored by Raman spectroscopy. Bovine dentine disks with adherent Enterococcus faecalis cells were exposed to test and control suspensions or buffered solutions for 1 h, 1 day and 1 week. Colony-forming units were counted and disks were inspected using scanning electron microscopy. Suspension supernatants and solutions were analysed for their pH, osmolarity, calcium and silicon content. RESULTS: Sodium containing glasses induced pH levels above 12, compared with less than pH 9 with sodium-free 77S. Calcium hydroxide, 45S5 and 28S5 killed all bacteria after 1 day and lysed them after 1 week. TCP caused the highest apatite induction and substantial calcification on bacteria adhering to dentine, but did not reduce viable counts. 77S achieved disinfection after 1 week without visible apatite formation, whilst the buffer solution at pH 9 caused only minimal reduction in counts. CONCLUSION: Bioactive glasses have a directly and an indirectly pH-related antibacterial effect. The effect not directly linked to pH is because of ion release rather than mineralization.

Antimicrobial effect of nanometric bioactive glass 45S5.

Most recent advances in nanomaterials fabrication have given access to complex materials such as SiO(2)-Na(2)O-CaO-P(2)O(5) bioactive glasses in the form of amorphous nanoparticles of 20- to 60-nm size. The clinically interesting antimicrobial properties of commercially available, micron-sized bioactive glass 45S5 have been attributed to the continuous liberation of alkaline species during application. Here, we tested the hypothesis that, based on its more than ten-fold higher specific surface area, nanometric bioactive glass releases more alkaline species, and consequently displays a stronger antimicrobial effect, than the currently applied micron-sized material. Ionic dissolution profiles were monitored in simulated body fluid. Antimicrobial efficacy was assessed against clinical isolates of enterococci from persisting root canal infections. The shift from micron- to nano-sized treatment materials afforded a ten-fold increase in silica release and solution pH elevation by more than three units. Furthermore, the killing efficacy was substantially higher with the new material against all tested strains.

Grain growth resistance and increased hardness of bulk nanocrystalline fcc cobalt prepared by a bottom-up approach.

Bulk nanocrystalline cobalt was prepared from reducing flame-spray-derived cobalt nanopowders exclusively in the face-centred cubic (fcc) modification. Compacts of approximately 60% density were obtained from uniaxial compression at room temperature and showed a strong resistance towards grain growth upon subsequent sintering to 90% relative density. The nanocrystalline structure remained stable well above 1000 degrees C and resulted in a pore-rich metal with about 10(15) nanovoids cm(-3). These sintered compacts displayed an up to three times higher bulk hardness if compared to conventional cobalt and local ductility as evidenced from scanning electron microscopy and nanoindentation. The strong grain-growth resistance and consequent increase in material hardness are discussed in respect of the presence of nanovoids, twin boundaries and material contamination.

Clinical outcomes of gamma-irradiated sterile cornea in aqueous drainage device surgery: a multicenter retrospective study.

PurposeThe purpose of the study was to evaluate the safety and efficacy of gamma-irradiated sterile cornea (GISC) for covering the tube in aqueous drainage device (ADD) surgery in a retrospective, multicenter case series.Patients and methodsParticipants included 297 patients (321 procedures) who had undergone ADD surgery for the first time using GISC patch at three clinic centers in the United States between April 2009 and July 2012. The medical records of those consecutive patients were reviewed. Preoperative, intraoperative, and postoperative parameters about GISC were collected and analyzed. The main outcome measures were patch graft failure (PGF) and postoperative complications related to GISC.ResultsThree hundred and nineteen eyes in 295 patients were included in the current analysis. Ten out of the 319 eyes experienced PGF with a mean follow-up of 15.4±9.8 (SD) months. The overall cumulative PGF proportion from Kaplan-Meier analysis was 2.6% (95% CI: 0.6-4.7%) at 18 months. We detected two cases of presumed endophthalmitis related to PGF.ConclusionsGISC appears to have a reasonable success rate for preventing tube exposure related to PGF over an 18-month period. This success rate, in combination with other features of GISC (transparency and storage at room temperature), makes it a viable choice for patch graft material during ADD.

The safety and efficacy of combined phacoemulsification and trabeculectomy with releasable sutures.

AIM: To evaluate the combined phacoemulsification with acrylic intraocular lens implantation and trabeculectomy with releasable sutures, without the use of antimetabolites, performed by one surgeon. METHODS: A retrospective analysis was performed on 251 eyes of 198 patients after a combined phaco/trabeculectomy procedure, followed up for an average of 16 months for vision, intraocular pressure (IOP), and number of glaucoma medications. Cox proportional hazard regression analysis provided measures of relative risk (RR) or hazards ratios and 95% confidence intervals (CIs) associated with the occurrence of several conditions or complications of surgery. RESULTS: Mean preoperative variables were best corrected vision, 0.6 (SD 0.4) logMAR (20/80 Snellen); IOP, 18.7 (4.9) mm Hg; and number of glaucoma medications, 1.7 (0.8). Mean postoperative results were best corrected vision, 0.2 (0.4) logMAR (20/32 Snellen); IOP, 15.1 (3.2) mm Hg; and number of glaucoma medications, 0.3 (0.6). Postoperatively, the mean decreases from baseline were 19.2% for IOP and 84.1% for number of glaucoma medications; 85% of eyes required fewer glaucoma medications and 78% of eyes were medication free, with IOP control. There were no significant operative or postoperative complications. CONCLUSION: Combined phacoemulsification and trabeculectomy with releasable sutures, in the absence of antimetabolites, is a safe, effective, and stable alternative for patients with cataracts and glaucoma.

A new donor cornea harvesting technique for posterior lamellar keratoplasty.

AIMS: To describe a technique for posterior lamellar keratoplasty donor preparation. METHODS: In an experimental study eight human donor research corneas were mounted onto an artificial anterior chamber and deep stromal pockets dissected. Four corneas were mounted in the standard endothelial side down orientation and dissected using standard instruments (group 1). Another four corneas were mounted endothelial side up and dissected using a flat spatula (group 2). Trephined lamellar graft thickness was assessed by ultrasound pachymetry. The grafts were also analysed using vital staining of the endothelium and standard histological preparation. RESULTS: Achieved posterior graft thickness was 118 (SD 32) microm (group 1) and 92 (23) microm (group 2) (p=0.324). Percentage of devitalised endothelial cells was 0.86% (1.48%) (group 1) and 3.9% (2.9%) (group 2) (p=0.185). The dissections using both harvesting techniques remained in plane and were smooth. CONCLUSIONS: A blunt spatula and endothelium side up orientation on an artificial anterior chamber can be used to create posterior lamellar dissections without compromising endothelial cell number or planarity when compared to standard endothelium side down harvest.

Silica particles with encapsulated DNA as trophic tracers.

Ecological networks such as food webs are extremely complex and can provide important information about the robustness and productivity of an ecosystem. In most cases, it is not feasible to observe trophic interactions between predators and prey directly and with the available methods, it is difficult to quantify the connections between them. Here, we show that submicron-sized silica particles (100-150 nm) with encapsulated DNA (SPED) enable accurate food and organism labelling and quantification of specific animal-to-animal transfer over more than one trophic level. We found that SPED were readily transferable and quantifiable from the bottom to the top of a two-level food chain of arthropods. SPED were taken up in the gut system and remained persistent in an animal over several days. When uniquely labelled SPED were applied at predefined ratios, we found that information about their relative abundance was reliably conserved after trophic level transfer and over time. SPED were also applied to investigate the flower preference of fly pollinators and proved to be a fast and accurate analysis method. SPED combine attributes of DNA barcoding and stable isotope analysis such as unique labelling, quantification via real-time PCR and exact backtracking to the tracer source. This improves and simplifies the analysis and monitoring of ecological networks.

Base-free Knoevenagel condensation catalyzed by copper metal surfaces.

For the first time Knoevenagel condensation has been catalyzed by elemental copper with unexpected activity and excellent isolated yields. Inexpensive, widely available copper powder was used to catalyze the condensation of cyanoacetate and benzaldehyde under mild conditions. To ensure general applicability, a wide variety of different substrates was successfully reacted.

Stable dispersions of azide functionalized ferromagnetic metal nanoparticles.

Ferromagnetic nanoparticles are covalently modified in order to enhance the dispersion stability as well as the antifouling properties. Insertion of an azide moiety allows "click"-reaction of a relevant tag molecule. This and the high saturation magnetization of the presented nanocomposite offer a promising platform for magnetic biosensors.

The association of lymphocytotoxic antibodies with corneal allograft rejection in high risk patients. The Collaborative Corneal Transplantation Studies Research Group.

The Collaborative Corneal Transplantation Studies are a pair of multicenter prospective clinical trials evaluating the effectiveness of histocompatibility matching in high risk keratoplasty patients. The antigen matching study (AMS) evaluated HLA matching in patients without circulating lymphocytotoxic antibody to HLA antigens and the cross-match study (CS) evaluated the effect of using cross-match-negative donors in patients with identified circulating lymphocytotoxic antibodies to HLA antigens. Sera from 510 patients considered for enrollment in the studies were screened preoperatively for the presence of anti-class I lymphocytotoxic antibodies (LA). The 42 patients (8%) found to have detectable LA entered the CS. The 468 patients found not to have detectable LA preoperatively entered the AMS. Fifteen of the 37 transplanted CS patients were found to have donor-specific anti-class I antibody (before or after surgery). These patients were also screened for anti-class II LA and 25 had anti-class II panel reactive antibody > or = 5%. Forty-nine of the 419 transplanted AMS patients (12%) were found to have produced anti-class I LA after surgery, and in 19 patients, antibody specificities were those of donor HLA antigens. There was a significant association between the number of mismatched class I antigens and the number of donor-specific LA produced. The production of LA by AMS patients was significantly associated with reaction episodes; eighty-two percent of patients (40 of 49) with LA had reaction, compared with 63% of patients (230 of 365) without LA (P = 0.02). Likewise, production of donor-specific LA was significantly associated with immune-mediated graft failure (P = 0.025). For CS patients, there was no correlation between the production of donor-specific anti-class I or nonspecific anti-class II antibodies and graft outcome. However, the CS patients had poorer graft survival than did AMS patients at 3 years (57% vs. 66%, P = 0.01). These data demonstrate that LA, especially directed against donor class I HLA antigens following corneal transplantation in high risk patients, are associated with immune graft rejection and can be an indicator of allograft rejection.

In vitro immunological function of human corneal fibroblasts.

Gamma-interferon-induced HLA-DR-like antigens on normal human fibroblasts were compared functionally to HLA-DR antigens on peripheral blood mononuclear leucocytes (PBML) from the same individual. Irradiated DR+ fibroblasts did not stimulate allogeneic PBML to divide in a mixed lymphocyte fibroblast culture (MLFC) as determined by 3H-thymidine incorporation, although lymphocytes autologous to the fibroblasts stimulated normally. Co-culture with interleukin 1 (IL-1), IL-2, 2-mercaptoethanol and indomethacin had no effect on MLFC. Pretreatment of stimulator fibroblasts with neuraminidase to modify sialoglycoproteins increased responses only modestly. Both DR+ and DR- fibroblasts inhibited a normal mixed lymphocyte culture (MLC) response in a dose-dependent fashion. These results suggest that the induction of HLA-DR-like molecules on fibroblasts is not sufficient to initiate host sensitization and that fibroblasts may inhibit normal lymphoproliferation.