Facilitation of adhesion and spreading of endothelial cells on silicone oxide-coated dacron material by microwave-excited low-pressure plasma

Objectives Autologous transplants are still the means of choice for bypass surgery. In addition to good tolerability, there is a reduced thrombogenicity and fewer neointima hyperplasia compared to artificial materials. However, since viable transplants are limited, attempts are being made to improve existing artificial vascular prosthesis material. Next to the reduction of thrombogenicity, a rapid endothelialization of the vascular graft should reduce intimal hyperplasia and thus prevent stenoses. The effect of newly developed silicon oxide coatings on the growth of endothelial cells was therefore the goal of this work in a cell culture study. Methods A woven, uncoated polyethylene terephthalate (PET) vessel prosthesis was used. The coating process was carried out in a low-pressure plasma reactor in a multi-step process. After preparation of the vacuum chamber hexamethyldisiloxane (HDMSO) with oxygen was evaporated using argon plasma. By this an approx. 1 nm thin adhesion promoter layer was separated from plasma and HMDSO. The silicone oxide barrier layer was applied to the PET vessel samples. The carbon content of the layer could be selectively altered by changing the HMDSO oxygen flow ratio, resulting in coatings of 100 nm, 500 nm, and 1,000 nm. In addition, two different oxygen-to-HMDSO ratios were used. To achieve a carbon coating as low as possible, the ratio was set to 200:1. A carbon-rich layer was obtained with the 1:1 setting. The various coatings were then examined for their surface texture by scanning electron microscopy (SEM) as well as by cell culture experiments for cell viability and growth using EA.hy 926 cells. Results SEM showed no changes in the surface morphology; however a layer thickness of 1,000 nm showed peeled off coating areas. Alamar blue assays showed a significantly higher metabolic activity (p=0.026) for the coating 500 nm, ratio 200:1 compared to untreated control samples and a significantly lower metabolic activity (p=0.037) of the coating 500 nm, ratio 1:1 compared to the coating 500 nm, ratio 200:1. This underlines the apparent tendency of the 1:1 coating to inhibit the metabolic activity of the cells, while the 200:1 coating increases the activity. Fluorescence microscopy after calcein acetoxymethyl ester (AM) staining showed no significant difference between the different coatings and the uncoated PET material. However, a tendency of the increased surface growth on the coating 500 nm, ratio 200:1, is shown. The coatings with the ratio 1:1 tend to be less densely covered. Conclusions The results of this work indicate a great potential in the silicon coating of vascular prosthesis material. The plasma coating can be carried out easy and gently. Cell culture experiments demonstrated a tendency towards better growth of the cells on the 200:1 ratio coating and a poorer growth on the carbon-rich coating 1:1 compared to the uncoated material. The coating with silicon oxide with a thickness of 500 nm and an oxygen-HMDSO ratio of 200:1, a particularly low-carbon layer, appears to be a coating, which should therefore be further investigated for its effects on thrombogenicity and intimal hyperplasia.

The use of dietary fibre to optimize microbial gut function in pigs, with particular consideration of dietary cereal grains and legumes

This chapter examines interactions of dietary fibre components of pig diets with GIT microbiota, emphasizing cereals and legumes fed to pigs. Carbohydrate composition of these feedstuffs are described, and their relationship to metabolic activity of the porcine intestinal microbiota and interactions with the host. Fermentable carbohydrates which act as substrates for microbial metabolism are described, followed by an assessment of cereals and legumes as potential modulators of intestinal microbiota. Past work focussed on purified extracts, but attention is now focussing on whole grains or their fractions such as brans, in terms of effects on microbial populations. Such studies are showing the positive consequences of mixtures of DF in the form of complex plant cellular structures, rather than single refined ingredients, to achieve beneficial health outcomes. Further work is also needed to define appropriate quantities and types of DF to achieve desired effects whilst minimising negative outcomes.

Regulatory Effect of Irresistin-16 on Competitive Dual-Species Biofilms Composed of Streptococcus mutans and Streptococcus sanguinis

Based on the ecological plaque hypothesis, suppressing opportunistic pathogens within biofilms, rather than killing microbes indiscriminately, could be a biofilm control strategy for managing dental caries. The present study aimed to evaluate the effects of irresistin-16 (IRS-16) on competitive dual-species biofilms, which consisted of the conditional cariogenic agent Streptococcus mutans (S. mutans) and oral commensal bacteria Streptococcus sanguinis (S. sanguinis). Bacterial growth and biofilm formation were monitored using growth curve and crystal violet staining, respectively. The microbial proportion was determined using fluorescence in situ hybridization. A 2, 5-diphenyltetrazolium bromide assay was used to measure the metabolic activity of biofilms. Bacterial/extracellular polysaccharide (EPS) dyeing, together with water-insoluble EPS measurements, were used to estimate EPS synthesis. A lactic acid assay was performed to detect lactic acid generation in biofilms. The cytotoxicity of IRS-16 was evaluated in mouse fibroblast L929 cells using a live/dead cell viability assay and cell counting kit-8 assay. Our results showed that IRS-16 exhibited selective anti-biofilm activity, leading to a remarkable survival disadvantage of S. mutans within competitive dual-species biofilms. In addition, the metabolic activity, EPS synthesis, and acid generation of dual-species biofilms were significantly reduced by IRS-16. Moreover, IRS-16 showed minimal cytotoxicity against mouse fibroblast L929 cells. In conclusion, IRS-16 exhibited remarkable regulatory effects on dual-species biofilms composed of S. mutans and S. sanguinis with low cytotoxicity, suggesting that it may have potential for use in caries management through ecological biofilm control.

Radiosensitizing effect of galunisertib, a TGF-ß receptor I inhibitor, on head and neck squamous cell carcinoma in vitro

SummaryBackground. Resistance to radiation therapy poses a major clinical problem for patients suffering from head and neck squamous cell carcinoma (HNSCC). Transforming growth factor ß (TGF-ß) has emerged as a potential target. This study aimed to investigate the radiosensitizing effect of galunisertib, a small molecule TGF-ß receptor kinase I inhibitor, on HNSCC cells in vitro. Methods. Three HNSCC cell lines were treated with galunisertib alone, or in combination with radiation. Of those three cell lines, one has a known inactivating mutation of the TGF-ß pathway (Cal27), one has a TGF-ß pathway deficiency (FaDu) and one has no known alteration (SCC-25). The effect on metabolic activity was evaluated by a resazurin-based reduction assay. Cell migration was evaluated by wound-healing assay, clonogenic survival by colony formation assay and cell cycle by FACS analysis. Results. Galunisertib reduced metabolic activity in FaDu, increased in SCC-25 and had no effect on CAL27. Migration was significantly reduced by galunisertib in all three cell lines and showed additive effects in combination with radiation in CAL27 and SCC-25. Colony-forming capabilities were reduced in SCC-25 by galunisertib and also showed an additive effect with adjuvant radiation treatment. Cell cycle analysis showed a reduction of cells in G1 phase in response to galunisertib treatment. Conclusion. Our results indicate a potential antineoplastic effect of galunisertib in HNSCC with intact TGF-ß signaling in combination with radiation.

Metabolic activity of Hordeum vulgare, Brassica napus and Vicia faba in Worm and Root type Biopore Sheaths

Aims Biopores offer favorable chemical, biological and physical properties for root growth in untilled soil layers. There they are considered as nutrient “hotspots” with preferential root growth. However, the literature lacks a quantification of metabolic activity due to nutrient acquisition of main crops while growing in the biopore sheath. Methods A pot experiment was performed to map the metabolic activity of roots, as indicated by pH change. The roots of spring barley (Hordeum vulgare L.), spring oilseed rape (Brassica napus L.) and faba bean (Vicia faba L.) were growing through the biopore sheath influenced by an earthworm (Lumbricus terrestris L.) or a taproot (Cichorium intybus L.), in comparison to subsoil without a pore (bulk soil). pH sensitive planar optodes were applied in order to image a planar section of the sheath, while preserving an intact biopore sheath during the experiment. Results Roots were first found in the field of view in worm biopore then root biopore and bulk soil. At time of the first measurement the pH value was highest in worm biopore sheath (LS-Mean±SEM: 7.16a±0.11), followed by root biopore sheath (6.99ab±0.12) and bulk soil (6.61b±0.12). In spring oilseed rape a significant alkalization (+0.80 Δ pH) was found over time in bulk soil. Faba bean significantly acidified the root biopore sheath (-0.73 Δ pH). Spring barley showed no significant pH changes. Conclusions The results of the current study reveal a trend of faster root growth through biopores and a higher initial pH value in the biopore sheaths compared to the bulk soil. Biopores serve not only as an elongation path for roots, but their sheaths also provide an environment for root activity in the subsoil.

Effects of Dietary Cereal and Protein Source on Fiber Digestibility, Composition, and Metabolic Activity of the Intestinal Microbiota in Weaner Piglets

This study aimed to investigate the effect of fiber-rich rye and rapeseed meal (RSM) compared to wheat and soybean meal (SBM) on fiber digestibility and the composition and metabolic activity of intestinal microbiota. At weaning, 88 piglets were allocated to four feeding groups: wheat/SBM, wheat/RSM, rye/SBM, and rye/RSM. Dietary inclusion level was 48% for rye and wheat, 25% for SBM, and 30% for RSM. Piglets were euthanized after 33 days for collection of digesta and feces. Samples were analyzed for dry matter and non-starch-polysaccharide (NSP) digestibility, bacterial metabolites, and relative abundance of microbiota. Rye-based diets had higher concentrations of soluble NSP than wheat-based diets. RSM-diets were higher in insoluble NSP compared to SBM. Rye-fed piglets showed a higher colonic and fecal digestibility of NSP (p < 0.001, p = 0.001, respectively). RSM-fed piglets showed a lower colonic and fecal digestibility of NSP than SBM-fed piglets (p < 0.001). Rye increased jejunal and colonic concentration of short-chain fatty acids (SCFA) compared to wheat (p < 0.001, p = 0.016, respectively). RSM-fed pigs showed a lower jejunal concentration of SCFA (p = 0.001) than SBM-fed pigs. Relative abundance of Firmicutes was higher (p = 0.039) and of Proteobacteria lower (p = 0.002) in rye-fed pigs compared to wheat. RSM reduced Firmicutes and increased Actinobacteria (jejunum, colon, feces: p < 0.050), jejunal Proteobacteria (p = 0.019) and colonic Bacteroidetes (p = 0.014). Despite a similar composition of the colonic microbiota, the higher amount and solubility of NSP from rye resulted in an increased fermentative activity compared to wheat. The high amount of insoluble dietary fiber in RSM-based diets reduced bacterial metabolic activity and caused a shift toward insoluble fiber degrading bacteria. Further research should focus on host–microbiota interaction to improve feeding concepts with a targeted use of dietary fiber.