Terpenoids are natural plant-derived products that are applied to treat a broad range of human diseases, such as airway infections and inflammation. However, pharmaceutical applications of terpenoids against bacterial infection remain challenging due to their poor water solubility. Here, we produce invasomes encapsulating thymol, menthol, camphor and 1,8-cineol, characterize them via cryo transmission electron microscopy and assess their bactericidal properties. While control- and cineol-invasomes are similarly distributed between unilamellar and bilamellar vesicles, a shift towards unilamellar invasomes is observable after encapsulation of thymol, menthol or camphor. Thymol- and camphor-invasomes show a size reduction, whereas menthol-invasomes are enlarged and cineol-invasomes remain unchanged compared to control. While thymol-invasomes lead to the strongest growth inhibition of S. aureus, camphor- or cineol-invasomes mediate cell death and S. aureus growth is not affected by menthol-invasomes. Flow cytometric analysis validate that invasomes comprising thymol are highly bactericidal to S. aureus. Notably, treatment with thymol-invasomes does not affect survival of Gram-negative E. coli. In summary, we successfully produce terpenoid-invasomes and demonstrate that particularly thymol-invasomes show a strong selective activity against Gram-positive bacteria. Our findings provide a promising approach to increase the bioavailability of terpenoid-based drugs and may be directly applicable for treating severe bacterial infections such as methicillin-resistant S. aureus.
New ecological trends and changes in consumer behavior are known to favor biofilm formation in household appliances, increasing the need for new antimicrobial materials and surfaces. Their development requires laboratory-cultivated biofilms, or biofilm model systems (BMS), which allow for accelerated growth and offer better understanding of the underlying formation mechanisms. Here, we identified bacterial strains in wildtype biofilms from a variety of materials from domestic appliances using matrix-assisted laser desorption/ionization-time of flight mass spectroscopy (MALDI-TOF-MS). Staphylococci and pseudomonads were identified by MALDI-TOF-MS as the main genera in the habitats and were analyzed for biofilm formation using various in vitro methods. Standard quantitative biofilm assays were combined with scanning electron microscopy (SEM) to characterize biofilm formation. While Pseudomonas putida, a published lead germ, was not identified in any of the collected samples, Pseudomonas aeruginosa was found to be the most dominant biofilm producer. Water-born Pseudomonads were dominantly found in compartments with water contact only, such as in detergent compartment and detergent enemata. Furthermore, materials in contact with the washing load are predominantly colonized with bacteria from the human.
The COVID-19 Pandemic leads to an increased worldwide demand for personal protection equipment in the medical field, such as face masks. New approaches to satisfy this demand have been developed, and one example is the use of 3D printing face masks. The reusable 3D printed mask may also have a positive effect on the environment due to decreased littering. However, the microbial load on the 3D printed objects is often disregarded. Here we analyze the biofilm formation of Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli on suspected antimicrobial Plactive™ PLA 3D printing filaments and non-antimicrobial Giantarm™ PLA. To characterize the biofilm-forming potential scanning electron microscopy (SEM), Confocal scanning electron microscopy (CLSM) and colony-forming unit assays (CFU) were performed. Attached cells could be observed on all tested 3D printing materials. Gram-negative strains P. aeruginosa and E. coli reveal a strong uniform growth independent of the tested 3D filament (for P. aeruginosa even with stressed induced growth reaction by Plactive™). Only Gram-positive S. aureus shows strong growth reduction on Plactive™. These results suggest that the postulated antimicrobial Plactive™ PLA does not affect Gram-negative bacteria species. These results indicate that reusable masks, while better for our environment, may pose another health risk.
Polyacrylonitrile (PAN) nanofiber mats are typical precursors for carbon nanofibers. They can be fixed or even elongated during stabilization and subsequent carbonization to gain straight, mechanically robust carbon nanofibers. These processes necessitate additional equipment or are—if the nanofiber mats are just fixed at the edges—prone to resulting in the specimens breaking, due to an uneven force distribution. Hence, we showed in a previous study that electrospinning PAN on aluminum foils and stabilizing them fixed on these substrates, is a suitable solution to keep the desired morphology after stabilization and incipient carbonization. Here, we report on the influence of different metallic and semiconductor substrates on the physical and chemical properties of the nanofiber mats after stabilization and carbonization at temperatures up to 1200 °C. For stabilization on a metal substrate, an optimum stabilization temperature of slightly above 240 °C was found, approached with a heating rate of 0.25 K/min. Independent from the substrate material, SEM images revealed less defect fibers in the nanofiber mats stabilized and incipiently carbonized on a metal foil. Finally, high-temperature carbonization on different substrates is shown to allow for producing metal/carbon nano-composites.
In the light of the SARS-CoV-2 pandemic and growing numbers of bacteria with resistance to antibiotics, the development of antimicrobial coatings is rising worldwide. Inorganic coatings are attractive because of low environmental leakage and wear resistance. Examples for coatings are hot metal dipping or physical vapor deposition of nanometer coatings. Here, magnetron sputtering of various transition metals, such as gold, ruthenium and tantalum, was investigated. Metal films were characterized by scanning electron microscopy (SEM), atomic force microscopy (AFM) and energy dispersive X-ray spectroscopy (EDX). We investigated the growth of Pseudomonas aeruginosa isolated from household appliances on different sputter-coated metal surfaces. The fine-grained nanometric structure of these metal coatings was between 14 nm (tantalum) and 26 nm (gold) and the roughness was in a range of 164 pm (ruthenium) to 246 pm (gold). Antibacterial efficacy of metal surfaces followed the order: gold > tantalum > ruthenium. Interestingly, gold had the strongest inhibitory effect on bacterial growth, as analyzed by LIVE/DEAD and CFU assay. High-magnification SEM images showed dead bacteria characterized by shrinkage induced by metal coatings. We conclude that sputtering might be a new application for the development of antimicrobial surfaces on household appliances and or surgical instruments.
The performance of plastic components in water-bearing parts of industrial and household appliances, often in the presence of harsh environments and elevated temperatures, critically relies on the mechanical and thermal polymer stability. In this light, the precise knowledge of aging properties of polymers formulated with dedicated antiaging additive packages as well as various fillers is crucial for long-time device warranty. We investigated and analysed the time-dependent, polymer-liquid interface aging of different industrial performance polypropylene samples in aqueous detergent solution at high temperatures (95 °C). Special emphasis was put on the disadvantageous process of consecutive biofilm formation that often follows surface transformation and degradation. Atomic force microscopy, scanning electron microscopy, and infrared spectroscopy were used to monitor and analyse the surface aging process. Additionally, bacterial adhesion and biofilm formation was characterised by colony forming unit assays. One of the key findings is the observation of crystalline, fibre-like growth of ethylene bis stearamide (EBS) on the surface during the aging process. EBS is a widely used process aid and lubricant enabling the proper demoulding of injection moulding plastic parts. The aging-induced surface-covering EBS layers changed the surface morphology and promoted bacterial adhesion as well as biofilm formation of Pseudomonas aeruginosa.