Space biofilms - An overview of the morphology of Pseudomonas aeruginosa biofilms grown on silicone and cellulose membranes on board the international space station.

Microorganisms' natural ability to live as organized multicellular communities - also known as biofilms - provides them with unique survival advantages. For instance, bacterial biofilms are protected against environmental stresses thanks to their extracellular matrix, which could contribute to persistent infections after treatment with antibiotics. Bacterial biofilms are also capable of strongly attaching to surfaces, where their metabolic by-products could lead to surface material degradation. Furthermore, microgravity can alter biofilm behavior in unexpected ways, making the presence of biofilms in space a risk for both astronauts and spaceflight hardware. Despite the efforts to eliminate microorganism contamination from spacecraft surfaces, it is impossible to prevent human-associated bacteria from eventually establishing biofilm surface colonization. Nevertheless, by understanding the changes that bacterial biofilms undergo in microgravity, it is possible to identify key differences and pathways that could be targeted to significantly reduce biofilm formation. The bacterial component of Space Biofilms project, performed on the International Space Station in early 2020, contributes to such understanding by characterizing the morphology and gene expression of bacterial biofilms formed in microgravity with respect to ground controls. Pseudomonas aeruginosa was used as model organism due to its relevance in biofilm studies and its ability to cause urinary tract infections as an opportunistic pathogen. Biofilm formation was characterized at one, two, and three days of incubation (37 °C) over six different materials. Materials reported in this manuscript include catheter grade silicone, selected due to its medical relevance in hospital acquired infections, catheter grade silicone with ultrashort pulsed direct laser interference patterning, included to test microtopographies as a potential biofilm control strategy, and cellulose membrane to replicate the column and canopy structure previously reported from a microgravity study. We here present an overview of the biofilm morphology, including 3D images of the biofilms to represent the distinctive morphology observed in each material tested, and some of the key differences in biofilm thickness, mass, and surface area coverage. We also present the impact of the surface microtopography in biofilm formation across materials, incubation time, and gravitational conditions. The Space Biofilms project (bacterial side) is supported by the National Aeronautics and Space Administration under Grant No. 80NSSC17K0036 and 80NSSC21K1950.

Design of a spaceflight biofilm experiment.

Biofilm growth has been observed in Soviet/Russian (Salyuts and Mir), American (Skylab), and International (ISS) Space Stations, sometimes jeopardizing key equipment like spacesuits, water recycling units, radiators, and navigation windows. Biofilm formation also increases the risk of human illnesses and therefore needs to be well understood to enable safe, long-duration, human space missions. Here, the design of a NASA-supported biofilm in space project is reported. This new project aims to characterize biofilm inside the International Space Station in a controlled fashion, assessing changes in mass, thickness, and morphology. The space-based experiment also aims at elucidating the biomechanical and transcriptomic mechanisms involved in the formation of a "column-and-canopy" biofilm architecture that has previously been observed in space. To search for potential solutions, different materials and surface topologies will be used as the substrata for microbial growth. The adhesion of bacteria to surfaces and therefore the initial biofilm formation is strongly governed by topographical surface features of about the bacterial scale. Thus, using Direct Laser-Interference Patterning, some material coupons will have surface patterns with periodicities equal, above or below the size of bacteria. Additionally, a novel lubricant-impregnated surface will be assessed for potential Earth and spaceflight anti-biofilm applications. This paper describes the current experiment design including microbial strains and substrata materials and nanotopographies being considered, constraints and limitations that arise from performing experiments in space, and the next steps needed to mature the design to be spaceflight-ready.

Wear behavior of dental Y-TZP ceramic against natural enamel after different finishing procedures.

OBJECTIVE: The aim of this in vitro study was to evaluate the influence of different finishing procedures on the wear behavior of zirconia against natural enamel. METHODS: 64 quadratic specimens (10 mm × 10 mm × 2 mm) were cut from a commercial hipped dental Y-TZP ceramic. Four different groups with 16 specimens each were formed according to the following finishing procedures: PZ (polished), RR (fine-grit diamond), GR (coarse-grit diamond), GZ (glazed). Polished specimens of a leucite-reinforced glass ceramic (Empress CAD) were used as a control (GC). The materials were subjected to the Ivoclar wear method (Willytec chewing simulator, 120,000 cycles, 5kg weight) with 80 natural caries-free cusps of first upper molars as antagonists. Wear was analyzed for both the enamel cusps and test specimens by measurement of the vertical substance loss with a laser scanner. Surface roughness was measured by means of a white-light interferometer. RESULTS: The surface roughness was significantly different among the polished, diamond-finished, and glazed ceramic specimens (ANOVA, post hoc Bonferroni p<0.05). The results of the one-way ANOVA indicated that the finishing technique significantly affected enamel wear (p<0.05). The post hoc test indicated that the specimens finished with the coarse diamond caused significantly higher antagonist wear than the polished ones. Polished zirconia showed the lowest wear of the antagonist enamel, with a mean value of 171.74 (SD = 121.68), and resulted in enamel wear that was not significantly different from that of the glass ceramic control group. No significant linear correlation could be found between pre-testing surface roughness and abrasive wear. SIGNIFICANCE: If zirconia is used without veneering material for crowns and fixed dental prostheses (FDPs), the surface must be well-polished if occlusal adjustments with coarse diamonds are performed. The polishing step reduces the wear of the opposing enamel.

Subcritical crack growth behavior and life data analysis of two types of dental Y-TZP ceramics.

OBJECTIVES: The aim of this study was to evaluate the mechanical properties and the subcritical crack growth behavior of a presintered dental Y-TZP (Kavo Everest ZS) and a hot isostatic pressed Y-TZP (Kavo Everest ZH) and to perform life data analysis. METHODS: For each material 150 bending bars were produced. The initial fracture strength was determined in a four-point bending test. The subcritical crack growth parameters n and A were determined in a dynamic fatigue method at four decreasing loading rates from 110MPa/s to 0.11MPa/s in distilled water at 38°C. For each loading rate Weibull statistics were performed and the Weibull moduli m and characteristic strengths σ(0) were calculated. Using these data, strength-fracture probability-life time (SPT) predictions were derived for 1 day, 1 year, 5 years and 10 years, based on a static crack growth mechanism. RESULTS: The "hipped" Y-TZP ceramic exhibited a higher initial strength (σ(c)=1618.18), characteristic strength (σ(0)=837.15) and fracture toughness (K(IC)=4.52MPa/m(1/2)) than the pre-sintered ceramic (σ(c)=1431, σ(0)=745.46 and K(IC)=3.17MPa/m(1/2), respectively). Fatigue parameters, n and A, were 28.5 and 7.97×10(-24) for Everest ZH and 30.15 and 5.47×10(-20) for Everest ZS. The predicted fracture stress at 5% failure probability for a lifetime of 10 years was 259.34MPa for Everest ZH and 263.2MPa for Everest ZS. CONCLUSIONS: Although the "hipped" Y-TZP showed favorable initial mechanical properties, no significant difference could be found in the susceptibility of both ceramics to subcritical crack growth and their long-term strength.

Laser ablation patterning by interference induces directional cell growth.

Laser-patterning by interference is a method to introduce micropatterns on the surface of TXL and TXB, which were shown to have an effect on the L929 growth. In this experiment, we have produced collagen-coated and laser-patterned TXL and TXB with different dimensions; the groove width of the line patterns varied approximately from 1.2 microm to 9.7 microm, ridge depth varied from 0.4 microm to 1.3 microm, and the groove depth varied between 0.4 microm and 1.3 microm. Therefore, a homogeneous smooth surface was achieved, and that L929 growth was only affected by the different dimensions of the line patterns. All the laser-patterned TXL and TXB have shown inducing different degrees of directional growth of L929 that the cells grew in the direction aligning the microgrooves. However, the different widths of the microgrooves were demonstrated to play an important role in determining cell morphology and growth orientation. For example, cells were elongated when they grew on the narrower widths, which were 1.26 microm, 1.91 microm, and 5.04 microm while cells tended to be triangular when grew on wider width about 9.76 microm. In addition, L929 might grow only on the top of the laser-patterns attaching the ridges when the groove widths were narrow, but might grow into the microgrooves when the width went beyond 5.04 microm.