Functional Surfaces for Passive Fungal Proliferation Control: Effect of Surface Micro- and Nanotopography, Material, and Wetting Properties.

Fungal proliferation can lead to adverse effects for human health, due to the production of pathogenic and allergenic toxins and also through the creation of fungal biofilms on sensitive surfaces (i.e., medical equipment). On top of that, food spoilage from fungal activity is a major issue, with food losses exceeding 30% annually. In this study, the effect of the surface micro- and nanotopography, material (aluminum, Al, and poly(methyl methacrylate), PMMA), and wettability against Aspergillus awamori is investigated. The fungal activity is monitored using dynamic conditions by immersing the surfaces inside fungal spore-containing suspensions and measuring the fungal biomass growth, while the surfaces with the optimum antifungal properties are also evaluated by placing them near spore suspensions of A. awamori on agar plates. Al- and PMMA-based superhydrophobic surfaces demonstrate a passive-like antifungal profile, and the fungal growth is significantly reduced (1.6-2.2 times lower biomass). On the other hand, superhydrophilic PMMA surfaces enhance fungal proliferation, resulting in a 2.6 times higher fungal total dry weight. In addition, superhydrophobic surfaces of both materials exhibit antifouling and antiadhesive properties, whereas both superhydrophobic surfaces also create an "inhibition" zone against the growth of A. awamori when tested on agar plates.

A Diagnostic Chip for the Colorimetric Detection of Legionella pneumophila in Less than 3 h at the Point of Need.

Legionella pneumophila has been pinpointed by the World Health Organization as the highest health burden of all waterborne pathogens in the European Union and is responsible for many disease outbreaks around the globe. Today, standard analysis methods (based on bacteria culturing onto agar plates) need several days (~12) in specialized analytical laboratories to yield results, not allowing for timely actions to prevent outbreaks. Over the last decades, great efforts have been made to develop more efficient waterborne pathogen diagnostics and faster analysis methods, requiring further advancement of microfluidics and sensors for simple, rapid, accurate, inexpensive, real-time, and on-site methods. Herein, a lab-on-a-chip device integrating sample preparation by accommodating bacteria capture, lysis, and DNA isothermal amplification with fast (less than 3 h) and highly sensitive, colorimetric end-point detection of L. pneumophila in water samples is presented, for use at the point of need. The method is based on the selective capture of viable bacteria on on-chip-immobilized and -lyophilized antibodies, lysis, the loop-mediated amplification (LAMP) of DNA, and end-point detection by a color change, observable by the naked eye and semiquantified by computational image analysis. Competitive advantages are demonstrated, such as low reagent consumption, portability and disposability, color change, storage at RT, and compliance with current legislation.

Durable, Ultrathin, and Antifouling Polymer Brush Coating for Efficient Condensation Heat Transfer.

Heat exchangers are made of metals because of their high heat conductivity and mechanical stability. Metal surfaces are inherently hydrophilic, leading to inefficient filmwise condensation. It is still a challenge to coat these metal surfaces with a durable, robust, and thin hydrophobic layer, which is required for efficient dropwise condensation. Here, we report the nonstructured and ultrathin (∼6 nm) polydimethylsiloxane (PDMS) brushes on copper that sustain high-performing dropwise condensation in high supersaturation. Due to the flexible hydrophobic siloxane polymer chains, the coating has low resistance to drop sliding and excellent chemical stability. The PDMS brushes can sustain dropwise condensation for up to ∼8 h during exposure to 111 °C saturated steam flowing at 3 m·s-1, with a 5-7 times higher heat transfer coefficient compared to filmwise condensation. The surface is self-cleaning and can reduce the level of bacterial attachment by 99%. This low-cost, facile, fluorine-free, and scalable method is suitable for a great variety of heat transfer applications.

A super liquid-repellent hierarchical porous membrane for enhanced membrane distillation.

Membrane distillation (MD) is an emerging desalination technology that exploits phase change to separate water vapor from saline based on low-grade energy. As MD membranes come into contact with saline for days or weeks during desalination, membrane pores have to be sufficiently small (typically <0.2 µm) to avoid saline wetting into the membrane. However, in order to achieve high distillation flux, the pore size should be large enough to maximize transmembrane vapor transfer. These conflicting requirements of pore geometry pose a challenge to membrane design and currently hinder broader applications of MD. To address this fundamental challenge, we developed a super liquid-repellent membrane with hierarchical porous structures by coating a polysiloxane nanofilament network on a commercial micro-porous polyethersulfone membrane matrix. The fluorine-free nanofilament coating effectively prevents membrane wetting under high hydrostatic pressure (>11.5 bar) without compromising vapor transport. With large inner micro-porous structures, the nanofilament-coated membrane improves the distillation flux by up to 60% over the widely used commercially available membranes, while showing excellent salt rejection and operating stability. Our approach will allow the fabrication of high-performance composite membranes with multi-scale porous structures that have wide-ranging applications beyond desalination, such as in cleaning wastewater.

Cold-atmospheric-plasma activated-ice as a cooling medium with antimicrobial properties: Case study on fish fillet preservation.

The efficacy and applicability of Plasma Activated Ice (PAI) -produced by cold atmospheric plasma (CAP) technology- on microorganisms and quality characteristics of perishable fresh sea bream (Sparus aurata) fillets, were evaluated. The changes in microbiological load and quality characteristics of fish fillets were investigated during storage with ice from deionized water (Control), PAI and ice from artificially produced water (Artificial) of H2O2 concentrations equal to those of PAI. Fresh sea bream fillets were packed under ice flakes (produced from PAI or Artificial or Control) on layers (as typically done in the relevant industry) and stored at 0.5 °C for 27 days. PAI application inhibited significantly the growth of microbial load of the fillets resulting in reduced growth rates while simultaneously significantly retarded the quality deterioration compared to the other disinfectant media. The use of PAI (with 10 mg/L H2O2) led to a 11-day and 6-day extension, i.e., 2-fold and a âˆ¼ 1.5-fold extension, of the fillets shelf-life compared to the samples treated with Control and Artificial ice, respectively. The results proved the efficiency of PAI in extending the shelf-life of perishable foods during storage (or/and transportation), by validating its antimicrobial properties and cooling capacity.

Plasma-Induced Superhydrophobicity as a Green Technology for Enhanced Air Gap Membrane Distillation.

Superhydrophobicity has only recently become a requirement in membrane fabrication and modification. Superhydrophobic membranes have shown improved flux performance and scaling resistance in long-term membrane distillation (MD) operations compared to simply hydrophobic membranes. Here, we introduce plasma micro- and nanotexturing followed by plasma deposition as a novel, dry, and green method for superhydrophobic membrane fabrication. Using plasma micro- and nanotexturing, commercial membranes, both hydrophobic and hydrophilic, are transformed to superhydrophobic featuring water static contact angles (WSCA) greater than 150° and contact angle hysteresis lower than 10°. To this direction, hydrophobic polytetrafluoroethylene (PTFE) and hydrophilic cellulose acetate (CA) membranes are transformed to superhydrophobic. The superhydrophobic PTFE membranes showed enhanced water flux in standard air gap membrane distillation and more stable performance compared to the commercial ones for at least 48 h continuous operation, with salt rejection >99.99%. Additionally, their performance and high salt rejection remained stable, when low surface tension solutions containing sodium dodecyl sulfate (SDS) and NaCl (down to 35 mN/m) were used, showcasing their antiwetting properties. The improved performance is attributed to superhydrophobicity and increased pore size after plasma micro- and nanotexturing. More importantly, CA membranes, which are initially unsuitable for MD due to their hydrophilic nature (WSCA ≈ 40°), showed excellent performance with stable flux and salt rejection >99.2% again for at least 48 h, demonstrating the effectiveness of the proposed method for wetting control in membranes regardless of their initial wetting properties.

How Different Are Fog Collection and Dew Water Harvesting on Surfaces with Different Wetting Behaviors?

As the clean water shortage becomes a serious problem for mankind, atmospheric water harvesting has emerged as a viable solution. Two main approaches to collect water from the atmosphere exist: the first is to capture it from fog, whereas the second is through condensation of vapor on surfaces with a temperature below the dew point. The water collection mechanism in these two modes is completely different. In this work, we develop a deeper understanding of the effect of surface wettability on gravity-assisted atmospheric water harvesting and a comparative study of the two collection modes (fog and dew). First, we present theoretical estimates for the maximum water mass available in each mode and introduce an efficiency factor η which enables the direct comparison among surfaces in different setups and modes. Then we fabricate a series of micronanostructured surfaces with different surface wetting properties from hydrophilic to superhydrophobic. Our results demonstrate that drop mobility, derived from the surface superhydrophobic properties and micronanotopography, is the most important factor affecting fog collection: superhydrophobic surfaces show 40-65% higher fog collection rates compared to flat hydrophilic surfaces, with the more mobile among superhydrophobic surfaces (hysteresis 2°, and air-liquid fraction fA-L > 0.9) showing higher water collection. On the other hand, dew harvesting efficiency depends on the combination of drop mobility and nucleation rate, with superhydrophobic surfaces exhibiting 40% higher water collection rate compared to the flat hydrophilic or hydrophobic surfaces due to their low hysteresis as well as high surface area available for nucleation.

Food container employing a cold atmospheric plasma source for prolonged preservation of plant and animal origin food products.

Cold Atmospheric Plasma is a non-thermal processing technology with great potential for application to food products as it can effectively reduce the microbial load, leading to substantial shelf-life extension. Herein, we present an easy-to-build and cost-effective Surface Dielectric Barrier Discharge (SDBD) plasma source adjusted to the plastic lid of a common commercial food container made of transparent glass. Implementation and evaluation of plasma treatment in real perishable food products such as sea bream fillets, fresh-cut rocket salads and fresh whole strawberries showed that such device might be efficiently used in-storage for the extension of their shelf-life.•Easy-to-build and cost-effective SDBD plasma source adjusted in a food container for generation of antimicrobial RONS in proximity to treated food product•Treatment of perishable food products by reducing their initial microbial load•In-storage treatment efficient for perishable food products shelf-life extension.

Poly-L-histidine coated microfluidic devices for bacterial DNA purification without chaotropic solutions.

We present a disposable polymeric microfluidic device capable of reversibly binding and purifying Salmonella DNA through solid phase extraction (SPE). The microfluidic channels are first oxygen plasma treated and simultaneously micro-nanotextured, and then functionalized with amine groups via modification with L-histidine or poly-L-histidine. L-Histidine and poly-L-histidine bind on the plasma treated chip surface, and are not detached when rinsing with DNA purification protocol buffers. A pH-dependent protocol is applied on-chip to purify Salmonella DNA, which is first bound on the protonated amines at a pH (5.0) lower than their pKa of surface amine-groups which is 6.0 and then released at a pH higher than the pKa value (10.5). It was found that modification with poly-L-histidine resulted in higher surface density of amine groups onto microfluidic channel. Using the chip modified with poly-L-histidine, high recovery efficiency of at least 550 ng of isolated Salmonella DNA as well as DNA purification from Salmonella cell lysates corresponding to less than 5000 cells or 0.026 ng of Salmonella DNA was achieved. The protocol developed does not require ethanol or chaotropic solutions typically used in DNA purification, which are known inhibitors for downstream operations such as polymerase chain reactions (PCR) and which can also attack some polymeric microfluidic materials. Therefore, the microfluidic device and the related protocol hold promise for facile incorporation in microfluidics and Lab-on-a-chip (LOC) platforms for pathogen detection or in general for DNA purification.

Three-dimensional (3D) hierarchical oxygen plasma micro/nanostructured polymeric substrates for selective enrichment of cancer cells from mixtures with normal ones.

The enrichment of cancer cell population when in mixtures with normal ones is of great importance for cancer diagnosis. In this work, poly(methyl methacrylate) films have been processed applying different oxygen plasma conditions to fabricate surfaces with structure height ranging from 22 to more than 2000 nm. The surfaces were then evaluated with respect to adhesion and proliferation of both normal and cancer human cells. In particular, normal skin and lung fibroblasts, and four different cancer cell lines, A431 (skin cancer), HT1080 (fibrosarcoma), A549 (lung cancer), and PC3 (prostate cancer), have been employed. It was found that adhesion and proliferation of cancer cells was favored when cultured onto the hierarchical micro/nanostructured surfaces as compared to untreated ones with the maximum values obtained for substrates treated at -100 V for 3 min. On the other hand, although the adhesion of normal fibroblasts was not influenced by the micro/nanostructured surfaces, their morphology and proliferation was significantly impaired, especially after 3-day culture on these surfaces. The reduced proliferation rate of adherent fibroblasts was linked to reduced focal points formation, as it was verified through vinculin staining, and not to apoptosis. The micro/nanostructured surfaces prepared with plasma treatment at -100 V for 3 min (hierarchical topography with mean height of ∼800 nm) were selected as substrates for normal and cancer cell co-culture experiments. It was found that 25-80 times enrichment of cancer over the normal cells was achieved on the nanostructured surfaces after 3-day culture, while it was 5-8 times lower on the untreated ones. It should be noticed that this is the first time such high enrichment ratios are achieved without implementing surfaces modified with binding molecules specific for cancer cells. Thus, the nanostructured surfaces hold a strong promise as culture substrates for separation and enrichment of cancer cells from mixtures with normal ones that should find application in cancer diagnostics.

3D Plasma Nanotextured® Polymeric Surfaces for Protein or Antibody Arrays, and Biomolecule and Cell Patterning.

Plasma micro-nanotexturing is a generic technology for topographical and chemical modification of surfaces and their implementation in microfluidics and microarrays. Nanotextured surfaces with desirable chemical functionality (and wetting behavior) have shown excellent biomolecule immobilization and cell adhesion. Specifically, nanotextured hydrophilic areas show (a) strong binding of biomolecules and (b) strong adhesion of cells, while nanotextured superhydrophobic areas show null adsorption of (a) proteins and (b) cells. Here we describe the protocols for (a) biomolecule adsorption control on nanotextured surfaces for microarray fabrication and (b) cell adhesion on such surfaces. 3D plasma nanotextured® substrates are commercialized through Nanoplasmas private company, a spin-off of the National Centre for Scientific Research Demokritos.

Micro-nano-bio acoustic system for the detection of foodborne pathogens in real samples.

The fast and efficient detection of foodborne pathogens is a societal priority, given the large number of food-poisoning outbreaks, and a scientific and technological challenge, given the need to detect as little as 1 viable cell in 25 gr of food. Here, we present the first approach that achieves the above goal, thanks to the use of a micro/nano-technology and the detection capability of acoustic wave sensors. Starting from 1 Salmonella cell in 25 ml of milk, we employ immuno-magnetic beads to capture cells after only 3 h of pre-enrichment and subsequently demonstrate efficient DNA amplification using the Loop Mediated Isothermal Amplification method (LAMP) and acoustic detection in an integrated platform, within an additional ½ h. The demonstrated 4 h sample-to-analysis time comes as a huge improvement to the current need of few days to obtain the same result. In addition, the work presents the first reported Lab-on-Chip platform that comprises an acoustic device as the sensing element, exhibiting impressive analytical features, namely, an acoustic limit of detection of 2 cells/µl or 3 aM of the DNA target and ability to detect in a label-free manner dsDNA amplicons in impure samples. The use of food samples together with the incorporation of the necessary pre-enrichment step and ability for multiple analysis with an internal control, make the proposed methodology highly relevant to real-world applications. Moreover, the work suggests that acoustic wave devices can be used as an attractive alternative to electrochemical sensors in integrated platforms for applications in food safety and the point-of-care diagnostics.

Hydrophobic and superhydrophobic surfaces fabricated using atmospheric pressure cold plasma technology: A review.

Hydrophobic surfaces are often used to reduce wetting of surfaces by water. In particular, superhydrophobic surfaces are highly desired for several applications due to their exceptional properties such as self-cleaning, anti-icing, anti-friction and others. Such surfaces can be prepared via numerous methods including plasma technology, a dry technique with low environmental impact. Atmospheric pressure plasma (APP) has recently attracted significant attention as lower-cost alternative to low-pressure plasmas, and as a candidate for continuous rather than batch processing. Although there are many reviews on water-repellent surfaces, and a few reviews on APP technology, there are hardly any review works on APP processing for hydrophobic and superhydrohobic surface fabrication, a topic of high importance in nanotechnology and interface science. Herein, we critically review the advances on hydrophobic and superhydrophobic surface fabrication using APP technology, trying also to give some perspectives in the field. After a short introduction to superhydrophobicity of nanostructured surfaces and to APPs we focus this review on three different aspects: (1) The atmospheric plasma reactor technology used for fabrication of (super)hydrophobic surfaces. (2) The APP process for hydrophobic surface preparation. The hydrophobic surface preparation processes are categorized methodologically as: a) activation, b) grafting, c) polymerization, d) roughening and hydrophobization. Each category includes subcategories related to different precursors used. (3) One of the most important sections of this review concerns superhydrophobic surfaces fabricated using APP. These are methodologically characterized as follows: a) single step processes where micro-nano textured topography and low surface energy coating are created at the same time, or b) multiple step processes, where these steps occur sequentially in or out of the plasma. We end the review with some perspectives in the field. We aspire to address scientists, who will get involved in the fields of (super)hydrophobicity and/or in atmospheric pressure plasma processing.

How to Achieve Reversible Electrowetting on Superhydrophobic Surfaces.

Collapse (Cassie to Wenzel) wetting transitions impede the electrostatically induced reversible modification of wettability on superhydrophobic surfaces, unless a strong external actuation (e.g., substrate heating) is applied. Here we show that collapse transitions can be prevented (the droplet remains suspended on the solid roughness protrusions) when the electrostatic force, responsible for the wetting modification, is smoothly distributed along the droplet surface. The above argument is initially established theoretically and then verified experimentally.

Three-dimensional (3D) plasma micro-nanotextured slides for high performance biomolecule microarrays: Comparison with epoxy-silane coated glass slides.

Glass slides coated with a poly(methyl methacrylate) layer and plasma micro-nanotextured to acquire 3D topography (referred as 3D micro-nanotextured slides) were evaluated as substrates for biomolecule microarrays. Their performance is compared with that of epoxy-coated glass slides. We found that the proposed three-dimensional (3D) slides offered significant improvements in terms of spot intensity, homogeneity, and reproducibility. In particular, they provided higher spot intensity, by a factor of at least 1.5, and significantly improved spot homogeneity when compared to the epoxy-silane coated ones (intra-spot and between spot coefficients of variation ranging between 5 and 15% for the 3D micro-nanotextured slides and between 25 and 85% for the epoxy-silane coated ones). The latter was to a great extent the result of a strong "coffee-ring" effect observed for the spots created on the epoxy-coated slides; a phenomenon that was severely reduced in the 3D micro-nanotextured slides. The 3D micro-nanotextured slides offered in addition higher signal to noise ratio values over a wide range of protein probe concentrations and shelf-life over one year without requirement for specific storage conditions. Finally, the protocols employed for protein probe immobilization were extremely simple.

Durable superhydrophobic and superamphiphobic polymeric surfaces and their applications: A review.

Wetting control is essential for many applications, such as self-cleaning, anti-icing, anti-fogging, antibacterial action as well as anti-reflection and friction control. While significant effort has been devoted to fabricate superhydrophobic/superamphiphobic surfaces (repellent to water and other low surface tension liquids), very few polymeric superhydrophobic/superamphiphobic surfaces can be considered as durable against various externally imposed stresses (e.g. application of heating, pressure, mechanical forces, chemical, etc.). Therefore, durability tests are extremely important for applications especially when such surfaces are made of "soft" materials. Here, we review the most recent and promising efforts reported towards the realization of durable, superhydrophobic/superamphiphobic, polymeric surfaces emphasizing the durability tests performed, and some important applications. We compare and put in context the scattered durability tests reported in the literature, and present conclusions, perspectives and challenges in the field.

Is There a Threshold in the Antibacterial Action of Superhydrophobic Surfaces?

The realization of antibacterial surfaces is an important scientific problem, which may be addressed by the use of superhydrophobic surfaces, reducing bacterial adhesion. However, there are several limitations and contradicting reports on the antibacterial efficacy of such surfaces. Moreover, achieving antibacterial action through minimization of adhesion does not ensure complete protection against bacteria. Here, we identify the important factors affecting antibacterial action on superhydrophobic surfaces, emphasizing the role of bacterial concentration, and observing an upper concentration threshold above which antibacterial action of any surface is compromised. Finally, we propose metal enriched, superhydrophobic surfaces, as the "ultimate" "hybrid" antibacterial surfaces for in vitro applications.

Plasma micro-nanotextured polymeric micromixer for DNA purification with high efficiency and dynamic range.

We present a polymeric microfluidic chip capable of purifying DNA through solid phase extraction. It is designed to be used as a module of an integrated Lab-on-chip platform for pathogen detection, but it can also be used as a stand-alone device. The microfluidic channels are oxygen plasma micro-nanotextured, i.e. randomly roughened in the micro-nano scale, a process creating high surface area as well as high density of carboxyl groups (COOH). The COOH groups together with a buffer that contains polyethylene glycol (PEG), NaCl and ethanol are able to bind DNA on the microchannel surface. The chip design incorporates a mixer so that sample and buffer can be efficiently mixed on chip under continuous flow. DNA is subsequently eluted in water. The chip is able to isolate DNA with high recovery efficiency (96± 11%) in an extremely large dynamic range of prepurified Salmonella DNA as well as from Salmonella cell lysates that correspond to a range of 5 to 1.9 × 108 cells (0.263 fg to 2 × 500 ng). The chip was evaluated via absorbance measurements, polymerase chain reaction (PCR), and gel electrophoresis.

Micro-Nano-Bio Diagnostic System for Food Pathogen Detection Revolutionizes Food Safety Management & Protects Consumers Health.

The development of integrated, fast and affordable platforms for pathogen detection is an emerging area where a multidisciplinary approach is necessary for designing microsystems employing miniaturized devices; these new technologies promise a significant advancement of the current state of analytical testing leading to improved healthcare. In this work, the development of a lab-on-chip microsystem platform for the genetic analysis of Salmonella in milk samples is presented. The heart of the platform is an acoustic detection biochip, integrated with a microfluidic module. This detection platform is combined with a micro-processor, which, alongside with magnetic beads technology and a DNA micro-amplification module, are responsible for performing sample pre-treatment, bacteria lysis, nucleic acid purification and amplification. Automated, multiscale manipulation of fluids in complex microchannel networks is combined with novel sensing principles developed by some of the partners. This system is expected to have a significant impact in food-pathogen detection by providing for the first time an integrated detection test for Salmonella screening in a very short time. Finally, thanks to the low cost and compact technologies involved, the proposed set-up is expected to provide a competitive analytical platform for direct application in field settings.

Superamphiphobic polymeric surfaces sustaining ultrahigh impact pressures of aqueous high- and low-surface-tension mixtures, tested with laser-induced forward transfer of drops.

Superamphiphobic, (quasi-)ordered plasma-textured surfaces, coated with a perfluorinated monolayer, exhibit extreme resistance against drop-pinning for both water-like and low-surface-tension mixtures (36 mN m(-1)). The highest values reported here are 36 atm for a water-like mixture, 5 times higher than previously reported in the literature, and 7 atm for a low-surface-tension mixture, the highest ever reported value for lotus-leaf-inspired surfaces.