Surface Wettability of Cellulose Sponges on Effective Oil Uptake.

Designing absorbents having specific wettability toward both oil and water is the key for selective and effective oil absorption and removal. For this purpose, establishing explicit correlations between surface tension of oils and surface wettability of absorbent is crucial. In this study, we modified common low-cost cellulose sponges with various organosilanes to achieve a range of hydrophobicity/oleophilicity and then assessed their oil uptake selectivity and capability. Oil uptake was followed as mass uptake versus time and analyzed based on the spreading coefficient (S) of a liquid over a solid surface. The results showed that sponges needed to be hydrophobic, not necessarily superhydrophobic, to selectively absorb oil from an oil/water mixture. To achieve a fast uptake and a high uptake capacity, an S ≥ 0 was necessary, that is, when the sponges were completely wet by the oil. Increasing the porosity of cellulose sponge led to a slight increase in oil uptake capacity, and a greater increase resulted when bacterial cellulose sponges that consisted of smaller and more uniform voids/pores were used. S ≥ 0 could be used as a criterion for evaluating effective and rapid oil uptake for porous absorbents, especially for those containing heterogeneous pore structures, such as common cellulose sponges.

Thermoresponsive Poly(vinyl methyl ether) (PVME) Retained by 3-Aminopropyltriethoxysilane (APTES) Network.

Thermoresponsive polymers (TRP)s have been widely used for various applications from controlling membrane fouling in separation to cell/cell sheet harvesting in regenerative medicine. While poly(N-isopropylacrylamide) (pNIPAAm) is the most commonly used TRP, less expensive and easily processed poly(vinyl methyl ether) (PVME) also shows a hydrophilic to hydrophobic transition at 32-35 °C, near physiological conditions. In this study, we investigated the processing conditions for retaining a stable layer of PVME thin film on silica surfaces via entrapment in a 3-aminopropyltriethoxysilane (APTES) network. In addition, the thermoresponsive behaviors (TRB) of the retained PVME films were evaluated. Blend thin films of PVME/APTES with 90:10 and 50:50 mass ratios were spin-coated from their solutions in ethanol under ambient conditions and then annealed in a vacuum oven at 40, 60, 80, or 120 °C for 1, 2, or 3 days. The annealed films were then thoroughly rinsed with room temperature water and then soaked in water for 3 days. Our results showed that annealing at a temperature of ≥40 °C was necessary for retaining a PVME film on the surface. The higher annealing temperature led to greater film retention, probably due to the formation of a tighter APTES network. Regardless of processing conditions, all retained PVME films showed TRB, determined by water contact angles below and above the transition temperature of PVME. Additionally, particle attachment and protein adsorption on retained PVME films showed lower attachment or adsorption at room temperature as compared to that at 37 °C, and a greater difference was observed for the 90:10 blend where more PVME was consisted. Furthermore, human mesenchymal stem cells attached and proliferated on the retained PVME surfaces at 37 °C and rapidly detached at room temperature. These results illustrated the potential applications of PVME surfaces as thermoresponsive supports for low-fouling applications and noninvasive cell harvesting.

Retention of poly(N-isopropylacrylamide) on 3-aminopropyltriethoxysilane.

Silane coupling agents are commonly employed to link an organic polymer to an inorganic substrate. One of the widely utilized coupling agents is 3-aminopropyltriethoxy silane (APTES). In this study, the authors investigated the ability of APTES to retain thermo-responsive poly(N-isopropylacrylamide) (pNIPAAm) on hydroxylated surfaces such as glass. For comparison purposes, the authors also evaluated the retention behaviors of (1) polystyrene, which likely has weaker van der Waals interactions and acid-base interactions (contributed by hydrogen-bonding) with APTES, on APTES as well as (2) pNIPAAm on two other silane coupling agents, which have similar structures to APTES, but exhibit less interaction with pNIPAAm. Under our processing conditions, the stronger interactions, particularly hydrogen bonding, between pNIPAAm and APTES were found to contribute substantially to the retention of pNIPAAm on the APTES modified surface, especially on the cured APTES layer when the interpenetration was minimal or nonexistent. On the noncured APTES layer, the formation of an APTES-pNIPAAm interpenetrating network resulted in the retention of thicker pNIPAAm films. As demonstrated by water contact angles [i.e., 7°-15° higher at 40 °C, the temperature above the lower critical solution temperature (LCST) of 32 °C for pNIPAAm, as compared to those at 25 °C] and cell attachment and detachment behaviors (i.e., attached/spread at 37 °C, above LCST; detached at 20 °C, below LCST), the retained pNIPAAm layer (6-15 nm), on both noncured and cured APTES, exhibited thermo-responsive behavior. The results in this study illustrate the simplicity of using the coupling/adhesion promoting ability of APTES to retain pNIPAAm films on hydroxylated substrates, which exhibit faster cell sheet detachment (≤30 min) as compared to pNIPAAm brushes (in hours) prepared using tedious and costly grafting approaches. The use of adhesion promoters to retain pNIPAAm provides an affordable alternative to current thermo-responsive supports for cell sheet engineering and stem cell therapy applications.

Cross-linked polystyrene sulfonic acid and polyethylene glycol as a low-fouling material.

A negatively charged hydrophilic low fouling film was prepared by thermally cross-linking a blend consisting of polystyrene sulfonic acid (PSS) and polyethylene glycol (PEG). The film was found to be stable by dip-washing. The fouling resistance of this material toward bacterial (Escherichia coli) and colloidal (polystyrene particles) attachment, non-specific protein (fibronectin) adsorption and cell (3T3 NIH) adhesion was evaluated and was compared with glass slides modified with polyethylene glycol (PEG) brushes, oxidized 3-mercaptopropyltrimethoxysilane (sulfonic acid, SA), and n-octadecyltrichlorosilane (OTS). The extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) theory and thermodynamic models based on surface energy were used to explain the interaction behaviors of E. coli/polystyrene particles-substrate and protein-substrate interactions, respectively. The cross-linked PSS-PEG film was found to be slightly better than SA and PEG toward resisting non-specific protein adsorption, and showed comparable low attachment results as those of PEG toward particle, bacterial and NIH-3T3 cells adhesion. The low-fouling performance of PSS-PEG, a cross-linked film by a simple thermal curing process, could allow this material to be used for applications in aqueous environments, where most low fouling hydrophilic polymers, such as PSS or PEG, could not be easily retained.

Dewetting based fabrication of fibrous micro-scaffolds as potential injectable cell carriers.

Although regenerative medicine utilizing tissue scaffolds has made enormous strides in recent years, many constraints still hamper their effectiveness. A limitation of many scaffolds is that they form surface patches, which are not particularly effective for some types of "wounds" that are deep within tissues, e.g., stroke and myocardial infarction. In this study, we reported the generation of fibrous micro-scaffolds feasible for delivering cells by injection into the tissue parenchyma. The micro-scaffolds (widths<100µm) were made by dewetting of poly(lactic-co-glycolic acid) thin films containing parallel strips, and cells were seeded to form cell/polymer micro-constructs during or post the micro-scaffold fabrication process. Five types of cells including rat induced vascular progenitor cells were assessed for the formation of the micro-constructs. Critical factors in forming fibrous micro-scaffolds via dewetting of polymer thin films were found to be properties of polymers and supporting substrates, temperature, and proteins in the culture medium. Also, the ability of cells to attach to the micro-scaffolds was essential in forming cell/polymer micro-constructs. Both in vitro and in vivo assessments of injecting these micro-scaffolding constructs showed, as compared to free cells, enhanced cell retention at the injected site, which could lead to improved tissue engineering and regeneration.

In vitro behaviors of rat mesenchymal stem cells on bacterial celluloses with different moduli.

Compressive moduli of bacteria-synthesized cellulose (BC) were altered by two drying techniques: ambient-air drying and freeze drying. While no significant differences in dry weight were found, their cross-sectional structures and thickness varied greatly. Freeze dried BCs had loose cross-sectional structures and a thickness of ~4.7 mm, whereas air dried BCs had more compacted cross-sectional structures and a thickness of ~0.1mm. The compressive moduli of the rehydrated freeze dried and rehydrated air dried BCs were measured to be 21.06±0.22 kPa and 90.09±21.07 kPa, respectively. When rat mesenchymal stem cells (rMSCs) were seeded on these BCs, they maintained a round morphology in the first 3 days of cultivation. More spread-out morphology and considerable proliferation on freeze dried BCs were observed in 7 days, but not on air-dried BCs. The cells were further grown for 3 weeks in the absence and presence of differentiation agents. Without using any differentiation agents, no detectable differentiation was noticed for rMSCs further cultivated on both types of BC. With differentiation inducing agents, chondrogenic differentiation, visualized by histological staining, was observed in some area of the rehydrated freeze dried BCs; while osteogenic differentiation was noticed on the stiffer rehydrated air dried BCs.

Effects of shear on initial bacterial attachment in slow flowing systems.

Initial bacterial attachment, likely affected by local shear, could influence biofilm formation. However, there are contradictory reports for the shear effects on attachment of different bacteria onto different surfaces. In this study, four bacteria, Staphylococcus epidermidis, Pseudomonas aeruginosa, Pseudomonas putida, and Escherichia coli, were examined for their attachment to glass and octadecyltrichlorosilane (OTS) modified glass under different shears. Polystyrene particles were used to verify that their shear dependent attachment on glass and OTS could be interpreted using an analysis based on the extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) theory. In particular, the critical shear force (F(c-shear)) could correlate with the maximum attractive force (F(MAX)(XDLVO)) toward the secondary energy minimum as F(c-shear)=cF(MAX)(XDLVO). For these particles, c of ~1 was obtained, the value was within the coefficient range (0.1-1) of substances sliding over glass. For S. epidermidis, E. coli and P. aeruginosa on glass, c was 0.3, <0.6 and 0.2, respectively. When considering potential protein adsorption on OTS during bacterial attachment, c of these species on OTS was slightly above 1. A greatly enhanced attachment of P. aeruginosa on OTS was also observed, probably due to the presence of flagella. For P. putida, the attachment first decreased slightly or maintained with shear and then increased. Such behaviors were probably caused by the increased secretion of extracellular polymeric substances (EPS) at higher shears by P. putida. The results from this study suggested that, without complications from surface features/EPS, the analysis based on the XDLVO theory could provide a basis for understanding shear effect on initial bacterial attachment.

Rapid cell sheet detachment using spin-coated pNIPAAm films retained on surfaces by an aminopropyltriethoxysilane network.

The ability to harvest cell sheets grown on thermoresponsive polymers, such as poly(N-isopropylacrylamide) (pNIPAAm), has been widely studied for use in tissue engineering applications. pNIPAAm is of special interest because of the phase change that it undergoes in a physiologically relevant temperature range. Two primary approaches have been adopted to graft pNIPAAm chains covalently onto tissue culture polystyrene dishes: electron beam irradiation and plasma polymerization. These approaches often involve non-easily accessible (e.g. e-beam) facilities and complicated procedures that have hindered most tissue culture laboratories in adopting this technology for their specific applications. In this study, we developed a simple and cost-effective approach to create thermoresponsive surfaces using commercially available pNIPAAm. Using a simple spin-coating technique, thermoresponsive thin films were deposited on glass slides or silicon wafers using pNIPAAm blended with a small amount of 3-aminopropyltriethoxysilane (APTES), which enhances the retention of pNIPAAm on the surface. We found that the thermoresponsive films created using our method support cell attachment and proliferation without additional adhesive proteins as well as cell sheet detachment within minutes.

Effects of rhamnolipids and shear on initial attachment of Pseudomonas aeruginosa PAO1 in glass flow chambers.

BACKGROUND, AIM, AND SCOPE: Solid surfaces in contact with water have been found to be biofouled due to the attachment of various organisms. For better understanding of the biofilm formation, the important initial stage of bacterial attachment was investigated with Pseudomonas aeruginosa PAO1 as a model microorganism. Effects of the biosurfactant rhamnolipids and the shear conditions were particularly examined. MATERIALS AND METHODS: A highly reproducible procedure was employed. The procedure involved monitoring and counting the number of attached cells on glass walls of the flow chambers, through which a PAO1 suspension was circulated and, subsequently, a saline solution was passed for washing. The experiments were made under different circulation rates (exerting different shear on the bacteria) and rhamnolipid concentrations. RESULTS AND DISCUSSIONS: Reproducibility of the procedure was confirmed. The velocity profiles near the flow chamber wall were determined. Rhamnolipids, even at a very low concentration of 13 mg/l, were found to deter the bacterial attachment substantially. Prewashing the cells with a 100 mg/l rhamnolipid solution, however, did not affect the attachment significantly. As for the effect of shear, the PAO1 attachment showed an increasing-then-decreasing trend in the range investigated, i.e., 1.0 to 26 mN/m(2) shear stresses at the chamber wall. The diffusion-limited transport of cells to the chamber wall might have contributed to, but could not fully explain, the increasing attachment observed in the very low shear range (up to 3.5-5.0 mN/m(2)). CONCLUSIONS: As compared to static systems, the flow chamber systems significantly improved the reproducibility of initial attachment results. Flow chamber systems were more suitable for experimental investigations of bacterial attachment to surfaces. Rhamnolipids were found to be potent antifoulants for PAO1 attachment on glass. The initial cell attachment increased with increasing shear at the very low shear range (up to 3.5-5.0 mN/m(2)), but the attachment could be minimized with further increase of the shear.

Porous polymer films templated by marangoni flow-induced water droplet arrays.

In this article, we report the development of a novel, simple, and cost-effective method for fabricating porous polymer films with controllable interpore distances, pore sizes, and arrangements using water droplets induced by Marangoni flow as templates. First, a spread-thin ethanol film on a partially water-wettable substrate is exposed to a humid airflow, facilitating the evaporation and recession of the ethanol film. Meanwhile, water in the airflow condenses on the ethanol film and accumulates near the receding contact line, which induces the formation of water fingers at the receding contact line and, finally, ordered arrays of water droplets after detachment. The formation of the hexagonal and square arrays of water droplets is due to the pinning and sliding of the water fingers on the silicone oxide (SiOx) and silicon (Si) substrates, respectively. By varying the thickness of the ethanol film spread on the Si substrate, the sliding velocity of water fingers can be tuned, subsequently leading to the fabrication of other arrangements. The interdroplet distance and droplet size can be dependently controlled by tuning the humidity of the airflow. The ordered arrays of water droplets on the substrate are then utilized to fabricate porous polymer films by dip-coating the substrate with a polystyrene solution. Films with hexagonal and square (and other arrangements) arrays of pores are fabricated on the silicon oxide (SiOx) and silicon (Si) substrates, respectively. The pore size can also be independently tuned by further condensation or evaporation of formed water droplets, leading to porous polymer films with both close- and non-close-packed arrays of pores. The ordered porous polymer films can be further used as templates for fabricating metal post patterns.

Marangoni flow-induced self-assembly of hexagonal and stripelike nanoparticle patterns.

We have developed a simple Marangoni flow-induced method for self-assembling nanoparticles (NPs) into both hexagonal and stripelike patterns. First, a NPs/ethanol suspension was spread on a slightly nonwettable and a wettable silicon oxide substrate. The Marangoni flow, induced by simultaneous evaporation of ethanol and condensation of water, leads to the formation of the corresponding hexagonal distributed circular NP rings and dotted stripes. The inter-ring spacing and ring size of the hexagonal patterns can be tuned by varying the relative humidity of the N2 stream blown over the slightly nonwettable substrate. Hexagonal patterns of circular NP patches can also be fabricated by lowering the evaporation of the condensed water droplets. On the wettable substrate, complex patterns result when the humidity of the N2 stream changes.

Evaluation of toxicity of capsaicin and zosteric acid and their potential application as antifoulants.

The toxicity of two natural product antifoulants, capsaicin and zosteric acid, was evaluated using the Microtox assay and a static toxicity test. The EC50 values obtained from the Microtox assay for capsaicin and zosteric acid were 11.75 +/- 1.02 and 442 +/- 100 mg/L, respectively. The static toxicity test, conducted with freshwater organisms, yielded capsaicin EC50 values of 5.5 +/- 0.5 and 23 +/- 2.0 mg/L for P. putida and Lake Erie bacteria, respectively. Zosteric acid EC50 values were 167 +/- 3.9 and 375 +/- 10 mg/L for P. putida and Lake Erie bacteria, respectively. Tests with marine organisms resulted in capsaicin EC50 values of 6.9 +/- 0.2 and 15.6 +/- 0.4 mg/L for V. natriegens and V. parahaemolyticus, respectively; whereas zosteric acid EC50 values were 7.4 +/- 0.1 and 18 +/- 0.6 mg/L for V. natriegens and V. parahaemolyticus, respectively. These results indicate that zosteric acid is much less toxic than capsaicin and that both are substantially less toxic than the currently used antifoulants, such as TBT (EC50 < 0.01 ppb). Their effectiveness as natural antifoulants was demonstrated by preliminary attachments studies. As the aqueous antifoulant concentration increased, significant inhibition of bacteria attachment or prevention of biofilm formation was achieved. Hence, both capsaicin and zosteric acid could be attractive alternatives as new antifouling compounds.

Self-assembling of polymer-enzyme conjugates at oil/water interfaces.

Interface-binding enzymes are desirable for biphasic reactions in that they offer simultaneous access to substrates dissolved in both phases across the interface. It has been shown that conjugating water-soluble enzymes with hydrophobic polymers facilitated the assembling of enzymes at oil/water interfaces. In this work, the interfacial assembling of alpha-chymotrypsin conjugated with polystyrene, poly(methyl methacrylate), and poly(l-lactic acid) was examined using the pendant drop method. The interface-assembling process of the conjugates from the organic phase followed a similar pattern of that of native alpha-chymotrypsin from the aqueous buffer phase, i.e., the interfacial tension decreased gradually with time. However, when the conjugates were dispersed in the form of particulates in the aqueous phase, in which the conjugate was insoluble, the assembling occurred faster and the interfacial tension quickly approached zero. It was suspected that the assembling in this case involved two steps, i.e., the adsorption of the particulates and the subsequent rearrangement, dissociation, and redispersion of the conjugates at the interface. The effect of other factors, including the polarity of organic solvent and pH and ionic strength of the aqueous phase, was evaluated. It was found that the polar solvent slightly facilitated the assembling, whereas pH and ionic strength showed minimal effects.