Biomimetic Hydrophobic Surfaces with Low or High Adhesion Based on Poly(vinyl alcohol) and SiO2 Nanoparticles

Superhydrophobic surfaces are often found in nature,such as plant leaves and insect wings.Inspired by superhydrophobic phenomenon of the rose petals and the lotus leaves,biomimetic hydrophobic surfaces with high or low adhesion were prepared with a facile drop-coating approach in this paper.Poly(vinyl alcohol) (PVA) was used as adhesive and SiO2 nanoparticles were used to fabricate surface micro-structure.Stearic acid or dodecafluoroheptyl-propyl-trimethoxysilane (DFTMS) were used as low surface energy materials to modify the prepared PVA/SiO2 coating surfaces.The effects of size of SiO2 nanoparticles,concentration of SiO2 nanoparticle suspensions and the modifications on the wettability of the surface were investigated.The morphology of the PVA/SiO2 coating surfaces was observed by using scanning electron microscope.Water contact angle of the obtained superhydrophilic surface could reach to 3°.Stearic acid modified PVA/SiO2 coating surfaces showed hydrophobicity with high adhesion.By mixing the SiO2 nanoparticles with sizes of 40 nm and 200 nm and modifying with DFTMS,water contact angle of the obtained coating surface could be up to 155° and slide angle was only 5°.This work provides a facile and useful method to control surface wettability through changing the roughness and chemical composition of a surface.     

Cloth bioreactor containing yeast cells immobilized on cotton cloth using polythylenimine

Summary Yeast cells were immobilized by adhesion to cotton cloth using polyethylenimine. Yeast cells could be adhered to cotton cloth by coating either the yeast cells or cotton cloth or both with polyethylenimine. Adhered cells were not desorbed by washing with 1 M KCl or 50% (v/v) ethylene glycol or 0.1 M buffers pH 3.6–8.0. Also presence of salt (1 M) in the cell suspension was not found to alter the binding capacity of the cells. The yeast cell bound cloth was used in a specially designed frame reactor (2.3 L) for the repeated inversion of sucrose in 16 batches over a period of 3 weeks still retaining about 80% of the original invertase activity.     

Design of a thermostable cell adhesion protein

The design principle of a thermostable functional protein has been proposed by demonstrating genetic engineering synthesis of a thermostable cell adhesion protein. The cell adhesive peptide sequence, Arg-Gly-Asp (RGD), was incorporated into the elastin-based polyhexapeptide, whose repeating unit is Ala-Pro-Gly-Val-Gly-Val (APGVGV). The resulting protein possesses cell adhesion activity approximately 80% of fibronectin. After autoclaving at 120°C for 20 min, the protein retained over 90% of cell adhesion activity, while the activity of autoclaved fibronectin decreased to 50%.     

Immobilization of Bacteria on to Polyester Cloth

Escherichia coli 3300 (constitutive for -galactosidase) had a greater adhesion (assayed by -galactosidase) to polyester cloth than to cotton cloth. Adhered bacteria developed immobilized cells on the cloth which can generate progeny efficiently. Similarly, four other Gram-negative bacteria and three Gram-positive bacteria had greater adhesion to polyester cloth than to cotton cloth. Since coating of polyester cloth with polyvinyl alcohol greatly decreased the bacterial adhesion, hydrophobic interaction is likely responsible for the bacterial adhesion to polyester cloth.     

The Influence of Biosurfactants Released by S. mitis BMS on the Adhesion of Pioneer Strains and Cariogenic Bacteria

The influence of Streptococcus mitis BMS biosurfactants on the adhesion of eight pioneer and four cariogenic oral bacterial strains was, for a first screening, examined in a microtiter plate assay. The adhesion to pellicle-coated wells of three cariogenic strains was inhibited > 70% by the biosurfactants, while only one pioneer strain showed > 70% reduction. The reduction for the other strains did not exceed 50%. Subsequently, adhesion of Streptococcus mutans ATCC 25175 and Streptococcus sobrinus HG 1025, both cariogenic strains, and Actinomyces naeslundii T14V-J1 and Streptococcus oralis J22, two pioneer strains, to biosurfactants-coated enamel with and without a salivary pellicle was studied in a parallel plate flow chamber. A biosurfactants coating to enamel with or without a pellicle caused a reduction in the number of adhering cariogenic organisms, although no such reduction was observed for the pioneer strains. Consequently, it is concluded that S. mitis BMS biosurfactants may play a protective role against adhesion of cariogenic bacteria.     

Temporal and spatial variations in macrofouling of silicone fouling‐release coatings

Nontoxic, low surface free energy silicone coatings having reduced biofouling adhesion strength have been developed as an alternative to antifouling paints. Silicone coatings permit macrofouling to adhere; however, fouling can be removed easily by water pressure or light scrubbing. One of the current methods used to evaluate the performance of non‐toxic silicone fouling‐release coatings relies heavily on fouling coverage. The organismal community structure as well as total coverage can affect the ease of fouling removal from these coatings. This paper explores fouling coverage and organismal adhesion over time. Long‐term fouling coverage data were collected at four sites (in Massachusetts, Hawaii and Florida) using static immersion panels coated with silicone and oil‐amended silicone systems. Inter‐site differences in fouling coverage and community structure were observed for each coating. Intra‐site variation and temporal change in coverage of fouling was minimal, regardless of coating formulation. The extent of coverage was affected by the duration of immersion and the local environmental conditions; these factors may also have an impact on the foul‐release capability of the silicone coatings. Organismal adhesion data was collected in Hawaii and Florida. These adhesion measurements were used as a tool to discriminate and rank fouling release coatings.     

Effect of nitrogen addition on the performance of microbial fuel cell anodes

Carbon cloth anodes were modified with 4(N,N-dimethylamino)benzene diazonium tetrafluoroborate to increase nitrogen-containing functional groups at the anode surface in order to test whether the performance of microbial fuel cells (MFCs) could be improved by controllably modifying the anode surface chemistry. Anodes with the lowest extent of functionalization, based on a nitrogen/carbon ratio of 0.7 as measured by XPS, achieved the highest power density of 938 mW/m². This power density was 24% greater than an untreated anode, and similar to that obtained with an ammonia gas treatment previously shown to increase power. Increasing the nitrogen/carbon ratio to 3.8, however, decreased the power density to 707 mW/m². These results demonstrate that a small amount of nitrogen functionalization on the carbon cloth material is sufficient to enhance MFC performance, likely as a result of promoting bacterial adhesion to the surface without adversely affecting microbial viability or electron transfer to the surface.     

Effect of flagella expression on adhesion of Achromobacter piechaudii to chalk surfaces

Aims: To examine flagella role and cell motility in adhesion of Achromobacter piechaudii to chalk. Methods and Results: Transmission electron microscopy revealed that stationary cells have thicker and longer flagella than logarithmic cells. SDS‐PAGE analysis showed that flagellin was more abundant in stationary cells than logarithmic ones. Sonication or inhibition of flagellin synthesis caused a 30% reduction in adhesion to chalk. Preincubation of chalk with flagella extracts reduced adhesion, by 50%. Three motility mutants were isolated. Mutants 94 and 153 were nonmotile, expressed normal levels of flagellin, have regular flagella and exhibited reduced adhesion. Mutant 208 expressed low levels of flagellin, no flagella and a spherical cell shape but with normal adhesion capacity. Conclusions: Multiple cell surface factors affect the adhesion efficiency to chalk. Flagella per se through physical interaction and through cell motility contribute to the adhesion process. The adhesion behaviour of mutant 208 suggests that cell shape can compensate for flagellar removal and motility. Significance and Impact of the Study: Physiological status affects bacterial cell surface properties and hence adhesion efficiency to chalk. This interaction is essential to sustain biodegradation activities and thus, remediation of contaminated chalk aquifers.     

Improving biomaterial properties of collagen films by chemical modification

Films of bovine collagen were chemically modified with the goal of improving their biomaterial properties. The modified films were investigated with respect to their affinity to fibroblast and endothelial cells, as well as their antibacterial properties tested by adhesion of Staphylococcus aureus. Modifications that only change the net charge of collagen, such as acetylation, succinylation, and treatment with glutaraldehyde (all increase the negative charge), and amination with ethylenediamine (EDA), N,N-dimethyl-EDA (DMEDA), or butylamine (all increase the positive charge), did not dramatically alter the mammalian cell attachment to the film. In contrast, derivatization of collagen using methoxypoly(ethylene glycol) (PEG) diminished the attachment of fibroblasts by 98 +/- 1% and of endothelial cells by more than 99% compared to unmodified collagen. Moreover, the rate of growth of fibroblasts dropped by 97 +/- 1% and that of endothelial cells by 88 +/- 3% as a result of PEGylation of collagen. Adhesion of S. aureus cells also plummeted by 93 +/- 2% as a result of this PEGylation. With these antifouling properties, PEG-collagen may be a promising coating material for coronary stents. Subsequent derivatization of PEG-collagen with EDA or DMEDA abolished its mammalian cell-repelling ability, whereas bacterial cell repulsion was partially retained: for example, DMEDA-modified PEG-collagen exhibits up to a 5-fold lower bacterial adhesion than collagen. It is worth noting that a material that allows mammalian cell attachment but reduces bacterial adhesion could be useful as an implant or coating.     

A Glycoprotein Expressed by Human Fibrous Astrocytes is a Hyaluronate-Binding Protein and a Member of the CD44 Family

We have isolated and characterized an antigen from normal human brain called p80, so called because it migrated with an Mr of 80 kDa on SDS PAGE. The Mr of 80 kDa consists of a protein of about 55-60 kDa and carbohydrate (20-25 kDa). The carbohydrate is almost entirely of the N-linked type, although a small amount of O-linked carbohydrate was detected. Cross-reactivity with monoclonal antibodies A3D8 and A1G3 showed that p80 could therefore be considered an isoform of the CD44 adhesion molecules. In addition, specific binding to hyaluronate which was not competed for by proteoglycan demonstrated that it involved different sites than the proteoglycan binding sites. We also observed that fucoidan and dextran sulphate increased the binding by 200-250% while chondroitin sulphate C also increased the binding but to a lesser extent. Heparin, heparan sulphate and chondroitin sulphates A and B did not have such an effect. The binding of p80 to hyaluronate was pH dependent with a maximum at pH 6.4. We concluded that p80 was an astrocyte specific adhesion molecule.     

Inhibition of Escherichia coli and Proteus mirabilis adhesion and biofilm formation on medical grade silicone surface

Silicone has been utilized extensively for biomedical devices due to its excellent biocompatibility and biodurability properties. However, its surface is easily colonized by bacteria which will increase the probability of nosocomial infection. In the present work, a hydrophilic antimicrobial carboxymethyl chitosan (CMCS) layer has been grafted on medical grade silicone surface pre-treated with polydopamine (PDA). The increase in hydrophilicity was confirmed from contact angle measurement. Bacterial adhesion tests showed that the PDA-CMCS coating reduced the adhesion of Escherichia coli and Proteus mirabilis by ≥ 90%. The anti-adhesion property was preserved even after the aging of the functionalized surfaces for 21 days in phosphate-buffered saline (PBS), and also after autoclaving at 121°C for 20 min. Both E. coli and P. mirabilis readily form biofilms on the pristine surface under static and flow conditions but with the PDA-CMCS layer, biofilm formation is inhibited. The flow experiments indicated that it is more difficult to inhibit biofilm formation by the highly motile P. mirabilis as compared to E. coli. No significant cytotoxicity of the modified substrates was observed with 3T3 fibroblasts.     

A novel approach in the assessment of polymeric film formation and film adhesion on different pharmaceutical solid substrates

The purpose of this study was to evaluate the nature of film formation on tablets with different compositions, using confocal laser scanning microscopy (CLSM), and to measure film adhesion via the application of a novel “magnet probe test”. Three excipients, microcrystalline cellulose (MCC), spray-dried lactose monohydrate, and dibasic calcium phosphate dihydrate, were individually blended with 0.5% magnesium stearate, as a lubricant, and 2.5% tetracycline HCl, as a fluorescent marker, and were compressed using a Carver press. Tablets were coated with a solution consisting of 7% hydroxypropyl methylcellulose (HPMC) phthalate (HP-55), and 0.5% cetyl alcohl in acetone and isopropanol (11:9). The nature of polymer interaction with the tablets and coating was evaluated using CLSM and a designed magnet probe test. CLSM images clearly showed coating efficiency, thickness, and uniformity of film formation, and the extent of drug migration into the film at the coating interfaces of tablets. Among the excipients, MCC demonstrated the best interface for both film formation and uniformity in thickness relative to lactose monohydrate and dibasic calcium phosphate dihydrate. The detachment force of the coating layers from the tablet surfaces, as measured with the developed magnet probe test, was in the order of MCC>lactose monohydrate>dibasic calcium phosphate dihydrate. It was also shown that the designed magnet probe test provides reliable and reproducible results when used for measurement of film adhesion and bonding strength.     

拟目乌贼受精卵孵化及室内附卵基附卵效果的研究

在实验室条件下,对福建沿海的拟目乌贼受精卵进行了孵化实验,研究结果表明,在水温为21℃~30℃,盐度为24~33条件下,受精卵均能孵化。在盐度为30的条件下,最适温度为24℃,孵化率达（93.3±2.9）%,孵化高峰期为23~24 d;在温度为24℃的条件下,最适盐度为30,孵化率达（93.3±2.9）%,孵化高峰期为23~24 d。不同附卵基的附卵效果研究结果表明：不同附卵基的附卵效果差异显著（P〈0.05）,网目为27 mm的聚乙烯网片附卵效果最佳,平均附卵量为193.4粒,平均附卵率32.51%;其次是15 mm的网片,平均附卵量为137.5粒,平均附卵率23.14%。直径为8 mm和10 mm的尼龙绳附卵效果也较好,平均附卵量分别为119.6粒和120.0粒,平均附卵率分别为20.09%和20.13%;直径为2 mm和4 mm的尼龙绳以及网目为2.5 mm的网片的附卵效果明显较差,平均附卵率分别为0.92%、1.45%和1.76%,基本不附卵。无论是尼龙绳还是网片,都是以直径或网目较大的附卵效果较好。     

Pemphigus vulgaris antigen, a desmoglein type of cadherin, is localized within keratinocyte desmosomes

Pemphigus vulgaris antigen (PVA) is a member of the desmoglein subfamily of cadherin cell adhesion molecules. Because autoantibodies in this disease cause blisters due to loss of epidermal cell adhesion, and because desmoglein is found in the desmosome cell adhesion junction, we wanted to determine if PVA is also found in desmosomes. By immunofluorescence, PV IgG bound, in a dotted pattern, to the cell surface of cultured human keratinocytes induced to differentiate with calcium, suggesting junctional staining. However, by preembedding, immunogold electron microscopic studies only slight labeling could be detected in desmosomes, presumably because of difficulty in gold penetration of intact desmosomes. We therefore treated the keratinocytes with 0.01% trypsin in 1 mM calcium, conditions known to preserve cadherin antigenicity but that caused slight separation of desmosomes, before immunogold staining. In this case there was extensive labeling of the extracellular part of desmosomes but not of the interdesmosomal cell membrane which was stained with anti-beta 2- microglobulin antibodies. To confirm the specificity of this binding we showed that antibodies raised in rabbits against the extracellular portions of PVA also bound desmosomes in these cultures. In intact mouse epidermis we could also show slight, but specific, immunogold desmosomal labeling with PV IgG. Furthermore, neonatal mice injected with PV IgG affinity purified on PVA showed desmosomal separation with the IgG localized to desmosomal cores. These results indicate that PVA is organized and concentrated within the desmosome where it presumably functions to maintain the integrity of stratifying epithelia.     

Production of cellulase by Aspergillus niger biofilms developed on polyester cloth

AIMS: To compare cellulase production by Aspergillus niger ATCC 10864 biofilms on polyester cloth and freely suspended cultures in shaken flasks and microbioreactors of bubble column type. METHODS AND RESULTS: Both shaken flasks and oxygenated microbioreactors containing 40 ml of production medium were used to compare cellulase secretion by free mycelium and biofilm cultures. Free mycelium cultures grew better in flasks than in microbioreactors producing compact and fluffy pellets, respectively, while the opposite was found for biofilm cultures without any visible change in biofilm morphology. Cellulase activities and volumetric productivities attained by biofilms in flask cultures were 70% higher than that produced by free mycelium cultures and threefold higher when biofilms were grown in microbioreactors. CONCLUSIONS: Fungal biofilms developed on polyester cloth in both flasks and microbioreactors produce higher cellulase yields and volumetric productivities than free mycelium cultures at lower biomass levels. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of the present study are of commercial and biological interest. All productivity parameters revealed that fungal biofilms may be used for the production of cellulase and other proteins in various types of bioreactors. Moreover, they may be used as model systems to study differential gene expression related to cell adhesion.     

Binding of ovarian cancer cells to immobilized hyaluronic acid

Ovarian cancer has the highest mortality rate of any gynaecological malignancy. This is caused by metastatic deposits obstructing the intestinal tract. Very little is known about the molecules involved in the initial attachment of the metastatic tumour cells to the peritoneal mesothelial lining. Previously, we showed that many ovarian tumour lines express the adhesion molecule, CD44, on their cell surface. The major ligand for CD44 is the extracellular matrix glycosaminoglycan, hyaluronic acid (HA). Because mesothelial cells have a pericellular cost that contains large amounts of HA, it was postulated that the CD44/HA interaction is an important stage in ovarian cancer spread. However, it was difficult to demonstrate this interaction in an in vitro adhesion assay with mesothelial cells as most of the HA, and presumably the bound tumour cells, were lost from the mesothelial cells during the washing steps of the assay. In order to try and clarify the situation, the adhesion of six ovarian tumour lines to immobilized HA was measured. Four lines expressed high levels of CD44 and two lines expressed negligible amounts. Preliminary experiments were carried out with one of the CD44-expressing lines. After coating a plate overnight with 3 mg ml−1 HA, the 5 min adhesion of this line varied between 2% and 73% according to the type of plate that was used. Falcon Micro Test III flexible plates gave the highest adhesion and was used for further experiments. Plates were coated with concentrations of HA between 0.001 mg ml−1 and 3 mg ml−1. All CD44 expressing lines adhered to HA, but the maximum adhesion and the adhesion strength varied with the line studied and was not closely related to the total CD44 expression. These results suggest that CD44 on ovarian tumour cells binds to HA on mesothelial cells. As much of the HA can be very easily lost from the mesothelial cell surface, additional factors such as the strength of the CD44/HA interaction, and the formation of bonds by the tumour cells with other membrane adhesion molecules, such as integrins, are also important in promoting tumour spread. This revised version was published online in November 2006 with corrections to the Cover Date.     

Binding of ovarian cancer cells to immobilized hyaluronic acid

Ovarian cancer has the highest mortality rate of any gynaecological malignancy. This is caused by metastatic deposits obstructing the intestinal tract. Very little is known about the molecules involved in the initial attachment of the metastatic tumour cells to the peritoneal mesothelial lining. Previously, we showed that many ovarian tumour lines express the adhesion molecule, CD44, on their cell surface. The major ligand for CD44 is the extracellular matrix glycosaminoglycan, hyaluronic acid (HA). Because mesothelial cells have a pericellular cost that contains large amounts of HA, it was postulated that the CD44/HA interaction is an important stage in ovarian cancer spread. However, it was difficult to demonstrate this interaction in an in vitro adhesion assay with mesothelial cells as most of the HA, and presumably the bound tumour cells, were lost from the mesothelial cells during the washing steps of the assay. In order to try and clarify the situation, the adhesion of six ovarian tumour lines to immobilized HA was measured. Four lines expressed high levels of CD44 and two lines expressed negligible amounts. Preliminary experiments were carried out with one of the CD44-expressing lines. After coating a plate overnight with 3 mg ml-1 HA, the 5 min adhesion of this line varied between 2% and 73% according to the type of plate that was used. Falcon Micro Test III flexible plates gave the highest adhesion and was used for further experiments. Plates were coated with concentrations of HA between 0.001 mg ml−1 and 3 mg ml−1. All CD44 expressing lines adhered to HA, but the maximum adhesion and the adhesion strength varied with the line studied and was not closely related to the total CD44 expression. These results suggest that CD44 on ovarian tumour cells binds to HA on mesothelial cells. As much of the HA can be very easily lost from the mesothelial cell surface, additional factors such as the strength of the CD44/HA interaction, and the formation of bonds by the tumour cells with other membrane adhesion molecules, such as integrins, are also important in promoting tumour spread. This revised version was published online in November 2006 with corrections to the Cover Date.     

Subtilisin Carlsberg in complex with sodium dodecyl sulfate--an effective catalyst for the solid phase segment coupling of peptides on polyvinyl alcohol cryogel

Subtilisin Carlsberg (SK) was shown to catalyze the solid phase segment coupling of peptides in complex with sodium dodecyl sulfate (SDS) in an organic medium on Aminosilochrom and polyvinyl alcohol (PVA) cryogel activated with glutaraldehyde or divinylsulfone. Diamines of different lengths with a general formula NH2-(CH2)n-NH2 (n = 2, 4, and 6) were used as spacers between the PVA cryogel and the peptide. A model reaction of enzymatic attachment of the Dnp-Ala-Ala-Leu-OMe tripeptide to the PVA cryogel was carried out by treatment with the SDS-SK complex in a mixture of anhydrous ethanol and DMSO (7: 3, v/v) using a tenfold excess of the carboxyl component. The molar enzyme-substrate ratio was 1 : 88. The effect of the method of matrix activation, length of a spacer, and reaction time on the coupling efficiency was studied. Hexamethylenediamine was found to be the most effective spacer for the enzymatic coupling on the PVA cryogel activated with glutaraldehyde (the reaction proceeded with the highest yield of 60%). The reaction efficiency was considerably lower in the case of ethylenediamine and tetramethylenediamine (10 and 15%, respectively). The best results were obtained on the PVA cryogel activated by divinylsulfone with hexamethylenediamine as a spacer. A two-step condensation of tripeptides was carried out on this supportsupport. The second step of condensation was shown to proceed better (in 85% yield) in comparison with the first step (37% yield).     

Identification of a new adhesin‐like protein from Lactobacillus mucosae ME‐340 with specific affinity to the human blood group A and B antigens

Aims: To identify and characterize a new adhesin‐like protein of probiotics that show specific adhesion to human blood group A and B antigens. Methods and Results: Using the BIACORE assay, the adhesion of cell surface components obtained from four lactobacilli strains that adhered to blood group A and B antigens was tested. Their components showed a significant adhesion to A and B antigens when compared to the bovine serum albumin (BSA) control. The 1 mol l^?1 GHCl fraction extracted from Lactobacillus mucosae ME‐340 contained a 29‐kDa band (Lam29) using SDS–PAGE. The N‐terminal amino acid sequence and homology analysis showed that Lam29 was 90% similar to the substrate‐binding protein of the ATP‐binding cassette (ABC) transporter from Lactobacillus fermentum IFO 3956. The complete nucleotide sequence (858 bp) of Lam29 was determined and encoded a protein of 285 amino acid residues. Phylogenetic analysis and multiple sequence alignments indicated this protein may be related to the cysteine‐binding transporter. Conclusions: The adhesion of ME‐340 strain to blood group A and B antigens was mediated by Lam29 that is a putative component of ABC transporter as an adhesin‐like protein. Significance and Impact of the Study: Lactobacillus mucosae ME‐340 expressing Lam29 may be useful for competitive exclusion of pathogens via blood group antigen receptors in the human gastrointestinal mucosa and in the development of new probiotic foods.     

Nanofibers from blends of polyvinyl alcohol and polyhydroxy butyrate as potential scaffold material for tissue engineering of skin

Nanofibers were prepared by electrospinning from pure polyvinyl alcohol (PVA), polyhydroxybutyrate (PHB), and their blends. Miscibility and morphology of both polymers in the nanofiber blends were studied by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and differential scanning calorimetry (DSC), revealing that PVA and PHB were miscible with good compatibility. DSC also revealed suppression of crystallinity of PHB in the blend nanofibers with increasing proportion of PVA. The hydrolytic degradation of PHB was accelerated with increasing PVA fraction. Cell culture experiments with a human keratinocyte cell line (HaCaT) and dermal fibroblast on the electrospun PHB and PVA/PHB blend nanofibers showed maximum adhesion and proliferation on pure PHB. However, the addition of 5 wt % PVA to PHB inhibited growth of HaCaT cells but not of fibroblasts. On the contrary, adhesion and proliferation of HaCaT cells were promoted on PVA/PHB (50/50) fibers, which inhibited growth of fibroblasts.