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Preparation and physicochemical properties of compression-molded keratin films.

The S-sulfo keratin was extracted from wool and was then spray-dried to give S-sulfo keratin powder. Differential scanning calorimetry analysis showed that the glass transition temperature of S-sulfo keratins became lowered with the increase of moisture content, while perfectly dried S-sulfo keratin powder did not give thermal transition in the temperature range 30-130 degrees C. The compression molding of the S-sulfo keratin powder supplemented with one-tenth weight of water afforded a plastic-like transparent proteinous film above the glass transition temperature. The film obtained from the powder without water addition or compression molded below glass transition temperature partly remained powdery. The film compression molded at 120 degrees C gave the maximum ultimate strength and Young's modulus, 27.8 +/- 2.9 and 1218 +/- 80 MPa, respectively. Obtained film was insoluble and slightly swelled in water, but, in the presence of reducing agent, the film significantly swelled at pH 7.0 and even dissolved at pH 9.0, suggesting the relevance of abundant disulfide linkage. The film supported the mammalian cell adhesion and proliferation, demonstrating the biocompatibility of S-sulfo keratin films.

Animals↗

Drug release from starch-acetate microparticles and films with and without incorporated alpha-amylase.

Acetylation of starch considerably decreases its swelling and enzymatic degradation. Thus, starch-acetate (SA) based delivery systems may be suitable for controlled drug delivery. The aim of the present study was to evaluate drug release from the SA microparticles (SA mps) and SA films. The average degree of acetyl substitution (DS) per glucose residue in the starch was either 1.9 (SA DS 1.9) or 2.6 (SA DS 2.6). Timolol (mw 332), calcein (mw 623) and bovine serum albumin (BSA, mw 68,000) were used as model drugs. A continuous timolol release from the both SA mps was observed in phosphate buffer solution (PBS) pH 7.4 (50-days incubation). The release of timolol was faster from the SA DS 1.9 mps than from the SA DS 2.6 mps. Calcein release from both SA mps was continuous in PBS pH 7.4 (5-days incubation). But, calcein release profile from the SA DS 2.6 film in PBS pH 7.4 showed discontinuities. However, the release of calcein from both SA films was continuous in human serum in vitro during the 7-day incubation, i.e. enzymes enhanced calcein release. Thus, alpha-amylase was incorporated into the SA films in order to enhance drug release from the films. However, the effects of incorporation of alpha-amylase on the model macromolecule (BSA) release from the SA films were modest. In conclusion, this study demonstrates the achievement of slow release of different molecular weight model drugs from the SA mps and films as compared to fast release from the native starch preparations. DS of SA, physicochemical properties of a drug and the presence of enzymes can all affect drug release profiles from SA based preparations.

Absorption↗

The inflammatory responses to silk films in vitro and in vivo.

Silks have a long history of biomedical use as sutures. Silk can be purified, chemically modified to attach RGD sequences and processed into highly porous scaffolds for tissue engineering. We report biocompatibility studies of silk films (with or without covalently bound RGD) that were seeded with bone-marrow derived mesenchymal stem cells (MSC) and (a) cultured in vitro with human MSC or (b) seeded with autologous rat MSC and implanted in vivo. Controls for in vitro studies included tissue culture plastic (TCP; negative control), TCP with lipopolysaccharide (LPS) in the cell culture medium (positive control), and collagen films; controls for in vivo studies included collagen, PLA and TCP. After 9 h of culture, the expression of the pro-inflammatory Interleukin 1 beta (IL-1beta) and inflammatory cyclooxygenase 2 (COX-2) in human MSC were comparable for silk, collagen and TCP. After 30 and 96 h, gene expression of IL-1beta and COX-2 in MSC returned to the baseline (pre-seeding) levels. These data were corroborated by measuring IL-1beta and prostaglandin E2 levels in culture medium. The rate of cell proliferation was higher on silk films than either on collagen or TCP. In vivo, films made of silk, collagen or PLA were seeded with rat MSCs, implanted intramuscularly in rats and harvested after 6 weeks. Histological and immunohistochemical evaluation of silk explants revealed the presence of circumferentially oriented fibroblasts, few blood vessels, macrophages at the implant-host interface, and the absence of giant cells. Inflammatory tissue reaction was more conspicuous around collagen films and even more around PLA films when compared to silk. These data suggest that (a) purified degradable silk is biocompatible and (b) the in vitro cell culture model (hMSC seeded and cultured on biomaterial films) gave inflammatory responses that were comparable to those observed in vivo.

Animals↗

Bone tissue engineering on patterned collagen films: an in vitro study.

This study aimed at guiding osteoblast cells from rat bone marrow on chemically modified and patterned collagen films to study the influence of patterns on cell guidance. The films were stabilized using different treatment methods including crosslinking with carbodiimide (EDC) and glutaraldehyde, dehydrothermal treatment (DHT), and deposition of calcium phosphate on the collagen membrane. Mesenchymal osteoprogenitor cells were differentiated into osteoblasts and cultured for 7 and 14 days on micropatterned (groove width: 27 microm, groove depth: 12 microm, ridge width: 2 microm) and macropatterned (groove width: 250 microm, groove depth: 250 microm, ridge width: 100 microm) collagen films to study the influence of pattern dimensions on osteoblast alignment and orientation. Fibrinogen was added to the patterned surfaces as a chemical cue to induce osteoblast adhesion. Cell proliferation on collagen films was determined using MTS assay. Deposition of calcium phosphate on the surface of the film increased surface hydrophilicity and roughness and allowed a good cell proliferation. Combined DHT and EDC treatment provided an intermediate wettability, and also promoted cell proliferation. Glutaraldehyde crosslinking was found to lead to the lowest cell proliferation but fibrinogen adsorption on glutaraldehyde treated film surfaces increased the cell proliferation significantly. Macropatterns were first tested for alignment and only microscopy images were enough to see that there is no specific alignment. As a result of this, micropatterned samples with the topography that affect cell alignment and guidance were used. Osteoblast phenotype expression (ALP activity) was observed to be highest in calcium phosphate deposited samples, emphasizing the effect of mineralization on osteoblast differentiation. In general ALP activity per cell was found to decrease from day 7 to day 14 of incubation. SEM and fluorescence microscopy revealed good osteoblast alignment and orientation along the axis of the patterns when micropatterned films were used. This study shows that it is possible to prepare cell carriers suitable for tissue engineering through choice of appropriate surface topography and surface chemistry. Presence of chemical cues and micropatterns on the surface enhance cell orientation and bone formation.

Animals↗

A new anti-adhesion film synthesized from polygalacturonic acid with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide crosslinker.

The most commonly used anti-adhesion device for separation and isolation of wounded tissues after surgery is the polymeric film. In this study, a new anti-adhesion membrane based on polygalacturonic acid (PGA) has been synthesized, and its biocompatibility and anti-adhesion capabilities evaluated. The PGA film was reacted with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) to obtain a cross-linked PGA film with an 86% gel content and a 47% water content when immersed in aqueous solution. This PGA-EDC film did not show any evidence of cytotoxic effects since it did not induce any significant increase in cytoplasmic LDH release from the L929 cells in contact with it. When implanted into rats, the PGA-EDC film exhibited a most promising anti-adhesion potential with only 1 out of 21 rats operated not forming any tissue adhesion. This anti-adhesion potency is significantly higher than that found for Seprafilm and untreated rats where 11 out of 21 and 18 out of 21 operated rats, respectively, formed tissue adhesions. The implanted PGA-EDC film did not elicit any acute inflammatory reaction based on the results of histological examination and peritoneal fluid leukocytes analysis. The newly developed PGA-EDC film thus has a great potential for future use in clinical applications.

Absorbable Implants↗

Multifunctional Ti-(Ca,Zr)-(C,N,O,P) films for load-bearing implants.

Films of Ti-Ca-P-C-O-(N), Ti-Ca-C-O-(N) and Ti-Zr-C-O-(N) were deposited by DC magnetron sputtering or ion implantation-assisted magnetron sputtering of composite targets TiC0.5 + 10%Ca10(PO4)6(OH)2, TiC0.5 + 20%(CaO + TiO2) and TiC0.5 + 10%ZrO2 in an Ar atmosphere or reactively in a gaseous mixture of Ar + 14%N2. The microstructure, elemental and phase composition of films were studied by means of X-ray diffraction, transmission electron microscopy, scanning force microscopy, X-ray photoelectron spectroscopy and energy-dispersive X-ray spectroscopy. The films were characterized in terms of their hardness, Young's modulus, elastic recovery, adhesion strength, and friction and wear both in air and under physiological solution. Particular attention was paid to the analysis of deformation and fracture for various film/substrate systems during scratch testing. The biocompatibility of the films was evaluated by both in vitro and in vivo experiments. In vitro studies involved the investigation of adhesion, spreading, and proliferation of MC3T3-E1 osteoblasts and IAR-2 epithelial cells, morphometric analysis, actin cytoskeleton, focal contacts staining, alkaline phosphatase activity and von Kossa staining of osteoblastic culture. Cell culture experiments demonstrated an increase of osteoblastic proliferation on Ca- and P-incorporated films. In vivo studies were fulfilled by subcutaneous implantation of Teflon plates coated with the tested films in mice and analysis of the population of adherent cells on their surfaces. The results obtained show that multicomponent nanostructured Ti-(Ca, Zr)-(C, N, O, P) films possess a combination of high hardness, wear resistance and adhesion strength, reduced Young's modulus, low friction coefficient and high biocompatibility.

3T3 Cells↗

Disassembly of layer-by-layer films of plasmid DNA and reducible TAT polypeptide.

This paper reports the disassembly of layer-by-layer (LbL) films of plasmid DNA and a reducible cationic polypeptide. To utilize a reducing microenvironment of cellular plasma membrane as a potential trigger, LbL films are assembled to contain both DNA and the TAT-based polypeptide (PTAT) with reducible disulfide bonds in the backbone. The assembly and disassembly processes are monitored by goniometry, ellipsometry, and atomic force microscopy (AFM). The structure of the PTAT films is compared with that of non-reducible poly(L-lysine) (PLL) films. Both PTAT and PLL films exhibit exponential growth but with the contact angle alternating between characteristic values. Ellipsometry and AFM show a gradual and complete disassembly of the PTAT but not the PLL films in a 24h period in the reducing environment in vitro. This study suggests a potential of using reducible LbL films for controlled DNA delivery.

DNA↗

Thin collagen film scaffolds for retinal epithelial cell culture.

Collagen films have been used in biological implantation and surgical grafts. The development of thin collagen films on the order of 10 microm thick that ensure a planar distribution of implanted cells is a necessary step towards surgical grafts for treatment of age-related macular degeneration (AMD). Here, collagen films were manufactured on a Teflon support to a thickness of 2.4+/-0.2 microm, comparable to that of native Bruch's membrane. Because one important function of Bruch's membrane is allowing the flow of nutrients and waste to and from the retinal pigment epithelium the diffusion properties of the collagen films were studied using blind-well chambers. The diffusion coefficient of the collagen film was determined to be 4.1 x 10(-10)cm(2)/s for 71,200 Da dextran molecules. Viability studies utilizing the blind-well chambers also confirmed that nutrient transport through the films was sufficient to sustain retinal pigment epithelial (RPE) cells. The films were bioassayed in a RPE cell culture model to confirm cell attachment and viability. RPE cells were shown to form an epithelial phenotype and were able to phagocytize photoreceptor outer segments.

Biocompatible Materials↗

The electrochemical preparation of FAD/ZnO with hemoglobin film-modified electrodes and their electroanalytical properties.

Flavin adenine dinucleotide (FAD)-modified zinc oxide self-assembly films were prepared using repeated cyclic voltammetry. The electrochemical reaction of the hemoglobin with the FAD/ZnO self-assembly film-modified electrodes and their electrocatalytic properties were investigated. This paper describes the successful loading of the electrochemically active molecules of hemoglobin and FAD along with ZnO by electrochemical method. In addition to the cyclic voltammetry, an electrochemical quartz crystal microbalance was used to study the growth mechanism and the properties of the films. The FAD/zinc oxide films exhibited a single redox couple, which corresponded to the FAD redox couple. The electrocatalytic properties of the O2, H2O2, trichloroacetic acid and SO(3)2- were studied by the FAD/zinc oxide films in the absence or in the presence of hemoglobin. The electrocatalytic reduction current had been developed from the cathodic peak of the FAD/zinc oxide redox couple. The electrocatalytic process involved an interaction of hemoglobin and FAD/GC film-modified electrode to increase the electrocatalytic reduction current. The electrocatalytic reduction of O2 using the FAD/zinc oxide films was investigated by cyclic voltammetry and rotating ring-disk electrode methods.

Biosensing Techniques↗

Carbon nanotubes-polymer-redox mediator hybrid thin film for electrocatalytic sensing.

Electrocatalytic sensing of NADH using a hybrid thin film derived from multi-wall carbon nanotubes (CNTs), Nafion (Nf) polymer and electrogenerated redox mediator is described. The redox mediator was electrochemically generated by the oxidation of serotonin on the hybrid thin film modified glassy carbon electrode (GC/Nf-CNT). Controlled potential electrolysis of serotonin at 0.1 V in neutral solution results in the generation of the redox mediator 5,5'-dihydroxy-4,4'-bitryptamine (DHB) on the hybrid thin film. The electrogenerated DHB has redox active quinone-imine structure and was electrochemically characterized by studying the pH dependent redox response. DHB on the hybrid thin film exhibits reversible redox peak at -0.05 V and the formal potential shifts by -55 mV while increasing the solution pH by 1 unit. The quinone-imine structure of DHB efficiently catalyzes the oxidation of NADH with a decrease in the overpotential of about 500 mV compared to the unmodified electrode. The CNTs of the hybrid thin film facilitates the mediated electrocatalytic oxidation of NADH. The hybrid thin film modified electrode exhibits stable amperometric response and it linearly responds to NADH (0.5-400 microM). This hybrid thin film modified electrode could detect NADH as low as 0.1 microM at -0.05 V with a sensitivity of 11.1 nA/microM in physiological pH.

Biosensing Techniques↗

Amperometric glucose biosensor based on layer-by-layer assembly of multilayer films composed of chitosan, gold nanoparticles and glucose oxidase modified Pt electrode.

A new strategy for fabricating glucose biosensor was presented by layer-by-layer assembled chitosan (CS)/gold nanoparticles (GNp)/glucose oxidase (GOD) multilayer films modified Pt electrode. First, a cleaned Pt electrode was immersed in poly(allylamine) (PAA), and then transferred to GNp, followed by the adsorption of GOD (GOD/GNp/PAA/Pt). Second, the GOD/GNp/PAA/Pt electrode was immersed in CS, and then transferred to GNp, followed by the adsorption of GOD (GOD/GNp/CS/GOD/GNp/PAA/Pt). Third, different layers of multilayer films modified Pt electrodes were assembled by repeating the second process. Film assembling and characterization were studied by quart crystal microbalance, and properties of the resulting glucose biosensors were measured by electrochemical measurements. The results confirmed that the assembling process of multilayer films was simple to operate, the immobilized GOD displayed an excellent catalytic property to glucose, and GNp in the biosensing interface efficiently improved the electron transfer between analyte and electrode surface. The amperometric response of the biosensors uniformly increased from one to six layers of multilayer films, and then reached saturation after the seven layers. Among the resulting biosensors, the biosensor based on the six layers of multilayer films was best. It showed a wide linear range of 0.5-16 mM, with a detection limit of 7.0 microM estimated at a signal-to-noise ratio of 3, fast response time (within 8s). Moreover, it exhibited good reproducibility, long-term stability and interference free. This method can be used for constructing other thin films, which is a universal immobilization method for biosensor fabrication.

Biosensing Techniques↗

Flexible chitin films: structural studies.

Chitin gels were transformed into thin, flexible chitin films with minimal dimensional shrinkage and maximum flexibility and thickness in the range of 25-80 microm by a cold-press process. Solvent residue was removed by heating the films at 50 degrees C for 12 h, followed by rinsing in 95% ethanol. The crystallinity and mechanical properties of the flexible chitin films were found to be a function of the amount of shrinkage from the gel to the final film that was obtained. For 28-microm thick films with 30% shrinkage, transparency of up to 90% was found. X-ray diffractometry (XRD) showed that the number of diffraction peaks appearing at 2theta;=23 degrees and 2theta;=27 degrees became increasingly sharper with shrinkage. Topographical information obtained from scanning electron microscopy (SEM) and atomic force microscopy (AFM) attributed the structural morphology of the films to the formation of sub-microscopic micelles. Scanning transmission electron microscopy (STEM) showed that shrinkage resulted in coarser microstructure, affecting tensile properties, where the ductility and toughness were proportional to the amount of shrinkage. These flexible chitin films have potential as wound dressing materials.

Bandages↗

The enhanced PC and PEC oxidation of formic acid in aqueous solution using a Cu-TiO2/ITO film.

The photocatalytic (PC) and photoelectrocatalytic (PEC) activity of Cu-TiO2/ITO films for degrading formic acid in aqueous solution was investigated in this study. Compared with a TiO2/ITO film, the degradation efficiency of formic acid on the Cu-TiO2 films increased markedly in both the PC and PEC oxidation processes. However, it was found that the photodeposited Cu metal on the Cu-TiO2 films could electrochemically dissolute during the PEC reaction, while an electrical bias with the voltage higher than 1.48 V was applied. It is believed this is a common problem occurred for several metals deposition on the TiO2 films, which results in a poor stability of the metal-deposited TiO2 electrode in PEC processes. To improve the stability of the Cu-TiO2 electrode, an alternative process between PC and PEC reactions was investigated. It was found that the dissolute Cu metal during the PEC process was re-deposited on the Cu-TiO2 film again during the PC process. The experiments with repeated runs demonstrated that this alternative process could not only overcome the loss of Cu, but also enhance the PEC oxidation efficiency of the Cu-TiO2 films.

Catalysis↗

Comparison of catalytic activity of two platinised TiO2 films towards the oxidation of organic pollutants.

Two types of platinised TiO2 films, i.e., Pt-TiO2/ITO and Pt(TiO2)/ITO, were prepared by a procedure of dip-coating and subsequent photo-deposition, and photo-deposition and subsequent dip-coating, respectively. They were well characterized by DRS, XRD spectra, SEM microscopy and photoelectrochemical measurement. Their photocatalytic, dark catalytic and photoelectrocatalytic activities were investigated using formic acid as a model organic pollutants. Compared with pure TiO2/ITO film, the photocatalytic activity of the platinised TiO2 films were apparently improved. However, the improvement was considerably dependent on the preparation method of these films. Pt-TiO2/ITO not only possessed higher photocatalytic activity but also showed a dark catalytic activity towards HCOOH degradation. As a sequence, it was first emphasized that the dark catalytic effect of Pt-TiO2/ITO was partly responsible for degradation of formic acid in the photocatalytic oxidation process. Although the Pt(TiO2)/ITO film does not exhibit the dark catalytic activity, its photocatalytic degradation efficiencies towards HCOOH are higher than that of Pt-TiO2/ITO film. Therefore, in view of enhanced photocatalytic activity, the Pt(TiO2)/ITO was more favored than Pt-TiO2/ITO film.

Catalysis↗

Films coated with molecular imprinted polymers for the selective stir bar sorption extraction of monocrotophos.

The flexibility and simplicity of stir bar sorption extraction (SBSE) have been combined with the selectivity of molecularly imprinted polymers (MIP). Stir bars were coated reproducible with a 180 microm film formed from a formic acid solution of nylon-6 polymer either nonimprinted or imprinted with monocrotophos. Time sorption profiles were measured for the extraction of monocrotophos from dichloromethane at the concentration of 10-200 micromol/L levels with both types of films in order to compare extraction characteristics. The results indicated that the MIP coated layer showed remarkable high affinity toward monocrotophos and equilibrium adsorption was attained rapidly (60 min) in contrast to free standing molecularly imprinted polymer in which equilibrium adsorption was normally attained after several hours. The stir bars coated with MIP films were capable of extracting four structural analogues of monocrotophos from dichloromethane solution, which suggests that both the amino group and PO part of these molecules is responsible for interaction with the imprinted polymer. Evidence was also presented by FT-IR analysis that the amide-hydrogen-bonding interaction between the MIP-coated films and monocrotophos was originated for monocrotophos recognition. To achieve selective extraction of monocrotophos from sample, stir bars coated with MIP films were washed with 10% (v/v) acetic acid/methanol and methanol. Clean extracts and yields of 95% were obtained, demonstrating the suitability of stir bar coated with MIP films for the analysis of environmental and biological samples. Compared with traditional MIP and SBSE, the MIP-coated film showed not only the high selectivity but also the rapid equilibrium adsorption.

Caprolactam↗

Optical storage and surface-relief gratings in azobenzene-containing nanostructured films.

This paper brings an overview of photoisomerization-derived properties in azobenzene-containing nanostructured films produced with the Langmuir-Blodgett (LB) and layer-by-layer (LbL) methods. Emphasis was placed on the optical storage and formation of surface-relief gratings (SRGs), where the distinctive properties of the nanostructured films were highlighted. For optical storage, in particular, a discussion was made of the higher birefringence induced in LB films from azopolymers due to their organized nature, and of the strong effects from ionic interactions on the photoisomerization of azochromophores in LbL films. The molecular-level control of film properties in the LbL films is described as being responsible for the varied behavior observed in the photoinscription of SRGs, which may involve considerable level of photodegradation depending on the film fabrication conditions and materials used.

Azo Compounds↗

In-situ monitoring of the dielectric and electrostrictive properties of anodised thin films for biochip applications.

Further improving the performance of capacitance-based DNA microsensors will depend on the ability to synthesize simply, at a low cost, thin films of high dielectric permittivity with a well-controlled growth morphology. Anodisation of the valve metals Ti, Zr and Ta is a promising route for preparing such films. In this paper, we propose a new method for following the evolution of the dielectric properties of such anodised metallic oxide thin films in-situ during their growth. The technique relies on measuring the evolution of the electrostatic stress developing in the film. Measurements have been carried out on anodic TiO(2) films grown galvanostatically in HNO(3) at 1 and 3 mA/cm(2) and in H(3)PO(4) at 0.5 mA/cm(2). The dielectric permittivity of the films was observed to decrease during the initial stage of anodisation, characterised by a steady increase of the cell voltage with time. The permittivity then reached a constant value when the oxide entered a second growth regime, characterised by a stabilisation of the cell voltage. The origin of the observed permittivity evolution is interpreted in terms of the evolution of the growth morphology of the films during anodisation.

Alloys↗

Degradation of raw or film-incorporated beta-cyclodextrin by enzymes and colonic bacteria.

beta-cyclodextrin (beta-CD) is a suitable excipient for peroral use, which improves the solubility of lipophilic drugs, as well as for colon-specific drug release when it is mixed with coating polymers. The first aim of this work was to examine the suitability of various enzymes as a simple in vitro model for the glycolytic activity in the human colon. alpha-Amylase (source Aspergillus oryzae) and taka diastase (source A. oryzae) showed remarkable degradation capacity of free beta-CD, whereas other alpha-amylases (sources Bacillus subtilis or Hog pancreas) were found to be unsuitable. The next aim was to find out if film-incorporated beta-CD is also degraded by these enzymes. Therefore, diffusion studies of 5-aminosalicylic acid (5-ASA) through Eudragit RS or Eudragit NE films containing beta-CD were performed with taka diastase present in the buffer medium. Pronounced diffusion of the drug through the Eudragit RS film was found only when swelling excipients like crosslinked sodium carboxymethylcellulose (CMC-CL sodium) or polyvinylpyrrolidone (PVP 25) were present in the film, indicating enhanced accessibility of beta-CD by the enzyme. Films containing CMC-CL without beta-CD showed even higher permeability, which also points to enzymatic degradation of CMC-CL. Permeabilization by taka diastase of Eudragit NE films without swelling agents correlated with the beta-CD content, whereas control films containing talcum remained impermeable upon enzyme action. Furthermore, the beta-CD degradation capacity of colonic bacteria like Escherichia fergusonii, Serratia odorifera or Proteus mirabilis was examined with beta-CD coatings on tablets, which contained bisoprolol as a model drug. Tablets with beta-CD-containing Eudragit RS coatings showed the highest drug release upon incubation with P. mirabilis. The moderate drug release by E. fergusonii could be increased almost to the same level when the bacteria were pre-incubated for 24 h in medium containing 2.5 mg/ml beta-CD, indicating the induction of glycolytic enzymes by beta-CD in this colonic bacteria strain.

Acrylic Resins↗