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Inhibition of surface spoilage bacteria in processed meats by application of antimicrobial films prepared with chitosan.

A study was undertaken to evaluate the feasibility of using antimicrobial films, designed to slowly release bacterial inhibitors, to improve the preservation of vacuum-packaged processed meats during refrigerated storage. The antimicrobial films were prepared by incorporating acetic or propionic acid into a chitosan matrix, with or without addition of lauric acid or cinnamaldehyde, and were applied onto bologna, regular cooked ham, or pastrami. At various times during storage, packages were opened and the amounts of antimicrobial agents remaining in the chitosan matrix were measured. Regardless of film composition or meat product type, propionic acid was nearly completely released from the chitosan matrix within 48 h of application, whereas release of acetic acid was more limited, with 2-22% of the acid remaining in chitosan after 168 h of storage. Addition of lauric acid, but not cinnamaldehyde, to the chitosan matrix generally reduced the release of acetic acid significantly (P < or = 0.05) and the release was more limited onto bologna than onto ham or pastrami. In addition, the efficacies of the various films for inhibiting bacterial growth were tested against indigenous lactic acid bacteria and Enterobacteriaceae, and against Lactobacillus sakei or Serratia liqueficiens, surface-inoculated onto the meat products. Whereas lactic acid bacteria were not affected by the antimicrobial films under study, the growth of Enterobacteriaceae and S. liquefaciens was delayed or completely inhibited as a result of film application. Strongest inhibition was observed on drier surfaces (bologna), onto which acid release was slower, and with films containing cinnamaldehyde, as a result of its greater antimicrobial activity under these conditions.

Acetic Acid↗

Sustained release of human growth hormone (hGH) from collagen film and evaluation of effect on wound healing in db/db mice.

Collagen films containing human growth hormone (hGH) were prepared and the release of hGH from these films and their effect on healing of full-thickness wounds in db/db mice were evaluated. The release profiles of hGH from the collagen films varied with composition and preparation conditions. The film prepared by air-drying of the mixture of hGH and collagen solution released hGH continuously over 3 days both in vitro and in vivo. By application of collagen film containing 3 mg of hGH twice at an interval of 6 days to wounds, area of wounds on day 21 was significantly reduced compared with that of non-treated wounds. Application of hGH alone at the same dose had no significant effect on wound healing. The maximum serum hGH concentration after single administration of the hGH collagen film was lower than that with hGH alone, and hGH persisted in serum over 3 days. These results suggest that hGH collagen film may be a useful topical formulation for the treatment of wounds.

Animals↗

Adsorption of serum albumin to thin films of poly(lactide-co-glycolide).

Protein adsorption has been implicated in the variability of drug release from biodegradable microspheres. We used optical reflectometry to measure the extent and kinetics of bovine serum albumin (BSA) adsorption to smooth spin-cast films prepared from two poly(lactide-co-glycolide) (PLG) samples that have different end-groups, one being a hydrophilic carboxylic end group and the other a hydrophobic ester end group. One of us has previously shown that these end-groups influence microsphere degradation (Tracy et al. , 1998, Factors affecting the degradation rate of poly(lactide-co-glycolide) microspheres in vivo and in vitro. Biomaterials: submitted for publication.). Both films were moderately hydrophobic, and their wettability was independent of the type of end-group. BSA adsorbed readily to both native PLG films, attaining as much as 50% surface coverage by area and was insensitive to the type of end-group. Aging the films in water for 24 h prior to BSA exposure decreased the hydrophobicity of the films and this in turn correlated with a significant decrease in the initial BSA adsorption rate. This was consistent with the often-observed trend that surface hydrophobicity favors protein adsorption. In spite of the lower adsorption affinity revealed by this decreased initial adsorption rate, the final adsorbed amounts on the aged films exceeded those attained on native films, presumably due to the increase in total surface area produced by partial PLG erosion.

Adsorption↗

Direct voltammetry and electrocatalytic properties of catalase incorporated in polyacrylamide hydrogel films.

The direct voltammetry and electrocatalytic properties of catalase (Cat) in polyacrylamide (PAM) hydrogel films cast on pyrolytic graphite (PG) electrodes were investigated. Cat-PAM film electrodes showed a pair of well-defined and nearly reversible cyclic voltammetry peaks for Cat Fe(III)/Fe(II) redox couples at approximately -0.46 V vs. SCE in pH 7.0 buffers. The electron transfer between catalase and PG electrodes was greatly facilitated in the microenvironment of PAM films. The apparent heterogeneous electron transfer rate constant (k(s)) and formal potential (E degrees ') were estimated by fitting square wave voltammograms with non-linear regression analysis. The formal potential of Cat Fe(III)/Fe(II) couples in PAM films had a linear relationship with pH between pH 4.0 and 9.0 with a slope of -56 mV pH(-1), suggesting that one proton is coupled with single-electron transfer for each heme group of catalase in the electrode reaction. UV-Vis absorption spectroscopy demonstrated that catalase retained a near native conformation in PAM films at medium pH. The embedded catalase in PAM films showed the electrocatalytic activity toward dioxygen and hydrogen peroxide. Possible mechanism of catalytic reduction of H(2)O(2) at Cat-PAM film electrodes was proposed.

Acrylic Resins↗

Electroactive hemoglobin-surfactant-polymer biomembrane-like films.

Polyions complex 2C12N+ PSS- was prepared by reacting poly(sodium styrenesulfonate) (Na+ PSS-) with didodecyldimethylammonium bromide (2Cl2N+ Br-). Stable thin films made from 2C12N+ PSS- with incorporated redox protein hemoglobin (Hb) on pyrolytic graphite (PG) electrodes were then characterized by electrochemistry and other techniques. Cyclic voltammetry (CV) of Hb-2C12N+ PSS- films showed a pair of well-defined and nearly reversible peaks for HbFe(III)/Fe(II) couple at about -0.17 V vs. saturated calomel electrode (SCE) in pH 5.5 buffers. The electron transfer rate between Hb and PG electrode was greatly facilitated in microenvironment of 2C12N+ PSS- films. Positions of Soret absorption band suggest that Hb keeps its secondary structure similar to its native state in 2C12N+ PSS- films at the medium pH. The results of X-ray diffraction and differential scanning calorimetry (DSC) suggest synthesized lipid 2C12N+ PSS- films have an ordered bilayer structure intercalated between PSS- polyion layers, and the incorporated Hb expands the layer spacing of the films. HbFe(I), a highly reduced form of Hb, might also be produced in these films at about -1.09 V, and could be used to catalytically reduce organohalide pollutants.

Calorimetry, Differential Scanning↗

Photoinduced transformation of 14-F-bacteriorhodopsin gelatin films based on both wild type and D96N mutant.

Spectral and kinetic transformations were studied in gelatin films made with 14-F wild type (WT) bacteriorhodopsin (BR) and 14-F D96N mutant BR. Unlike the recent study of water suspensions of the same pigments, where a red shifted species at 660 nm was shown to form under the light in 14-F WT only, there are no drastic differences in photoinduced behavior between gelatin films based on 14-F WT and 14-F D96N. It is not observed any photoinduced formation of red shifted species at 660 nm for both types of films as it is observed for corresponding pigments in water suspension. The observed results are explained in a terms of relationship between the rates of two photoinduced processes that occur in suspensions and films of corresponding pigments. Kinetic characteristics of the photoinduced processes for the films with chemical additives suggest that there are no advantages in using 14-F D96N films when compared to films based on 14-F WT.

Bacteriorhodopsins↗

Lift-off patterning of thin Au films on Si surfaces with atomic force microscopy

We studied a new lift-off process of thin Au film on silicon surfaces in nanometer-scale, combining anodic oxidation patterning with AFM, deposition of Au thin film on the patterned substrate and chemical etching processes of the Si oxide underneath the Au film. For Au films of thickness of 2-5 nm, the Au films on the Si oxide patterns were left unbroken and bent down to stick to Si surface after the removal of the oxide by the chemical etching. For an Au film of 1 nm in thickness, it was possible to lift-off the Au film on oxide patterns of the lines and dots in nanometer-scale using Si oxide as a sacrificial mask.

Journal Article↗

A TSM sensor investigation of low crystallinity cellulose films.

A thickness shear mode (TSM) quartz sensor has been used to characterize the substantivity, viscoelasticity, and mucoadhesive properties of low crystallinity cellulose (LCC) films. LCC is a novel pharmaceutical excipient that has been attributed with mucoadhesive properties. Thin films of LCC were deposited onto TSM sensors by a spin coating technique. The films were treated by passing water or 1.0% w/v mucin solution (pH 3.7 or 7.0) over the surface. Changes in the mass and viscosity of the film were observed by monitoring changes in the impedance spectra of the coated TSM sensors. Scanning electron micrographs (SEMs) of each film were used to assist the interpretation of the TSM sensor data. This study showed that LCC forms highly tenacious and viscoelastic films able to withstand prolonged (approximately 1 h) exposure to both water and mucin solution. Furthermore, these results indicate that the films may have mucoadhesive properties as LCC was found to bind significant (P<0.05) amounts of mucin in comparison with control measurements. Mucin binding to the LCC sensor was greater at pH 3.7 (P<0.05) than at pH 7.0, suggesting that the LCC formulation is mucoadhesive under these conditions.

Cellulose↗

Retention and transit of intestinal mucoadhesive films in rat small intestine.

The retention and transit characteristics of intestinal mucoadhesive film systems have been studied after intraduodenal administration in rats. Small size four layered film preparations, 0.5x0.5 mm, were prepared, where the backing layer (45.1+/-2.9 microm thick) was made of a water-insoluble polymer, ethylcellulose (EC), the surface layer was made of enteric pH-sensitive polymers, Eudragit L100, S100 or HP-55 and the middle layer was made of cellulose membrane. The surface layer was attached to the middle layer with an adhesive layer composed of carboxyvinyl polymer (Hiviswako(R) 103). After administration of ten films to the duodenum, the rats were sacrificed hourly and the distribution of the films in the whole small intestine was directly observed after abdominal incision. The HP-55, Eudragit L100 and S100 film systems were found to adhere to the upper, middle and lower part of the small intestine after 1, 2 and 4 h, respectively, for 2-3 h. Direct inspection study suggests that intestinal mucoadhesive film system has functions of: (1) pH-dependent intestinal adhesion site specificity; (2) adhesion to the intestinal wall; and (3) retention in the small intestinal adhesion site for at least 2 h. Intestinal mucoadhesive film system has been suggested to be a targeting system for drugs to the gastrointestinal tract.

Adhesiveness↗

Biphasic drug release: the permeability of films containing pectin, chitosan and HPMC.

The permeabilities of mixed films of pectin/chitosan/HPMC have been studied to assess their value in producing a dosage form with biphasic drug release characteristics. The inclusion of chitosan enhanced the properties of the films, rendering them stable at all physiological pH values. Pectin/HPMC films were soluble at pH values above 3.0. All pectin/chitosan/HPMC films were permeable to a model drug, paracetamol. HPMC initially increased the permeability of the films and subsequently reduced it at higher concentrations. The minimum permeability was obtained at pH 3 and at an HPMC level of 5% where the potential for polyelectrolyte complex formation between pectin and chitosan exists. The permeabilities of the films increased when they were exposed to pectinolytic enzymes, a system designed to mimic conditions in the colon. The film formulation thus show the potential for biphasic delivery with an initial, controllable slow phase that can be manipulated by changes in the formulation followed by a faster phase under conditions pertaining in the colon.

Acetaminophen↗

Polymerized rosin: novel film forming polymer for drug delivery.

Polymerized rosin (PR) a novel film forming polymer is characterized and investigated in the present study for its application in drug delivery. Films were produced by a casting/solvent evaporation method from plasticizer free and plasticizer containing solutions. Films prepared from different formulations were studied for their mechanical (tensile strength, percent elongation and Young's modulus), water vapour transmission and moisture absorption characteristics. Neat PR films were slightly brittle and posed the problem of breaking during handling. Hydrophobic plasticizers, dibutyl sebacate and tributyl citrate, improved the mechanical properties of free films with both the plasticizers showing significant effects on film elongation. Release of diclofenac sodium (model drug) from coated pellets was sustained with high coating levels. Concentration of plasticizer was found to affect the release profile. PR films plasticized with hydrophobic plasticizers could therefore be used in coating processes for the design of oral sustained delivery dosage forms.

Chemistry, Pharmaceutical↗

Enhanced stability of rubbery amylose-rich maize starch films plasticized with a combination of sorbitol and glycerol.

Well known aging problems with rubbery starch films are the migration of plasticizer and increased crystallinity leading to embrittlement. The effects of a combination of sorbitol and glycerol used as plasticizers on mechanical, moisture permeability and solid-state properties of rubbery amylose maize starch (Hylon VII) films were studied. The films were prepared by casting and were exposed to conditions of 25 degrees C/60% relative humidity (RH) and 40 degrees C/75% RH for 9 months. The starch films plasticized with a combination of sorbitol and glycerol (1:1) at equal amount to the polymer weight, were shown to be the most stable alternative of the studied films during the 9 months storage period. The water vapor transmission (WVTR) of the films did not change during the period of storage and neither did the elongation at break, but the tensile strength increased. X-ray diffraction (XRD) results showed that during storage no crystallization had occurred. The combination of sorbitol and glycerol prevented the migration of the plasticizer molecules out of the film.

Amylose↗

Periodontal delivery of ipriflavone: new chitosan/PLGA film delivery system for a lipophilic drug.

The aim of the present work was to design a film dosage form for sustained delivery of ipriflavone into the periodontal pocket. For this purpose, monolayer composite systems made of ipriflavone loaded poly(D,L-lactide-co-glycolide) (PLGA) micromatrices in a chitosan film form, were obtained by emulsification/casting/evaporation technique. Multilayer films, made of three layers of polymers (chitosan/PLGA/chitosan), were also prepared and compared to monolayer films for their "in vitro" characteristics. Morphology and physico-chemical properties of the different systems were evaluated. The influence of pH, ionic strength and enzymatic activity on film degradation, was also investigated. Significant differences in swelling, degradation and drug release were highlighted, depending on film structure and composition. In vitro experiments demonstrated that the composite micromatricial films represent a suitable dosage form to prolong ipriflavone release for 20 days.

Chitin↗

Hydrolytic degradation of poly(lactide-co-glycolide) films: effect of oligomers on degradation rate and crystallinity.

Oligomers are thought to accelerate the hydrolytic degradation of devices prepared from poly(lactide-co-glycolide), PLGA, due to their increased number of carboxylic end groups. To experimentally verify this hypothesis, two D,L-lactic acid oligomers having molecular weights close to their critical limit of solubility were synthesized and incorporated into PLGA films in three concentrations (0, 10, and 30% w/w). All films were translucent, rather flexible and initially amorphous. With increasing oligomer concentration the glass transition temperature (T(g)) and the molecular weight of films decreased prior to erosion. The degradation studies show that initial mass loss and water absorption are increased in oligomer-containing films as a function of average molecular weight and oligomer concentration. However, the incorporation of oligomers does not accelerate the degradation of films. By contrast, oligomer-containing films show extended lag phase until onset of polymer erosion. This was shown to be related to crystallization. Moreover, it was found that crystallization occurs earlier in oligomer-containing films and that the degree of crystallization is related to the average molecular weight of the oligomer. These findings bring new insight into the role of oligomers in the degradation process and can be used to explain why erosion in massive polymer devices occurs from the center to the surface.

Biotransformation↗

Sustained effect of an intervention to limit ordering of emergency department lumbosacral spine films.

Attempts at modification of test-ordering practices among housestaff demonstrate reversion to prior behavior after relaxation of surveillance. To test the hypothesis that use of pre-specified criteria for obtaining emergency department (ED) lumbosacral spine (LSS) films would reduce LSS utilization, we designed a cross-sectional observational study, with 1-year follow-up. The primary endpoint was estimate of efficacy, expressed as a proportionate change in LSS films from 1982 to 1992, adjusted for ED volume. The secondary endpoint was estimate of safety of this protocol. We found a 28% proportionate decrease in LSS films from 1982 to 1992. Among 520 patients with back complaints not meeting criteria for films, 4 had serious back pathology on follow-up that may have been detected on the index ED visit if LSS films had been ordered. All four were HIV(+), three of whom initially denied, then subsequently admitted, to active intravenous drug use. We conclude that use of a simple requisition form is associated with a quantitatively and statistically significant sustained reduction in utilization of ED LSS radiography. This protocol appears to provide a safe, cost-efficient means of limiting LSS film ordering by housestaff, with the possible exception of HIV(+) individuals. Because the association between HIV and radiographic abnormalities may be confounded by intravenous drug use, we cannot determine, on the basis of our data, whether HIV(+) status should be considered an independent criterion for obtaining LSS films.

Adult↗

The influence of multivalent phosphate structure on the properties of ionically cross-linked chitosan films for controlled drug release.

The aim of this paper was to investigate the electrostatic interactions between multivalent phosphates (phosphate (Phos), pyrophosphate (Pyro) and tripolyphosphate (TPP)) and chitosan, as well as the influence of electrostatic interactions on the properties of chitosan films ionically cross-linked by the above mentioned phosphates. The charge number of Phos was too low to interact with chitosan, while Pyro and TPP with more negative charges showed a significant ability to ionically cross-link chitosan. Solution pH played an important role on the charge numbers carried by Pyro, TPP and chitosan, especially for Pyro/chitosan. For instance, at pH less than 2.0 the interaction between Pyro and chitosan disappeared, while for TPP/chitosan even in solutions at pH less than 0.5 it still existed. Media pH and ionic strength also had a significant influence on the properties of cross-linked chitosan film with multivalent phosphates. Usually these films swelled and drug was released quickly in acidic conditions (such as in simulated gastric fluid) while under neutral conditions (such as in simulated intestinal fluid) they remained in a shrinkage state and drug was released slowly. Compared to TPP/chitosan films, Pyro/chitosan films exhibited much better pH-sensitive swelling and controlled release properties due to their relatively weak electrostatic interaction. The same reasoning was used to explain the significant acceleration of Pyro/chitosan film swelling and model drug release observed on adding sodium chloride. These films may be promising for site-specific drug delivery in the stomach.

Anions↗

The formation of colonic digestible films of amylose and ethylcellulose from aqueous dispersions at temperatures below 37 degrees C.

The film forming properties of a commercial aqueous ethylcellulose dispersion (Surelease) mixed with a range of ratios of an amylose/butanol complex in the presence of a range of concentrations of a plasticiser has been studied by measuring the minimum film forming temperature (MFFT). Contrary to what was to be anticipated from the literature, it was found that an additional 4% of the plasticiser (dibutyl sebacate), normally present in the standard formulation of the ethyl cellulose dispersion, was sufficient to lower the MFFT to allow the formation of films at 35 degrees C. This was confirmed by assessment of the glass transition temperature of free films prepared by casting and drying at 35 degrees C by the application of dynamic mechanical analysis. This technique also demonstrated that the ethylycellulose and the amylose were not miscible. The ability of faecal slurry to digest the films formed at low temperatures was confirmed by the use of a batch fermenter. The extent of digestion was directly related to the amylose content of the films, ensuring the potential to provide films, which could function as colon specific coatings.

Amylose↗

The formation and permeability of drugs across free pectin and chitosan films prepared by a spraying method.

This study has investigated the permeation of drugs through free films made of pectin and chitosan. The background for this study is the intended use of the films as coating material in a colon-specific drug delivery device. The factors that varied when making the films were the pectin source and grade of the pectin, degree of deacetylation of the chitosan and ratio between pectin and chitosan. The permeability of the model drug in 0.1 M HCl was low with an average drug release of 1.3 x 10(-3)%/cm. The films containing high content of chitosan showed exponential kinetics while the films containing high content of pectin showed 0-order kinetics. The release of drug in phosphate buffer pH 6.8 showed 0-order kinetics. The lowest permeability was obtained for a film consisting of a high content of pectin to chitosan, chitosan with a high degree of deacetylation and non-amidated low methoxylated citrus pectin. The permeation of paracetamol for this combination was 9.4 x 10(3)%/cm. This film combination had a combined diffusion of only 0.046%/cm after 1 h in 0.1 M HCl and 4 h in phosphate buffer pH 6.8.

Chitin↗