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PubMed1997

Calcium alginate dressings promote healing of split skin graft donor sites.

A prospective controlled trial was carried out to assess the healing efficacy of calcium alginate and paraffin gauze on split skin graft donor sites. Thirty patients were randomised to the calcium alginate group and 21 to the paraffin gauze group. The donor sites were assessed at 10 days post harvesting to determine if they were completely healed (100%) or not. Twenty one of the 30 patients dressed with calcium alginate were completely healed at day 10, while only 7/21 in the paraffin gauze group were healed (p < 0.05). There were two infections in the study, both occurring in the alginate group while there was no difference in dressing slippage between the two groups. Calcium alginate dressings provide a significant improvement in healing split skin graft donor sites.

Adolescent↗

PubMed1997

Preparation and characterization of antibacterial alginate film containing DNA as a carrier of silver ion.

Silver ion was effectively bound to alginate film by the aid of DNA. Amounts of Ag(I) bound to the films (18 mm x 18 mm) were 4.2 micrograms and 24.7 micrograms in the absence and presence of DNA, respectively. The Ag(I)-induced films showed antibacterial activity for both of Escherichiacoli (E. coli) and Staphylococcus aureus (S. aureus), and the activity of DNA-alginate film was obviously larger than alginate film. These results indicate that Ag(I) induced in the DNA-alginate film keeps the potent antibacterial activity, and so DNA is very useful as a carrier of silver ion in the alginate film.

Alginates↗

PubMed1998

A fibre-free alginate dressing in the treatment of split thickness skin graft donor sites.

BACKGROUND: Alginate dressings are gaining acceptance in would management although comparative published data with conventional treatment is inconclusive. AIMS: The aim of this randomised controlled study was to compare a fibre-free alginate dressing (Comfeel SeaSorb) with conventional treatment of standardised split-thickness skin graft donor sites in 17 patients regarding initial absorption of blood and healing. RESULTS: The alginate dressing absorbed 40% (P < 0.05) more blood, measured as total iron content of used dressings, during the first 10 post-wounding minutes than fine mesh gauze, resulting in less subsequent bleeding. Light microscopic examination of punch biopsies obtained from 10 wounds on post-operative day 6 demonstrated that nine wounds treated with the alginate dressing compared with seven wounds treated conventionally with paraffin-impregnated gauze (Jelonet) were completely epithelialized, a statistically non-significant difference (P = 0.46). CONCLUSIONS: In conclusion, the fibre-free alginate dressing showed increased initial blood absorption resulting in quicker haemostasis but showed no greater beneficial effect on epithelialisation of split-thickness skin graft donor sites compared with conventional topical treatment.

Absorption↗

PubMed2006

Purification and characterization of an alginate lyase from marine Bacterium Vibrio sp. mutant strain 510-64.

Marine Vibrio sp. 510 was chosen as a parent strain for screening high producers of alginate lyase using the complex mutagenesis of Ethyl Methanesulphonate and UV radiation treatments. The mutant strain Vibrio sp. 510-64 was selected and its alginate lyase activity was increased by 3.87-fold (reaching 46.12 EU/mg) over that of the parent strain. An extracellular alginate lyase was purified from Vibrio sp. 510-64 cultural supernatant by successive fractionation on DEAE Sepharose FF and two steps of Superdex 75. The purified enzyme yielded a single band on SDS-PAGE with the molecular weight of 34.6 kDa. Data of the N-terminal amino acid sequence indicated that this protein might be a novel alginate lyase. The substrate specificity results demonstrated that the alginate lyase had the specificity for poly G block.

Amino Acid Sequence↗

PubMed2000

Influence of dissolved oxygen tension and agitation speed on alginate production and its molecular weight in cultures of Azotobacter vinelandii*

The alginate production by Azotobacter vinelandii, as well as the molecular weight of the polymer, are strongly influenced by the dissolved oxygen tension (DOT) and stirring speed of the culture. Under high DOT (5% of air saturation), the bacteria produced more alginate (4.5 g/l) than that obtained at low (0.5%) oxygen tension (1.0 g/l) in cultures conducted at 300 rpm. On the other hand, under constant DOT (3%), the higher the stirring speed (from 300 to 700 rev./min), the higher the specific growth rate and the alginate production rate. However, low agitation speed (300 rev./min) lead the culture to produce a polymer of high molecular weight (680 000 g/g mol) whereas a low molecular weight (352 000 g/g mol) alginate was isolated from cultures conducted at high (700 rev./min) stirring speed. At 700 rev./min, the MMW increased to a plateau between 1 and 3% DOT and then decreased to a minimum of 0.11 x 10(6) g/g mol at 7%. Microscopic observations revealed the presence of cell aggregates (one order of magnitude larger than individual cells) when the culture was conducted at 300 rev./min. Oxygen gradients occurring within the aggregates could be responsible of this phenomenon. At high agitation rate, the MMW of the alginate dropped towards the end of the culture in all conditions evaluated. Alginase activity was detected, which would be responsible for this phenomenon.

Journal Article↗

PubMed2005

Effects of surfactant and temperature on rheological and structural properties of semidilute aqueous solutions of unmodified and hydrophobically modified alginate.

The dynamic and structural perturbations that result from the interactions between the anionic surfactant sodium dodecyl sulfate (SDS) and the hydrophobically modified biopolymer alginate (HM-alginate) have been studied with the aid of rheological methods, turbidimetry, and small-angle neutron scattering (SANS). The rheological results for a semidilute HM-alginate solution in the presence of SDS disclose strong interactions between HM-alginate and SDS at a low level of surfactant addition, and this feature is accompanied by enhanced turbidity. At higher surfactant concentrations the association complexes are disrupted. A strong temperature effect of the viscosity is observed in HM-alginate solutions at moderate SDS concentrations, where an elevated temperature leads to enhanced chain mobility, which promotes a breakup of the association complexes. The SANS results reveal a pronounced peak in the plot of scattered intensity versus wavevector q at intermediate q values for SDS concentrations above the critical micelle concentration (cmc). With contrast-matching conditions, using deuterated SDS instead of SDS, no interaction peak appears but an "upturn" in the scattered intensity is observed at small q value. The magnitude of this effect decreases with increasing surfactant concentration, showing clearly that SDS is capable of breaking up the large aggregates created.

Journal Article↗

PubMed2006

Alginate lyase enhances antibiotic killing of mucoid Pseudomonas aeruginosa in biofilms.

Once mucoid (alginate-producing) strains of Pseudomonas aeruginosa have become established in the respiratory tracts of cystic fibrosis patients they can rarely be eliminated by antibiotic treatment alone; we have investigated, in an in vitro biofilm system, the putative role of co-administration of alginate lyase with antibiotic. Biofilms were maintained in continuous flow culture in a medium resembling sputum from CF patients. Antibiotics and/or alginate lyase were added to some of the cultures. Biofilms of two mucoid CF strains of P. aeruginosa were, in most cases, not eradicated by a one-week course of treatment with 64 microg/ml of gentamicin; the same concentration of gentamicin, under the same conditions, led to the apparent elimination of all biofilms of non-mucoid derivatives of these strains. When alginate lyase and gentamicin were administered together the apparent elimination of mucoid bacteria from biofilms was achieved, whereas the mucoid bacteria in most control biofilms treated only with gentamicin persisted. Ceftazidime treatment of biofilms was more effective against those containing the non-mucoid strains than those with mucoid strains. These studies support the view that co-administration of antibiotics with alginate lyase, which degrades the exopolysaccharide produced by mucoid strains of P aeruginosa, might benefit CF patients by increasing the efficacy of antibiotic in the respiratory tract.

Anti-Bacterial Agents↗

PubMed1998

Method To immobilize the aphid-pathogenic fungus erynia neoaphidis in an alginate matrix for biocontrol

Erynia neoaphidis is an important fungal pathogen of aphid pests worldwide. There have been few reported attempts to formulate this natural agent for use in biocontrol. In the current study, factors involved in the immobilization of E. neoaphidis hyphae in an alginate matrix were investigated. Hyphae of two isolates cultured in liquid medium were 220 to 620 &mgr;m in length and 7 to 19 &mgr;m in diameter with a 74 to 83% cytoplasmic content. The optimal concentration of low-viscosity sodium alginate for production of conidia from entrapped hyphae was 1.5% (wt/vol), and 0.1 and 0.25 M calcium chloride were equally suitable for use as the gelling solution. Alginate beads were rinsed with 10% sucrose after gelling. However, beads should not be left for longer than 40 min in 0.1 M calcium chloride or 10% sucrose to prevent a 10% loss in conidial production. A 40% (vol/vol) concentration of fungal biomass produced significantly more conidia than either 20% or the standard concentration of 10%. This effect persisted even after beads were dried overnight in a laminar flow hood and stored at 4 degreesC for 4 days. Conidia from freshly produced alginate beads caused 27 to 32% infection in Pea aphids as determined by standardized laboratory bioassays. This finding was not significantly different from infections in aphids inoculated with fresh mycelial mats or plugs from Petri dish cultures. In conclusion, algination appears to be a promising technique for utilizing E. neoaphidis in the biocontrol of aphid pests.

Journal Article↗

PubMed2001

Gene expression by human articular chondrocytes cultured in alginate beads.

Culture of articular chondrocytes in alginate beads offers several advantages over culture in monolayer; cells retain their phenotype for 8 months or longer. Earlier studies of chondrocytes cultured in alginate concentrated on collagen and proteoglycan synthesis. However, gene expression by in situ hybridization (ISH) has not been investigated. The purposes of the present study on human chondrocytes were (a) to modify the ISH procedure for the alginate beads to examine the mRNA expression of alpha1 (II) procollagen, aggrecan, and two matrix metalloproteinases (MMP-3 and MMP-8) thought to be involved in cartilage matrix degradation, and (b) to compare expression in cultured chondrocytes with that in chondrocytes of intact human cartilage. The modifications made for ISH include the presence of CaCl2 and BaCl2 in the fixation and washing steps and exclusion of cetyl pyridinium chloride. By ISH we show that aggrecan, MMP-3, and MMP-8 are continuously expressed during 8 months of culture. The alpha1 (II) procollagen gene is expressed only during the first 2 months of culture and after 3 months its expression is undetectable, which is consistent with its absence in adult articular cartilage. By Western blotting, Type II collagen protein had been synthesized and deposited in both the cell-associated and further-removed matrix compartments at 7 and 14 days of culture. These data indicate that chondrocytes cultured in alginate beads could be preserved for immunohistochemistry and ISH and that culture of human chondrocytes in alginate beads may serve as a good model for studying cartilage-specific phenotype as well as factors that influence cartilage matrix turnover.

Adult↗

PubMed2001

Analysis and expression of algL, which encodes alginate lyase in Pseudomonas syringae Pv. syringae.

Pseudomonas syringae produces alginate, an exopolysaccharide that contributes to the virulence and epiphytic fitness of this phytopathogenic bacterium. P. syringae also produces the algL-encoded alginate lyase, which cleaves the alginate biopolymer via a beta-elimination reaction. The algL gene from P. syringae maps to a 1134 bp region within the alginate biosynthetic operon, and is similar to algL from Halomonas marina, P. aeruginosa, Azotobacter chroococcum, and A. vinelandii. algL from P. syringae was over expressed in Escherichia coli; two periplasmic forms of AlgL were overproduced (40 and 37 kDa). Both forms were enzymatically active and recognized by antibodies raised against AlgL from P. aeruginosa. Analysis of the regions flanking algL revealed significant homology to algX and algI, genes previously identified in the biosynthetic operon of other alginate-producing bacteria.

Amino Acid Sequence↗

PubMed1998

DNA protection from extracapsular nucleases, within chitosan- or poly-L-lysine-coated alginate beads.

DNA was immobilized within alginate matrix using an external or an internal calcium source, and then membrane coated with chitosan or poly-L-lysine. Membrane thickness increased with decreasing polymer molecular weight and increasing degree of deacetylation (chitosan). Beads were exposed to a 31,000 molecular weight nuclease to determine the levels of DNA protection offered by different membrane and matrix combinations. Almost total hydrolysis of DNA was observed in alginate beads following nuclease exposure. Less than 1% of total double-stranded DNA remained unhydrolyzed within chitosan- or poly-L-lysine-coated beads, corresponding with an increase in DNA residuals (i.e. double- and single-stranded DNA, polynucleotides, bases). Chitosan membranes did not offer sufficient DNA protection from DNase diffusion since all of the double-stranded DNA was hydrolyzed after 40 min of exposure. Both chitosan and poly-L-lysine membranes reduced the permeability of alginate beads, shown by enhanced retention of DNA residuals after DNase exposure. The highest level of DNA protection within freshly prepared beads was obtained with high molecular weight (197,100) poly-L-lysine membranes coated on beads formed using an external calcium source, where over 80% of the double-stranded DNA remained after 40 min of DNase exposure. Lyophilization and rehydration of DNA beads also reduced permeability to nucleases, resulted in DS-DNA recoveries of 60% for chitosan-coated, 90% for poly-L-lysine-coated, and 95% for uncoated alginate beads.

Alginates↗

PubMed1999

In vivo evaluation of a novel alginate dressing.

Alginate dressings are currently used in the management of epidermal and dermal wounds, and provide a moist environment that leads to rapid granulation and reepithelialization. However, a cytotoxic effect on proliferation of fibroblasts and residual material with inflammation in healing wounds have been reported recently. We have developed a new alginate dressing (AGA-100), which does not have an inhibitory effect on proliferation of fibroblasts. The purpose of this study was to evaluate the new alginate dressing with respect to wound healing in full- and partial-thickness pig wounds and with respect to biodegradation following implantation into rabbit muscle. Kaltostat and Sorbsan, both well-established commercial dressings, were used as control. The closure rate of full-thickness wounds treated with AGA-100 was significantly higher on day 15 compared with that with Kaltostat and Sorbsan. Reepithelialization rate of partial-thickness wounds treated with Sorbsan was statistically significantly lower on day 3 than those with the other two dressings. As to dressing debris remained in the healing wound, a large amount of foreign debris was noted in all the full-thickness wounds treated with Kaltostat or Sorbsan, while only about one-third of wounds treated with AGA-100 showed a little dressing debris. AGA-100 implanted into the muscle of rabbits was bioresorbed completely within 3 months. Therefore, dressing residue in AGA-100-treated full-thickness wounds might be fully absorbed in a few months. In conclusion, it is shown that our newly developed AGA-100 possesses superior properties compared with typical alginate dressings.

Alginates↗

PubMed2001

Osmotic pressure test: a simple, quantitative method to assess the mechanical stability of alginate microcapsules.

Implantation of microencapsulated, nonautologous cells and tissues is an effective method to deliver therapeutic proteins in vivo. Its success depends on the maintenance of the immunoisolating barrier provided by the microcapsule. Thus, one goal in the development of this technology is to create mechanically stable microcapsules. We have developed an osmotic pressure test to quantify the strength of microcapsules by exposing alginate microcapsules to a graded series of hypotonic solutions and quantifying the percentage of broken microcapsules. The test was validated by confirming the relative strengths of different types of alginate capsules, previously known from implantation in dogs to have differing mechanical stability in vivo. Thus, solid alginate microcapsules crosslinked with Ba(2+) were shown to be stronger than those crosslinked with Ca(2+), which in turn were shown to be stronger than the corresponding hollow alginate microcapsules. The incorporation of cells was demonstrated to reduce the mechanical stability of the microcapsules significantly. Hence, this test provides a simple and quantitative method for rapidly determining the strength of a large number of microcapsules. Thus, it is suitable for monitoring the mechanical stability of various types of microcapsules, predicting the performance of microcapsules in vivo, and for quality control of microcapsules during scale-up productions.

Alginates↗

PubMed2001

The effect of host factors and capsule composition on the cellular overgrowth on implanted alginate capsules.

Microencapsulation of islets of Langerhans in alginate/poly-L-lysine (PLL)/alginate capsules may provide a method for transplantation in the absence of immunosuppression. The aim of this study was to investigate the problem of overgrowth on implanted capsules with regard to the composition of the capsules and host factors such as cytokine and nitric oxide production. Empty capsules were implanted to C57BL/6 mice for 1, 3, 7, or 28 days. Glucose oxidation rates showed the metabolic activity of the cellular overgrowth on retrieved capsules. DNA content, histological score, and retrieval rates were also measured to assess the overgrowth. It was noted that the pericapsular host reaction arose by day 7 and had not increased further by day 28. Capsules of varying alginate compositions and different concentrations of PLL were implanted for 7 days to either C57BL/6 or Balb/c mice. Capsules were also implanted to mice lacking the inducible nitric oxide synthase enzyme. Glucose oxidation rates, DNA content, and histological score were positively correlated to each other and negatively correlated to retrieval rates. The pericapsular reaction was reduced if PLL was omitted from the capsule or if a high mannuronic acid alginate was used. Balb/c mice had reduced cellular overgrowth on implanted capsules and had reduced mRNA expression of interleukin-1 beta and tumor necrosis factor-alpha in their peritoneal macrophages. The capsular overgrowth seemed more severe in animals lacking inducible nitric oxide synthase compared with wild-type controls. It is concluded that alginate composition, PLL, and recipient factors such as nitric oxide production and cytokine expression affect the cellular overgrowth on implanted alginate capsules.

Alginates↗

PubMed2001

Degradation of phenol by Trichosporon sp. LE3 cells immobilized in alginate.

The degradation of phenol by freely suspended cells of Trichosporon sp. LE3 and alginate-immobilized cells was studied in batch culture. The alginate concentration (2 or 4%) and the cross-linking salt used (BaCl2 or CaCl2) affected the rate and percentage of phenol degradation. The highest values were obtained for immobilized cells at 2% calcium alginate, although complete degradation of 15 and 18 mM phenol was not observed. When the cell concentrations in the assays were doubled, the 2% calcium alginate-immobilized cells were able to degrade up to 30 mM phenol in less than 120 hours, although the free cells did not completely degrade phenol at concentrations above 20 mM. The maximum phenol degradation rate was a strong function of initial phenol concentrations, being the highest values being observed for 20 mM phenol.

Alginates↗

PubMed2004

Gelation conditions and transport properties of hollow calcium alginate capsules.

The diameter, membrane thickness, and compression intensity of hollow Ca-alginate capsules were measured at different gelation conditions, such as the reactant concentration, dropping velocity, and gelation time. The optimum operation conditions for preparing capsules were determined at 100 g/L CaCl(2), 10 g/L sodium alginate (Na-alginate), a dropping velocity of 150 droplets/min, and a gelation time of 10 min. Diffusion of some saccharide and amino acid from bulk solution into capsules was investigated, and the diffusion coefficients were calculated by the developed mathematical model. All the tested substances can diffuse easily into the capsules. The combined diffusion coefficients of the capsule D(m) are 92-99% as large as their diffusion coefficients in pure water, while the diffusion coefficients in the capsule membrane D(1) are 60-95% as large as those. By employing polyethylene glycol (PEG) and bovine serum albumin (fraction V) (BSA(V)), the molecular weight cut-off of the capsule was determined. For linear macromolecules, hollow Ca-alginate capsules have a molecular weight cut-off of 4000. No diffusion of BSA(V) into the capsules was observed.

Alginates↗

PubMed1990

Clinical trial of calcium alginate haemostatic swabs.

The influence of a new haemostatic material on surgical bleeding was evaluated in 100 patients who were prospectively randomized to either normal surgical gauze or calcium alginate swabs used throughout cholecystectomy (n = 40), simple mastectomy (n = 18) or inguinal hernia repair (n = 42). Overall, median (range) blood loss was 91 (3-329) ml for gauze and was significantly reduced by calcium alginate swabs to 72 (2-181) ml (P less than 0.05). Unexpectedly, operation times were also shortened from 45 (17-95) min for gauze to 35 (13-70) min with calcium alginate swabs (P less than 0.02). This reduction in blood loss and operating time was greatest for mastectomy, was still statistically significant for cholecystectomy, but was unimportant in inguinal hernia repair. Calcium alginate haemostatic swabs may become routine in major surgery, particularly where blood loss leads to the need for transfusion.

Alginates↗

PubMed2002

Chitosan-alginate PEC membrane as a wound dressing: Assessment of incisional wound healing.

Flexible, thin, transparent, novel chitosan-alginate polyelectrolyte complex (PEC) membranes, cast from aqueous suspensions of chitosan-alginate coacervates with CaCl(2), were evaluated as potential wound-dressing materials. MTT and NR assays suggested that the chitosan-alginate PEC membranes and their aqueous extracts were nontoxic towards mouse and human fibroblast cells. Cell growth was also not hindered by co-incubation with the membranes. Compared to conventional gauze dressing, the PEC membranes caused an accelerated healing of incision wounds in a rat model. Wounds closed at 14 days postoperatively, and histological observations showed mature epidermal architecture with keratinized surface of normal thickness and a subsided inflammation in the dermis. This was followed by an excellent remodeling phase with organized thicker collagen bundles and mature fibroblasts at 21 days postoperative. Control wounds continued to show signs of an active inflammatory phase under scab on Day 21. Closure rate and appearance of PEC membrane-treated wounds were comparable with Opsite(R)-treated wounds. On the basis of its biocompatibility and wound-healing efficacy, the chitosan-alginate PEC membrane can be considered for wound-dressing applications.

Alginates↗