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Effect of cecal cultures encapsulated in alginate beads or lyophilized in skim milk and dietary lactose on Salmonella colonization in broiler chicks.

Cultures of cecal bacteria that were grown under anaerobic conditions were prepared as lyophilized powder or encapsulated and lyophilized in alginate beads and compared with broth cultures for control of Salmonella typhimurium enteric colonization and for ease of delivery in four experiments. Day-old broiler chicks were provided a standard corn-soybean diet with: 1) no culture, no lactose (control); 2) dietary lactose; 3) broth culture by crop gavage; 4) culture in lyophilized powder; or 5) culture encapsulated and lyophilized in alginate beads. Two percent lactose (wt/wt) was included in the diet of chicks in Groups 2 to 5. The lyophilized powder was provided by mixing with the feed. Alginate beads were force-fed or top-dressed on the feed. All groups were challenged on Day 3 with 10(4) cfu of S. typhimurium per chick. Culture by gavage and culture in alginate beads and dietary lactose significantly (P < .05) reduced mean Salmonella colony-forming units in cecal contents by 3.4 to 5.3 log10 units at 10 d of age. Likewise, the number of cecal-culture-positive chicks in the groups provided culture by gavage or in beads was significantly less than controls. Eighty to 90% of the beads provided were consumed by the chicks within 2 h. The results indicated that cultures of cecal bacteria encapsulated and lyophilized in alginate beads were as effective as culture by crop gavage for the reduction of Salmonella colonization in broiler chicks and provide an effective and practical method of delivery.

Alginates↗

The effects of poly(ethyleneimine) (PEI) molecular weight on reinforcement of alginate hydrogels.

Alginate hydrogels are widely used for cell encapsulation and transplantation, and they are frequently surface reinforced with secondary polymers to enhance their mechanical rigidity and stability. We hypothesized that the molecular weight (MW) of the polymer utilized to reinforce alginate would be an important factor in their stability, particularly when the gel network was homogeneously reinforced with the polymer. This hypothesis was investigated with alginate hydrogels cross-linked with Ca2+, and reinforced throughout the bulk of the gel with poly(ethyleneimine) (PEI) having different MWs. Interactions between the two polymers became significant following gelation, leading to higher elastic moduli (E) than gels with no PEI. The decrease in E of gels incubated in isotonic salt solutions over time, utilized as an indication of gel break down, was ameliorated with an increase in the MW of the PEI. In addition, the dependencies of the moduli and viscoelasticity on the temperature also became smaller with the use of high MW PEI. This is likely due to the limited mobility of high MW PEI, leading to a higher energy for dissociation. The stable interactions between the alginate and PEI prevented alterations of the pore structure in the gels, and slowed the deterioration of gel properties even under continuous agitation in a bioreactor. The results of this study will likely be useful in designing alginate encapsulation strategies for various applications.

Alginates↗

Cryopreservation of alginate-encapsulated recombinant cells for antiangiogenic therapy.

The potential benefit of continuous local administration of antiangiogenic proteins to CNS tumors in vivo has recently been demonstrated using endostatin-producing recombinant cells encapsulated in alginate beads. Due to the treatment potential of transplanted alginate-encapsulated cells producing therapeutic proteins, we describe a successful method of cryopreservation (CP) of such beads, in which cellular viability, alginate structure, and protein secretion were maintained. Alginate beads containing human embryonic kidney cells (HEK 293 cells) stably transfected with the gene encoding for endostatin were cryopreserved in dimethyl sulfoxide (DMSO) using a slow freezing procedure. Briefly, the DMSO concentration was gradually increased prior to the freezing procedure. The cryotubes were further supercooled to -7.5 degrees C and nucleated. Thereafter, the samples were cooled at a rate of 0.25 degrees C/min and stored in liquid nitrogen. The viability of the encapsulated cells was assessed using confocal microscopy quantification (CLSM) technique and a MTS assay. The cell cycle distribution inside the beads was assessed by DNA flow cytometry and endostatin production was determined by an endostatin-specific ELISA assay, both prior to and after CP. CLSM measurements showed sustained esterase activity in the beads after thawing, with only a slight transient decrease 24 h after CP. The MTS assay verified these findings by displaying similar variations of intracellular dehydrogenase activity. Flow cytometric analyses revealed no cryorelated disturbances in cellular ploidy. Furthermore, ELISA measurements showed a well-preserved endostatin production after CP. In conclusion, this work describes the successful CP of alginate-encapsulated recombinant cells secreting a therapeutic protein. Together with previous published reports, these results further substantiate the feasibility and potential of cell encapsulation therapy in the treatment of malignant tumors.

Alginates↗

Hyaluronate-alginate gel as a novel biomaterial: mechanical properties and formation mechanism.

With the aim of producing a biomaterial for surgical applications, the alginate-hyaluronate association has been investigated to combine the gel-forming properties of alginate with the healing properties of hyaluronate. Gels were prepared by diffusion of calcium into alginate-hyaluronate mixtures, with an alginate content of 20 mg/mL. The hyaluronate source was shown to have significant effect on the aspect and the properties of the gels. The gels have viscoelastic behaviour and the transient measurements carried out in creep mode could be interpreted through a Kelvin-Voigt generalised model: experimental data led to the steady state hardness and a characteristic viscosity of the gel. Gels prepared from Na rooster comb hyaluronate with weight ratio up to 0.50 have satisfactory mechanical properties, and fully stable gels are obtained after a few days; on the contrary, use of lower molecular weight hyaluronate led to loose gels for hyaluronate contents over 0.25. Gel formation was investigated by measurements of the exchange fluxes between the calcium chloride solution and the forming gel, which allowed thorough investigations of the occuring diffusion phenomena of water, calcium ion and hyaluronate. Strong interactions of water with hyaluronate reduce significantly the rate of weight loss from the gel beads and allows higher water content in steady-state gels. Calcium content in the gel samples could be correlated to the actual alginate concentration, whatever the nature and the weight ratio of hyaluronate.

Alginates↗

Effects of a silicone-coated polyamide net dressing and calcium alginate on the healing of split skin graft donor sites: a prospective randomised trial.

An open randomised prospectively controlled trial was performed to assess the healing efficacy, slippage rate and degree of discomfort on removal of calcium alginate and a silicone-coated polyamide net dressing on split skin graft donor sites. Sixteen patients were randomised to the calcium alginate group and 14 to the silicone-coated group. The donor sites were assessed at days 7, 10, 14 and up to day 21. The mean time to healing in the calcium alginate group was 8.75 +/- 0.78 days (range 7 to 14 days) compared to 12 +/- 0.62 days (range 7 to 16 days) for the silicone-coated group (p < 0.01). Although more silicone-coated dressings slipped (5 versus 1), the difference was not statistically significant. Pain during the first dressing change was assessed using a visual analogue pain scale. Although no significant differences were found between the groups, it was necessary to change the dressing protocol in the silicone-coated arm of the trial after entering the first two patients. Overlaid absorbent gauze adhered to the donor site through the fenestrations in the dressing necessitating the placement of paraffin gauze between the experimental dressing and the overlying cotton gauze. There was one infection in the study, occurring in the alginate group. Based on these results we recommend calcium alginate as the dressing of choice for split skin graft donor sites.

Adult↗

[Effect of alginate purity on microencapsulated hepatocyte overgrowth in peritoneal transplantation].

OBJECTION: To study the effect of alginate purity on microcapsule overgrowth in peritoneal transplantation. METHODS: Alginate was purified by filtration and chloroform/butanol extraction. Using purified and crude alginate to make two kinds of microencapsulated hepatocytes (MHs), we transplanted 5 ml of each kind of MHs into the peritoneal cavities of rats. After 2 weeks, 1, 2, and 6 months, MHs were washed out to study overgrowth of MH, histology, enzyme and albumin of hepatocytes. Human lymphocytes transformation was measured by 3H-TdR incorporation with two kinds of alginate solutions. RESULTS: The amount of retrieved MHs was larger in the purified than in the crude ( P < 0.01). In the purified groups, most hepatocytes had normal features of morphology, enzymatic histochemistry, and albumin immunohistochemistry staining after one month of transplantation, but were damaged in the crude groups only 2 weeks later. Microcapsules were smooth, regular and had no overgrowth during 6 months in the purified but overgrowth one month later after transplantation in the crude. The liquid scintillation counting (cpm) was lower in the purified than in the crude. CONCLUSION: The purified alginate could reduce the overgrowth of MH and improve the function of hepatocytes significantly.

Alginates↗

[The effect of levels of alkali salts in Alginate-LF-250 on physico-mechanical properties of materials used in making impressions based on irreversible colloids].

The aim of this paper is the examination of physical-mechanical properties of alginate composition regarding the change of alkali salt alginate composition in the limits of the presence from 8 to 20%, and after the standard ISO 1563, 1978 (E). The combinations make homogenized mixtures of micronized powders of the basic component, retarders, accelerators, system stabilizators, the means for colloides and constituents lowering, which are prepared in the relation of 16 gr/40ml. The change of alkali salt alginate concentration from 8 to 20% does not effect the working time and the time of alginate mash binding, while percentage of permanent deformation of the binding material is larger than the allowed standard after ISO 1563, 1978 (E) in case when alginate alkali salts in the composition are present in the smaller quantity than 12%.

Alginates↗

Use of calcium alginate to pack abscess cavities: a controlled clinical trial.

Many conventional dressings are painful when removed, and may be detrimental to healing. In a pilot study ten consecutive abscesses, requiring incision and drainage, were packed with a calcium alginate dressing: this was well tolerated, its removal causing minimal pain. No adverse effects were attributable to its use. A controlled trial was therefore carried out to compare calcium alginate with the more traditional saline-soaked gauze for packing abscess cavities, following incision and drainage. Patients were randomized to receive either calcium alginate (16 patients) or gauze dressing (18 patients). At the first dressing change the patient marked on a linear analogue scale the pain experienced; the nurse noted similarly the ease of removal of the dressing. Calcium alginate was significantly less painful to remove after operation (P less than 0.01), and also easier to remove (P less than 0.01) than gauze dressings. If abscess cavities are packed after incision and drainage, calcium alginate appears to be an improvement on conventional dressings.

Abscess↗

[Studies on the influencing factors on the drug release from sodium alginate matrices].

AIM: To investigate the in vitro influencing factors on drug release from matrices with sodium alginate as the hydrophilic polymer. METHODS: Sodium alginate hydrophilic matrix tablets were prepared by direct compression method with theopylline as a model drug. The in vitro influencing factors on drug release behavior from matrices were studied by investigating the swelling, water uptake and erosion characteristics of pure sodium alginate matrices. RESULTS: The results showed that drug release rate and drug release mechanism were both related to the viscosity of sodium alginate used in matrices, pH values and ionic strength of dissolution media and rotation speeds. CONCLUSION: Sodium alginate can be tailor-made to suit the demands of applicants in sustained delivery systems as a good candidate of hydrophilic polymer.

Alginates↗

[Alginate, elastase and alkaline protease production of Pseudomonas aeruginosa strains isolated from various body sites].

In this study, alginate, elastase and alkaline protease production of Pseudomonas aeruginosa, which are accepted as virulence factors, have been investigated in the strains isolated from 60 lower respiratory tract samples of cystic fibrosis patients, together with the strains isolated from 59 lower respiratory tract, 62 urine and 32 blood samples of non-cystic fibrosis patients, in order to evaluate the relationship between the virulence factors and infected body sites. Our results indicated that all isolates including the ones from cystic fibrosis patients produced similar levels of alginate ranging between 1.830+/-1.666 and 1.305+/-1.690 microg/ml with the exception of urinary isolates for which the alginate amount was 0.9655+/-1.386 (p<0.05). Elastase levels exhibited no statistically significant difference among groups of isolates; and finally alkaline protease levels, although not statistically significant, was greater in urine isolates than blood and respiratory isolates; however, it was significantly lower in cystic fibrosis isolates compared to the remaining three groups (p<0.05). It can be concluded that alginate, elastase and alkaline protease levels do not differ when different body sites are considered. However, it was observed that alkaline protease was the least produced virulence factor in terms of different body sites. Alginate was found to be the most commonly produced virulence factor in respiratory tract isolates, regardless of presence of cystic fibrosis.

Alginates↗

[A study on the chondrogenesis of the compound of alginate gelatin and bone marrow stromal cells in vivo].

OBJECTIVE: To investigate the chondrogenisis by alginate gelatin and rats' bone marrow stromal cells (BMSCs) chondrogenicly induced in vitro. METHODS: Thirty-two male adult SD rats were assigned randomly to experimental and control groups. In experimental group, bone marrow was obtained from the right tibias of all the rats. After expanding and culturing 3 passages, induced BMSCs by chondrogenic culture medium for 10 days. Suspended induced cells in alginate gelatin, and injected the complex into the hypodermic tissue of the backs of rats autogenously. In control group only alginate gelatins were injected. The grafts were taken out for examinations 4 and 8 weeks after the operations. RESULTS: Considerable cartilage appeared in experimental group 8 weeks after operations. Regular HE staining and alcian blue staining showed a great deal of cartilage holding chondrocyte masses surrounded by abundant matrix. Alginate gelatin decompounded obviously, and the rest distributed among newly formed cartilage. No cartilage appeared in control group all through. CONCLUSION: BMSCs and alginate gelatin have a beautiful future in cartilage tissue engineering.

Alginates↗

A comparative study of Cu (II) biosorption on Ca-alginate and immobilized live and inactivated Cladosporium sp.

Spores of Cladosporium sp. were immobilized into Ca-alginate beads via entrapment. The alginate beads and both entrapped live and inactivated spores of Cladosporium sp. were used for comparison of biosorptive capacity from aqueous solutions. The factors affecting the adsorption ability on Cu (II), such as the contact time, initial pH, temperature were investigated. The results showed that the Ca-alginate beads containing live spores of Cladosporium sp. had the maximum biosorptive capacity. The biosorption equilibrium was established in about 3 h. The maximum biosorption of Cu (II) on Ca-alginate entrapping spores and no spores were obtained between pH 4.0 and 3.5. Temperature over the range of 15-45 degrees C had no significant effect on the biosorption capacity. The biosorptive capacity increased with initial concentrations in the concentration range of 30-800mg/l. The equilibrium was well described by Langmuir biosorption isotherms. The Ca-alginate beads could be regenerated using 0.1M HCl, The biosorbents were reused in three biosorption-desorption cycles with negligible decrease in biosorptive capacity.

Adsorption↗

Molecular weight characteristics of alginate produced by Azotobacter vinelandii in a membrane bioreactor.

A membrane bioreactor which combines a continuous fermentor and a cross-flow microfiltration module enabling continuous cell recycling and removal of the fermentation products freed of microbial cells was tested for the production of alginate by Azotobacter vinelandii. The molecular weight characteristics of the polysaccharide produced were investigated. The macromolecular characteristics of alginate recovered by first filtering batch fermentation broths through different microporous ceramic membranes with pore sizes varying from 0.45 to 1.4 microm seemed to be independent of the MPS. The 0.8-microm membrane pore size (MPS) was selected for continuous alginate production in the membrane bioreactor. A mean yield value of 0.21 g alginate g(-1) saccharose was obtained and an increase in alginate polydispersity as incubation proceeded was observed, probably due to enzymatic degradation of the polysaccharide.

Alginates↗

[Joint resurfacing using allograft chondrocytes embedded in alginate gel].

OBJECTIVE: To explore the feasibility of repairing the articular cartilage with allo-articular chondrocytes embedded in alginate gel. METHODS: Allo-articular chondrocytes were isolated from three adult New zealand rabbits. The cells were cultured in DMEM/F-12 supplemented with 20% fetal bovine serum (FBS). Chondrocytes of 2nd - 3rd passage were harvested and were diluted to 5.0 x 10(7) cells/ml with 1.2% alginate. Then alginate gel was formed by 102 mM CaCl(2). The gels were cultured subsequently for 1 week and then transferred to the full-thickness defects in the femoral condyles of adult rabbits. In control group the defects were left untreated. The animals were sacrificed in the 3rd and 6th month after operation respectively. The specimens were decalcified with 50% formic acid. The histologic sections were stained with safranin O-fast green, hematoxylin-eosin (H&E) and picric acid-Sirius red and immunohisto-stained for type II collagen and aggrecan. The repairing efficiency was evaluated according to Wakitani scoring. RESULT: In the experiment group all 8 defects acquired repair, 7/8 were repaired with mature hyaline cartilage tissue, and 1/8 was with fibrocartilage tissue for less cell-gel inputted. The thick of repaired tissues were closed to the normal and the tissue integrated smoothly with cartilage around the defects. Safranin O staining of the matrix acted in accordance with the normal and immunostaining for type II collagen and aggrecan showed positive. Picric acid-Sirius red staining showed that the chondrocytes lined in lines and the collagen aligned like Gothic architecture structure by polarization microscopy. There was no evidence of residue of alginate and inflammation in 3rd month specimens and no obvious deterioration at 6th month. But in control group, only a small amount of fibrous, fibrocartilage, or hyaline-like tissue was seen on the surface of the defects. Wakitani scoring showed 1.75 points for the experiment group and 7.65 for the control group. CONCLUSION: It is a promising way to repair the articular cartilage with homogeneous articular chondrocytes embedded in alginate gel.

Alginates↗

Absorption of fluoride following inhalation and ingestion of alginate impression materials.

The fluoride (F) contents of ten alginate impression materials in powder form, were found to range from 0.44% to 2.42%. Plasma F concentrations showed no appreciable changes in five dental personnel exposed to aerosols of these powders during routine clinical work. Elevated plasma F levels (120-158 ng/ml) were recorded after ingesting 2 g (approximately 10 mg F) of Zelgan normal-set alginate. The bioavailability of F from the alginate was about 55% of the total F in the administered dose. It is concluded that routine clinical exposure to alginate does not cause any important change in plasma F levels, whereas accidental swallowing of alginate raises the plasma F level significantly.

Adult↗

[Alginates of Pseudomonas aeruginosa: a complex regulation of the pathway of biosynthesis].

Pseudomonas aeruginosa is an opportunistic pathogen causing severe infections, especially in lungs of patients with cystic fibrosis. Environmental conditions induce the production by the bacteria of a viscous mucoid exopolysaccharide, called alginate, which is one of the most important factor of virulence of P. aeruginosa. Alginate is a linear polymer of beta-1, 4-linked L-guluronic acid and D-mannuronic acid. The alginate biosynthetic pathway involves genes called alg which are clustered at the 34 min region of chromosomal DNA of P. aeruginosa. The key enzyme of alginate biosynthesis, the GDP-mannose dehydrogenase is encoded by the gene algD. Expression of algD is positively controlled by several proteins, especially AlgU, a putative sigma factor homologous to sigma E of E. coli, AlgR and AlgP, a transactivator and an histone-like respectively. Here, a scheme of alginate biosynthetic pathway and a model for the alg genes regulation are described from results published in literature and from our own interpretation.

Alginates↗

Healing properties of calcium alginate dressings.

In a prospective, randomised, controlled trial of 92 patients with full-thickness pressure ulcers, the efficacy of an alginate wound dressing was compared to that of an established local treatment with dextranomer paste. During treatment, a minimal 40% reduction in wound area was obtained in 74% of the patients in the alginate group and in 42% of those in the dextranomer group. The median time taken to achieve this goal was four weeks with alginate and more than eight weeks in the control group. Mean surface area reduction per week was 2.39 cm2 (sd 3.54) and 0.27 cm2 (sd 3.21) in the alginate and dextranomer groups respectively (p = 0.0001). This difference was still highly significant when the sub-groups of almost completely healed subjects at the end of the study were considered. This striking healing efficacy of an alginate dressing suggests it possesses pharmacological properties which require further investigation.

Aged↗

[Protein isolation by means of complexing agents. 2. Formation of insoluble complexes of sunflower albumins with alginate or pectin].

The formation of insoluble complexes of sunflower seed albumin and alginate or pectin is studied by means of turbidimetric titration and by determining the pH-dependent precipitability of protein. The complex formation that is based on electrostatic interaction is a function of the pH value and the protein-polyanion ratio. Consequently, it is affected by the neutral salt content of the solutions. 90% and more of the dissolved protein may be precipitated if the proportion of the precipitant amounts to 20%. A sodium chloride content of 0.6% reduces the precipitability by alginate to 74%. In the presence of 0.3% sodium chloride, at most 55% of protein are still precipitated by pectin. The difference in strength between the albumin-alginate and the albumin-pectin complex is also expressed by the dye-binding power. Albumin-pectin complexes bind the same amount of amido black as free protein. On the contrary, albumin-alginate complexes exhibit reduced dye-binding power due to stronger binding of the protein to the polyanion. The results obtained by turbidimetric titration of model systems can, in principle, be extrapolated to the precipitation of albumins from protein extracts. In accordance with the heterogenicity of the protein, the turbidimetric titration of the albumin-alginate and the albumin-pectin complexes exhibits two maxima.

Albumins↗