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PubMed1994

Isolation and characterization of alginate-derived oligosaccharides with root growth-promoting activities.

Lytic digestion of poly(mannuronate), poly(guluronate), and alginate with an alginate lyase from Alteromonas macleodii was used to prepare mixtures of unsaturated oligosaccharides. Four oligosaccharides isolated from the alginate lyase-lysate by anion-exchange chromatography on Q-Sepharose were found to be the major components of the root growth-promoting lysate. The oligosaccharides were analyzed by NMR and SIMS and identified as di- and tri-saccharides having O-(4-deoxy-L-erythro-hex-4-enopyranosyluronic acid)-1--> at the nonreducing terminus. The trisaccharides from the lysate were found to have root growth-promoting activity in a barley bioassay.

Alginates↗
PubMed1992

Intraoral vesicles occurring after alginate impressions.

In 47 of 227 dental students intraoral vesicles developed after multiple alginate impressions. The lesions were generally solitary and clear, and appeared within 24 to 48 hours after the impression. They were most frequently located inside the vermilion border of the lips and resolved spontaneously in 2 to 5 days. The purpose of this study was to determine the cause of these reactions. Histopathologically one lesion was suggestive of a contact allergy. Cutaneous patch tests, which proved negative, were performed on 14 students to determine whether an allergy to the alginate flavoring existed. The surface of three lesions were cultured and the organisms identified. Contamination studies were carried out on seven unopened containers of the alginate powder and resulted in the isolation of some organisms similar to the mucosal cultures; however, no relationship can be proved. These findings indicate that the cause of the vesicles remains unknown, and further studies are necessary to establish the cause.

Alginates↗
PubMed1995

The relation between thermal and pH changes in alginate impression materials.

OBJECTIVES: The purpose of this study was to determine if pH changes are related to thermal changes during setting in alginate impression materials, and to investigate the possibility of establishing a mathematical model between thermal and pH changes. METHODS: Thermal and pH changes were recorded during the 20 min gelation period following the mixing of five commercial alginate materials. The data obtained were plotted and statistically analyzed using a student's T-test, multiplicative, linear and reciprocal analyses at a significance level of p<0.05. These analyses were used to determine the differences between various materials and to establish regression models between thermal and pH changes for each of the materials tested. RESULTS: For all samples, the pH values fell within the first minutes from the alkaline range into the neutral and even into the acidic range, while the initial sharp fall in temperature was followed by a subsequent gradual increase. Differences in the final gel structures of the alginate materials were shown to be related to the differences in thermal and pH changes during setting. This could be expected from their various chemical compositions such as the variety of components, the chemical forms of principal components and the proportion of each component. SIGNIFICANCE: This work contributes to the development of a mathematical model relating pH changes and thermal changes to form a basis for the correlation between chemical properties and characteristics in practical use.

Alginates↗
PubMed1994

A new enzyme immobilization procedure using copper alginate gel: application to a fungal phenol oxidase.

A new procedure was developed for enzyme immobilization by entrapment in copper alginate gel. The mechanical properties of the copper alginate gel were characterized and compared with those of the most widely used calcium alginate. The system was applied to the immobilization of a fungal phenol oxidase. Optimal conditions for enzyme immobilization were set up: the system immobilized 85% of the enzyme, and the remaining 15% was recovered in the aqueous immobilization medium. The stability and activity of the immobilized enzyme were studied. After immobilization, the enzyme was active in a wider pH range, the temperature of its optimal activity was shifted to lower values, and the possibility of storage at 4 degrees C was greatly improved. The immobilized enzyme generally increased the rate of oxidation of various substrates. The results indicate a potential use of this system for the construction of bioreactors to be used in the detoxification of polluted waste waters.

Agaricales↗
PubMed1994

11 alpha-hydroxylation of progesterone using modified alginate-immobilized cells.

The major disadvantage of using alginate-immobilized Aspergillus ochraceus for 11 alpha-hydroxylation of progesterone is its instability in phosphate solutions. In this study, different gel-hardening methods were used to improve the stability of alginate gels. Addition of Fe3+ and Fe2+ ions increased the activity of hydroxylation, but Fe2+ was not a good gelling agent. Replacement of CaCl2 by FeCl3 as the gelling agent increased the activity of the immobilized cells by about 33%. Hardening the alginate-immobilized cells with polyacrylamide (PAA) increased their stability in a 0.1 M phosphate buffer from 0.35 h to over 36 h. The productivity was increased by a factor of 1.8 compared to those without hardening, and the maximum yield was also increased from 84.3% to 90.8%. The physical properties and biocatalytic characteristics of PAA-hardened, immobilized cells were found to be feasible for application in a steroid bioconversion system.

Acrylic Resins↗
PubMed1987

Acute tissue reactions to potassium alginate with and without colour/flavour additives.

The acute tissue reactions to potassium alginate, locally applied to a microvascular bed, were studied with the use of the vital microscopic hamster cheek pouch model combined with correlative histology. This experimental model permitted us to study microvascular permeability, blood flow, vessel diameters and leucocyte adhesion to vessel walls intravitally, and leucocyte migration and mastcell degranulation histologically. Deionized water alone and potassium alginate with flavour and colour mixed in saline was found to cause severe microvascular alterations, while potassium alginate, without flavour and colour, but mixed in saline and applied to the microvasculature resulted in a minor inflammatory reaction.

Alginates↗
PubMed1986

Chronic (89-week) feeding study with hydroxypropyl distarch phosphate, starch acetate, lactose and sodium alginate in mice.

A chronic feeding study was carried out in mice with two chemically modified potato starches, hydroxypropyl distarch phosphate (HP-starch) and starch acetate (AC-starch), and with lactose and sodium alginate. Each of the materials was fed to a group of 75 male and 75 female mice for 89 wk. The dietary level of the test products was gradually increased until the diets contained (by weight) 55% HP-starch, 55% AC-starch, 55% lactose or 25% alginate. The control diet contained 55% pregelatinized potato starch. Each of the four test materials caused increased water consumption, distinct caecal and colonic enlargement, a slightly increased incidence of intratubular nephrosis and, with the exception of AC-starch, also slightly lower body weights. An increased incidence of gastric trichobezoars was observed in mice fed either the modified starches or lactose. The occurrence of concrements in the renal pelvis along with slight urinary changes, such as increased amounts of amorphous material in the urine and increased urinary Ca content, in mice fed HP-starch, AC-starch or lactose was regarded as an effect of little, if any, toxicological significance. Alginate fed at 25% (w/w) of the diet was nephrotoxic to mice, as shown by extremely high water consumption, high urine production, urinary incontinence, high pH and low specific gravity of the urine, increased level of blood urea nitrogen, increased kidney weights, distension of the renal calyx and the high incidence of dilated distal tubules. Caecal and colonic enlargement and changes in urinalysis were found to be reversible and had completely or largely disappeared within 2-5 wk of the cessation of the treatment in wk 87. The incidence of intratubular calcinosis or of concrements in the pelvic space was not reduced during the recovery period. The study did not provide any evidence of carcinogenicity of the products tested.

Alginates↗
PubMed1991

Influence of mixing disinfectant solutions into alginate on working time and accuracy.

Possible transfer of pathogens from dental surgery to dental laboratory is of concern to the profession. This study investigates changes in the working time and accuracy of an alginate impression material when mixed with solutions of selected disinfectants rather than water. The results indicate that certain disinfectant solutions, when used as a water substitute, do not modify the working time of the alginate investigated beyond acceptable limits and have no significant effect on the dimensional accuracy of that material. The use of a 0.2 per cent solution of chlorhexidene gluconate is recommended as a water substitute for the alginate investigated.

Alginates↗
PubMed1992

Occurrence of alginate gene sequences among members of the pseudomonad rRNA homology groups I-IV.

Total genomic DNA of 13 pseudomonads representing rRNA homology groups I-IV were screened for sequences homologous to four Pseudomonas aeruginosa alginate (alg) genes by Southern hybridization. Biotinylated probes for three structural genes (algA, algC and algD) and one regulatory gene (algR1) were prepared. Genomic DNA of strains representing group I (P. syringae pv. glycinea, P. viridiflava and P. corrugata) hybridized with all four gene probes. Hybridizing fragments were of differing sizes, indicating that evolutionary divergence among group I members has occurred. P. corrugata has not been reported to synthesize alginate. Genomic DNA from representatives of groups II-IV gave no or very weak hybridization with the probes except for algC. This study indicates that the ability to produce alginic acid as an exopolysaccharide among the pseudomonads is restricted to members of rRNA homology group I in agreement with earlier physiological studies.

Alginates↗
PubMed1993

Sequence of the alg8 and alg44 genes involved in the synthesis of alginate by Pseudomonas aeruginosa.

The alg8 and alg44 genes, which are required for alginate biosynthesis by Pseudomonas aeruginosa, are located in the alginate biosynthetic gene cluster between the algD and algE genes. The nucleotide sequence of these two genes is presented. Although the functions of the Alg8 and Alg44 proteins are not known, we believe that they may be involved in the polymerization of mannuronic acid residues to form alginate.

Alginates↗
PubMed1995

Peptide immobilization on calcium alginate beads: applications to antibody purification and assay.

Two different procedures were developed for the non-covalent immobilization of peptide antigens on calcium alginate beads. The antigenic peptide is first synthesized in a multimeric form starting from a polydentate lysine core, and then immobilized on alginate beads (average volume 0.05 ml) by entrapment or simply by non-covalent adsorption. Coupling yields, as monitored by RP-HPLC analysis of the immobilization time course and/or by amino acid analysis of derivatized beads, were close to 1-2 mg of peptide per ml of gel. Immobilization yields were not dependent on the peptide net charge, hydrophobicity or length, but mainly on the extent of peptide multimerization. After immobilization on alginate gel, peptide antigenic properties were fully retained, as clearly demonstrated by the batchwise micropreparative purification of anti-peptide antibodies in good yields and with a high degree of purity, directly from crude sera in a single adsorption-elution step. Derivatized beads were sufficiently stable towards repeated washing-equilibration procedures, allowing very limited peptide leakage from the matrix. Peptide beads were also successfully used for the development of solid-phase immunoassays in test-tubes to characterize the corresponding antibodies, with the immobilization yield and signal-to-noise ratio being greatly enhanced in comparison with other types of conventional supports.

Adsorption↗
PubMed1990

Clinical acceptability of orthodontic retainers fabricated from stored alginate impressions.

The purpose of this study was to determine the clinical acceptability of orthodontic retainers fabricated from stored alginate impressions. In the laboratory, alginate impressions were made of a standard dentoform. In the clinic, maxillary alginate impressions were made of the teeth of 20 dentate adults. In each case, sets of stone models were made either immediately or after the impressions had been stored for 24 or 72 hours at high humidity. The linear dimensions of the three sets of dentoform models were compared with one another and with the standard dentoform. No differences were found. Claspless acrylic retainers were fabricated on the models from the adult subjects. The retainers were positioned in the subjects' mouths and evaluated for snap fit, stability, cervical margin fit, and self-retention. With the use of these criteria, 95% of the retainers made from impressions that were poured immediately were clinically acceptable. When impressions were stored 24 and 72 hours, the percentages of acceptable retainers were 80% and 90%, respectively. The differences in percentage of clinically acceptable retainers were not statistically significant. When impressions are stored at high humidity, a large percentage of clinically acceptable orthodontic retainers can be made from models poured as long as 72 hours after the impression is made.

Adult↗
PubMed2004

A protein delivery system: biodegradable alginate-chitosan-poly(lactic-co-glycolic acid) composite microspheres.

In the present study we developed alginate-chitosan-poly(lactic-co-glycolic acid) (PLGA) composite microspheres to elevate protein entrapment efficiency and decrease its burst release. Bovine serum albumin (BSA), which used as the model protein, was entrapped into the alginate microcapsules by a modified emulsification method in an isopropyl alcohol-washed way. The rapid drug releases were sustained by chitosan coating. To obtain the desired release properties, the alginate-chitosan microcapsules were further incorporated in the PLGA to form the composite microspheres. The average diameter of the composite microcapsules was 31+/-9microm and the encapsulation efficiency was 81-87%, while that of conventional PLGA microspheres was just 61-65%. Furthermore, the burst releases at 1h of BSA entrapped in composite microspheres which containing PLGA (50:50) and PLGA (70:30) decreased to 24% and 8% in PBS, and further decreased to 5% and 2% in saline. On the contrary, the burst releases of conventional PLGA microspheres were 48% and 52% in PBS, respectively. Moreover, the release profiles could be manipulated by regulating the ratios of poly(lactic acid) to poly(glycolic acid) in the composite microspheres.

Alginates↗
PubMed2004

A rapid-curing alginate gel system: utility in periosteum-derived cartilage tissue engineering.

In this study, we have developed a rapid-curing alginate gel system and demonstrated its utility as a scaffold for periosteum-derived chondrogenesis for articular cartilage tissue engineering applications. A homogeneous mechanically stable gel was formulated by inducing gelation of a 2% (w/v) solution of a high G content alginate (65-75% G) with a 75 mM solution of CaCl(2). The gel exhibited near-elastic behavior at low levels of deformation (15%, R(2)=0.996), Young's modulus of 0.17+/-0.01 MPa, and rapid gelation kinetics (<1 min to completion). The in vitro cell culture of chondrocytes in the gel yielded alginate/cell constructs that lacked the continuous, interconnected collagen/proteoglycan network of hyaline cartilage. In addition, we have demonstrated that this gel system is capable of supporting periosteum-derived chondrogenesis. We observed that when whole-tissue explants of periosteum were cultured in vitro within the gel, after 6 weeks, significant quantities (>50%) of the total area of the periosteal explants was composed of cartilage that was hyaline-like in appearance and contained cartilage-specific proteoglycans and type-II collagen. It is envisioned that such explants could be transplanted or regenerated in vivo within the biodegradable alginate matrix for the treatment of partial or full-thickness defects in articular cartilage. Importantly, the injectable delivery of the gel could be used in filling complex defects in the articular surface via minimally invasive procedures.

Alginates↗
PubMed2005

Deletion of the tissue response against alginate-pll capsules by temporary release of co-encapsulated steroids.

Transplantation of encapsulated living cells is a promising approach for the treatment of a wide variety of diseases. Large-scale application of the technique, however, is hampered by inflammatory responses against the capsules. In the present study, we investigate whether tissue responses against alginate-PLL-alginate capsules can be modulated by co-encapsulation and temporary release of immunomodulating factors such as dexamethasone. Such an approach may be mandatory in order to increase the function and survival of encapsulated tissue since it has been shown that the tissue response can be caused by many, insurmountable factors. In an in vitro assay, we demonstrated an antiproliferative effect of dexamethasone-containing capsules on L929-mouse-fibroblasts. Subsequently, capsules prepared of purified alginate with or without solved dexamethasone were implanted in the peritoneal cavity of rats and retrieved one month later for histological evaluation. Most of the capsules without dexamethasone proved to be overgrown and adherent to the abdominal organs whereas with co-encapsulated dexamethasone the majority of the capsules were found freely floating in the peritoneal cavity without overgrowth. We conclude that co-encapsulation of dexamethasone has a profound effect on fibroblasts and macrophages adherence to immunoisolating capsules.

Alginates↗
PubMed2005

Effects of growth regulation on conditionally-transformed alginate-entrapped insulin secreting cell lines in vitro.

The ability to control cell growth is an issue of critical importance for the use of transformed beta-cell lines within a bioartificial pancreas. Such control can be achieved either by entrapping the cells in a biomaterial that can inhibit cell proliferation or by genetically modifying the cells to regulate growth. Integrating tetracycline-off or -on operon systems into murine insulinoma cell lines (betaTC-tet and R7T1, respectively) allows cell growth regulation upon exposure to tetracycline (TC) or its derivative doxycycline (Dox), respectively. However, the effects of this regulatory approach on the long-term phenotypic metabolic and secretory stability of alginate-entrapped cells have yet to be thoroughly investigated. In this study, cultures of betaTC-tet and R7T1 cells entrapped in alginate beads were allowed to grow freely, or were growth-regulated, either at the onset, or after 20 days of growth. The data show that growth regulation of alginate-entrapped cells is achievable with chronic administration of the regulatory compound in a concentration-dependent manner. However, as these cultures age, the amount of insulin released does not always reflect the metabolic and histological characteristics of the cultures. This change, coupled with a loss of glucose stimulated insulin secretion in the Dox treated R7T1 cell line, indicate a phenotypic shift of cells with an activated tet-operon. These observations have implications on the selection and long-term function of three-dimensional bioartificial pancreatic constructs that include conditionally transformed beta-cell lines.

Alginates↗
PubMed2005

Evaluation of an in situ forming hydrogel wound dressing based on oxidized alginate and gelatin.

Wound dressings that can be formed in situ offer several advantages over the use of preformed dressings such as conformability without wrinkling or fluting in the wound bed, ease of application and improved patient compliance and comfort. Here we describe such an in situ forming hydrogel wound dressing from gelatin, oxidized alginate and borax. Periodate oxidized alginate rapidly cross-links proteins such as gelatin in the presence of borax to give in situ forming hydrogels that are both non-toxic and biodegradable. The composite matrix has the haemostatic effect of gelatin, the wound healing-promoting feature of alginate and the antiseptic property of borax to make it a potential wound dressing material. The hydrogel was found to have a fluid uptake of 90% of its weight which would prevent the wound bed from accumulation of exudates. The water vapour transmission rate (WVTR) of the hydrogel was found to be 2686+/-124 g/m2/day indicating that the hydrogel can maintain a moist environment over wound bed in moderate to heavily exuding wound which would enhance epithelial cell migration during the healing process. The wound healing efficacy of hydrogel was evaluated in experimental full thickness wounds using a rat model which demonstrated that within 2 weeks, the wound covered with gel was completely filled with new epithelium without any significant adverse reactions. These in situ forming hydrogels fulfil many critical elements desirable in a wound dressing material.

Alginates↗
PubMed2005

Biochemical consequences of alginate encapsulation: a NMR study of insulin-secreting cells.

In this study we explore the biochemical consequences of alginate encapsulation on betaTC3 cells. (13)C NMR spectroscopy and isotopomer analysis were used to investigate the effects of encapsulation on several enzymatic processes associated with the TCA cycle. Our data show statistically significant differences in various enzymatic fluxes related to the TCA cycle and insulin secretion between monolayer and alginate-encapsulated cultures. The principal cause for these effects was the process of trypsinization. Embedding the trypsinized cells in alginate beads did not have a compounded effect on the enzymatic fluxes of entrapped cells. However, an additional small but statistically significant decrease in insulin secretion was measured in encapsulated cells. Finally, differences in either enzymatic fluxes or glucose consumption as a function of bead diameter were not observed. However, differences in T(2), assessed by (1)H NMR microimaging, were observed as a function of bead diameter, suggesting that smaller beads became more organized with time in culture, while larger beads displayed a looser organization.

Alginates↗