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Effect of immersion disinfection with Perform-ID on alginate, an alginate alternative, an addition-cured silicone and resultant type III gypsum casts.

OBJECTIVE: This study investigated the effect of a commonly used immersion disinfectant upon three different impression materials and any subsequent effects on the abrasion resistance, hardness and surface detail reproduction of gypsum casts. DESIGN: A laboratory study. MATERIALS AND METHODS: Under standardised conditions a total of 120 impressions were made of a ruled test block using irreversible hydrocolloid (Alginoplast), an 'alginate alternative' addition-cure silicone (Position Penta) and a conventional addition-cure silicone (President). The impressions were examined for surface detail reproduction prior to and after disinfection with Perform-ID. The type III casts were evaluated for surface detail reproduction, surface hardness and abrasion resistance. RESULTS: (1) None of the disinfected alginate specimens could reproduce the 50 microm line. (2) Casts produced from the disinfected alginate were significantly less hard than from disinfected Position Penta and President (P <0.001). (3) Disinfection significantly affected the abrasion resistance of casts made from Position Penta (P = 0.029). (4) Disinfection did not significantly affect President or its subsequent casts (P >0.05). CONCLUSION: If disinfecting with Perform-ID, the impression should be made with a conventional addition-cured silicone if good surface detail reproduction of the impression material and a hard and abrasion resistant type III gypsum cast are required.

Alginates↗

An experimental comparison of hydrocolloid and alginate dressings, and the effect of calcium ions on the behaviour of alginate gel.

Gel-forming dressings, which absorb the wound exudate to form a non-adherent gel, accelerate wound healing in a moist environment. The most commonly used of these dressings are calcium alginate and hydrocolloid membranes. This study was designed to evaluate the physical properties of these dressings, including fluid retaining ability, gel formation, and the dissolution process to clarify the mechanisms that promote healing. A simple immersion method was used for measurement. This technique was useful for the quantitative assessment of gel-forming dressings and may help to evaluate potential application of these dressings. A comparative analysis showed that alginate dressings remained gelled longer than hydrocolloid dressings. This result was consistent with previously reported clinical features of these dressings. The absence of calcium in wound fluid induced rapid resolution of alginate gel while the presence of a certain concentration of calcium prevented degradation of the gel for as long as one month.

Alginates↗

Anaerobic production of alginate by Pseudomonas aeruginosa: alginate restricts diffusion of oxygen.

Pseudomonas aeruginosa produced alginate and elevated algD (encoding GDPmannose 6-dehydrogenase) transcription under strict anaerobic conditions, especially when using nitrate as a terminal electron acceptor. Purified alginate added to bacterial suspensions caused a decrease in growth, suggesting that alginate contributes to oxygen limitation for the organism and likely for patients afflicted with the inherited autosomal disease cystic fibrosis.

Alginates↗

Alginate lyase (AlgL) activity is required for alginate biosynthesis in Pseudomonas aeruginosa.

To determine whether AlgL's lyase activity is required for alginate production in Pseudomonas aeruginosa, an algLdelta::Gm(r) mutant (FRD-MA7) was created. algL complementation of FRD-MA7 restored alginate production, but algL constructs containing mutations inactivating lyase activity did not, demonstrating that the enzymatic activity of AlgL is required for alginate production.

Alginates↗

A novel method to enhance the stability of alginate-poly-L-lysine-alginate microcapsules.

Implantation of microencapsulated recombinant cells is an alternative approach to gene therapy. These genetically-engineered cells enclosed in microcapsules to deliver therapeutic recombinant products have been effective in treating several murine models of human diseases. However, the most commonly used microcapsules fabricated from alginate ionically cross-linked with calcium suffer from loss of long-term mechanical stability. We now report on a method to improve their stability by introducing additional polymers to provide covalent linkages via photopolymerization. Vinyl monomers and a photoinitiator were allowed to diffuse into the initially formed calcium-alginate microcapsules. In situ photopolymerization in the presence of sodium acrylate and N-vinylpyrrolidone substantially enhanced their mechanical strength. After four months of storage in saline, > 70% of these capsules remained intact in the osmotic pressure test, while the un-modified alginate microcapsules totally disintegrated. Tests of their permeability to polyethylene glycol of different molecular weight and their ability to support cell survival showed that these properties remained unaffected by the photopolymerization. Hence, these microcapsules modified by adding a network of vinyl polymers are promising candidates to use for long-term delivery of recombinant gene products in this cell-based method of gene therapy.

Alginates↗

Effect of alginate-polylysine-alginate microencapsulation on in vitro insulin release from rat pancreatic islets.

We investigated the effect of alginate-polylysine-alginate microencapsulation on glucose-induced insulin secretion by rat islets. Applying the encapsulation method originally described by Lim, we found severely reduced in vitro insulin release (expressed as picomoles of insulin.10 islets-1.45 min-1 when incubated in 16.5 mM glucose), because the insulin release with encapsulated islets was 1.42 +/- 0.49 compared to 13.58 +/- 0.80 with free control islets. This could not be explained by inadequate permeability of the capsule, because insulin release was also severely reduced (2.12 +/- 0.61) when islets were subjected to the procedure but without the membrane-forming polylysine step. Therefore, islets were tested after having been subjected separately to each of the steps of the procedure. Insulin release was not affected by either alginate or CaCl2 but was severely reduced after prolonged suspension in saline or treatment with citrate. When saline and citrate were replaced by Ca2(+)-free Krebs-Ringer bicarbonate buffer (KRBB) and 1 mM EGTA, respectively, insulin release improved significantly both with complete and with incomplete (no polylysine step) encapsulation. This outcome was verified in a set of experiments run in parallel with islets derived from the same isolation procedure. Insulin release was 1.20 +/- 0.23 from islets encapsulated with the method of Lim and 10.73 +/- 1.04 from free control islets. With the modified procedure, insulin release was 9.17 +/- 0.52 vs. 9.61 +/- 1.27 for complete versus incomplete encapsulation, respectively. We conclude that Ca2(+)-free KRBB instead of saline and EGTA instead of citrate should be used to obtain an adequate insulin response from encapsulated islets and that the capsule membrane as such has no influence on glucose and insulin diffusion.

Alginates↗

Immunological studies of SK2 hybridoma cells microencapsulated with alginate-poly(L)lysine-alginate (APA) membrane following allogeneic transplantation.

Microencapsulation of living cells or tissues has been proposed to prevent their immune destruction following transplantation. In this study, we examined whether SK2 hybridoma cells microencapsulated in an alginate-poly(L)lysine-alginate (APA) membrane (APA-SK2 cells) were immunoisolated from the allogeneic host's immune system using a cytotoxicity test. The APA membrane inhibited the activation of the host's cellular immune response, but did not prevent the production of cytotoxic antibodies against entrapped SK2 cells following allogeneic transplantation. However, the APA-SK2 cells remained vital in SK2 cell-immunized mice as well as in intact mice. We considered that complement regulatory factors which were present on cell membrane and had species-specific restriction blocked the complement-mediated cell lysis on allogeneic transplantation, since APA-SK2 cells were destroyed by rabbit anti-SK2 cell antiserum. Our results demonstrated that APA membrane could inhibit cell-cell contact between entrapped cells and the host's lymphocytes, but could not completely protect the entrapped cells from xenogeneic humoral immunity.

Adoptive Transfer↗

Cytomedical therapy for IgG1 plasmacytosis in human interleukin-6 transgenic mice using hybridoma cells microencapsulated in alginate-poly(L)lysine-alginate membrane.

Cytomedical therapy for human interleukin-6 transgenic mice (hIL-6 Tgm) was implemented by the intraperitoneal injection of alginate-poly(L)lysine-alginate (APA) membranes microencapsulating SK2 hybridoma cells (APA-SK2 cells) which secrete anti-hIL-6 monoclonal antibodies (SK2 mAb). IgG1 plasmacytosis in the hIL-6 Tgm was suppressed by a single injection of APA-SK2 cells, and the survival time of these mice was remarkably prolonged. The viable cell number and the SK2 mAb-secretion of APA-SK2 cells increased for at least one month both under culture conditions and in allogeneic recipients (in vivo). Moreover, SK2 mAb which were secreted from APA-SK2 cells injected into allogeneic recipients was detected in serum at high concentrations; 3-5 mg/ml from day 14 to day 50 post-injection. In contrast, the injection of free SK2 cells had no therapeutic effect on hIL-6 Tgm. These results strongly suggest that APA membranes microencapsulating cells which were modified to secrete molecules useful for the treatment of a disorder were effective as an in vivo long-term delivery system of bioactive molecules, as 'cytomedicine'.

Alginates↗

Prolongation of the effective duration of cytomedical therapy by re-injecting SK2 hybridoma cells microencapsulated within alginate-poly(L)lysine-alginate membranes into human interleukin-6 transgenic mice.

We previously reported that SK2 hybridoma cells that secreted anti-human interleukin-6 (hIL-6) monoclonal antibodies (SK2 mAb) were microencapsulated within alginate-poly(L)lysine-alginate (APA) membranes (APA-SK2 cells) for immunoisolation, and a single intraperitoneal injection of these APA-SK2 cells remarkably improved IgG1 plasmacytosis in hIL-6 transgenic mice (hIL-6 Tgm). However, the duration of the effectiveness of APA-SK2 cells as a cytomedicine was unfortunately limited. In this study, we attempted to re-inject APA-SK2 cells into hIL-6 Tgm for the purpose of prolonging the cytomedical therapy. In hIL-6 Tgm re-injected with APA-SK2 cells, the plasma IgG1 level did not show any increase in 37 week old mice, and their survival time was at least three times longer than those of untreated hIL-6 Tgm. These results suggest that re-injected APA-SK2 cells survived and secreted SK2 mAb in the allogeneic mice. Thus, the limited duration of the cytomedical effects of APA-SK2 cells was probably caused by the disappearance of the inner space of microcapsules for cell proliferation, not by the rejection of the host's immune system. Therefore, if we can regulate the proliferation of the cells microencapsulated within a semipermeable membrane, we may be able to develop a cytomedicine which will continue its function longer after a single injection.

Alginates↗

Application of alginate gel as a vehicle for liposomes. II. Erosion of alginate gel beads and the release of loaded liposomes.

The possibility of producing calcium-induced alginate gel beads as a vehicle for liposomes was explored. The maximal loading of egg phosphatidylcholine liposomes (ca. 26 nm in diameter) in a fully-cured bead (1.2 mm in radius, initial alginate concn. of 4%) was 2.9 x 10(-6) mol/bead or ca. 18%, and the size of the bead slightly increased with an increase in liposome loading. The liposomes were well maintained within both fully-cured and washed beads. The liposome release from the fully-cured bead was much slower than that from the corresponding washed bead in a pH 7.4 releasing medium. The greater the liposome loading, the faster the release of the vesicles. The liposome release was investigated in terms of liposome loading, swelling of the gel body, calcium discharge and gel erosion, using washed beads. The liposome loading did not affect the bead erosion or calcium discharge but did the initial swelling ratio and liposome release. The results suggest that the loaded liposomes are not uniformly distributed in the bead but are rather gradually concentrated to the center. Such an inhomogeneous distribution of liposomes is possibly due to the fact that the gelation occurred instantly on the surface of the droplets, and the resulting gel network or layer acts as semipermeable membrane for liposomes and forces the vesicles to move into deeper concentric sections as gelation proceeds to the interior. As the liposomes loading increases, the forced migration might be very limited because of concentrically decreasing extra room to accommodate the vesicles in the bead.

Alginates↗

[Theoretical calculation and experimental study of membrane thickness of alginate-(poly-L-lysine)-alginate microcapsules].

Alginate-(Poly-L-Lysine)-Alginate(APA) microcapsules were prepared by Electrostatic Droplet Generator(EDG) technique and the thickness of microcapsule membrane, which was composed by polyelectrolyte complex, were studied in this paper. The theoretical formula was given for the measurement of membrane thickness of APA microcapsules by element analysis of membrane and calculation. The membrane thickness was 7-10 microns by theoretical calculation. On the other hand, the thickness of membrane was measured by SEM and optical microscopy and the results were 7 microns and 12 microns, respectively. The results showed that theoretical calculation is in good accordance with experimental determoination of mermbrane thickness and the membrane thickness of APA microcapsule is about 7-10 microns. The optical microscopy is an easy way to measure membrane thickness.

Alginates↗

[Study on cytomedicine of alginate-poly(L) lysine-alginate microencapsulated hybridoma cells].

AIM: To study the cytomedicine of alginate-poly (L) lysine-alginate (APA) microencapsulated hybridoma cells and their characteristics. METHODS: The spleen cells taken from BALB/C mice immunized with purified human IgG1 kappa type were fused with mouse myeloma cells SP2/0. The hybridoma cell lines secreting monoclonal antibodies (mAb) against human IgG1 kappa type was named JY-A1. The APA microencapsulated JY-A1 cells were prepared with a high-voltage electrostatic system. Microencapsulation parameters were optimized and their morphology was studied. The mechanical strength and chemical intensity of microcapsules were measured. The mAb secrete from APA microencapsulated JY-A1 cells was determined by ELISA kit. The microcapsules injected into mice abdominal cavity previously were recovered at intervals. RESULTS: The microcapsules prepared in the same condition of the high-voltage electrostatic system were round and homogeneous. The mAb secreted by the microencapsulated JY-A1 cells were shown to permeate the membranes of APA microcapsules in vitro. After an intraperitoneal injection to mice, APA microcapsules were recovered on day 7, 14, 28, 56. The electron microscopy study revealed that the majority of recovered microcapsules were intact, and no evidence of immunological reaction in terms of fibrosis. CONCLUSION: APA microencapsulated hybridoma cells prepared by high-voltage electrostatic system have good mechanical strength and chemical intensity. The APA microencapsulated hybridoma cells can maintain physiological functions in vitro, and the microcapsules have good biocompatibility in vivo.

Alginates↗

Comparative study of alginate and non-alginate antacids concurrently administered with H2 antagonists in cases of duodenal ulcer.

In an open comparative study in endoscopically proven cases of duodenal ulcer, concomitant therapy of H2 antagonist was initiated in two randomly allocated, homogenous patient groups, ie, H2 A + RF alginate antacid (Riflux Forte, Sol Pharmaceuticals Ltd) or H2 A + S non-alginate antacid (Siloxogene, Searle Ltd), both Riflux Forte and Siloxogene having similar compositions of antacids. Overall clinical assessment, percentage improvement, assessment of individual clinical symptoms, global assessment and endoscopic assessment all showed that Riflux Forte and Siloxogene together with H2 antagonist are significantly effective in providing a progressive symptomatic relief in duodenal ulcer cases over an evaluable period of eight weeks. Although differences between Riflux Forte and Siloxogene were not statistically significant, a definite trend of more rapid and favourable responses was seen with Riflux Forte, demonstrating the role played by therapy in preventing reflux and stopping the acid pain cycle.

Adolescent↗

Release characteristics of diclofenac sodium from poly(vinyl alcohol)/sodium alginate and poly(vinyl alcohol)-grafted-poly(acrylamide)/sodium alginate blend beads.

In this study, acrylamide (AAm) was grafted onto poly(vinyl alcohol) (PVA) with UV radiation at ambient temperature. The graft copolymer (PVA-g-PAAm) was characterized by using Fourier transform infrared spectroscopy (FTIR), elemental analysis and differential scanning calorimetry (DSC). Polymeric blend beads of PVA-g-PAAm and PVA with sodium alginate (NaAlg) were prepared by cross-linking with glutaraldehyde (GA) and used to deliver a model anti-inflammatory drug, diclofenac sodium (DS). Preparation condition of the beads was optimized by considering the percentage entrapment efficiency, particle size, swelling capacity of beads and their release data. Effects of variables such as PVA/NaAlg ratio, acrylamide content, exposure time to GA and drug/polymer ratio on the release of DS were discussed at three different pH values (1.2, 6.8, 7.4). It was observed that, DS release from the beads decreased with increasing PVA/NaAlg (m/m) ratio, drug/polymer ratio (d/p) and extent of cross-linking. However, DS release increased with increasing acrylamide content of the PVA-g-PAAm polymer. The highest DS release was obtained to be 92% for 1/1 PVA-g-PAAm/NaAlg ratio beads. It was also observed from release results that DS release from the beads through the external medium is much higher at high pH (6.8 and 7.4) than that at low pH (1.2). The drug release from the beads mostly followed Case II transport.

Acrylamides↗

Immobilization of P. pictorum in open pore agar, alginate and polylysine-alginate microcapsules for serum cholesterol depletion.

P. pictorum depletes serum cholesterol levels upon induction of the involved enzymes. Analysis of the fermentation broth shows cholesterol esterase and cholesterol oxidase activity. P. pictorum was immobilized in alginate beads, polylysine microcapsules and open pore agar beads. The pore size of the former two was too small to allow a barrier free mass transfer of lipoproteins. However, open pore agar beads depleted serum cholesterol. The rates of depletion were similar to those of free P. pictorum, suggesting a barrier free mass transfer. Furthermore, open pore agar beads could effectively retain P. pictorum, and no leakage was detected.

Agar↗