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H Dautzenberg

Publications and source records attributed to H Dautzenberg.

16 recordsLinked to original sources
PubMed1999

Development of cellulose sulfate-based polyelectrolyte complex microcapsules for medical applications.

Microencapsulation, as a tool for immunoisolation for allogenic or xenogenic implants, is a rapidly growing field. However most of the approaches are based on alginate/polylysine capsules, despite this system's obvious disadvantages such as its pyrogenicity. Here we report a different encapsulation system based on sodium cellulose sulfate and polydiallyldimethyl ammonium chloride for the encapsulation of mammalian cells. We have characterized this system regarding capsule formation, strength and size of the capsules as well as viability of the cells after encapsulation. In addition, we demonstrate the efficacy of these capsules as a "microfactory" in vitro and in vivo. Using encapsulated hybridoma cells we were able to demonstrate long-term release of antibodies up to four months in vivo. In another application we could show the therapeutic relevance of encapsulated genetically modified cells as an in vivo activation center for cytostatic drugs during tumor therapy.

Alginates↗
PubMed1999

Nitrile hydratase from Rhodococcus erythropolis: metabolization of steroidal compounds with a nitrile group.

The progestin dienogest (17alpha-cyanomethyl-17beta-hydroxy-estra-4,9-dien-3-one) was metabolized by the nitrile hydratase-containing microorganism Rhodococcus erythropolis. An enzymatic hydrolysis of the nitrile group at the 17alpha-side chain was intended to obtain novel derivatives and to test them for progesterone receptor affinity. In contrast to the rapid enzymatic hydrolysis of nonsteroidal nitriles, the nitrile group of dienogest was cleaved very slowly. The dominant reaction was an aromatization of ring A. After prolonged fermentation, the 17alpha-acetamido derivatives of estradiol and of 9(11)-dehydroestradiol were formed. Three of the metabolites were also prepared synthetically. They were tested for hormonal activity by assessing their binding to progesterone and estrogen receptors in vitro. Neither the aromatized 17alpha-acetamido derivatives nor the dienogest derivative 17alpha-acetamido-17beta-hydroxy-estra-4,9-dien-3-one, which was prepared synthetically only, exhibited affinity for the progesterone receptor.

Animals↗
PubMed1996

Viscosimetric affinity assay.

Affinity ligands and/or affinity receptors may be quantified by a viscosimetric assay which can be carried out with a simple technique and has the potential of broad applications. The viscosimetric affinity assay is based on the high contribution of affinity bonds to the viscosity of an aqueous dispersion of a hydrocolloid that is bearing affinity ligands. In dispersions of such sensitive hydrocolloids at a concentration above the overlapping point, agglutination is not possible and the modulation of viscosity by the formation or dissociation of intercolloidal affinity bonds may be several orders of magnitude larger than the basic viscosity measurable in the absence of intercolloidal affinity bonds. If dispersions (30 g liter-1) of branched dextran with high molecular weight were used as reagent for concanavalin A (Con A), the Con A concentration necessary for a significant rise in viscosity was decreased with increasing colloid size. The viscosity of dispersions containing both a ligand-bearing high-molecular-weight dextran and an appropriate polyvalent receptor protein (lectin or antibody) showed a dependence on the concentration of free ligands (sugars or insulin) according to the law of mass action. In this competitive mode the viscosimetric affinity assay seems to be well adaptable to many analytical problems.

Antibodies↗
PubMed1996

Physicochemical studies on xylinan (acetan). I. Characterization by gel permeation chromatography on sepharose Cl-2B coupled with static light scattering and viscometry.

Laboratory-made samples of the polysaccharide xylinan (acetan) were fractionated on Sepharose Cl-2B using 0.1M NaCl as eluant. The weight average molar masses and intrinsic viscosities were estimated in the fractions by multiangle laser light scattering (off-line) and capillary viscometry, respectively. The Mark-Houwink-Sakurada plot was found to be indicative of semiflexible coils (a = 0.90). The angular dependence of scattered light was interpreted by fitting with theoretically calculated "Master Curves" in terms of a wormlike chain model. The ambiguity of the interpretation of scattering curves owing to the overlapping effects of chain stiffness and polydispersity is discussed in detail. The experimental data is found to be consistent with a persistence length of Lp = 100 nm. The main proportion consists of double-stranded chains (consistent with a robust double-helix), but single- and multistranded chains also are present. Our results suggest a fractionation according to the contour length rather than the molar mass.

Acetobacter↗
PubMed1996

Encapsulation of artificial tissues in polyelectrolyte complexes: preliminary studies.

The in vitro engineering of vital tissues from isolated cells requires primarily the synthesis of a new intercellular matrix. Structural components of the extracellular matrix are large molecules such as collagens and proteoglycans. To retain and accumulate new matrix molecules within three-dimensional cell cultures, chondrocyte-polymer constructs were encapsulated in polyelectrolyte complex membranes. Further, these membranes might also be relevant for other applications where cells or tissues have to be isolated from their environment by semipermeable structures.

Cells, Cultured↗
PubMed1996

Physico-chemical characterization of legumin-T from faba bean (Vicia faba L).

Legumin-T, the high-molecular mass product of limited tryptic hydrolysis of faba bean legumin, was investigated using hydrodynamic methods, static light scattering, fluorescence and ultraviolet spectroscopy. The following physico-chemical parameters were determined in a high-ionic strength buffer system: molecular mass, 2.4 x 10(5) g/mol; sedimentation coefficient, SO20 = 10.8 x 10(-13)Si; diffusion coefficient, DO20 = 4.1 x 10(-7) cm2 s-1; intrinsic viscosity, [eta] = 3.51 mL/g; partial specific volume, v = 0.719 mL/g; frictional ratio, f/f0 = 1.22; shape factor, beta = 2.17 x 10(6). Conformational changes during the formation of legumin-T can be deduced from the fluorescence emission and UV spectra.

Fabaceae↗
PubMed1993

Prolonged biochemical and morphological stability of encapsulated liver cells--a new method.

In this work a new type of polyelectrolyte complex capsules is introduced as an artificial housing for liver cells. Male Wistar rat hepatocytes were encapsulated using cellulose sulphate and polydimethyldialyllammonium chloride as polyelectrolytes. Amino acid metabolism rate and urea synthesis of the cells increased over the investigation period in contrast to the decrease observed in control monolayer cultures. The encapsulated cells were morphologically characterized. The described procedure represents a sufficient method for the cultivation of living cells in mechanically stable semipermeable microcapsules.

Amino Acids↗
PubMed1991

A new method for the encapsulation of mammalian cells.

A new encapsulation method was developed for the cultivation of mammalian cells. The capsules were produced using a solution of sodium cellulose sulphate (CS)(1.5%) and poly-dimethyl-diallyl-ammonium chloride (PDMDAAC). When CS droplets fell into the precipitation bath consisting of a 2% solution of PDMDAAC, immediately a membrane at the interphase was built up. The influences of varying encapsulation process parameters on capsule characteristics, cell growth, and monoclonal antibody production were tested. This new method showed advantages when compared to other methods mainly due to time simplicity of the whole process.

Animals↗
PubMed1991

Immobilization of invertase by encapsulation in polyelectrolyte complexes.

Free and polystyrene-bound invertase from Saccharomyces cerevisiae were encapsulated within symplex membranes which were composed of cellulose sulfate as the polymeric anion and poly(dimethyldiallylammonium chloride) as the polymeric cation. The kinetics and the performance of the encapsulated enzyme preparations have been compared to the free enzyme employing the hydrolysis of sucrose. The pH and temperature optima were only slightly affected by the encapsulation. The kinetic constants, however, were changed by the encapsulation as a result of diffusional limitation. Encapsulated invertase showed a high storage stability and a high operational stability if low substrate concentrations were applied. The coimmobilization of invertase with living cells, which are not capable of utilizing sucrose, in the described capsules, opens many possibilities in fermentation technology.

Enzyme Stability↗
PubMed1990

[An improved method for the microencapsulation of liver microsomes for use in extracorporeal detoxication].

Rat liver microsomes were microencapsulated in a pure aqueous medium by means of a new technique. The wall of the microcapsules consists of a semipermeable simplex membrane which is stabilized mainly by electrostatic interactions between a polymeric polyanion (sodium cellulose sulphate) and a polymeric polycation (polydimethyldiallylammonium chloride). The metabolic as well as the mechanic parameters of the microcapsules could be markedly improved by separating the metabolic (liver microsomes) from the membrane component (sodium cellulose sulphate) in such a way that two distinct compartments are formed during the preparation of the microcapsules.

Animals↗
PubMed1986

[The release of immobilized substances from Symplex capsules].

A new procedure of microencapsulation was studied with regard to substance release and quantification of diffusion processes on the capsule membrane. The permeability behaviour on the capsule membrane was especially studied in metabolites, which are essential for immobilized biological objects (i.g. preimplantative mammal embryos). Peptide and proteohormones, cyanmethemoglobin and proteins were enclosed in simple and multiple Symplex Capsules. All substances examined are able to pass the Symplex membrane. The speed of release is influenced by the size of the capsule, the ion force, temperature, concentration of immobilized substances as well as their linear and globular structur. Compared with simple capsules the release of substances from multiple capsules was delayed. Corresponding to the results found under the experimental design described the Symplex membrane can be considered as coating for the compartmentation of cells, that allows the passage of essential substances for the immobilized objects. The method of microencapsulation used and described has various ways of application.

Capsules↗
PubMed1985

The encapsulation of pancreatic islets. Investigation of insulin secretion and content in vitro.

Polyelectrolyte complex capsules from cellulose sulphate can be formed by precipitation in a polycation bath. The application of this new method for encapsulation of pancreatic islets requires investigations whether and to what extent cellulose sulphate injures viability and functionality of the pancreatic islets. Islets cultures in the presence of 2% cellulose sulphate for up to 3 weeks are characterized by unchanged insulin content, secretion and biosynthesis when compared to appropriate controls.

Animals↗
PubMed1983

[Immobilization of proteins and cell fragments using a new method of microencapsulation].

Cytochrome c, hemoglobin, urease and liver microsomes were microencapsulated in aqueous solution by use of a new method. The membrane of the microcapsules consist of a symplex, which is formed predominantly by electrostatic interactions between a polymeric polyanion and a polymeric polycation. The membrane is characterized by high mechanical stability and is a barrier for globular substances with molecular weights larger than 12000, but permeable for substances with lower molecular weight. The microcapsules contain the protein or microsomes and additionally the polymeric polyanion in the liquid interior. Structure and function of microencapsulated proteins and microsomes are preserved. The developed method of microencapsulation opens new possibilities for the application of biomacromolecules, particularly for extracorporal detoxification.

Animals↗