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Biomedical subjects

J Vienken

Publications and source records attributed to J Vienken.

At least 19 recordsLinked to original sources

[Clinical study of biocompatibility of dialysis membranes made from non-modified and modified cellulose].

Basic biocompatibility parameters of dialysis membranes made of non-substituted regenerated cellulose (NRC) and cellulose membranes with hydroxyl groups substituted, to a higher (H) or lower (L) degree, by dl-ethyl-amino-ethyl groups (DEAE), or by acetate (CA) were investigated in a 16-week clinical study, involving 10 long-term haemodialysis patients. In the 15th minute of dialysis, the decrease in blood leukocyte count, while using NRC (0.24 +/- 0.03 of baseline value, arithmetic mean +/- SEM) was deeper compared with that seen in DEAE-L (0.88 +/- 0.10, p < 0.001), in DEAE-H (0.79 +/- 0.10, p < 0.01), and in CA (0.73 +/- 0.05. p < 0.05). In the 15th minute of the procedure, C5a concentrations, reflecting complement activation, were higher in NRC (4.4 +/- 0.51 micrograms/L) than in DEAE-L (1.41 +/- 0.22, p < 0.001), in DEAE-H (1.68 +/- 0.47, p < 0.01), and in CA (1.68 +/- 0.22, p < 0.01). Activated clotting times were, in the 10th minute of the procedure, significantly longer in NRC (2.94 +/- 0.37 of baseline value) than in DEAE-H (1.74 +/- 0.10, p < 0.05) and, by the end of dialysis, the difference between these membranes (NRC: 1.47 +/- 0.21, DEAE-H: 0.85 +/- 0.08, p = 0.07) was close to the level of statistical significance. The authors conclude: 1. Substitution of the hydroxyl groups of regenerated cellulose reduces the decrease in leukocyte count and complement activation in the initial phase of haemodialysis. 2. At the same time, substitution by DEAE groups may raise thrombogenicity, as indicated by the shorter activated clotting times.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Systemic levels of tumor necrosis factor alpha during hemodialysis with cellulosic membranes: no effect of the sterilization procedure.

Extractable constituents of dialyzer membranes (e.g., monomers and beta-glucans) may induce the production of cytokines in vitro. We therefore studied circulating tumor necrosis factor alpha (TNF alpha) levels in 23 stable hemodialysis patients during treatment with dry Cuprophan membranes (ETO-sterilized n = 10, steam-sterilized n = 13) longitudinally over a period of 4 weeks. After 4 weeks, those 5 patients of each group showing the highest TNF alpha levels were switched to steam-sterilized, wet Cuprophan membranes. No significant increase in plasma TNF alpha was observed during hemodialysis with either ETO- or steam-sterilized dry Cuprophan membranes. A substantial TNF alpha increase (> or = 100% compared to pre-HD values), however, was observed during 14 of 84 treatment sessions. In 5 selected patients with ETO-sterilized, dry Cuprophan dialyzers, TNF alpha rose from (mean +/- SEM) 17.2 +/- 3.0 (pre-HD) to 20.9 +/- 6.2 (120 min) and 21.9 +/- 4.5 pg/ml (240 min). Corresponding levels in patients with steam-sterilized, dry Cuprophan were 16.2 +/- 5.4 (pre-HD), 21.9 +/- 6.8 (120 min), and 16.0 +/- 3.7 pg/ml (240 min), respectively. There was no difference between ETO- and steam-sterilized dialyzers. No significant reduction in mean TNF alpha plasma levels or in frequency of elevated peak levels was achieved when these patients were switched to wet Cuprophan dialyzers for another 4 weeks. It is suggested that an induction of elevated TNF alpha levels during hemodialysis is possible but is not observed regularly during treatment with Cuprophan membranes.(ABSTRACT TRUNCATED AT 250 WORDS)

Cellulose

Variables associated with the assessment of systemic tumor necrosis factor alpha levels during hemodialysis.

Conflicting results have been published concerning the systemic induction of the cytokine tumor necrosis factor alpha (TNF alpha) during hemodialysis (HD). We therefore evaluated in vitro TNF alpha production in whole blood as well as in vivo variability of TNF alpha levels in patients on long-term HD. Whole blood was incubated at room temperature (RT) with or without exogenously added endotoxin (ET), and plasma-TNF alpha was measured after 5, 30, 120, 240, and 960 min by specific enzyme immunoassay. Additionally, plasma-TNF alpha before and after 120 and 240 min HD was studied longitudinally once a week over a period of 4 weeks in 36 patients on Cuprophan (CU, n = 23) or polysulfone-F60 (PSu, n = 13) HD. Mean plasma TNF alpha levels in vitro rose from (mean) 8 pg/ml after 5 min to 12 pg/ml (120') and 32 pg/ml (960') even without ET addition, and to 18 pg/ml (after 120') and 88 pg/ml (after 960') when 0.1 microgram/ml ET were added. Pre-dialytic as well as intra-dialytic TNF alpha levels in patients showed high intra-individual variability. A substantial (> 100%) increase in plasma TNF alpha was observed during only 14 out of 84 treatments with CU and 20 out of 47 with PSu, however, the increase in TNF alpha was not statistically significant in either group. We conclude that the sampling procedure, if not carefully standardized, is a potential source of artifacts with regard to "systemic" TNF alpha levels. The high intra and inter-individual variability of plasma TNF alpha suggests that results of cross-sectional studies are questionable.

Cellulose

Detection of charges and their distribution on dialysis membranes with cationic/anionic dyes using confocal laser scanning microscopy.

Methods for the detection of positive or negative charges on the surface of biomaterials/membranes and inside a membrane are important for the characterisation of such materials. We tested different dyes and optimized staining procedures. Under standardized conditions negatively charged membranes were stained with cationic triarylmethane compounds such as crystal violet and positively charged membranes with the anionic anthraquinone dye anthralan blue B. There was no staining of uncharged cellulose membranes. The applicability of these methods was demonstrated on membranes coated to varying degrees with charged compounds such as heparin, these changes in charge being detectible quantitatively by photometry. The distribution of charges inside a membrane was detected by optical sectioning across the stained (FITC labelled poly-L-lysine) membrane using confocal laser scanning microscopy (LSM). LSM offers a completely new application possibility in biomaterial and biocompatibility research.

Biocompatible Materials

Biocompatibility and membrane development.

Development of new biocompatible membranes for clinical application needs the expertise of various specialists, chemists, chemical engineers, and clinicians. As the biocompatibility of a membrane cannot be considered in terms of a single biochemical pathway, due to the interrelation between the complement, coagulation, and inflammatory systems, it is difficult to achieve optimal results with one single polymer. A compromise between different approaches has therefore to be found. Membrane development also needs sophisticated test systems, which simulate the clinical situation as closely as possible. Adequate results are achieved with the 'ex vivo' model, which represents open-loop haemodialysis. The ex vivo model gives good results which are close to those from clinical experiments.

Biocompatible Materials

Gas supply across membranes in bioreactors for hepatocyte culture.

The conditions required for hepatocyte cultures is a main topic in the development of bioreactors for hybrid liver support systems. The detoxification of ammonia and the synthesis of urea due to primary isolated hepatocytes was measured in order to compare two different models of gas supply in bioreactors: (a) indirect medium oxygenation and (b) direct membrane-contact oxygenation of the hepatocytes using polypropylene membranes. Increasing oxygen pressure promoted cell function. At day 6 of culture, urea synthesis was 0.8 +/- 0.3 mM in 21% of O2 cultures and 1.5 +/- 0.1 mM in oxygenated cultures. Alkalosis due to CO2 loss decreased ammonia metabolism. The direct membrane-contact oxygenation resulted in enhanced cell metabolism in comparison to medium oxygenation: urea synthesis at day six was 1.42 +/- 0.2 mM in 21% O2 cultures. Polypropylene oxygenation membranes proved to be sufficient for hepatocyte adhesion. Two functions can be integrated in one element in liver support systems using the investigated polypropylene membrane and the direct membrane-contact oxygenation: oxygenation with physiological oxygen pressure in bioreactors due to gas supply across the membrane and adhesion of hepatocytes in bioreactors on the membrane.

Animals

Assembly of terminal SC5b-9 complement complexes: a new index of blood-membrane interaction.

Activation of the complement system during the course of hemodialysis was recognized more than 20 years ago and since then the generation of C3a and C5a desarg has been used as parameters of blood-membrane interaction. More recently, determination of terminal C5b-9 complement complexes has become feasible. In the present study we determined plasma concentrations of C5b-9 complexes during hemodialysis using Cuprophan or Hemophan membranes. As early as 10 min into dialysis, Cuprophan membranes led to higher arterial plasma concentrations of C5b-9 complexes in comparison to Hemophan-containing devices. With Cuprophan, systemic arterial peak values of 237 +/- 27 U/ml were reached 45 min after the onset of dialysis, while corresponding peak values using Hemophan were only 58 +/- 16 U/ml. Venous concentrations of C5b-9 complexes, measured at the outlet of the dialyzer, were 489 +/- 102 U/ml with Cuprophan and 77 +/- 19 U/ml with Hemophan dialyzers. As an index of red cell lysis, plasma levels of free hemoglobin were evaluated. There was hemolysis with both membranes. Free hemoglobin levels increased threefold with Cuprophan and only twofold with Hemophan membranes. Taken together, plasma concentrations of C5b-9 complexes clearly discern between dialysis membranes of high or low compatibility. The fact that there is simultaneous lysis of red cells might indicate that deposition of C5b-9 complexes on innocent cells occurs which would lead subsequently to an array of diverse pathophysiological reactions.

Adult

Computer aided time-lapse video analysis of hepatocyte morphology during adhesion to cellulose membranes.

An investigation was performed to demonstrate that time-lapse cinematography and computer aided video analysis of cell morphology is suitable to study and compare the characteristics of hepatocytes during the adhesion process to membranes. We chose to compare ordinary cellulose Cuprophan membranes and membranes coated with collagen or fibronectin. Striking differences between uncoated cellulose and fibronectin or collagen coating were seen in the cell count per square millimeter and adhesion behaviour. On the investigated uncoated Cuprophan the hepatocytes were found to attach but not to spread whilst on collagen coated Cuprophan most of the cells spread spherically, and on fibronectin coated membranes most of the cells flattened spherically or polygonally. Time-lapse video microscopy seems to be a valuable technique for assessing the morphologic behaviour of cells in a detailed and quantitative manner in order to improve the hepatocyte culture technique in bioreactors for hybrid systems.

Animals

Cellulosic versus synthetic membranes: a reasonable comparison?

Of the two main classes of dialysis membranes, cellulosic and synthetic, the former represents the standard membrane used in dialysis therapy. The disputed properties of cellulosic membranes related to biocompatibility have inspired a number of authors to compare these two membrane classes from a variety of perspectives. However, such a strict categorization as "synthetic" or "cellulosic" is of doubtful value from the point of view of polymer chemistry. Here, biocompatibility and performance properties of these two membrane classes are compared with the aim of investigating the validity of this categorization. The biocompatibility parameters studied are complement and leukocyte activation together with activation of the coagulation cascade. Analysis of a variety of both cellulosic and synthetic membranes with different degrees of biocompatibility showed that biocompatibility can be achieved by both classes of membranes and is, therefore, not a particular property of one class only. Furthermore, performance and beta 2-microglobulin removal properties of the two classes of membranes do not particularly favor one of these classes. Therefore, differences between cellulosic and synthetic membranes are not manifested in parameters like biocompatibility, hydraulic permeability, and overall performance.

Biocompatible Materials

Clinical evaluation of a new high-flux cellulose acetate membrane.

One major goal of dialysis therapy has become the removal of beta 2-microglobulin (beta 2-m). The interdialytic elimination of beta 2-m was studied using a newly developed high-flux cellulose acetate (CA) membrane. The results show that high-flux CA dialyzers offer better biocompatibility than classical Cuprophan or high-flux Cuprophan devices, with regard to leukopenia, C3a desarg generation, and elastase release from polymorphonuclear (PMN) leukocytes. Compared to high-flux CA membranes, high-flux PMMA membranes induce less C3a desarg formation but comparable leukopenia. High-flux PMMA membranes, however cause greater leukocyte stimulation than CA as demonstrated by more PMN elastase release during hemodialysis. Using high-flux CA or high-flux PMMA membranes, serum beta 2-m levels decreased 32% during dialysis. Serum beta 2-m dropped 10% with high-flux Cuprophan membranes, but remained unchanged with conventional Cuprophan dialyzers. Sieving coefficients for beta 2-microglobulin (beta 2-m) were virtually zero with classical Cuprophan and 0.66 with high-flux cellulose acetate membranes. High-flux membranes made of Cuprophan and PMMA gave coefficients of 0.25 and 0.45, respectively. This indicates the high removal capacity of the new CA-membrane for substances with high molecular weight. This high-flux CA membrane thus appears to combine a good degree of biocompatibility with a high capacity for beta 2-m removal.

Adult

Ex vivo model.

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Biocompatible Materials

Electro-acoustic fusion of erythrocytes and of myeloma cells.

Mammalian cells can be concentrated in a sound field. A method is introduced, which combines the reversible aggregation of cells in a sound field with the electrical breakdown of cell membranes to fuse cells, which are in contact. Human red blood cells and mouse myeloma cells are fused by means of that procedure.

Animals

An improved electrofusion technique for production of mouse hybridoma cells.

An experimental procedure is described for the reproducible production of hybridoma cells using the electrofusion technique. High yields can be obtained when fusion is performed in isotonic inositol solutions containing Ca2+ and Mg2+ in a ratio of 1:5 in the millimolar range. The hybridoma cells are transferred 10 min after the field pulse application into a balanced salt solution for 30 min at 37 degrees C.

Animals