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High density culture of anchorage-dependent animal cells by polyurethane foam packed-bed culture systems.

Monkey kidney cells (Vero) and Chinese hamster ovary cells (CHO-K1) attached to the internal surface of polyurethane foam (PUF) and grew to a high cell density (1.1 x 10(8) cells/cm3 PUF and 4.2 x 10(7) cells/cm3 PUF, respectively) in a PUF-plates packed-bed culture system. This density of Vero cells was twice that obtained previously with a PUF-particles packed-bed culture system. A maximum cell density of 6.7 x 10(7) cells/cm3 culture vessel volume was obtained in a PUF-disc packed-bed culture of Vero cells. From the cell density of CHO-K1, growing in a monolayer on the surface of PUF and a petri dish, per bulk volume of PUF, we estimated that a surface area to volume ratio of PUF plates effective for cell growth was about 109 cm2/cm3.

Animals↗

Characterization of immobilized enzymes in polyurethane foams in a dynamic bed reactor.

beta-D-Galactosidase (E 3.2.1.23) from Aspergillus oryzae was immobilized with polyurethane foam (PUF). Among several immobilization methods attempted in this work, the immobilized enzyme preparation by in-situ co-polymerization between enzyme and prepolymer HYPOL 3000 showed the highest activity. The intrinsic kinetics of PUF-immobilized enzyme was determined in a dynamic bed reactor, used to increase transport rates. The immobilization mechanism in PUF was studied by measurements of immobilized enzyme kinetics and by using scanning electron microscopy combined with immuno-gold labeling techniques. The results showed that immobilization was predominantly by covalent bonding between primary amino groups of beta-D-galactosidase and isocyanate groups of the prepolymers. Entrapment in the PUF micropores assisted the immobilization of enzymes, and adsorption on the surface of macropores was not important for immobilization. The bicinchoninic acid method was applied for the determination of PUF loading capacity and specific enzyme activity and used to determine enzyme deactivation during immobilization.

Adsorption↗

High albumin production by multicellular spheroids of adult rat hepatocytes formed in the pores of polyurethane foam.

Adult rat hepatocytes formed spherical multicellular aggregates (spheroids) when they were cultured in the pores of polyurethane foam (PUF). The diameter of the spheroids was within the range 100-200 microns. These spheroids partly attached and immobilized in the PUF pores for at least 2 weeks. The albumin production rate by the spheroids increased up to 17.0 micrograms/10(6) nuclei per day during the first 6 days and maintained at a high level for 2 weeks. In contrast, the albumin production rate by the monolayer markedly decreased after 3 days. The spheroid culture using PUF seems to be a convenient and simple method for maintaining some differentiated functions of hepatocytes and for making a bioreactor using the function of spheroids.

Albumins↗

Mutagenicity of aerosols from the oxidative thermal decomposition of rigid polyurethane foam.

The aerosol fraction of the oxidative thermal decomposition products (700 degrees C) of rigid polyurethane foam was collected on glass fiber filters and fractionated into either-soluble neutral, acidic, and basic fractions and water-soluble compounds. The fractions showed mutagenic activity in a bacterial fluctuation test with Salmonella typhimurium TA98 or Escherichia coli CM891 as the tester strains. All the fractions induced mutations in both strains after metabolic activation with rat liver S-9 mix. The basic and the water-soluble fractions were mutagenic for S. typhimurium TA 98 even without activation. Thin-layer chromatography showed the presence of several primary aromatic amines in the aerosol. Polycyclic aromatic hydrocarbons were not detected by glass capillary gas chromatogaphy.

Aerosols↗

Symptoms and lung function in low-exposure to TDI by polyurethane foam manufacturing.

A cross-sectional investigation was performed in seven polyurethane foam manufacturing factories. Lung function tests were performed at the factories by spirometric and nitrogen wash-out methods. Sixty-seven persons were investigated before and after the work shift, and exposure to toluene diisocyanate (TDI) was evaluated by subject-carried sampling equipment on the same day. The day mean exposure to TDI was 0.008 mg/m3 (0.001 ppm). In the non-smokers there was an increased frequency of symptoms from the airways reported in the exposed subjects as compared to the unexposed controls. There were, however, no signs of acute or chronic effects on the lung function measured by spirometric and nitrogen wash-out methods, which could not be explained by smoking.

Adult↗

Short-term respiratory changes in polyurethane foam workers exposed to low MDI concentration.

Our study included 27 polyurethane foam workers exposed to MDI only at low concentrations (ranging from 0.0005 to 0.001 ppm) and 27 clerks from the same factory matched by age. Respiratory function tests were performed on a Monday and Friday of the same week at shift onset, 4 h later and at shift end. The subjects under study were asymptomatic for asthma. The two groups had quite similar spirometric values with minimal functional impairment. A statistical analysis was carried out by Student's t-test for matched pairs and two-way analysis of variance (ANOVA), in order to take into account both occupational exposure and smoking habits. No significant differences between the two groups were observed with paired t-tests in the respiratory parameter trend during both the Monday and Friday work shift. Nor were differences observed within the two groups when Friday's and Monday's results were compared. No significant differences between the two groups were found in paired comparisons between Friday and Monday for respiratory parameters. ANOVA demonstrated that the FEV1 and FEF25-75 reduction present on Friday, when compared to Monday, was related to smoking and not to occupational exposure. In conclusion our findings showed no short-term respiratory changes in subjects exposed to low MDI concentration.

Analysis of Variance↗

MDA in plasma as a biomarker of exposure to pyrolysed MDI-based polyurethane: correlations with estimated cumulative dose and genotype for N-acetylation.

The object of this study was to investigate whether exposure of pipe-layers to thermal degradation products of diphenylmethane diisocyanate (MDI) could be assessed by analysing 4,4-methylenedianiline (MDA) in hydrolysed plasma and urine, and whether the genotype for N-acetylation affected these biomarker levels. Blood and urine samples were drawn from 30-pipe-layers who had been welding polyurethane (PUR) insulated pipes during the preceding 3 months. MDA in hydrolysed plasma and urine was determined with a gas chromatography-mass spectrometry technique, and genotype for N-acetylation was analysed with a polymerase chain reaction technique. MDA in plasma was detected in 18 of the 30 pipe-layers. Their plasma concentrations of MDA varied from 0.05 to 8.48 micrograms/l. There was a significant negative correlation between time since last welding of PUR-insulated pipes and P-MDA (rs = 0.50, P = 0.005). There was also a significant positive correlation between the estimated number of welded PUR-insulated pipes during the preceding 3 months and P-MDA (rs = 0.68, P = < 0.001). No significant association between genotype of N-acetylation and P-MDA was observed in a multiple regression analysis when adjustment was made for the estimated cumulative exposure to thermal degradation products of MDI. MDA in urine was detected in only four of the 30 pipe-layers. These four subjects had been welding PUR pipes on the same day as the sampling, or on the day before. The present results indicate the spot plasma samples analysed for MDA may give a rather good estimate of exposure to MDI during the preceding months. P-MDA, but not U-MDA, therefore seems to be a useful biomarker of long-term exposure to MDI. The individual N-acetylation capacity did not affect the plasma levels of MDA.

Acetylation↗

Four-year follow-up of effects of toluene diisocyanate exposure on the respiratory system in polyurethane foam manufacturing workers. I. Study design and results of the first cross-sectional observation.

A 4-year cohort study was designed to assess the exposure-effect relationship of working in polyurethane foam (PF) manufacturing factories with exposure to toluene diisocyanate (TDI) and its effects on the respiratory system. This paper describes the results of the first cross-sectional observations. The study population included 90 male workers who had been working in PF factories for 0.5-25 years (mean 13.3 years) (PF workers) and 44 reference workers in the same factories. The mean exposure concentration of TDI calculated from 129 personal samples was 3.2 ppb. Peak exposure excursions above 20 ppb occurred in 16 of 129 samples. Pulmonary function and its change during the working day as assessed by examining the forced expiratory flow-volume curve, respiratory impedance, and airway resistance and specific airway conductance were not different in the PF workers from those in the reference workers. Chest X-radiographs did not show any noteworthy radiological changes. Prevalences of "phlegm in winter," "nasal stuffiness or discharge in winter," and "irritation of eye and throat mucous membranes" were significantly higher in the PF workers. The findings indicate that TDI exposure at levels around 3 ppb may not adversely affect the pulmonary function over many years of exposure of those who are not hypersensitive to TDI. The causal chemicals inducing some respiratory and irritative symptoms could not be specifically identified since the PF workers were exposed not only to TDI but also to other irritative agents in the PF manufacturing processes.

Adolescent↗

Four-year follow-up of effects of toluene diisocyanate exposure on the respiratory system in polyurethane foam manufacturing workers. II. Four-year changes in the effects on the respiratory system.

Fifty-seven polyurethane foam manufacturing workers (PF workers) and 24 reference workers were followed for 4 years to clarify the effects on pulmonary function of working in PF factories with exposure to toluene diisocyanate (TDI). No significant differences in the average annual losses (AALs) of pulmonary function for 4 years were observed among the 28PF workers whose TDI exposure levels were very low (mean = 0.1 ppb, group L), the remaining 29 PF workers with mean TDI exposure of 5.7 ppb (group H), and the reference workers. However, 15 PF workers in group H who had experienced peak exposure excursions to 30 ppb or above with a mean concentration of 8.2 ppb showed significantly larger AALs in percentage maximal mid-expiratory flow, forced expiratory volume in 1 s ratio to vital capacity (FVC), and forced expiratory flow at 25% of FVC than expected, and significantly larger AALs in some obstructive pulmonary function indices than those of the 14 remaining PF workers in group H whose peak exposure excursion levels were 3-14 ppb with a mean time-weighted average (TWA) of 1.7 ppb, group L, and the reference workers. These findings suggest that the peak exposure excursion level of TDI might be important in inducing obstructive pulmonary function changes in the PF workers rather than the TWA exposure levels, though further comparative studies of the AAL in those who are exposed to different peak exposure excursion levels but the same mean exposure levels are necessary. From the standpoint of prevention, the proposition that peak exposure excursion levels exceeding 20 ppb should be avoided is reasonable.

Adult↗

Tissue plasminogen activator (t-PA) production by a high density culture of weakly adherent human embryonic kidney cells using a polyurethane-foam packed-bed culture system.

Weakly adherent cells of the 293 line attached well to the internal surface of polyurethane foam (PUF) and grew to the high density of 6.83 x 10(7) cells/cm3 PUF in stationary culture. The maximum productivity of tissue plasminogen activator (t-PA) was 0.158 IU/10(6) cells per day. The productivity decreased at the stationary phase of cell growth, so we designed a PUF-plate packed-bed culture system for high density culture and continuous production of t-PA. A maximal cell density of 3.24 x 10(7) cells/cm3 PUF and a t-PA productivity of 0.326 IU/10(6) cells per day were obtained in 25-day perfusion cultures. Although the cell density decreased to half that in PUF stationary culture, the t-PA productivity increased twofold and was maintained for 25 culture days.

Cell Division↗

Correction of established capsular contractures with polyurethane implants.

Twelve patients with severe and recurrent capsular contractures following breast augmentation and reconstruction were improved with polyurethane-covered implants. It still has not been established whether these implants prevent or merely delay the onset of contractures, but some of these patients have been followed for over three years and all 12 were improved.

Adult↗

Polyurethane-covered silicone gel mammary prosthesis for successful breast reconstruction.

A polyurethane-covered silicone gel implant has been used by the author in 150 breast reconstructions and augmentations in the past 9 years. The results have been most gratifying with regard to breast softness, breast compressibility, and esthetics. Only 4 patients have developed a unilateral capsule contracture and firm breast. The reasons are postulated for these satisfying results. Complications have been few and very minor.

Adult↗

Stored blood microfiltration. Evaluation of micro-aggregate filter composed of polyurethane foam and nylon wool.

Aggregates of amorphous material which develop during storage of banked blood have been implicated as a cause of pulmonary micro-embolism in man following massive transfusion. Such pulmonary micro-embolism may be a causal factor in the development of post traumatic pulmonary insufficiency. At present several microaggregate filters for use in massive transfusion are commercially available, and on of these is the Fenwal filter. It is composed of a screen filter which removes microclots of 250 microns and higher, a layer of polyurethane foam and a layer of nylon wool. The resistance of this device is very acceptable and the filter may be used for several blood units, but its efficiency seems less than that of Dacron wool filter.

Blood Platelets↗

A rapidly polymerizing polyurethane for transcatheter embolization.

A polyurethane prepolymer was evaluated as a transcatheter embolizing agent. Low viscosity permits injection through small catheters. Polymerization is initated on contact with blood and completed within 10 seconds. Downstream propagation is better then that of cyanoacrylate and comparable to that of silicone rubber. No systemic toxicity was observed in acute animal experiments. However, dissolution of arterial walls and extravasation out of the vascular bed of chronically embolized organs suggest significant local tissue toxicity and make further evaluation necessary before clinical testing.

Abscess↗

Thermoplastic polyurethane--the material used for the Erlanger silver catheter.

The Erlanger silver catheter consists of a thermoplastic polyurethane elastomer (TPU). To achieve an antimicrobial effect, silver particles at a concentration of 0.5-1.0% by weight are incorporated into the basic material. The good mechanical properties were shown to remain present after the incorporation of the silver. The characteristic mechanical data of blends made of a soft and a hard TPU type approximated closely to a linear relationship with the ratio of the mixture. The buckling stability of catheters as a function of their cross section could be described by Euler's formula. This enables the buckling stability to be calculated when material constants and catheter geometry are known.

Anti-Infective Agents↗

Use of semipermeable polyurethane hollow fibers for pituitary organ culture.

A new model for organ culture of endocrine tissue is described. Rat anterior pituitary fragments were cultured for 4 wk within semipermeable polyurethane isocyanate hollow fibers. Growth hormone and prolactin, two of the anterior pituitary hormones, were released into the medium during the entire culture period. Electron microscopy of the pituitary fragments after 2 wk in culture showed a rim of viable tissue in all specimens examined. Individual cells, from this outer rim, exhibited excellent organelle preservation and numerous secretory granules. Experiments involving potassium depolarization and 10(-6) M dopamine provided evidence for the normal responsiveness of the cultured pituitary tissue to both stimulatory and inhibitory factors. These studies illustrate the potential utility of the described organ culture system for further investigations of endocrine physiology.

Animals↗

Immobilization of amyloglucosidase using two forms of polyurethane polymer.

Amyloglucosidase was covalently immobilized using two hydrophilic prepolymers: Hypol FHP 2002 (creates foams) and Hypol FHP 8190H (creates gels). The foamable prepolymer was superior as a support for enzyme immobilization. The percent activity immobilized in the polyurethane foams was 25 +/- 1.5%. Large substrates (greater than 200,000 daltons in mol wt) were hydrolyzed as effectively as smaller ones by the immobilized enzyme. The Km value of the foam-immobilized enzyme increased from 0.76 mg/mL (free) to 0.86 mg/mL (immobilized), whereas the Vmax dropped from 90.9 (free) to 12.4 nmol glucose/min/mL (immobilized). The long-term (2 mo) storage stability of amyloglucosidase was enhanced by immobilization in foams (70% activity retained; free enzyme only retained 50%). Immobilization also improved the enzyme stability to various denaturing agents (sodium chloride, urea, and ethanol). The immobilized enzyme exhibited increased stability compared to the free enzyme at high temperatures (95 degrees C). Both glycogen and starch could be utilized by the immobilized enzyme, indicating that this technique could prove useful for starch hydrolysis.

Enzyme Stability↗

The chromatographic behavior of group (IIB) metal ions on polyurethane foam functionalized with 8-hydroxyquinoline.

Polyurethane foam functionalized with 8-hydroxyquinoline has been prepared by coupling the foam matrix with 8-hydroxyquinoline (oxine) through an azo spacer. The oxine-bonded foam (Ox PUF) was characterized by use of different tools (UV-Vis spectra, IR spectra, density, and stability). Ox PUF was found to be very suitable for separation and preconcentration of trace metals, e.g. Zn(II), Cd(II), and Hg(II) ions, from wastewater in the pH ranges 2-12, 9-12, and 3-6, respectively. Various conditions influencing the sorption of these metal ions on to Ox PUF were optimized. Extraction of the metal ions was accomplished in 15 to 20 min. Study of the variation of the sorption of the tested metal ions with temperature yielded average values for DeltaH, DeltaS, and DeltaG of 41.99, 158.23, and -5.1 kJ mol(-1), respectively. The capacities of the foam material were 0.27, 0.16, and 0.09 mmol g(-1) for Zn(II), Cd(II), and Hg(II), respectively. Preconcentration factors >50 were achieved (RSD approximately 6.18). The quantitative results were obtained from experiments performed using certified reference materials.

Cadmium↗