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A biomaterial named P558 is a new austenitic stainless steel (SS) with a negligible amount of Ni (<0.20%). In previous in vitro and in vivo studies it was compared with conventional SS and Ti6Al4V and shown to be a promising material in orthopedics. Because osteoporosis is a type of pathology very often encountered in implanted patients and can be studied with in vitro models, the purpose of the present study was to evaluate P558 in vitro through comparison of normal (nOB) with osteopenic (oOB) bone-derived primary rat osteoblasts. Osteoblasts were cultured directly on P558 and polystyrene as controls for 72 h. Osteoblast proliferation, adhesion, and activity (ALP, OC, TGF-beta1, and IL-6) were evaluated at 24 and 72 h. Results demonstrated that the growth of nOB and oOB cultured on P558 was not affected negatively when compared to control. Cells on P558 did not show any alteration in terms of adhesion, proliferation, and metabolic marker production in nOB and oOB cultures, and a significant increase in ALP, OC, and TGF-beta1 production was observed. SEM images revealed no alteration in cell morphology. The current findings demonstrate that P558 promotes osteoblast proliferation, activation, and differentiation not only in normal bone, but also in osteopenic bone-derived osteoblasts.
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PMID:A new austenitic stainless steel with a negligible amount of nickel: an in vitro study in view of its clinical application in osteoporotic bone. 1536 25

Aramid fibers from aromatic polyamide group are plastic materials with high mechanical resistance to breaking, small elongation and low mass. They have obtained numerous technical usage for products carrying high mechanical forces and resistance to consumption. Application of aramid in alloplasty would satisfy the grafts with high mechanical resistance. However, the biological influence and biostability of aramid is unknown and univocal. Research of the degree of biocompatibility of aramid fibres in comparison with commonly used polyester fibres with known and accepted biological reaction was the purpose of the study. The research included in the first phase: physicochemic properties, hemolytic action, cytotoxity degree, intracutaneous reactivity of aqueous extracts, in the second phase: estimation of tissue reaction after implantation the early and distant period, estimation of systemic and local induction of proinflammatory cytokines IL-1beta ad IL-6 after implantation. In physicochemic studies of aqueous extracts from fibers proper electric conductivity and dry residue were determined. The research of toxic action was carried out on frozen sperm of a bull estimating the survival time of sperm cells in the tested aqueous extracts. The estimation of hemolytic was conducted on human erythrocytes according to the method based on photometric measurement of blood supernatant and aqueous extracts. The research of irritating action measured with the changes of colour and size reaction after intracutaneous injection of aqueous extracts was carried out on rabbits. The research of the local reaction of soft tissue after implanting of aramid and polyester fibers into back muscles and into the peritoneal cavity was carried on 90 rats. Estimation was performed in early period, that is to 90th, and distant period, that is to the 360th day after implantation. The estimation of the local reaction of cartilaginous tissue was performed on 24 rabbits after implantation on 60, 90, 180, 270 and 360 days of aramid and polyester fibers into xiphoid process of sternum. The quantitative microscopic estimation of the local reaction of tissue after implantation of fibers was performed using punctual estimation of tissue reaction. The research of proinflammatory cytokines IL-1beta and IL-6 induction after implantation of aramid and fibers into peritoneal cavity was performed on 85 mice Balb/c. the level IL-1beta and IL-6 was determined in the serum of peripheral blood of mouse and in peritoneal fluid in 3rd, 7th, 14th and 21st day after implantation. The results were compared with control of operative and spontaneous production of proinflammatory cytokines in non-operated animals. A rubber medical drain constituted the control of the tested cytokines induction. The determinations were made with ELISA method. The tests of resistance to biocorrosion in electronic scanning microscope were performed after implantation of fibers into peritoneal cavity of rats for 180 and 360 days. Biostability of mechanical properties of fibers after remaining for 360 days in the application of the tested breaking force in the knot and in the dynamic system of cyclic strain of fibers (with strength 20N and 1Hz). In the tests of aqueous extracts from fibers comparative pH (aramid--6.42 and polyester 6.35), four-time higher proper electric conductivity (aramid 45.5; polyester 11.0) and five-time higher-dry mass of residue after vaporization of extracts from aramid fibers in comparison with polyester fibers (aramid--10.4 mg, polyester--2.1 mg) were observed. In biological tests of aqueous extracts toxic, hemolytic or irritating action was not observed. The local reaction soft tissues after implantation of aramid and polyester fibers was similar. Macroscopically it was characterized with producing on the third day thin, transparent membrane which in distant period, was whitish, thicker and strongly connected with the implanted fibers and surrounding tissues. In the microscopic tests the reaction was characterized with a short-lasting exudative phase less intense around the aramid fibers and a proliferating phase led to producing a capsule from newly created areolar tissue sharply separated from muscles undergoing collagenization in the cicatrization phase. The produced connective tissue capsule from the side of the graft both around aramid and polyester had the cellular character with the presence of single giant multinucleated cells till the 360th day after implantation. From the side of the muscles the connective tissue streak had more dense character and in distant period consisted mainly of collagenous fibers with hyalinization features. The reaction of cartilaginous tissue after implantation both kinds of fibers from the 90th to the 360th day was similar. It was characterized with producing cartilaginous tissue and locally fibrous tissue with a large amount of intercellular substance. Connective tissue showed hyalinization features and was transformed into cartilaginous tissue. According to the punctual estimation of reaction the tested fibers caused minimal reaction. In the testes of level of proinflammatory cytokines IL-1beta and IL-6 their constant presence on the low level in the serum of mouse blood and in the peritoneal fluid was observed. aramid fibers did not induce an increase of the level IL-1beta and IL-6, whereas polyester fibers in the 3rd day after surgery stimulated locally their moderate increase. The influence of surgery on the local induction IL-1beta and IL-6 was statistically essential in the 3rd day. The rubber medical drain was strong inductor of mediators of inflammation IL-1beta and IL-6. In physiocomechanical fatigue-testing and in SEM in the period to 360th day after implantation, biocorrosion of aramid and polyester fibers was not observed. The value of breaking force of aramid fibers was twice higher and the resistance to cyclic fatigue was over two hundred times higher in comparison to polyester fibers. Those differences were statistically significant (p < 0.001). On the basis of the performed tests it is possible observed that aramid fibers cause minimal tissue reaction compared with the observed one around polyester fibers, they do not cause local nor systemic stimulation of proinflammatory cytokines IL-1beta and IL-6. aramid fibers show essentially higher statistic and dynamic mechanical resistance in comparison with polyester fibers; they not undergo biocorosion and they can be useful to increase mechanical resistance of medical materials or as independent biomaterial resistant to high mechanical forces.
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PMID:[Estimation of biocompatibility of fibers with large mechanical resistance]. 1563 Nov 54

In vitro studies suggest that extracellular nucleotides and nucleosides may be important regulators of inflammatory and immune responses. Most studies with adenosine 5'-triphosphate (ATP) have been performed in cell lines, which are remote from the human situation. The purpose of the present study was to determine the effects of ATP on TNF-alpha, IL-6 and IL-10 release in stimulated whole blood. Blood samples were drawn from healthy volunteers and incubated with ATP and lipopolysaccharide (LPS) + phytohemagglutinin (PHA) for 24 h. Contrary to expectations, ATP at 100 microM and 300 microM induced a reduction in TNF-alpha secretion by 32+/-8% (mean +/- SEM) and 65+/-4%, respectively. Furthermore, these ATP concentrations induced an increase in IL-10 secretion by 48+/-5% and 62+/-7% in whole blood. The ATP analogue adenosine 5'-O-(3-thiotriphosphate) (ATP-gamma-S) and adenosine 5'-diphosphate (ADP) also inhibited TNF-alpha release, but only ADP showed a stimulatory effect on IL-10. Co-treatment with adenosine deaminase did not reverse the ATP effect on TNF-alpha and IL-10. These results show, for the first time, that ATP inhibits the inflammatory response in stimulated whole blood as indicated by inhibition of TNF-alpha and stimulation of IL-10 release and that this effect is predominantly mediated by ATP and not by adenosine.
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PMID:Immunoregulatory effects of adenosine 5'-triphosphate on cytokine release from stimulated whole blood. 1571 72

The aim of this study was to compare the serum cytokine profiles of Plasmodium vivax malaria patients who presented with and without hepatic dysfunction. This is a retrospective analysis of 74 consecutive cases of P. vivax malaria seen at 3 military hospitals near the Demilitarized Zone in South Korea from 1999 to 2000. All patients studied were adult active duty servicemen. On admission, the mean (+/- SEM) age of the patients who presented with (n = 36) and without hepatic dysfunction (n = 38) was 21.6 +/- 0.24 and 22.5 +/- 0.44 years, respectively (P = 0.72). On admission, there was no significant difference between the 2 patient populations in terms of mean temperature, haemoglobin level, haematocrit, total white blood cell count, platelet count, parasite index, and serum concentration of transforming growth factor-beta. Plasmodium vivax malaria patients who presented with hepatic dysfunction had significantly higher mean serum concentrations of soluble Fas ligand, interleukin (IL)-l, IL-4, IL-6, IL-10, tumor necrosis factor-alpha, and interferon-gamma than those without hepatic dysfunction, suggesting the involvement of these cytokines in the development of hepatic dysfunction. The mean serum concentration of IL-12 was significantly lower in patients with hepatic dysfunction. The mean body temperature was not significantly different between the 2 patient populations.
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PMID:Serum cytokine profiles in patients with Plasmodium vivax malaria: a comparison between those who presented with and without hepatic dysfunction. 1611 64

The maternal syndrome preeclampsia is characterized by a generalized inflammatory response with activation of circulating leukocytes and altered levels of inflammatory cytokines. We hypothesized that one potential source of inflammatory cytokines during preeclampsia is the circulating maternal monocytes. By using flow cytometry, we found that the spontaneous intracellular synthesis of IL-1beta, IL-6, and IL-8 in monocytes of preeclamptic women was higher than in normal pregnant and non-pregnant women. The highest levels of cytokines were detected in women with the most abnormal laboratory values. When stimulated with lipopolysaccharide (LPS), the percentage of IL-1beta+ monocytes was lower in preeclampsia (72.6% +/- 8.2 SEM) than in normal pregnancy (90.7% +/- 2 SEM) (P = 0.03) and non-pregnant women (92.5% +/- 1.4 SEM) (P = 0.04) suggesting that monocytes from preeclamptic patients cannot be further stimulated. These results indicate that maternal circulating monocytes represent a source of inflammatory cytokines during preeclampsia.
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PMID:Monocytes of preeclamptic women spontaneously synthesize pro-inflammatory cytokines. 1633 93

PFAPA syndrome is characterized by periodic episodes of high fever, aphthous stomatitis, pharyngitis, and/or cervical adenitis. It is of unknown etiology and manifests usually before 5 years of age. We determined serum and intracellular cytokine levels in six PFAPA patients (4 males, 2 females, mean age 8 years (+/- 1.2 SEM), range 4-13) during the symptom-free period as well as 6-12 hours and 18-24 hours after fever onset. Values were compared to age-matched, healthy controls. Febrile PFAPA attacks led to a significant increase in IL-6 and IFN-gamma serum concentrations compared to symptom-free periods and to controls, with IL-1beta, TNF-alpha and IL-12p70 levels being significantly higher than in controls. Lymphocytic IFN-gamma and CD8+ IL-2 production was consistently significantly elevated compared to healthy children. During the asymptomatic period, serum concentrations of IL-1beta, IL-6, TNF-alpha and IL-12p70 were significantly increased compared to controls. Intracellular TNF-alpha synthesis was not elevated at any time point. Soluble TNFRp55 levels were even lower in between febrile episodes, reaching values comparable to controls during attacks, whereas soluble TNFRp75 levels increased during attacks compared to healthy children. Anti-inflammatory IL-4 in serum was at all times lower in PFAPA patients compared to controls with no difference in levels of intracellular IL-4 and IL-10 or serum IL-10. The observed increase of pro-inflammatory mediators, even between febrile attacks, suggests a dysregulation of the immune response in PFAPA syndrome, with continuous pro-inflammatory cytokine activation and a reduced anti-inflammatory response.
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PMID:Cytokine profile in PFAPA syndrome suggests continuous inflammation and reduced anti-inflammatory response. 1684 27

High linoleic acid (LA) intakes have been suggested to reduce alpha-linolenic acid [ALA, 18:3(n-3)] metabolism to eicosapentaenoic acid [EPA, 20:5(n-3)] and docosahexaenoic acid [DHA, 22:6(n-3)], and favor high arachidonic acid [ARA, 20:4(n-6)]. We used a randomized cross-over study with men (n = 22) to compare the effect of replacing vegetable oils high in LA with oils low in LA in foods, while maintaining constant ALA, for 4 wk each, on plasma (n-3) fatty acids. Nonvegetable sources of fat, except fish and seafoods, were unrestricted. We determined plasma phospholipid fatty acids at wk 0, 2, 4, 6, and 8, and triglycerides, cholesterol, serum CRP, and IL-6, and platelet aggregation at wk 0, 4, and 8. LA and ALA intakes were 3.8 +/- 0.12% and 1.0 +/- 0.05%, and 10.5 +/- 0.53% and 1.1 +/- 0.06% energy with LA:ALA ratios of 4:0 and 10:1 during the low and high LA diets, respectively. The plasma phospholipid LA was higher and EPA was lower during the high than during the low LA diet period (P < 0.001), but DHA declined over the 8-wk period (r = -0.425, P < 0.001). The plasma phospholipid ARA:EPA ratios were (mean +/- SEM) 20.7 +/- 1.52 and 12.9 +/- 1.01 after 4 wk consuming the high or low LA diets, respectively (P < 0.001); LA was inversely associated with EPA (r = -0.729, P < 0.001) but positively associated with ARA:EPA (r = 0.432, P < 0.001). LA intake did not influence ALA, ARA, DPA, DHA, or total, LDL or HDL cholesterol, CRP or IL-6, or platelet aggregation. In conclusion, high LA intakes decrease plasma phospholipid EPA and increase the ARA:EPA ratio, but do not favor higher ARA.
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PMID:Decreasing linoleic acid with constant alpha-linolenic acid in dietary fats increases (n-3) eicosapentaenoic acid in plasma phospholipids in healthy men. 1737 59

To study the correlation between the degree of histopathological changes in the liver and selected biochemical tests after bile duct ligation (BDL) in male and female Wistar rats. Rats were divided into 4 groups. Laparotomized male rats (M-LAP), male rats with BDL (M-BDL), laparotomized female rats (F-LAP) and female rats with BDL (F-BDL). Liver bioptic samples were taken 7 and 14 days after operations and were cultivated 24 hours under standard conditions. TNF-alpha and IL-6 (pg/ml) concentrations were estimated in cell culture supernatants. Rats were sacrificed 14 days after operations, histological evaluation of liver tissue was performed, and serum activity of alanine-transaminase (ALT) and gamma-glutamyl transferase (GMT) (microkat/l) and estradiol concentration (EST) (pmol/1) were estimated. Results are presented as median (cytokines, estradiol) or mean +/- SEM. IL-6 concentration in cell culture supernatants was 7 days after the operation 1495 pg/ml in M-BDL and 1050 pg /ml in F-BDL, but were not detected in M-LAP and F-LAP. TNF-alpha concentration in supernatants was detected in all groups and was not significantly higher in male rats in comparison with female rats. M-BDL group had higher activity of ALT (1.5+/-0.1) and GMT (1.24+/-0.38) in comparison with F-BDL (ALT - 1.24+/-0.38, GMT - 0.98+/-0.38). Estradiol concentrations were detected in BDL groups, male rats 175 pmol/l and female rats 543 pmol/l. Although ALT and GMT activities and TNF-alpha and IL-6 concentrations were higher in male rats histopathological findings have shown slightly more advanced fibrotic and inflammatory changes in female rats.
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PMID:The differences in selected biochemical markers and histological findings after bile duct ligation in male and female rats. 1758 39

We developed a novel tissue adhesive consisting of human serum albumin (HSA) and tartaric acid derivative (TAD). Four different concentrations of TAD namely, 0.05 mM, 0.1 mM, 0.2 mM and 0.3 mM were mixed with 40%, 42% and 44% HSA individually and were made in the form of disks. J774A.1 mouse macrophage cells were seeded on top of these disks. The disks were pre-treated with sterile water and Eagle's medium before every seeding. All the seeding was incubated from 1 day to 3 days before making any investigations on it. SEM images were recorded and it was observed that these cells adhered to these materials very well. Mouse IL-6 cytokine expressions were studied using ELISA. It was seen from the cytokine expression results that the release of IL-6 was minimum at 0.3 mM TAD concentrations with 44% HSA disks. No significant difference was observed in the cytokine expressions of IL-6 at 42% and 44% HSA at all concentrations of TAD studied in this work. mRNA gene expressions of IL-6 were investigated using RT-PCR technique. In 40% HSA, the gene expression level of IL-6 gene did not change during 3-day-culture in the range of TAD concentration of 0.05 mmol to 0.2 mmol. However, 0.3 mM TAD suppressed the gene expression at all concentration of HSA. In 42% HSA, although 0.05 mM and 0.1 mM TAD did not affect the gene expression, 0.2 mM and 0.3 mM TAD induced the expression level with incubation time. In 44% HSA, all the concentration of TAD increased the expression level even though the cytokine expression levels were quite low. Hence it could be thought that the expression at the cytokine level is quite insignificant where as it is to be considered at the gene expression level. On the whole, 0.3 mM TAD with 44% HSA could be considered as a challenging material as a tissue adhesive material for use in the field of tissue engineering.
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PMID:Investigations on the interaction of tartaric acid derivative/human serum albumin tissue adhesive with J774A.1 mouse macrophage cells through SEM, IL-6 cytokine and gene expression techniques. 1761 Dec 96

A prototype in-line filtration/adsorption device has been developed using novel synthetic pyrolysed carbon monoliths with controlled mesoporous domains of 2-50nm. Porosity was characterized by SEM and porosimetry. Removal of inflammatory cytokines TNF, IL-6, IL-1beta and IL-8 was assessed by filtering cytokine spiked human plasma through the walls of the carbon modules under pressure. The effect of carbon filtration on plasma clotting response and total plasma protein concentration was also assessed. Significant removal of the cytokines IL-6, IL-1beta and IL-8 was observed. Initially marked TNF removal diminished over time. The coagulation studies indicated that the carbon device does not exacerbate the propensity of blood plasma to clot. The total plasma protein concentration remained constant. The device offers a broader approach to the treatment of systemic inflammatory response syndrome (SIRS) by the removal of inflammatory mediators central to its progression.
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PMID:Inflammatory cytokine removal by an activated carbon device in a flowing system. 1820 34


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