Apoptosis and cytokine release in human monocytes cultured on polystyrene and fibrinogen-coated polystyrene surfaces.
The effects of polystyrene (PS) material surface preadsorption with fibrinogen (3 mg/ml) and a low concentration of lipopolysaccharide (LPS; 10 ng/ml) and polystyrene particles (PS; 10(5)/ml) on human monocyte adhesion, viability and cytokine release were studied during 24h culture in vitro. LPS caused an upregulation of CD14 in adherent cells. In comparison with unstimulated cells on uncoated polystyrene surfaces, LPS did not alter the number of adherent cells but caused a markedly increased release of the proinflammatory cytokines (IL-1alpha and TNF-alpha) and the down-regulating IL-10. The expression of indicators of various stages of cell death, TdT, annexin-V, propidium iodide (PI) and lactate dehydrogenase (LDH), were unaltered, decreased, decreased and increased, respectively, after LPS stimulation. PS particles (3 microm psi) caused an increased DNA fragmentation but had a reduced proportion of annexin-V and PI positive cells in comparison with unstimulated cells on uncoated PS. In contrast, 1microm psi particles had a similar proportion of TdT, annexin-V and PI expressing cells as unstimulated controls. Cultures stimulated with particles (irrespective of size), had a similar concentration of proinflammatory cytokines as unstimulated controls, whereas a higher level of IL-10 was detected. Precoating of PS with fibrinogen revealed an enhanced cell adhesion and a concomitant reduction of CD14 expression. irrespective of stimulation with various agonists. The proportions of TdT, annexin-V and PI positive cells were unaltered or reduced on fibrinogen-coated PS in both unstimulated and agonist-challenged cultures. However, depending on the presence and type of agonist, fibrinogen mediated either a markedly increased (LPS) or equivalent (particles and unstimulated) IL-1alpha and TNFalpha release. Further, in comparison with uncoated substrates, fibrinogen was associated with a reduction of IL-10 release, irrespective of the type of stimuli. These observations, using low concentrations of bacterial and material products, indicate that fibrinogen modulates cell material interactions and up- and down-regulates specific events depending on the nature/ type of immediate stimuli.
Gretzer
C
C
Institute of Anatomy and Cell Biology, Göteborg University, Sweden. gretzerc@anatcell.gu.se
Werthén
M
M
Thomsen
P
P
D017304:Annexin A5 / Q000378:metabolism / Q000494:pharmacology; D017209:Apoptosis; D002448:Cell Adhesion; D002470:Cell Survival; D002478:Cells, Cultured; D004396:Coloring Agents / Q000494:pharmacology; D016207:Cytokines / Q000378:metabolism*; D005340:Fibrinogen / Q000378:metabolism*; D006801:Humans; D020287:In Situ Nick-End Labeling; D007375:Interleukin-1 / Q000378:metabolism; D016753:Interleukin-10 / Q000096:biosynthesis / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D018950:Lipopolysaccharide Receptors / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000378:metabolism; D009000:Monocytes / Q000378:metabolism*; D011137:Polystyrenes / Q000737:chemistry*; D011419:Propidium / Q000494:pharmacology; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D017304:Annexin A5; D004396:Coloring Agents; D016207:Cytokines; D007375:Interleukin-1; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D011137:Polystyrenes; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D011419:Propidium; D005340:Fibrinogen; D007770:L-Lactate Dehydrogenase