interleukin-8 and Cadaver

Corneal limbal stem cell deficiency (LSCD) may be treated using ex vivo limbal epithelial stem cells (LESCs) derived from cadaveric donor tissue. However, continuing challenges exist around tissue availability, inflammation, and transplant rejection. Lipopolysaccharide (LPS) or recombinant human IL-1β stimulated primary human keratocyte and LESC models were used to investigate the anti-inflammatory properties of a short chain, IL-1 receptor antagonist peptide for use in LESC sheet growth to control inflammation. The peptide was characterized using mass spectroscopy and high performance liquid chromatography. Peptide cytotoxicity, patterns of cell cytokine expression in response to LPS or IL-1β stimulation, and peptide suppression of this response were investigated by MTS/LDH assays, ELISA, and q-PCR. Cell differences in LPS stimulated toll-like receptor 4 expression were investigated using immunocytochemistry. A significant reduction in rIL-1β stimulated inflammatory cytokine production occurred following LESC and keratocyte incubation with anti-inflammatory peptide and in LPS stimulated IL-6 and IL-8 production following keratocyte incubation with peptide (1 mg/mL) (P < 0.05). LESCs produced no cytokine response to LPS stimulation and showed no TLR4 expression. The peptide supported LESC growth when adhered to a silicone hydrogel contact lens indicating potential use in improved LESC grafting through suppression of inflammation.

Topics: Cadaver; Cell Proliferation; Cells, Cultured; Cornea; Corneal Diseases; Corneal Keratocytes; Epithelium, Corneal; Humans; Inflammation; Interleukin-1beta; Interleukin-8; Lipopolysaccharides; Peptides; Receptors, Interleukin-1; Recombinant Proteins; Stem Cells; Toll-Like Receptor 4

Topics: Cadaver; Humans; Interleukin-8; Kidney; Kidney Transplantation; Tissue Donors

To investigate the longterm effects (12 days) of nonsteroidal antiinflammatory drugs [NSAID: aceclofenac (ACECLO), sodium diclofenac (DICLO), indomethacin (INDO), nimesulide (NIM), rofecoxib (ROFE), celecoxib (CELE), piroxicam (PIROX), and ibuprofen (IBUP)] on the metabolism of human chondrocytes cultured in alginate beads.. Enzymatically isolated osteoarthritic (OA) chondrocytes were cultured in alginate beads in a well defined culture medium for 12 days. The DNA content was measured according to a fluorimetric method and cell proliferation was determined by the incorporation of 3H-thymidine in the newly synthesized DNA. Interleukin 6 (IL-6) and IL-8, stromelysin [matrix metalloproteinase-3 (MMP-3)], and aggrecan (AGG) production were assayed by specific enzyme amplified sensitivity immunoassays, and prostaglandin E2 (PGE2) production by specific radioimmunoassay. All NSAID were tested at the mean peak plasma concentration (Cmax) obtained after oral administration of a therapeutic dose.. In alginate beads, chondrocytes synthesized high amounts of AGG, which were largely (98%) immobilized in the alginate matrix. A large amount (43%) of the IL-8 produced was stored in the alginate beads, whereas almost all IL-6 production (94%) was released in the culture supernatant. At the therapeutic concentration, all NSAID tested fully blocked PGE2 production. ACECLO, DICLO, INDO, NIM significantly inhibited basal and IL-1beta stimulated IL-6 production; CELE and IBUP only inhibited IL-1beta stimulated IL-6 production; and ROFE and PIROX had no significant effects. No NSAID showed significant effects on basal and IL-1beta stimulated IL-8 production, except CELE and IBUP, which slightly increased basal IL-8 production. ACECLO and INDO increased AGG content by 25% in the alginate beads, while the other NSAID were without significant effect. No NSAID were able to modify the inhibitory effect of IL-1beta on AGG production. NSAID did not modify MMP-3 production.. The mechanism of action of NSAID seems to be multifactorial and not limited to the inhibition of cyclooxygenases. Further, in our culture conditions, at the Cmax and by comparison with other NSAID, ACECLO and INDO show an advantageous activity profile. They fully blocked PGE2 production, inhibited IL-6 synthesis, and increased aggrecan synthesis. These effects appear advantageous for the longterm treatment of chronic joint diseases such as osteoarthritis.

Topics: Aggrecans; Alginates; Anti-Inflammatory Agents, Non-Steroidal; Cadaver; Cartilage, Articular; Cell Culture Techniques; Cell Division; Cell Separation; Cells, Cultured; Chondrocytes; DNA; Extracellular Matrix Proteins; Glucuronic Acid; Hexuronic Acids; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Lectins, C-Type; Matrix Metalloproteinase 3; Osteoarthritis; Proteoglycans

Topics: Adolescent; Adult; Aged; Biomarkers; Biopsy; C-Reactive Protein; Cadaver; Creatinine; Drug Therapy, Combination; Female; Graft Rejection; Humans; Immunosuppressive Agents; Interleukin-6; Interleukin-8; Kidney Transplantation; Living Donors; Male; Middle Aged; Reoperation; Sensitivity and Specificity; Tissue Donors; Tumor Necrosis Factor-alpha

We investigated the role of interleukin 8 (IL-8) in the pathogenesis of proliferative vitreoretinopathy (PVR). The IL-8 level in the vitreous and the blood of 20 patients with PVR was measured by a specific enzyme-linked immunoassay method. Vitreous from cadaver eyes (n = 20) was used as the control. The IL-8 level in the vitreous was between 31.3 and 65 pg/ml in 40% of eyes with PVR. IL-8 was detected neither in the blood of the patients with PVR nor in the vitreous from cadaver eyes. IL-8 levels in the vitreous did not correlate with either the grade of PVR or the duration of symptoms. These results suggest that IL-8 may play a role in the pathogenesis of PVR.

Topics: Adult; Cadaver; Female; Humans; Interleukin-8; Male; Middle Aged; Time Factors; Vitreoretinopathy, Proliferative; Vitreous Body