title: Inhibitory effects of serum from sepsis patients on epithelial cell migration in vitro: a case control study.
abstract: Sepsis delays wound re-epithelialization. In this study we explored the effect of human sepsis sera as well as the effects of cytokines, growth factors and exosomes of sepsis sera treated normal fibroblasts (NF) on keratinocyte migration and proliferation in vitro.
  
  
  
  Serum samples were taken on days 1, 4, and 9 from 44 patients diagnosed with severe sepsis, and from 14 matching healthy controls. We evaluated the effects of sepsis serum with or without TNF-α, EGF, EGF receptor inhibitor or exosomes of sepsis sera treated NF on human keratinocyte (HaCaT) proliferation (BrdU assay), viability (MTT assay), and migration (horizontal wound healing model). Cytokine levels of sepsis and healthy sera were measured by multiplex assay. Comparisons between groups were carried out using SPSS statistics and P < 0.05 was considered significant.
  
  
  
  Severe-sepsis sera collected on days 1, 4, and 9 reduced keratinocyte proliferation by 6% (P = 0.005), 20% (P = 0.001), and 18% (P = 0.002), respectively, compared to control sera. Cell viability in cultures exposed to sepsis sera from days 4 and 9 was reduced by 38% (P = 0.01) and 58% (P < 0.001), respectively. Open-surface wounds exposed to sepsis sera from days 1 and 4 were larger than those exposed to sera from healthy controls (60 vs. 31%, P = 0.034 and 66 vs. 31%, P = 0.023, respectively). Exosomes of sepsis or healthy sera treated NF inhibited keratinocyte migration. We detected higher serum levels of cytokines TNF-α (5.7 vs. 0.7 pg/ml, P < 0.001), IL-6 (24.8 vs. 3.8 pg/ml, P < 0.001), IL-10 (30.0 vs. 11.9 pg/ml, P = 0.040), and VEGF (177.9 vs. 48.1 pg/ml, P = 0.018) in sepsis sera. Levels of EGF were significantly lower in sepsis sera than in that of healthy controls (6.5 vs. 115.6 pg/ml, P < 0.001). Sepsis serum supplemented with EGF 5 ng/ml and TNF-α in all concentrations improved keratinocyte migration.
  
  
  
  Keratinocyte viability, proliferation and migration were reduced in severe sepsis in vitro. Exosomes from NF added in healthy or sepsis serum media inhibited keratinocyte migration. Decreased levels of EGF in sepsis sera may partially explain the delay of wound healing with severe-sepsis patients. Increased levels of TNF-α in sepsis sera do not explain diminished keratinocyte migration.
authors.affiliation: Research Group of Surgery, Anesthesia and Intensive Care, Oulu University Hospital, P. O. Box 21, 90029, Oulu, Finland. henna.jaurila@fimnet.fi.|Cancer and Translational Medicine Research Unit, Faculty of Medicine, Medical Research Center Oulu, University of Oulu, P.O. Box 5281, 90014, Oulu, Finland. henna.jaurila@fimnet.fi.
authors.forename: Henna
authors.identifier: 0000-0003-0952-9819
authors.initials: H
authors.lastname: Jaurila
authors.affiliation: Research Group of Surgery, Anesthesia and Intensive Care, Oulu University Hospital, P. O. Box 21, 90029, Oulu, Finland.
authors.forename: Vesa
authors.initials: V
authors.lastname: Koivukangas
authors.affiliation: Research Group of Surgery, Anesthesia and Intensive Care, Oulu University Hospital, P. O. Box 21, 90029, Oulu, Finland.
authors.forename: Marjo
authors.initials: M
authors.lastname: Koskela
authors.affiliation: Research Group of Surgery, Anesthesia and Intensive Care, Oulu University Hospital, P. O. Box 21, 90029, Oulu, Finland.
authors.forename: Fiia
authors.initials: F
authors.lastname: Gäddnäs
authors.affiliation: Research Group of Biomedicine, Faculty of Biochemistry and Molecular Medicine, University of Oulu, P. O. Box 5000, Oulu, 90014, Finland.
authors.forename: Sirpa
authors.initials: S
authors.lastname: Salo
authors.affiliation: Cancer and Translational Medicine Research Unit, Faculty of Medicine, Medical Research Center Oulu, University of Oulu, P.O. Box 5281, 90014, Oulu, Finland.
authors.forename: Johanna
authors.initials: J
authors.lastname: Korvala
authors.affiliation: Cancer and Translational Medicine Research Unit, Faculty of Medicine, Medical Research Center Oulu, University of Oulu, P.O. Box 5281, 90014, Oulu, Finland.
authors.forename: Maija
authors.initials: M
authors.lastname: Risteli
authors.affiliation: Research Unit of Biomedicine, Faculty of Medicine and Biocenter of Oulu, University of Oulu, P.O. Box 5000, Oulu, 90014, Finland.
authors.forename: Toni
authors.initials: T
authors.lastname: Karhu
authors.affiliation: Research Unit of Biomedicine, Faculty of Medicine and Biocenter of Oulu, University of Oulu, P.O. Box 5000, Oulu, 90014, Finland.|Department of Gastroenterology and Metabolism, Poznan University of Medical Sciences, Collegium Maius, Fredry 10, 61-701, Poznan, Poland.
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Herzig
authors.affiliation: Cancer and Translational Medicine Research Unit, Faculty of Medicine, Medical Research Center Oulu, University of Oulu, P.O. Box 5281, 90014, Oulu, Finland.|Research Group of Oral Health Sciences, Oulu University Hospital, Medical Research Center Oulu, University of Oulu, P. O. Box 5000, Oulu, 90014, Finland.
authors.forename: Tuula
authors.initials: T
authors.lastname: Salo
authors.affiliation: Research Group of Surgery, Anesthesia and Intensive Care, Oulu University Hospital, P. O. Box 21, 90029, Oulu, Finland.
authors.forename: Tero I
authors.initials: TI
authors.lastname: Ala-Kokko
mesh_terms: D000328:Adult; D000368:Aged; D016022:Case-Control Studies; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D016207:Cytokines / Q000097:blood; D003710:Demography; D004847:Epithelial Cells / Q000473:pathology*; D055354:Exosomes / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000473:pathology; D006801:Humans; D015603:Keratinocytes / Q000473:pathology; D008297:Male; D008875:Middle Aged; D018805:Sepsis / Q000097:blood* / Q000473:pathology*
chemical_list: D016207:Cytokines
keywords: EGF; Exosome; HaCaT; Human serum; In vitro; Migration; Sepsis; TNF-α