title: Hydroxylated fullerene: a potential antiinflammatory and antioxidant agent for preventing mouse preterm birth.
abstract: OBJECTIVE
  Intrauterine infection such as by Escherichia coli and Ureaplasma spp induce placental inflammation and are one of the leading causes of preterm birth. Here we evaluated hydroxylated fullerene (C60[OH]44) for its in vitro antiinflammatory and antioxidant effects against host cellular responses to the ureaplasma toll-like receptor 2 (TLR2) ligand, UPM-1. In addition, we investigated the preventative effects of C60(OH)44 in vivo in a mouse preterm birth model that used UPM-1.
  
  
  METHODS
  TLR2-overexpressing cell lines and the primary cultures of mouse peritoneal macrophages were pretreated with C60(OH)44. After UPM-1 addition to the cell lines, the activation of the nuclear factor kappa-light chain-enhancer of activated B cells (NF-kappaB) signaling cascade and the production of reactive oxygen species were monitored. The levels of expression of inflammatory cytokines of interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF)-α, and the production of reactive oxygen species were quantified after stimulation with UPM-1. The in vivo preventative effects of C60(OH)44 on mice preterm birth were evaluated by analyzing the preterm birth rates and fetal survival rates in the preterm birth mouse model with placental histological analyses.
  
  
  RESULTS
  Pretreatment with C60(OH)44 significantly suppressed UPM-1-induced NF-kappaB activation and reactive oxygen species production in TLR2-overexpressing cell lines. In the primary culture of mouse peritoneal macrophages, UPM-1-induced production of reactive oxygen species and the expression of inflammatory cytokines such as IL-6, IL-1β, and TNF-α were significantly reduced by pretreatment with C60(OH)44. In the UPM-1-induced preterm birth mouse model, the preterm birth rate decreased from 72.7% to 18.2% after an injection of C60(OH)44. Placental examinations of the group injected with C60(OH)44 reduced the damage of the spongiotrophoblast layer and reduced infiltration of neutrophils.
  
  
  CONCLUSIONS
  C60(OH)44 was effective as a preventative agent of preterm birth in mice.
authors.affiliation: Department of Obstetrics and Gynecology, Graduate School of Medicine, Osaka University, Osaka, Japan; Department of Developmental Medicine, Research Institute, Osaka Medical Center for Maternal and Child Health, Osaka, Japan.
authors.forename: Tetsu
authors.initials: T
authors.lastname: Wakimoto
authors.affiliation: Department of Developmental Medicine, Research Institute, Osaka Medical Center for Maternal and Child Health, Osaka, Japan.
authors.forename: Kaoru
authors.initials: K
authors.lastname: Uchida
authors.affiliation: Department of Obstetrics and Gynecology, Graduate School of Medicine, Osaka University, Osaka, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Mimura
authors.affiliation: Department of Obstetrics and Gynecology, Graduate School of Medicine, Osaka University, Osaka, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kanagawa
authors.affiliation: Department of Obstetrics and Gynecology, Graduate School of Medicine, Osaka University, Osaka, Japan; Department of Developmental Medicine, Research Institute, Osaka Medical Center for Maternal and Child Health, Osaka, Japan.
authors.forename: Tzvetozar Roussev
authors.initials: TR
authors.lastname: Mehandjiev
authors.affiliation: Vitamin C60 BioResearch Corporation, Osaka, Japan.
authors.forename: Hisae
authors.initials: H
authors.lastname: Aoshima
authors.affiliation: Division of Applied Chemistry, Graduate School of Engineering, Osaka University, Osaka, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Kokubo
authors.affiliation: Department of Maternal-Fetal Medicine, Osaka Medical Center for Maternal and Child Health, Osaka, Japan.
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Mitsuda
authors.affiliation: Department of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Vaccine Creation Project, BIKEN Innovative Vaccine Research Alliance Laboratories, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Yoshioka
authors.affiliation: Department of Toxicology and Safety Science, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Tsutsumi
authors.affiliation: Department of Obstetrics and Gynecology, Graduate School of Medicine, Osaka University, Osaka, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Kimura
authors.affiliation: Department of Developmental Medicine, Research Institute, Osaka Medical Center for Maternal and Child Health, Osaka, Japan. Electronic address: itaruy@mch.pref.osaka.jp.
authors.forename: Itaru
authors.initials: I
authors.lastname: Yanagihara
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000378:metabolism; D000975:Antioxidants / Q000378:metabolism; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D037741:Fullerenes / Q000737:chemistry / Q000627:therapeutic use*; D007150:Immunohistochemistry; D017737:Macrophages, Peritoneal / Q000378:metabolism; D051379:Mice; D018384:Oxidative Stress / Q000187:drug effects* / Q000502:physiology; D047928:Premature Birth / Q000517:prevention & control*; D017382:Reactive Oxygen Species / Q000378:metabolism; D051195:Toll-Like Receptor 2 / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D000975:Antioxidants; D016207:Cytokines; D037741:Fullerenes; D017382:Reactive Oxygen Species; C493486:Tlr2 protein, mouse; D051195:Toll-Like Receptor 2; D014409:Tumor Necrosis Factor-alpha
keywords: hydroxylated fullerene; nuclear factor-kappaB; preterm birth mouse; reactive oxygen species; ureaplasma membrane-1