title: A new application of cell-free bone regeneration: immobilizing stem cells from human exfoliated deciduous teeth-conditioned medium onto titanium implants using atmospheric pressure plasma treatment.
abstract: BACKGROUND
  Surface modification of titanium (Ti) implants promotes bone formation and shortens the osseointegration period. The aim of this study was to promote bone regeneration and stability around implants using atmospheric pressure plasma (APP) pretreatment. This was followed by immobilization of stem cells from human exfoliated deciduous teeth-conditioned medium (SHED-CM) on the Ti implant surface.
  
  
  METHODS
  Ti samples (implants, discs, powder) were treated with APP for 30 seconds. Subsequently, these were immobilized on the treated Ti surface, soaked and agitated in phosphate-buffered saline or SHED-CM for 24 hours at 37 °C. The surface topography of the Ti implants was observed using scanning electron microscopy with energy dispersive X-ray spectroscopy. In vivo experiments using Ti implants placed on canine femur bone were then conducted to permit histological analysis at the bone-implant boundary. For the in vitro experiments, protein assays (SDS-PAGE, Bradford assay, liquid chromatography-ion trap mass spectrometry) and canine bone marrow stromal cell (cBMSC) attachment assays were performed using Ti discs or powder.
  
  
  RESULTS
  In the in vitro study, treatment of Ti implant surfaces with SHED-CM led to calcium phosphate and extracellular matrix protein immobilization. APP pretreatment increased the amount of SHED-CM immobilized on Ti powder, and contributed to increased cBMSC attachment on Ti discs. In the in vivo study, histological analysis revealed that the Ti implants treated with APP and SHED-CM stimulated new bone formation around implants.
  
  
  CONCLUSIONS
  Implant device APP pretreatment followed by SHED-CM immobilization may be an effective application to facilitate bone regeneration around dental implants.
authors.affiliation: Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan. nud.omori000227@med.nagoya-u.ac.jp.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Omori
authors.affiliation: Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan. t-shuhei@med.nagoya-u.ac.jp.
authors.forename: Shuhei
authors.initials: S
authors.lastname: Tsuchiya
authors.affiliation: Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan. hara@med.nagoya-u.ac.jp.
authors.forename: Kenji
authors.initials: K
authors.lastname: Hara
authors.affiliation: EcoTopia Science Institute, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8502, Japan. kkuroda@numse.nagoya-u.ac.jp.
authors.forename: Kensuke
authors.initials: K
authors.lastname: Kuroda
authors.affiliation: Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan. hibihi@med.nagoya-u.ac.jp.
authors.forename: Hideharu
authors.initials: H
authors.lastname: Hibi
authors.affiliation: EcoTopia Science Institute, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8502, Japan. okido@numse.nagoya-u.ac.jp.
authors.forename: Masazumi
authors.initials: M
authors.lastname: Okido
authors.affiliation: Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan. mueda@med.nagoya-u.ac.jp.
authors.forename: Minoru
authors.initials: M
authors.lastname: Ueda
mesh_terms: D000818:Animals; D001274:Atmospheric Pressure; D001854:Bone Marrow Cells / Q000166:cytology; D001861:Bone Regeneration / Q000502:physiology*; D001842:Bone and Bones / Q000000981:diagnostic imaging / Q000473:pathology; D002448:Cell Adhesion; D002478:Cells, Cultured; D018914:Cells, Immobilized / Q000737:chemistry / Q000166:cytology; D002648:Child; D017077:Culture Media, Conditioned / Q000494:pharmacology; D015921:Dental Implants; D004285:Dogs; D006801:Humans; D008855:Microscopy, Electron, Scanning; D013052:Spectrometry, X-Ray Emission; D013234:Stem Cells / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects; D017154:Stromal Cells / Q000737:chemistry / Q000166:cytology; D014025:Titanium / Q000737:chemistry*; D014057:Tomography, X-Ray Computed; D014094:Tooth, Deciduous / Q000166:cytology*
chemical_list: D017077:Culture Media, Conditioned; D015921:Dental Implants; D014025:Titanium