title: Biomimetic approach on human periodontal ligament cells using synthetic oligopeptides.
abstract: BACKGROUND
  Periodontal ligament (PDL) cells, connecting root cementum with alveolar bone, are important for periodontal wound healing. In order to obtain a predictable periodontal regeneration, selective adhesion and proliferation of PDL cells are essential. The purpose of this study was to investigate the effects of synthetic peptides mimicking cell-binding domain of fibronectin (FN) on human PDL cells.
  
  
  METHODS
  Two types of oligopeptides, Gly3-Pro-His-Ser-Arg-Asn-Gly6-Arg-Gly-Asp-Gly (G3PHSRNG6RGDG) and Gly3-His-Pro-Asn-Arg-Ser-Gly6-Arg-Gly-Asp-Gly (G3HPNRSG6RGDG), were constructed using a solid-phase peptide synthesizer. Fibronectin type III ninth to tenth domain (FN III 9-10) and Arg-Gly-Asp-Ser (RGDS) were prepared for comparison with the effects of synthetic peptides. These peptides were coated onto 96-well cell culture plates with 0.001 approximately 100 microM concentrations. Cultured human PDL cells were then applied to the peptide-coated wells at a density of 1 x 10(4)/well. After 1 hour incubation at 37 degrees C, adhered cells were fixed, stained, and examined by phase contrast microscopy for cell spreading assay. Attached PDL cells were solubilized with 2% sodium dodecyl sulfate (SDS) for the cell attachment assay by measuring absorbance at 595 nm in microplate reader. Western blot analysis was performed to determine extracellular signal-regulated kinase (ERK1/2) activity.
  
  
  RESULTS
  Cell attachment and spreading assays revealed that G3PHSRNG6RGDG (> or = 10 microM) possesses similar adhesive behavior to FN III 9-10. G3PHSRNG6RGDG showed a comparable ERK1/2 activity when compared to FN III 9-10.
  
  
  CONCLUSIONS
  G3PHSRNG6RGDG enhanced an attachment and spreading of human PDL cells thereby increasing ERK1/2 activity. Taken together, it is anticipated that this peptide might be a potential tool for arranging a biologically attractive environment for PDL cells, which would enhance periodontal regeneration efficacy.
authors.affiliation: Department of Periodontology, College of Dentistry, Seoul National University, Chongno-Ku, Seoul, South Korea.
authors.forename: Tae-Il
authors.initials: TI
authors.lastname: Kim
authors.forename: Jun-Hyeog
authors.initials: JH
authors.lastname: Jang
authors.forename: Yong-Moo
authors.initials: YM
authors.lastname: Lee
authors.forename: In-Chul
authors.initials: IC
authors.lastname: Rhyu
authors.forename: Chong-Pyoung
authors.initials: CP
authors.lastname: Chung
authors.forename: Soo-Boo
authors.initials: SB
authors.lastname: Han
authors.forename: Sang-Mook
authors.initials: SM
authors.lastname: Choi
authors.forename: Young
authors.initials: Y
authors.lastname: Ku
mesh_terms: D000704:Analysis of Variance; D032701:Biomimetics; D015153:Blotting, Western; D002448:Cell Adhesion; D005353:Fibronectins / Q000627:therapeutic use*; D016557:Guided Tissue Regeneration, Periodontal / Q000379:methods*; D006801:Humans; D039081:Integrin alpha5beta1 / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009842:Oligopeptides / Q000138:chemical synthesis / Q000378:metabolism / Q000627:therapeutic use*; D010513:Periodontal Ligament / Q000378:metabolism / Q000502:physiology*; D012038:Regeneration / Q000187:drug effects*
chemical_list: D005353:Fibronectins; D039081:Integrin alpha5beta1; D009842:Oligopeptides; C046617:arginyl-glycyl-aspartyl-serine; D020928:Mitogen-Activated Protein Kinases