Biomimetic approach on human periodontal ligament cells using synthetic oligopeptides.
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.
Kim
Tae-Il
TI
Department of Periodontology, College of Dentistry, Seoul National University, Chongno-Ku, Seoul, South Korea.
Jang
Jun-Hyeog
JH
Lee
Yong-Moo
YM
Rhyu
In-Chul
IC
Chung
Chong-Pyoung
CP
Han
Soo-Boo
SB
Choi
Sang-Mook
SM
Ku
Young
Y
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*
D005353:Fibronectins; D039081:Integrin alpha5beta1; D009842:Oligopeptides; C046617:arginyl-glycyl-aspartyl-serine; D020928:Mitogen-Activated Protein Kinases