title: Bone marrow mesenchymal stem cells stimulated by bFGF up-regulated protein expression in comparison with periodontal fibroblasts in vitro.
abstract: OBJECTIVE
  The aim of this study was to evaluate, in vitro, the role of bFGF in the proliferation and expression of collagen type I and fibronectin of dog bone marrow mesenchymal stem cells (dBMMSCs) in comparison with the expression of the same proteins in dog periodontal fibroblasts (dPLFs).
  
  
  METHODS
  dBMMSCs from the iliac crest were cultivated in Dulbecco's Modified Eagle's Medium (DMEM). Flow cytometry analysis (FCA) was used to characterize dBMMSC. Cells were stimulated with bFGF (1, 5 and 10 ng/mL) after 24 and 48 h. Real time RT-PCR was performed to verify collagen type I and fibronectin expressions. MTT assay was used to confirm cellular proliferation. Statistical analyses were performed (ANOVA and Kruskal-Wallis tests; p<0.05).
  
  
  RESULTS
  FCA showed 55.98% of CD34+ and 32.67% of CD90+ after bone marrow aspiration; 3.33% of CD34+ and 33.0% of CD90+ before P1. After P2, 10.54% of dBMMSCs expressed CD90, whereas after P3, this number decreased to 1.58%. dPLFs presented 4.04% of CD90+ and 1.05% of CD34+ after P3. MTT evaluation showed increase in dBMSC proliferation with 5 ng/mL bFGF-stimulus after 24-h. Both collagen I and fibronectin expression were very similar between the two cells groups after 24-h stimulation with 1 ng/mL bFGF concentration. Fibronectin and collagen I expressions were higher after 24-h stimulation with 5 ng/mL bFGF.
  
  
  CONCLUSIONS
  dBMMSCs (1 ng/mL-bFGF stimulus after 24 h) are very similar to dPLFs as regards morphological and immunostaining characteristics, and collagen and/or fibronectin production. The dBMMSCs presented the highest protein expression rates with 5 ng/mL-bFGF stimulus after 24-h.
authors.affiliation: DDS, School of Dentistry, UNESP - Univ. Estadual Paulista, São Paulo, Brazil.
authors.forename: Renato
authors.initials: R
authors.lastname: Colenci
authors.affiliation: Department of Stomatology, School of Dentistry, Federal University of Paraná, UFPR, Paraná, Brazil.
authors.forename: Luciana Reichert
authors.initials: LR
authors.lastname: da Silva Assunção
authors.affiliation: Department of Clinics, Surgery and Animal Reproduction, School of Veterinary Medicine, UNESP - Univ. Estadual Paulista, São Paulo, Brazil.
authors.forename: Suely Regina
authors.initials: SR
authors.lastname: Mogami Bomfim
authors.affiliation: Laboratory of Flow Cytometry, Botucatu Blood Center, School of Medicine, UNESP - Univ. Estadual Paulista, São Paulo, Brazil.
authors.forename: Marjorie
authors.initials: M
authors.lastname: de Assis Golim
authors.affiliation: Laboratory of Cellular Engineering, Botucatu Blood Center, School of Medicine, UNESP - Univ. Estadual Paulista, São Paulo, Brazil.
authors.forename: Elenice
authors.initials: E
authors.lastname: Deffune
authors.affiliation: Department of Basic Sciences, School of Dentistry, UNESP - Univ. Estadual Paulista, São Paulo, Brazil. Electronic address: shpoliv@foa.unesp.br.
authors.forename: Sandra Helena
authors.initials: SH
authors.lastname: Penha Oliveira
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects; D024042:Collagen Type I / Q000378:metabolism*; D004285:Dogs; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D005347:Fibroblasts / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism*; D005434:Flow Cytometry; D066298:In Vitro Techniques; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D015854:Up-Regulation
chemical_list: D024042:Collagen Type I; D005353:Fibronectins; D016222:Fibroblast Growth Factor 2
keywords: Bone marrow; Fibroblasts; Mesenchymal stem cells; Periodontal ligament; bFGF