title: Osteogenic differentiation of mesenchymal stem cells in fibrin-hydroxyapatite matrix in a 3-dimensional mesh scaffold.
abstract: OBJECTIVE
  To explore the feasibility of culturing mesenchymal stem cells in an hydroxyapatite-fibrin matrix held by a mesh scaffold and inducing osteogenic differentiation of these cells. The aim was to obtain bone-material in vitro in a desired form.
  
  
  METHODS
  Rat mesenchymal stem cells were mixed with fibrin and nanocrystalline hydroxyapatite in tubular scaffolds constructed from a poly(L-lactic acid) mesh, and cultured under standard and osteogenic differentiating conditions. Cell viability, cytotoxicity and alkaline phosphatase activity were followed for 3 weeks. Living cells and the expression of bone markers were visualized by fluorescence staining and immunofluorescence staining, respectively. Attachment of cells to the scaffold mesh surface was examined by scanning electron microscopy.
  
  
  RESULTS
  Cell viability decreased and cytotoxicity increased rapidly during the first day of culture but stabilized gradually afterwards, indicating fast adaptation of the cells in the matrix-scaffold environment. From day 17, cytotoxicity started to decrease, paralleled by an increase in alkaline phosphatase activity, indicating osteogenic differentiation. A large number of living cells were visible in the matrix and on the mesh scaffold. Expression of collagen type I, osteoponin, osteocalcin and core binding factor 1 were evident under osteogenic differentiation conditions.
  
  
  CONCLUSIONS
  The three-dimensional construction of a fibrin-hydroxyapatite matrix in a biocompatible poly(L-lactic acid) as mesh-scaffold provides a promising carrier for producing bone-material in vitro in a desired form for applications in regenerative medicine.
authors: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany ole.tiberius.jung@gmail.com.
authors: Ole
authors: O
authors: Jung
authors: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Henning
authors: H
authors: Hanken
authors: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Ralf
authors: R
authors: Smeets
authors: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Philip
authors: P
authors: Hartjen
authors: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Reinhard E
authors: RE
authors: Friedrich
authors: Interdisciplinary Center for Clinical Research (IZKF) 'BIOMAT', RWTH Aachen University Hospital, Aachen, Germany.
authors: Bettina
authors: B
authors: Schwab
authors: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Alexander
authors: A
authors: Gröbe
authors: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Max
authors: M
authors: Heiland
authors: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Ahmed
authors: A
authors: Al-Dam
authors: Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Wolfgang
authors: W
authors: Eichhorn
authors: Department of Medical Biometry and Epidemiology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Susanne
authors: S
authors: Sehner
authors: Department of Oral and Maxillofacial Surgery, Technical University of Munich, Hospital rechts der Isar, Munich, Germany.
authors: Andreas
authors: A
authors: Kolk
authors: Interdisciplinary Center for Clinical Research (IZKF) 'BIOMAT', RWTH Aachen University Hospital, Aachen, Germany.
authors: Michael
authors: M
authors: Wöltje
authors: Department of Operative Dentistry, Periodontology and Preventive Dentistry, University Hospital Aachen, Aachen, Germany.
authors: Jamal M
authors: JM
authors: Stein
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002470:Cell Survival; D017886:Durapatite; D005337:Fibrin; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D010012:Osteogenesis / Q000502:physiology*; D051381:Rats; D023822:Tissue Engineering; D054457:Tissue Scaffolds
chemical_list: D005337:Fibrin; D017886:Durapatite; D000469:Alkaline Phosphatase
keywords: MSC; additives; bone regeneration; cell culture; differentiation; tissue engineering