title: Effect of dynamic three-dimensional culture on osteogenic potential of human periodontal ligament-derived mesenchymal stem cells entrapped in alginate microbeads.
abstract: OBJECTIVE
  Bioreactors are devices that efficiently create an environment that enables cell cultures to grow in a three-dimensional (3D) context mimicking in vivo conditions. In this study, we investigate the effect of dynamic fluid flow on the osteogenic potential of human mesenchymal stem cells obtained from periodontal ligament and entrapped in alginate microbeads.
  
  
  METHODS
  After proper immunophenotyping, cells were encapsulated in barium alginate, cultured in 3D static or 3D dynamic conditions represented by a bioreactor system. Calcein-AM/propidium iodide staining was used to assess cellular viability. Quantitative real-time polymerase chain reaction was used to analyze the expression of osteogenic markers (Runx2 and COL1). Alizarin Red S staining and the Fourier transform infrared spectroscopy were used to assess mineral matrix deposition.
  
  
  RESULTS
  Optimal encapsulation procedure, in terms of polymer pumping rate, distance from droplet generator to the gelling bath and atomizing airflow was assessed. Cell viability was not affected by encapsulation in alginate microbeads. Bioreactor cell exposure was effective in anticipating osteogenic differentiation and improving mineral matrix deposition.
  
  
  CONCLUSIONS
  For the first time human mesenchymal stem cells obtained from periodontal ligaments encapsulated in alginate microbeads were cultured in a bioreactor system. This combination could represent a promising strategy to create a cell-based smart system with enhanced osteogenic potential useful for many different dental applications.
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, Ferrara University, Ferrara, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Vecchiatini
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, Ferrara University, Ferrara, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Penolazzi
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, Ferrara University, Ferrara, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Lambertini
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, Ferrara University, Ferrara, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Angelozzi
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, Ferrara University, Ferrara, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Morganti
authors.affiliation: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Mazzitelli
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, Ferrara University, Ferrara, Italy.|Research Centre for the Study of Periodontal and Peri-implant Diseases, Ferrara University, Ferrara, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Trombelli
authors.affiliation: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Nastruzzi
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, Ferrara University, Ferrara, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Piva
mesh_terms: D050156:Adipogenesis / Q000502:physiology; D000464:Alginates / Q000737:chemistry; D001031:Apatites / Q000032:analysis; D001672:Biocompatible Materials / Q000737:chemistry; D019149:Bioreactors; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D020219:Chondrogenesis / Q000502:physiology; D024042:Collagen Type I / Q000032:analysis; D000090602:Collagen Type I, alpha 1 Chain; D050659:Core Binding Factor Alpha 1 Subunit / Q000032:analysis; D005109:Extracellular Matrix / Q000737:chemistry; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006801:Humans; D057446:Hydrodynamics; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D008863:Microspheres; D010012:Osteogenesis / Q000502:physiology*; D010513:Periodontal Ligament / Q000166:cytology*; D013499:Surface Properties; D054457:Tissue Scaffolds / Q000737:chemistry; D059631:Wharton Jelly / Q000166:cytology
chemical_list: D000464:Alginates; D001031:Apatites; D001672:Biocompatible Materials; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D050659:Core Binding Factor Alpha 1 Subunit; D006603:Hexuronic Acids; C493709:RUNX2 protein, human; D020723:Glucuronic Acid
keywords: 3D cell culture system; RCCS bioreactor; alginate microbeads; human periodontal ligament stem cells; osteogenic differentiation