title: A paper-based scaffold for enhanced osteogenic differentiation of equine adipose-derived stem cells.
abstract: OBJECTIVE
  We investigated the applicability of single layer paper-based scaffolds for the three-dimensional (3D) growth and osteogenic differentiation of equine adipose-derived stem cells (EADSC), with comparison against conventional two-dimensional (2D) culture on polystyrene tissue culture vessels.
  
  
  RESULTS
  Viable culture of EADSC was achieved using paper-based scaffolds, with EADSC grown and differentiated in 3D culture retaining high cell viability (>94 %), similarly to EADSC in 2D culture. Osteogenic differentiation of EADSC was significantly enhanced in 3D culture, with Alizarin Red S staining and quantification demonstrating increased mineralisation (p < 0.0001), and an associated increase in expression of the osteogenic-specific markers alkaline phosphatase (p < 0.0001), osteopontin (p < 0.0001), and runx2 (p < 0.01). Furthermore, scanning electron microscopy revealed a spherical morphology of EADSC in 3D culture, compared to a flat morphology of EADSC in 2D culture.
  
  
  CONCLUSIONS
  Single layer paper-based scaffolds provide an enhanced environment for the in vitro 3D growth and osteogenic differentiation of EADSC, with high cell viability, and a spherical morphology.
authors.affiliation: School of Biomedical Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia. gpetersen@csu.edu.au.
authors.forename: Gayle F
authors.initials: GF
authors.lastname: Petersen
authors.affiliation: School of Animal and Veterinary Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia. bhilbert@csu.edu.au.
authors.forename: Bryan J
authors.initials: BJ
authors.lastname: Hilbert
authors.affiliation: School of Animal and Veterinary Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia. gtrope@csu.edu.au.
authors.forename: Gareth D
authors.initials: GD
authors.lastname: Trope
authors.affiliation: School of Biomedical Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia. wkalle@csu.edu.au.
authors.forename: Wouter H J
authors.initials: WH
authors.lastname: Kalle
authors.affiliation: School of Biomedical Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia. pstrappe@csu.edu.au.
authors.forename: Padraig M
authors.initials: PM
authors.lastname: Strappe
mesh_terms: D017667:Adipocytes / Q000166:cytology*; D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology*; D006736:Horses; D010012:Osteogenesis / Q000502:physiology*; D010209:Paper; D013234:Stem Cells / Q000166:cytology*; D054457:Tissue Scaffolds / Q000737:chemistry*
keywords: Equine adipose-derived stem cells; Osteogenic differentiation; Paper-based scaffold; Three-dimensional culture; Two-dimensional culture