title: Osteogenic differentiation of equine cord blood multipotent mesenchymal stromal cells within coralline hydroxyapatite scaffolds in vitro.
abstract: OBJECTIVE
  To investigate the osteogenic differentiation potential of equine umbilical cord blood-derived multipotent mesenchymal stromal cells (CB-MSC) within coralline hydroxyapatite scaffolds cultured in osteogenic induction culture medium.
  
  
  METHODS
  Scaffolds seeded with equine CB-MSC were cultured in cell expansion culture medium (control) or osteogenic induction medium (treatment). Cell viability and distribution were confirmed by the MTT cell viability assay and DAPI nuclear fluorescence staining, respectively. Osteogenic differentiation was evaluated after 10 days using reverse transcription polymerase chain reaction, alkaline phosphatase activity, and secreted osteocalcin concentration. Cell morphology and matrix deposition were assessed by scanning electron microscopy (SEM) after 14 days in culture.
  
  
  RESULTS
  Cells showed viability and adequate distribution within the scaffold. Successful osteogenic differentiation within the scaffolds was demonstrated by the increased expression of osteogenic markers such as Runx2, osteopontin, osteonectin, collagen IA; increased levels of alkaline phosphatase activity; increased osteocalcin protein secretion and bone-like matrix presence in the scaffold pores upon SEM evaluation.
  
  
  CONCLUSIONS
  These results demonstrate that equine CB-MSC maintain viability and exhibit osteogenic potential in coralline hydroxyapatite scaffolds when induced in vitro . Equine CB-MSC scaffold constructs deserve further investigation for their potential role as biologically active fillers to enhance bone-gap repair in the horse.
authors.affiliation: Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada.
authors.forename: R J
authors.initials: RJ
authors.lastname: Figueroa
authors.forename: T G
authors.initials: TG
authors.lastname: Koch
authors.forename: D H
authors.initials: DH
authors.lastname: Betts
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002470:Cell Survival; D002516:Ceramics / Q000737:chemistry*; D004247:DNA / Q000096:biosynthesis; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005312:Fetal Blood / Q000166:cytology*; D005786:Gene Expression Regulation; D006736:Horses; D006882:Hydroxyapatites / Q000737:chemistry*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D008855:Microscopy, Electron, Scanning; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D015675:Osteocalcin / Q000378:metabolism; D010012:Osteogenesis / Q000502:physiology*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D023822:Tissue Engineering / Q000379:methods
chemical_list: D006882:Hydroxyapatites; C119955:coralline hydroxyapatite; D015675:Osteocalcin; D004247:DNA