title: Ligament Tissue Engineering Using a Novel Porous Polycaprolactone Fumarate Scaffold and Adipose Tissue-Derived Mesenchymal Stem Cells Grown in Platelet Lysate.
abstract: OBJECTIVE
  Surgical reconstruction of intra-articular ligament injuries is hampered by the poor regenerative potential of the tissue. We hypothesized that a novel composite polymer "neoligament" seeded with progenitor cells and growth factors would be effective in regenerating native ligamentous tissue.
  
  
  METHODS
  We synthesized a fumarate-derivative of polycaprolactone fumarate (PCLF) to create macro-porous scaffolds to allow cell-cell communication and nutrient flow. Clinical grade human adipose tissue-derived human mesenchymal stem cells (AMSCs) were cultured in 5% human platelet lysate (PL) and seeded on scaffolds using a dynamic bioreactor. Cell growth, viability, and differentiation were examined using metabolic assays and immunostaining for ligament-related markers (e.g., glycosaminoglycans [GAGs], alkaline phosphatase [ALP], collagens, and tenascin-C).
  
  
  RESULTS
  AMSCs seeded on three-dimensional (3D) PCLF scaffolds remain viable for at least 2 weeks with proliferating cells filling the pores. AMSC proliferation rates increased in PL compared to fetal bovine serum (FBS) (p < 0.05). Cells had a low baseline expression of ALP and GAG, but increased expression of total collagen when induced by the ligament and tenogenic growth factor fibroblast growth factor 2 (FGF-2), especially when cultured in the presence of PL (p < 0.01) instead of FBS (p < 0.05). FGF-2 and PL also significantly increased immunostaining of tenascin-C and collagen at 2 and 4 weeks compared with human fibroblasts.
  
  
  CONCLUSIONS
  Our results demonstrate that AMSCs proliferate and eventually produce a collagen-rich extracellular matrix on porous PCLF scaffolds. This novel scaffold has potential in stem cell engineering and ligament regeneration.
authors.affiliation: 1 Department of Orthopedic Surgery, Mayo Clinic , Rochester, Minnesota.
authors.forename: Eric R
authors.initials: ER
authors.lastname: Wagner
authors.affiliation: 1 Department of Orthopedic Surgery, Mayo Clinic , Rochester, Minnesota.
authors.forename: Dalibel
authors.initials: D
authors.lastname: Bravo
authors.affiliation: 2 Department of Orthopedic Surgery and Biomedical Engineering, Mayo Clinic College of Medicine , Rochester, Minnesota.
authors.forename: Mahrokh
authors.initials: M
authors.lastname: Dadsetan
authors.affiliation: 2 Department of Orthopedic Surgery and Biomedical Engineering, Mayo Clinic College of Medicine , Rochester, Minnesota.
authors.forename: Scott M
authors.initials: SM
authors.lastname: Riester
authors.affiliation: 2 Department of Orthopedic Surgery and Biomedical Engineering, Mayo Clinic College of Medicine , Rochester, Minnesota.
authors.forename: Steven
authors.initials: S
authors.lastname: Chase
authors.affiliation: 1 Department of Orthopedic Surgery, Mayo Clinic , Rochester, Minnesota.
authors.forename: Jennifer J
authors.initials: JJ
authors.lastname: Westendorf
authors.affiliation: 2 Department of Orthopedic Surgery and Biomedical Engineering, Mayo Clinic College of Medicine , Rochester, Minnesota.
authors.forename: Allan B
authors.initials: AB
authors.lastname: Dietz
authors.affiliation: 2 Department of Orthopedic Surgery and Biomedical Engineering, Mayo Clinic College of Medicine , Rochester, Minnesota.
authors.forename: Andre J
authors.initials: AJ
authors.lastname: van Wijnen
authors.affiliation: 2 Department of Orthopedic Surgery and Biomedical Engineering, Mayo Clinic College of Medicine , Rochester, Minnesota.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Yaszemski
authors.affiliation: 1 Department of Orthopedic Surgery, Mayo Clinic , Rochester, Minnesota.
authors.forename: Sanjeev
authors.initials: S
authors.lastname: Kakar
mesh_terms: D000273:Adipose Tissue / Q000166:cytology* / Q000502:physiology; D001792:Blood Platelets / Q000737:chemistry / Q000378:metabolism*; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D004867:Equipment Design; D019544:Equipment Failure Analysis; D048091:Guided Tissue Regeneration / Q000295:instrumentation; D008022:Ligaments / Q000166:cytology / Q000254:growth & development*; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology; D011091:Polyesters / Q000737:chemistry*; D016062:Porosity; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods; D054457:Tissue Scaffolds
chemical_list: D011091:Polyesters; C506476:poly(caprolactone fumarate)