title: Bilayered constructs aimed at osteochondral strategies: the influence of medium supplements in the osteogenic and chondrogenic differentiation of amniotic fluid-derived stem cells.
abstract: The development of osteochondral tissue engineered interfaces would be a novel treatment for traumatic injuries and aging associated diseases that affect joints. This study reports the development of a bilayered scaffold, which consists of both bone and cartilage regions. On the other hand, amniotic fluid-derived stem cells (AFSCs) could be differentiated into either osteogenic or chondrogenic cells, respectively. In this study we have developed a bilayered scaffolding system, which includes a starch/polycaprolactone (SPCL) scaffold for osteogenesis and an agarose hydrogel for chondrogenesis. AFSC-seeded scaffolds were cultured for 1 or 2 weeks in an osteochondral-defined culture medium containing both osteogenic and chondrogenic differentiation factors. Additionally, the effect of the presence or absence of insulin-like growth factor-1 (IGF-1) in the culture medium was assessed. Cell viability and phenotypic expression were assessed within the constructs in order to determine the influence of the osteochondral differentiation medium. The results indicated that, after osteogenic differentiation, AFSCs that had been seeded onto SPCL scaffolds did not require osteochondral medium to maintain their phenotype, and they produced a protein-rich, mineralized extracellular matrix (ECM) for up to 2 weeks. However, AFSCs differentiated into chondrocyte-like cells appeared to require osteochondral medium, but not IGF-1, to synthesize ECM proteins and maintain the chondrogenic phenotype. Thus, although IGF-1 was not essential for creating osteochondral constructs with AFSCs in this study, the osteochondral supplements used appear to be important to generate cartilage in long-term tissue engineering approaches for osteochondral interfaces. In addition, constructs generated from agarose-SPCL bilayered scaffolds containing pre-differentiated AFSCs may be useful for potential applications in regeneration strategies for damaged or diseased joints.
authors.affiliation: Wake Forest Institute for Regenerative Medicine, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA.
authors.forename: Márcia T
authors.initials: MT
authors.lastname: Rodrigues
authors.forename: Sang Jin
authors.initials: SJ
authors.lastname: Lee
authors.forename: Manuela E
authors.initials: ME
authors.lastname: Gomes
authors.forename: Rui L
authors.initials: RL
authors.lastname: Reis
authors.forename: Anthony
authors.initials: A
authors.lastname: Atala
authors.forename: James J
authors.initials: JJ
authors.lastname: Yoo
mesh_terms: D053674:Aggrecans / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D000653:Amniotic Fluid / Q000166:cytology*; D000880:Anthraquinones / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D018914:Cells, Immobilized / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D020219:Chondrogenesis / Q000187:drug effects*; D024043:Collagen Type II / Q000378:metabolism; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism; D003470:Culture Media / Q000494:pharmacology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D008855:Microscopy, Electron, Scanning; D010012:Osteogenesis / Q000187:drug effects*; D011091:Polyesters / Q000494:pharmacology; D012685:Sepharose / Q000494:pharmacology; D013052:Spectrometry, X-Ray Emission; D013194:Staining and Labeling; D013213:Starch / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D053674:Aggrecans; D000880:Anthraquinones; D024043:Collagen Type II; D050659:Core Binding Factor Alpha 1 Subunit; D003470:Culture Media; D011091:Polyesters; C492325:starch polycaprolactone; C010078:alizarin; D013213:Starch; D012685:Sepharose; D000469:Alkaline Phosphatase