title: Combinatorial Analysis of Growth Factors Reveals the Contribution of Bone Morphogenetic Proteins to Chondrogenic Differentiation of Human Periosteal Cells.
abstract: Successful application of cell-based strategies in cartilage and bone tissue engineering has been hampered by the lack of robust protocols to efficiently differentiate mesenchymal stem cells into the chondrogenic lineage. The development of chemically defined culture media supplemented with growth factors (GFs) has been proposed as a way to overcome this limitation. In this work, we applied a fractional design of experiment (DoE) strategy to screen the effect of multiple GFs (BMP2, BMP6, GDF5, TGF-β1, and FGF2) on chondrogenic differentiation of human periosteum-derived mesenchymal stem cells (hPDCs) in vitro. In a micromass culture (μMass) system, BMP2 had a positive effect on glycosaminoglycan deposition at day 7 (p < 0.001), which in combination with BMP6 synergistically enhanced cartilage-like tissue formation that displayed in vitro mineralization capacity at day 14 (p < 0.001). Gene expression of μMasses cultured for 7 days with a medium formulation supplemented with 100 ng/mL of BMP2 and BMP6 and a low concentration of GDF5, TGF-β1, and FGF2 showed increased expression of Sox9 (1.7-fold) and the matrix molecules aggrecan (7-fold increase) and COL2A1 (40-fold increase) compared to nonstimulated control μMasses. The DoE analysis indicated that in GF combinations, BMP2 was the strongest effector for chondrogenic differentiation of hPDCs. When transplanted ectopically in nude mice, the in vitro-differentiated μMasses showed maintenance of the cartilaginous phenotype after 4 weeks in vivo. This study indicates the power of using the DoE approach for the creation of new medium formulations for skeletal tissue engineering approaches.
authors.affiliation: 1 Tissue Engineering Laboratory, Skeletal Biology and Engineering Research Center , Katholieke Universiteit Leuven, Leuven, Belgium .|2 Prometheus, Division of Skeletal Tissue Engineering, Katholieke Universiteit Leuven , Leuven, Belgium .
authors.forename: Luis Filipe
authors.initials: LF
authors.lastname: Mendes
authors.affiliation: 1 Tissue Engineering Laboratory, Skeletal Biology and Engineering Research Center , Katholieke Universiteit Leuven, Leuven, Belgium .|2 Prometheus, Division of Skeletal Tissue Engineering, Katholieke Universiteit Leuven , Leuven, Belgium .
authors.forename: Wai Long
authors.initials: WL
authors.lastname: Tam
authors.affiliation: 1 Tissue Engineering Laboratory, Skeletal Biology and Engineering Research Center , Katholieke Universiteit Leuven, Leuven, Belgium .|2 Prometheus, Division of Skeletal Tissue Engineering, Katholieke Universiteit Leuven , Leuven, Belgium .
authors.forename: Yoke Chin
authors.initials: YC
authors.lastname: Chai
authors.affiliation: 2 Prometheus, Division of Skeletal Tissue Engineering, Katholieke Universiteit Leuven , Leuven, Belgium .|3 Biomechanics Research Unit, University of Liege , Liege, Belgium .|4 Department of Mechanical Engineering, Biomechanics Section, Katholieke Universiteit Leuven, Heverlee, Belgium .
authors.forename: Liesbet
authors.initials: L
authors.lastname: Geris
authors.affiliation: 1 Tissue Engineering Laboratory, Skeletal Biology and Engineering Research Center , Katholieke Universiteit Leuven, Leuven, Belgium .|2 Prometheus, Division of Skeletal Tissue Engineering, Katholieke Universiteit Leuven , Leuven, Belgium .
authors.forename: Frank P
authors.initials: FP
authors.lastname: Luyten
authors.affiliation: 1 Tissue Engineering Laboratory, Skeletal Biology and Engineering Research Center , Katholieke Universiteit Leuven, Leuven, Belgium .|2 Prometheus, Division of Skeletal Tissue Engineering, Katholieke Universiteit Leuven , Leuven, Belgium .|5 Institute of Orthopaedics and Musculoskeletal Science, Division of Surgery and Interventional Science, University College London , The Royal National Orthopaedic Hospital, London, United Kingdom .
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Roberts
mesh_terms: D000293:Adolescent; D000818:Animals; D019485:Bone Morphogenetic Proteins / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D020219:Chondrogenesis / Q000187:drug effects / Q000502:physiology*; D005260:Female; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D010521:Periosteum / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D014019:Tissue Donors; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D019485:Bone Morphogenetic Proteins; D036341:Intercellular Signaling Peptides and Proteins