title: Healing of a Large Long-Bone Defect through Serum-Free In Vitro Priming of Human Periosteum-Derived Cells.
abstract: Clinical translation of cell-based strategies for regenerative medicine demands predictable in vivo performance where the use of sera during in vitro preparation inherently limits the efficacy and reproducibility. Here, we present a bioinspired approach by serum-free pre-conditioning of human periosteum-derived cells, followed by their assembly into microaggregates simultaneously primed with bone morphogenetic protein 2 (BMP-2). Pre-conditioning resulted in a more potent progenitor cell population, while aggregation induced osteochondrogenic differentiation, further enhanced by BMP-2 stimulation. Ectopic implantation displayed a cascade of events that closely resembled the natural endochondral process resulting in bone ossicle formation. Assessment in a critical size long-bone defect in immunodeficient mice demonstrated successful bridging of the defect within 4 weeks, with active contribution of the implanted cells. In short, the presented serum-free process represents a biomimetic strategy, resulting in a cartilage tissue intermediate that, upon implantation, robustly leads to the healing of a large long-bone defect.
authors.affiliation: Tissue Engineering Laboratory, Skeletal Biology and Engineering Research Center, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium; Prometheus, Division of Skeletal Tissue Engineering, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium.
authors.forename: Johanna
authors.initials: J
authors.lastname: Bolander
authors.affiliation: Tissue Engineering Laboratory, Skeletal Biology and Engineering Research Center, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium; Prometheus, Division of Skeletal Tissue Engineering, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium.
authors.forename: Wei
authors.initials: W
authors.lastname: Ji
authors.affiliation: Tissue Engineering Laboratory, Skeletal Biology and Engineering Research Center, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium; Prometheus, Division of Skeletal Tissue Engineering, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium; Department of Developmental BioEngineering, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, Drienerlolaan 5, 7522NB Enschede, the Netherlands.
authors.forename: Jeroen
authors.initials: J
authors.lastname: Leijten
authors.affiliation: Tissue Engineering Laboratory, Skeletal Biology and Engineering Research Center, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium; Prometheus, Division of Skeletal Tissue Engineering, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium.
authors.forename: Liliana Moreira
authors.initials: LM
authors.lastname: Teixeira
authors.affiliation: Prometheus, Division of Skeletal Tissue Engineering, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium; Materials Technology TC, Campus Group T, KU Leuven, Andreas Vesaliusstraat 13, 3000 Leuven, Belgium.
authors.forename: Veerle
authors.initials: V
authors.lastname: Bloemen
authors.affiliation: Tissue Engineering Laboratory, Skeletal Biology and Engineering Research Center, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium; Prometheus, Division of Skeletal Tissue Engineering, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium.
authors.forename: Dennis
authors.initials: D
authors.lastname: Lambrechts
authors.affiliation: Tissue Engineering Laboratory, Skeletal Biology and Engineering Research Center, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium; Prometheus, Division of Skeletal Tissue Engineering, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium.
authors.forename: Malay
authors.initials: M
authors.lastname: Chaklader
authors.affiliation: Tissue Engineering Laboratory, Skeletal Biology and Engineering Research Center, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium; Prometheus, Division of Skeletal Tissue Engineering, KU Leuven, O&N 1, Herestraat 49, Box 813 13, 3000 Leuven, Belgium. Electronic address: frank.luyten@uzleuven.be.
authors.forename: Frank P
authors.initials: FP
authors.lastname: Luyten
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D015415:Biomarkers; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism; D001861:Bone Regeneration; D002454:Cell Differentiation; D020219:Chondrogenesis / Q000235:genetics; D004195:Disease Models, Animal; D050723:Fractures, Bone / Q000378:metabolism* / Q000473:pathology* / Q000628:therapy; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D010012:Osteogenesis / Q000235:genetics; D010521:Periosteum / Q000166:cytology*; D066329:Protein Aggregates; D015398:Signal Transduction; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D014945:Wound Healing
chemical_list: D018952:Antigens, CD34; C525718:BMP2 protein, human; D015415:Biomarkers; D055396:Bone Morphogenetic Protein 2; D066329:Protein Aggregates
keywords: bone morphogenetic proteins; cell-based constructs; critical bone fractures; fracture healing; fracture nonunion regenerative medicine; progenitor cells; serum free; stem cells