title: The effects of 1α, 25-dihydroxyvitamin D3 and transforming growth factor-β3 on bone development in an ex vivo organotypic culture system of embryonic chick femora.
abstract: Transforming growth factor-beta3 (TGF-β3) and 1α,25-dihydroxyvitamin D3 (1α,25 (OH) 2D3) are essential factors in chondrogenesis and osteogenesis respectively. These factors also play a fundamental role in the developmental processes and the maintenance of skeletal integrity, but their respective direct effects on these processes are not fully understood. Using an organotypic bone rudiment culture system the current study has examined the direct roles the osteotropic factors 1α,25 (OH)2D3 and TGF-β3 exert on the development and modulation of the three dimensional structure of the embryonic femur. Isolated embryonic chick femurs (E11) were organotypically cultured for 10 days in basal media, or basal media supplemented with either 1α,25 (OH) 2D3 (25 nM) or TGF-β3 (5 ng/mL & 15 ng/mL). Analyses of the femurs were undertaken using micro-computed tomography (μCT), histology and immunohistochemistry. 1α,25 (OH)2D3 supplemented cultures enhanced osteogenesis directly in the developing femurs with elevated levels of osteogenic markers such as type 1 collagen. In marked contrast organotypic femur cultures supplemented with TGF-β3 (5 ng/mL & 15 ng/mL) demonstrated enhanced chondrogenesis with a reduction in osteogenesis. These studies demonstrate the efficacy of the ex vivo organotypic embryonic femur culture employed to elucidate the direct roles of these molecules, 1α,25 (OH) 2D3 and TGF-β3 on the structural development of embryonic bone within a three dimensional framework. We conclude that 1α,25(OH)2D and TGF-β3 modify directly the various cell populations in bone rudiment organotypic cultures effecting tissue metabolism resulting in significant changes in embryonic bone growth and modulation. Understanding the roles of osteotropic agents in the process of skeletal development is integral to developing new strategies for the recapitulation of bone tissue in later life.
authors.affiliation: Bone & Joint Research Group, Human Development and Health, University of Southampton, Southampton, United Kingdom.
authors.forename: Emma L
authors.initials: EL
authors.lastname: Smith
authors.affiliation: Centre for Biomolecular Sciences, School of Pharmacy, University Park, University of Nottingham, Nottingham, United Kingdom.
authors.forename: Hassan
authors.initials: H
authors.lastname: Rashidi
authors.affiliation: Bone & Joint Research Group, Human Development and Health, University of Southampton, Southampton, United Kingdom.
authors.forename: Janos M
authors.initials: JM
authors.lastname: Kanczler
authors.affiliation: Centre for Biomolecular Sciences, School of Pharmacy, University Park, University of Nottingham, Nottingham, United Kingdom.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Shakesheff
authors.affiliation: Bone & Joint Research Group, Human Development and Health, University of Southampton, Southampton, United Kingdom.
authors.forename: Richard O C
authors.initials: RO
authors.lastname: Oreffo
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002642:Chick Embryo; D020219:Chondrogenesis / Q000187:drug effects*; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D005269:Femur / Q000166:cytology / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D015870:Gene Expression; D010012:Osteogenesis / Q000187:drug effects*; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics / Q000378:metabolism; D046509:Tissue Culture Techniques; D053782:Transforming Growth Factor beta3 / Q000494:pharmacology*; D014807:Vitamin D / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: D015415:Biomarkers; D024042:Collagen Type I; D018809:Proliferating Cell Nuclear Antigen; D053782:Transforming Growth Factor beta3; C118756:dihydroxy-vitamin D3; D014807:Vitamin D
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