title: Angiogenesis in the distal femoral chondroepiphysis of the rabbit during development of the secondary centre of ossification.
abstract: In the developing chondroepiphyses of long bones, the avascular cartilaginous anlage is invaded by numerous blood vessels, through the process of angiogenesis. The objective of this study was to investigate the chronology of this vascular invasion with the spontaneous calcification of the cartilaginous epiphysis during development of the secondary ossification centre in the rabbit distal femur. The time-course of chondroepiphyseal vascular invasion was determined histologically and standardized for eight gestational and four postnatal intervals by plotting kit body mass against crown-rump length. Similarly, microcomputed tomography (micro-CT) helped to visualize calcification at those same gestational and postnatal intervals. To confirm the angiogenic nature of the avascular chondroepiphysis, such samples were assayed on the chick chorio-allantoic membrane (CAM). Neovascular outgrowths from the CAM were apparent 48 h following introduction of an 18-day (gestational) chondroepiphyseal sample. Chondroepiphyseal samples were assayed for the potent developmental angiogenic factors bFGF and VEGF, with the mRNA expression for both these mediators being confirmed using RT-PCR. As angiogenesis and calcification during chondroepiphyseal development occur in a defined tissue environment initially devoid of blood vessels and mineral, those processes provided a unique opportunity to study their progression without complication of injury-related inflammation or extant vasculature and mineral. Furthermore, the discovery of angiogenic, angiostatic or mineral-regulating mediators specific to developing connective tissue may prove useful for analysing the regulation of vascular and mineral pathogenesis in articular tissues.
authors.affiliation: Department of Surgery, McCaig Centre for Joint Injury and Arthritis Research, Faculty of Medicine, University of Calgary, Alberta, Canada. doschak@ucalgary.ca
authors.forename: M R
authors.initials: MR
authors.lastname: Doschak
authors.forename: D M L
authors.initials: DM
authors.lastname: Cooper
authors.forename: C N
authors.initials: CN
authors.lastname: Huculak
authors.forename: J R
authors.initials: JR
authors.lastname: Matyas
authors.forename: D A
authors.initials: DA
authors.lastname: Hart
authors.forename: B
authors.initials: B
authors.lastname: Hallgrimsson
authors.forename: R F
authors.initials: RF
authors.lastname: Zernicke
authors.forename: R C
authors.initials: RC
authors.lastname: Bray
mesh_terms: D000199:Actins / Q000235:genetics; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002356:Cartilage / Q000098:blood supply / Q000000981:diagnostic imaging / Q000196:embryology*; D002642:Chick Embryo; D020219:Chondrogenesis / Q000502:physiology*; D016228:Endothelial Growth Factors / Q000235:genetics; D004838:Epiphyses / Q000098:blood supply / Q000000981:diagnostic imaging / Q000196:embryology; D005260:Female; D005269:Femur / Q000098:blood supply / Q000000981:diagnostic imaging / Q000196:embryology*; D016222:Fibroblast Growth Factor 2 / Q000235:genetics; D015870:Gene Expression; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry / Q000379:methods; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics; D008222:Lymphokines / Q000235:genetics; D018919:Neovascularization, Physiologic; D010012:Osteogenesis / Q000502:physiology; D011817:Rabbits; D011859:Radiography; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018180:Thrombomodulin / Q000032:analysis; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
chemical_list: D000199:Actins; D015415:Biomarkers; D016228:Endothelial Growth Factors; D036341:Intercellular Signaling Peptides and Proteins; D008222:Lymphokines; D018180:Thrombomodulin; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D016222:Fibroblast Growth Factor 2