title: Conditioned media from hypoxic-cultured human dental pulp cells promotes bone healing during distraction osteogenesis.
abstract: Distraction osteogenesis (DO) is a surgical procedure used to correct various skeletal disorders. Improving the technique by reducing the healing time would be of clinical relevance. The aim of this study was to determine the angiogenic and regenerative potential of conditioned media (CMs) collected from human dental pulp cells (hDPCs) grown under different culture conditions. CM collected from cells under hypoxia was used to improve bone healing and the DO procedure in vivo. The angiogenic potentials of CMs collected from hDPCs grown under normoxic (-Nor) and hypoxic (-Hyp) conditions were evaluated by quantitative PCR (VEGF-A, angiopoietin-1, angiopoietin-2, interleukin-6 (IL-6) and CXCL12), ELISA assays (VEGF-A, Ang-2), tube-formation and wound-healing assays, using human umbilical vein endothelial cells. The results demonstrated that hypoxic CM had significantly higher angiogenic potential than normoxic CM. Human fetal osteoblasts (hFOBs) were exposed to CM, followed by alizarin red staining, to assess the osteogenic potential. It was found that CM did not enhance the mineralization capacity of hFOBs. DO was performed in the tibiae of 30 mice, followed by a local injection of 20 µl CM (CM-Nor and CM-Hyp groups) or serum-free DMEM (control group) into the distraction zone every second day. The mice were sacrificed at days 13 and 27. The CM-Hyp treatment revealed a higher X-ray density than the control group (p < 0.05). Our study suggests that the angiogenic effect promoted by hypoxic culture conditions is dependent on VEGF-A and Ang-2 released from hDPCs. Furthermore, CM-Hyp treatment may thus improve the DO procedure, accelerating bone healing. © 2015 The Authors. Journal of Tissue Engineering and Regenerative Medicine published by John Wiley & Sons, Ltd.
authors.affiliation: Department of Clinical Dentistry, Centre for Clinical Dental Research, University of Bergen, Norway.|Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, Japan.
authors.forename: Masahito
authors.initials: M
authors.lastname: Fujio
authors.affiliation: Department of Clinical Dentistry, Centre for Clinical Dental Research, University of Bergen, Norway.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Xing
authors.affiliation: Department of Clinical Dentistry, Centre for Clinical Dental Research, University of Bergen, Norway.
authors.forename: Niyaz
authors.initials: N
authors.lastname: Sharabi
authors.affiliation: Department of Clinical Dentistry, Centre for Clinical Dental Research, University of Bergen, Norway.
authors.forename: Ying
authors.initials: Y
authors.lastname: Xue
authors.affiliation: Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, Japan.
authors.forename: Akihito
authors.initials: A
authors.lastname: Yamamoto
authors.affiliation: Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, Japan.
authors.forename: Hideharu
authors.initials: H
authors.lastname: Hibi
authors.affiliation: Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, Japan.
authors.forename: Minoru
authors.initials: M
authors.lastname: Ueda
authors.affiliation: Department of Clinical Dentistry, Centre for Clinical Dental Research, University of Bergen, Norway.
authors.forename: Inge
authors.initials: I
authors.lastname: Fristad
authors.affiliation: Department of Clinical Dentistry, Centre for Clinical Dental Research, University of Bergen, Norway.
authors.forename: Kamal
authors.initials: K
authors.lastname: Mustafa
mesh_terms: D000818:Animals; D015687:Cell Hypoxia; D017077:Culture Media, Conditioned / Q000494:pharmacology; D003782:Dental Pulp / Q000378:metabolism* / Q000473:pathology; D005260:Female; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism / Q000473:pathology; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology*; D051379:Mice; D019857:Osteogenesis, Distraction; D013977:Tibia / Q000293:injuries / Q000378:metabolism / Q000473:pathology; D014945:Wound Healing / Q000187:drug effects
chemical_list: D017077:Culture Media, Conditioned; D036341:Intercellular Signaling Peptides and Proteins
keywords: angiogenesis; bone; conditioned media; distraction osteogenesis; regeneration