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Am J Physiol Cell Physiol 282: C775-C785, 2002; doi:10.1152/ajpcell.00310.2001
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Vol. 282, Issue 4, C775-C785, April 2002

Effect of HUVEC on human osteoprogenitor cell differentiation needs heterotypic gap junction communication

F. Villars1, B. Guillotin1, T. Amédée2, S. Dutoya1, L. Bordenave1, R. Bareille1, and J. Amédée1

Institut National de la Santé et de la Recherche Médicale 1 U-443 and 2 U-394, 33076 Bordeaux Cedex, France

Bone development and remodeling depend on complex interactions between bone-forming osteoblasts and other cells present within the bone microenvironment, particularly vascular endothelial cells that may be pivotal members of a complex interactive communication network in bone. Our aim was to investigate the interaction between human umbilical vein endothelial cells (HUVEC) and human bone marrow stromal cells (HBMSC). Cell differentiation analysis performed with different cell culture models revealed that alkaline phosphatase activity and type I collagen synthesis were increased only by the direct contact of HUVEC with HBMSC. This "juxtacrine signaling" could involve a number of different heterotypic connexions that require adhesion molecules or gap junctions. A dye coupling assay with Lucifer yellow demonstrated a functional coupling between HUVEC and HBMSC. Immunocytochemistry revealed that connexin43 (Cx43), a specific gap junction protein, is expressed not only in HBMSC but also in the endothelial cell network and that these two cell types can communicate via a gap junctional channel constituted at least by Cx43. Moreover, functional inhibition of the gap junction by 18alpha -glycyrrhetinic acid treatment or inhibition of Cx43 synthesis with oligodeoxyribonucleotide antisense decreased the effect of HUVEC cocultures on HBMSC differentiation. This stimulation could be mediated by the intercellular diffusion of signaling molecules that permeate the junctional channel.

coupling; intercellular messenger; connexin43; osteoblast; endothelial cells


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