TY - JOUR
T1 - Structure and quantification of microvascularisation within mouse long bones
T2 - What and how should we measure?
AU - Roche, Bernard
AU - David, Valentin
AU - Vanden-Bossche, Arnaud
AU - Peyrin, Françoise
AU - Malaval, Luc
AU - Vico, Laurence
AU - Lafage-Proust, Marie Hélène
N1 - Funding Information:
This study was supported by the Agence Nationale de la Recherche (ANR AdapHyG, Adaptation to hypergravity during mouse development, 2010–12), by the CNES (Centre National d'Etudes Spatiales) and by a grant from the Société Française de Rhumatologie .
PY - 2012/1
Y1 - 2012/1
N2 - Bone marrow vascularisation is involved in both remodeling and hematopoïesis. Challenged mouse models often require imaging and quantitative assessment of blood vessels and bone cell activities for a better understanding of the role of the vascular system. In this study we compared images of mouse hind limb long bone vascularisation after infusion of either barium sulfate or lead chromate-loaded silicon. The images were then analyzed through histology as well as low-resolution and synchrotron-radiation microtomography. We show that barium sulfate infusion provides the best vessel images and furthermore, that it is compatible with staining procedures used in bone histomorphometry and CD31 immunohistochemistry. Bone marrow vascularisation displays large structural and spatial distribution heterogeneity, including large lobular clusters of sinusoids and an unexpectedly substantial amount of capillaries in the adipocytes-rich distal third of the tibia. For an unbiased assessment of bone vascular development/changes, these features must be taken into account. We describe the conditions under which the quantification of microvascularisation on histological sections of barium-infused long bones is reproducible, as applied to seven-month-old male C57/Bl6J and mixed CD1/129Sv/J mice, and we propose a nomenclature for the histological parameters measured. Finally, we validate our technique by studying the effect of ovariectomy on mouse tibial vascular density.
AB - Bone marrow vascularisation is involved in both remodeling and hematopoïesis. Challenged mouse models often require imaging and quantitative assessment of blood vessels and bone cell activities for a better understanding of the role of the vascular system. In this study we compared images of mouse hind limb long bone vascularisation after infusion of either barium sulfate or lead chromate-loaded silicon. The images were then analyzed through histology as well as low-resolution and synchrotron-radiation microtomography. We show that barium sulfate infusion provides the best vessel images and furthermore, that it is compatible with staining procedures used in bone histomorphometry and CD31 immunohistochemistry. Bone marrow vascularisation displays large structural and spatial distribution heterogeneity, including large lobular clusters of sinusoids and an unexpectedly substantial amount of capillaries in the adipocytes-rich distal third of the tibia. For an unbiased assessment of bone vascular development/changes, these features must be taken into account. We describe the conditions under which the quantification of microvascularisation on histological sections of barium-infused long bones is reproducible, as applied to seven-month-old male C57/Bl6J and mixed CD1/129Sv/J mice, and we propose a nomenclature for the histological parameters measured. Finally, we validate our technique by studying the effect of ovariectomy on mouse tibial vascular density.
KW - Baryum sulfate infusion
KW - Bone microvascularisation
KW - Histology
KW - Vascular quantification
KW - μCT
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U2 - 10.1016/j.bone.2011.09.051
DO - 10.1016/j.bone.2011.09.051
M3 - Article
C2 - 22019874
AN - SCOPUS:84455205646
VL - 50
SP - 390
EP - 399
JO - Bone
JF - Bone
SN - 8756-3282
IS - 1
ER -