Summary
A method for demineralization of bone, preserving the antigenicity of neuroactive peptides, was developed. In all parts of rat long bones, nerves immunoreactive to substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) and tyrosine hydroxylase (TH) were detected after immunohistochemical staining. The majority of nerves were vascular, although several non-vascular endings were observed at the growth plate and amidst marrow cells. An abundance of nerves were demonstrated near the epiphyseal plate and in the periosteum, regions of high osteogenic activity. The occurrence of different nerve types was analyzed at different stages of heterotopic osteogenesis, induced by allogeneic bone matrix. Nerve fibres immunoreactive to SP, CGRP, NPY and TH occurred amidst differentiating chondroblastic cells in the second week. They gradually increased in number during the ensuing eight weeks. In an in vitro study of osteoblastic cells (UMR 106-01, ROS 17/2.8, Saos-2, MC3T3-E1) receptors to CGRP, VIP, noradrenaline (NA) and NPY were demonstrated as assessed by analysis of cyclic AMP formation. In UMR cells, NPY inhibited the effects of NA and parathyroid hormone (PTH), which is the first demonstration of a receptor interaction between a local neuropeptide and a systemic calcium regulating hormone. The combined findings indicate a neuroendocrine influence on bone physiology.
Résumé
Une méthode de déminéralisation de l'os, préservant l'antigénicité des peptides neuro-actifs, a été mise au point. Après coloration immuno-histochimique, on a mis en évidence, à tous les niveaux des os longs du rat, des nerfs immuno-réactifs à la substance P (SP), à la calcitonine peptide «gene-related» (CPGR), au polypeptide intestinal vaso-moteur (PIV), au neuropeptide Y (NPY), à la tyrosine hydroxylase (TH) et à l'interleukine 1 (IL-1). La majorité des nerfs sont vascularisés, bien que nombre de terminaisons non vascularisées aient été observées au niveau du cartilage de croissance et parmi les cellules de la moelle. On a montré la présence d'un grand nombre de fibres nerveuses à proximité de la plaque épiphysaire et dans le périoste, c'est à dire dans les régions d'importante activité ostéogénique. La fréquence des divers types de nerfs a été étudiée aux différentes étapes de l'ostéogénèse hétérotopique induite par la matrice osseuse allogénique. Les fibres nerveuses immunoréactives à SP, CPGR, NPY, TH et IL-1 apparaissent parmi les cellules chondroblastiques en voie de différenciation au cours de la deuxième semaine. Leur nombre augmente progressivement pendant les huit semaines suivantes. Dans une étude in vitro des cellules ostéoblastiques (UMR 106-01, ROS 17/2.8, Saos-2), des récepteurs de CPGR, PIV, noradrénaline (NA) et NPY ont pu être mis en évidence par l'analyse de la formation de l'AMP cyclique. L'action de l'IL-1 n'a pas été étudiée. Dans les cellules UMR, le NPY inhibe les effets de la NA et de l'hormone parathyroïdienne, ce qui est la première démonstration de l'interaction entre un neuro-peptide local et une hormone réglant le métabolisme systémique du calcium. Cet ensemble de constatations est en faveur d'une influence neuro-endocrinienne sur la physiologie osseuse et éventuellement aussi sur le développement du squelette.
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Bjurholm, A. Neuroendocrine peptides in bone. International Orthopaedics 15, 325–329 (1991). https://doi.org/10.1007/BF00186871
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DOI: https://doi.org/10.1007/BF00186871