One of the prerequisites for the development of safe and effective protocols designed to prevent and treat disorders of bone metabolism, growth, and maturation is a thorough understanding of the cell and molecular biology of bone growth and remodeling. Bone development and remodeling involves interactions between bone cells, bone extracellular matrix molecules, the immune system, and systemic factors. Specific cell surface molecules are likely to play important roles in bone cell regulation. However, little is known about the osteoblast and osteoclast cell surface and less is known about plasma membrane modifications during development. Since bone growth and remodeling are dependent on the number and activity of bone resorbing osteoclasts the objective of the proposed studies is to gain additional information on the regulatory mechanisms involved in osteoclast recruitment, differentiation, and action. Specifically, we propose to identify, purify, and biochemically characterize osteoclast-specific cell surface molecules defined by a monoclonal antibody library raised against isolated osteoclasts. Included here is to determine the molecular weights, partial amino acid sequence, and carbohydrate composition of these molecules. Moreover, we plan to determine the functional importance of the osteoclastspecific molecules. In this context we will analyze the response mediated by the interaction between a 96K osteoclast-specific cell surface molecule and its 80K ligand. Functional studies will include monitoring changes in bone resorption activity, cell attachment, phosphorylation, motility, etc. Monoclonal antibodies and oligonucleotide probes generated from known amino acid sequence information will be used to screen an osteoclast cDNA library to identify osteoclastspecific genes. Along with monoclonal antibodies to osteoclasts and their precursors these molecular probes will be employed to trace osteoclast lineage and study osteoclast regulation. We will utilized isolated chick osteoclasts to purify and study osteoclast- specific proteins and determine their function. We have described cell culture systems in which monocyte and marrow nononuclear cells will fuse and form multinucleated giant cells. These cultures will be used to help identify osteoclast precursors. The results from these studies will help understand normal and inflammatory bone loss and aid in devising therapies for arthritic, diabetic, osteoporotic, and periodontal disease associated osteopenia.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR032927-08
Application #
3156444
Study Section
General Medicine B Study Section (GMB)
Project Start
1984-07-01
Project End
1992-06-30
Budget Start
1991-07-01
Budget End
1992-06-30
Support Year
8
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Washington University
Department
Type
Schools of Dentistry
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Collin-Osdoby, Patricia; Osdoby, Philip (2012) Isolation and culture of primary chicken osteoclasts. Methods Mol Biol 816:119-43
Collin-Osdoby, Patricia; Osdoby, Philip (2012) RANKL-mediated osteoclast formation from murine RAW 264.7 cells. Methods Mol Biol 816:187-202
Collin-Osdoby, Patricia; Yu, Xuefeng; Zheng, Hong et al. (2003) RANKL-mediated osteoclast formation from murine RAW 264.7 cells. Methods Mol Med 80:153-66
Collin-Osdoby, Patricia; Anderson, Fred; Osdoby, Philip (2003) Primary isolation and culture of chicken osteoclasts. Methods Mol Med 80:65-88
Yu, Xuefeng; Huang, Yuefang; Collin-Osdoby, Patricia et al. (2003) Stromal cell-derived factor-1 (SDF-1) recruits osteoclast precursors by inducing chemotaxis, matrix metalloproteinase-9 (MMP-9) activity, and collagen transmigration. J Bone Miner Res 18:1404-18
Collin-Osdoby, Patricia; Rothe, Linda; Bekker, Simon et al. (2002) Basic fibroblast growth factor stimulates osteoclast recruitment, development, and bone pit resorption in association with angiogenesis in vivo on the chick chorioallantoic membrane and activates isolated avian osteoclast resorption in vitro. J Bone Miner Res 17:1859-71
Collin-Osdoby, P; Rothe, L; Anderson, F et al. (2001) Receptor activator of NF-kappa B and osteoprotegerin expression by human microvascular endothelial cells, regulation by inflammatory cytokines, and role in human osteoclastogenesis. J Biol Chem 276:20659-72
Collin-Osdoby, P; Rothe, L; Bekker, S et al. (2000) Decreased nitric oxide levels stimulate osteoclastogenesis and bone resorption both in vitro and in vivo on the chick chorioallantoic membrane in association with neoangiogenesis. J Bone Miner Res 15:474-88
Collin-Osdoby, P; Li, L; Rothe, L et al. (1998) Inhibition of avian osteoclast bone resorption by monoclonal antibody 121F: a mechanism involving the osteoclast free radical system. J Bone Miner Res 13:67-78
Khalkhali-Ellis, Z; Collin-Osdoby, P; Li, L et al. (1997) A human homolog of the 150 kD avian osteoclast membrane antigen related to superoxide dismutase and essential for bone resorption is induced by developmental agents and opposed by estrogen in FLG 29.1 cells. Calcif Tissue Int 60:187-93

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