Part I: Cellular and Molecular Biology of Osteoarthritis and Osteoporosis
1. Isolation of Murine and Human Osteocyte
Matthew Prideaux, Amber Rath Stern, and Lynda F. Bonewald
2. Expansion and Chondrogenic Differentiation of Human Bone Marrow-Derived Mesenchymal Stromal Cells
Roberto Narcisi, Wendy J.L.M. Koevoet, and Gergo J.V.M. van Osch
3. A Novel Enzymatic Digestion Approach for Isolation and Culture of Rodent Bone Marrow Mesenchymal Progenitors
Leilei Zhong, Lutian Yao, and Ling Qin
4. Isolation of Nucleus Pulposus and Annulus Fibrosus Cells from the Intervertebral Disc
Guus G.H. van den Akker, Andy Cremers, Donatus A.M. Surtel, Willem Voncken, and Tim J.M. Welting
5. Engineering Cartilage Tissue by Co-Culturing of Chondrocytes and Mesenchymal Stromal Cells
Yao Fu, Carlo A. Paggi, Amel Dudakovic, Andre J. van Wijnen, Janine Post, and Marcel Karperien
6. Generation of Induced Pluripotent Stem Cells
David R. Deyle
7. Specimen Preparation for Single Cell Sequencing Analysis of Skeletal Cells
Shawon Debnath and Matthew Greenblatt
8. Mapping 5-Hydroxymethylcytosine (5hmC) Modifications in Skeletal Tissues Using High-Throughput Sequencing
Fiorella Carla Grandi and Nidhi Bhutani
9. Using FRAP to Quantify Changes in Transcription Factor Dynamics after Cell Stimulation: Cell Culture, FRAP, Data Analysis, and Visualization
Kannan Govindaraj and Janine N. Post
10. Quantitative Molecular Models for Biological Processes: Modeling of Signal Transduction Networks with ANIMO
Sakshi Khurana, Janet Huisman, Stefano Schivo, and Janine N. Post
Part II: In Vivo Models of Skeletal Tissue Injury, Degeneration, and Repair
11. Generation and Characterization of Mouse Models for Skeletal Disease
Gabrielle E. Foxa, Ye Liu, Lisa M. Turner, Alexander G. Robling, Tao Yang, and Bart O. Williams
12. Drill Hole Models to Investigate Bone Repair
Zhijun Li and Jill Helms
13. Generation and Experimental Outcomes of Closed Femoral Fracture in Mice
Joseph L. Roberts, Christopher W. Kinter, and Hicham Drissi
14. Mouse Models of Osteoarthritis: Surgical Model of Post-Traumatic Osteoarthritis Induced by Destabilization of the Medial Meniscus
Kirsty L. Culley, Purva Singh, Samantha Lessard, Mengying Wang, Brennan Rourke, Mary B. Goldring, and Miguel Otero
15. Immunostaining of Skeletal Tissues
Crystal Idleburg, Madelyn R. Lorenz, Elizabeth N. DeLassus, Erica L. Scheller, and Deborah J. Veis
16. Mapping Regional Cortical Bone Responses to Local Changes in Loading and Systemic Stimuli
Sara Windahl, Peter J. Delisser, and Gabriel L. Galea
17. Pain and Activity Measurements
David H.H. Molstad and Elizabeth W. Bradley
This detailed second edition provides key updates on well-established methods as well as coverage of recently emerging sophisticated techniques and methods that are essential for an in-depth and state-of-the-art understanding of skeletal development and homeostasis and the pathological mechanisms that cause skeletal degeneration. The first half of the book examines cell and molecular biological aspects of osteoarthritis and osteoporosis, while the remainder of the collection focuses on in vivo models of skeletal tissue injury, degeneration, and repair. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, Osteoporosis and Osteoarthritis, Second Edition aims to assist investigators concerned with research topics broadly related to osteoporosis, osteoarthritis, intervertebral disc degeneration, as well as other musculoskeletal disorders.