Osteoporosis, Methods and Protocols

In recent years, the study of bone cells and tissues at the cellular and molecular levels in a variety of models has revolutionized the field. In Osteoporosis: Methods and Protocols, leading scientists from around the world share their step-by-step laboratory protocols for studying bone biology. The topics covered in this volume include in vitro models, in vivo models utilized for drug testing, tissue engineering and osteoporosis studies in either gender, state-of-the-art molecular techniques to assess single genes or for global genomic analysis, powerful imaging techniques, and many more. As a volume in the highly successful Methods in Molecular Biology™ series, each chapter provides a brief introduction, a list of necessary materials, and a Notes section detailing tips on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, Osteoporosis: Methods and Protocols is the perfect guide for both new and experienced experimentalists attempting to study the devastating silent disease known as osteoporosis.

Table of contents

Table of Contents

(* indicates corresponding author)



In vitro models

1. Isolation and Culture of Rodent Osteoprogenitor Cells

Thomas A. Owen* and Lydia C. Pan

2. Osteoclast Culture and Resorption Assays

Elizabeth W. Bradley and Merry Jo Oursler*

3. Assessing New Bone Formation in Neonatal Calvarial Organ Cultures

Khalid S. Mohammad, John M. Chirgwin and Theresa Guise*

4. Detection of Apoptosis of Bone Cells In Vitro

Teresita Bellido*and Lilian I. Plotkin



In vivo rodent models

5. In Vivo Parathyroid Hormone Treatments and mRNA Isolation and Analysis

Xin Li, Ling Qin and Nicola C. Partridge*

6. Assessment of Bone Formation Capacity Using In Vivo Transplantation Assays: Procedure and Tissue Analysis

Badem M. Abdalleh, Nicholas Ditzel and Moustapha Kassem*

7. Ossicle and Vossicle Implant Model Systems

Glenda J. Pettway and Laurie K. McCauley*

8. Utility of the Ovariectomized (OVX) Rat as a Model for Human Osteoporosis In Drug Discovery

Yogendra P. Kharode*, Michael C. Sharp and Peter V. N. Bodine

9. Orchidectomy (ORX) Models of Osteoporosis

Stéphane Blouin, Hélène Libouban, Marie Françoise Moreau and Daniel Chappard*



Methods of gene delivery

10. Gene Delivery by Adenoviruses

Renny Franceschi* and Chunxi Ge

11. Lentiviral Delivery of shRNA Constructs into Osteoblasts

Marc N. Wein, Dallas C. Jones and Laurie H. Glimcher*

12. Gene Delivery by Retroviruses

Valerie Deregowski and Ernesto Canalis*

Molecular and Genetic Techniques

13. Chromatin Immunoprecipitation (ChIP) Assays: Application of ChIP-on-chip for Defining Dynamic Transcriptional Mechanisms in Bone Cells

Margaret van der Deen, Mohammad Q. Hassan, Jitesh Pratap, Nadiya M. Teplyuk, Daniel W. Young, Amjad Javed, Sayyed K. Zaidi, Jane B. Lian, Martin Montecino, Janet L. Stein, Gary S. Stein and Andre J. van Wijnen*

14. Identification of Transcription Factor Target Genes by ChIP Display (CD)

Artem Barski, Steve Pregizer and Baruch Frenkel*

15. Application of Laser Capture Microdissection (LCM) Technique for Molecular Definition of Skeletal Cell Differentiation In Vivo

Dafna Benayahu*, Rina Socher, and Irena Shur

16. Quantitative Trait Loci (QTL) Mapping

Dong-Hai Xiong, Jian-Feng Liu, Yan-Fang Guo, Yan Guo, Tie-Lin Yang, Shu-Gui Chen, Hui Jiang, Yuan Chen, Fang Yang, Robert R. Recker and Hong-Wen Deng*

Imaging Techniques

17. In Situ Nuclear Organization of Regulator Machinery

Shirwin M. Pockwinse, Sayyed K. Zaidi, Ricardo F. Medina, Rachit Bakshi, Krishna P. Kota, Syed A. Ali, Daniel W. Young, Jeffery A. Nickerson, Amjad Javed, Martin Montecino, Andre J. van Wijnen, Jane B. Lian, Janet L. Stein and Gary S. Stein*

18. Bioluminescent Imaging in Bone

Yoram Zilberman, Yossi Gafni, Gadi Pelled, Zulma Gazit, and Dan Gazit*

19. Micro Computed Tomography – A Method for the Nondestructive Evaluation of the Three-Dimensional Structure of Biological Specimens

Martin Stauber and Ralph Müller*

20. Fourier Transform Infrared (FT-IR) Microspectroscopy and Microscopic Imaging

Samuel Gourion-Arsiquaud, Paul A. West and Adele L. Boskey*

Mechanical Testing

21. Assessment of the In Vivo Adaptive Response to Mechanical Loading

Leanne K. Saxon* and Lance E. Lanyon

22. Determination of Bone’s Mechanical Matrix Properties by Nanoindentation

Engin Ozcivici, Suzanne Ferreri, Yi-Xian Qin and Stefan Judex*

23. Fluid Flow Assays

Ryan C. Riddle, Amanda F. Taylor and Henry J. Donahue*

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