Skeletal Research: An Experimental Approach

Skeletal Research, An Experimental Approach discusses experimental studies on bone cell and tissue biochemistry, biomechanics, isolation, and methods of analysis. Divided into four parts encompassing 21 chapters, this book describes the advantages and limitations of approaches and options available for bone laboratories.
The opening part of this book describes the isolation, characterization, and methods of analysis of bone and cartilage cells and their organ cultures. This description is followed by discussions on the intermediary metabolism of growth and articular cartilage; the isolation of plasma membranes from calcified connective tissues; and the aspects of lipidology as it applies to calcified tissue. The third part of this book focuses on the chemical, histochemical, and pathophysiological studies of bone as tissue. This text includes significant chapters on bone's biomechanics and bioelectricity; bone mineral composition; collagen biochemistry; calcium metabolism; and blood flow. Studies on bone and cartilage as circadian systems in rats and mice, emphasizing the importance of this aspect of experimental design, are covered in the concluding part of this book.
This text is an ideal resource to experienced researchers and young investigators who wish to expand their technical knowledge on bone research.

Contenu

List of Contributors
Preface
Part I Bone Cells, Cartilage Cells, and Organ Culture
1 / Isolated Bone Cells
I. Introduction
II. Important Considerations in the Study of Isolated Bone Cells
III. Method of Choice
IV. Characterization of Separated Bone Cells
References
2 / Organ Culture of Bone
I. Historical Overview
II. Scope of This Chapter
III. Aspects of Methodology
IV. Quantitative Methods for the Study of Bone Resorption in Organ Culture
V. Quantitative Methods for the Study of Bone Formation in Organ Culture
VI. Future Uses and Problems with Organ Culture Techniques
References
3 / Cartilage Cells and Organ Culture
I. Introduction
II. Organ and Expiant Cultures
III. Chondrocyte Cultures
IV. Species Considerations
V. Metabolism
VI. Regulation of Metabolism
References
Part II Cell and Tissue Biochemistry
4 / Studies on the Intermediary Metabolism of Growth Cartilage
I. Introduction
II. Important Considerations in the Use of Tissue Slices
III. Isolation of Mitochondria from Chick Tibial Epiphyseal Cartilage
References
5 / Partial Purification of Plasma Membranes from Calcified Connective Tissues
I. Introduction
II. Tissue Disruption
III. Particle Separation
IV. Fractionation Markers
V. Epiphyseal Cartilage Plasma Membranes
VI. Rat Calvaria and Osteosarcoma Plasma Membranes
References
6 / Isolation of Cartilage Matrix Vesicles
I. The Discovery and Function of Matrix Vesicles
II. Methods for Isolation of Cells and Subcellular Particles from Cartilage
III. Method for Isolation of Matrix Vesicles from Epiphyseal Cartilage
IV. Comments on the Procedure
V. Comparison with Other Modified Methods and Conclusions
References
7 / Lipid Techniques for Calcified Tissues
I. Introduction
II. Preparation of Tissue Samples
III. Extraction of Tissue Lipids
IV. Processing of Lipid Extracts
V. Separation of Lipid Classes
VI. Lipid Metabolic Studies
References
Part III Bone as Tissue
8 / Special Histochemical Methods
I. Methods for the Study of Enzymes
II. Immunohistochemical Methods
III. Methods for the Study of Matrix
References
9 / Bone Mineral Composition and Structure
I. Chemistry
II . X-Ray Diffraction and Related Techniques
III. Surface Chemistry
IV. Summary
References
10 / Noncollagenous Proteins
I. Introduction
II. Isolation of Noncollagenous Proteins
III. Analysis of Individual Noncollagenous Proteins
IV. Proteins Associated with Collagen after EDTA Demineralization
V. Types of Noncollagenous Proteins Isolated from Bone and Other Mineralized Tissues
VI. Conclusion: Current Investigations and Possible Trends
References
11 / Collagen Biochemistry
I. Introduction
II. Hydroxyproline and Proline
III. Lysine Hydroxylation and Collagen Carbohydrated Hydroxylysines
IV. Carboxymethyl Cellulose Chromatography of Denatured Collagen Chains
V. Polyacrylamide Electrophoresis of Collagen Components
VI. Cross-Link Analysis of Collagen from Collagenous Tissues (Reduction with [3H]NaBH4)
References
12 / Clinical Investigations of Calcium Metabolism
I. Introduction
II. The Input - Output Function
III. Systems Analysis of Mineral Metabolism
IV. Measuring the Skeletal Calcium Deposition Rate
V. Practical Considerations
VI. Applications
References
13 / Biomechanics of Bone
I. Introduction
II. Mechanical Testing Procedures
III. Whole-Bone Studies
IV. Summary
References
14 / Biophysical and Electromechanical Properties of Articular Cartilage
I. Introduction
II. Biomechanical Properties of Articular Cartilage
III. Summary
References
15 / Bioelectricity
I. Tissue Preparation
II. Electrode Systems
III. Measurement Systems
IV. Measurement Techniques
V. Analysis of Data
References
Critical Guide to References
16 / Bone Vascular Supply
I. Introduction
II. Infusion of Blood Vessels
III. Preparation of Tissue Specimens for Microangiography and Histology
IV. Critique and Summary
References
17 / Blood Flow and Ion Exchange in Bone
I. Introduction
II. Anatomic Basis for Blood - Tissue Exchange in Bone
III. Methods of Studying Bone Blood Flow
IV. Diffusional Processes in Bone
V. Summary
References
18 / Electron Microscope Techniques
I. Introduction
II. Preparation of Tissues for TEM
III. Scanning Electron Microscopy
IV. Electron Microscopy and X-Ray Microanalysis
V. Evaluation of Micrographs and Future Utilization
Selected Readings
19 / Bone Tracers: Radionuclide Imaging and Related Techniques
I. Introduction
II. Radionuclide Imaging
III. Mechanisms of Radiopharmaceutical Localization in Bone
IV. Bone-Seeking Radiopharmaceuticals
V. Clinical and Research Applications of Radionuclide Bone Imaging
References
20 / Bone Tracers: Cell and Tissue Level Techniques
I. Introduction
II. Radioactive Tracers
III. Non Radioactive Tracers
IV. Summary
References
Part IV Experimental Design
21 / Experimental Design and the Implication of Circadian Skeletal Rhythmicity
I. Introduction
II. Endochondral Ossification
III. Cortical Bone (Diaphysis)
IV. Articular Cartilage
V. Synovium
VI. Teeth
VII. Environmental Synchronizers
VIII. Timing of Laboratory Studies
IX. Chronopharmacology
X. An Overview
References
Index