Skeletal Research: An Experimental Approach

1st Edition - January 28, 1979
Editor: David J. Simmons
Language: English
eBook ISBN:
9 7 8 - 0 - 3 2 3 - 1 4 1 5 9 - 8

Skeletal Research, An Experimental Approach discusses experimental studies on bone cell and tissue biochemistry, biomechanics, isolation, and methods of analysis. Divided into four… Read more

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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.

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