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Full Description
This second edition volume expands on the previous edition with a discussion of the latest methods used to explore collagen biology from molecular synthesis and folding to biomechanical analysis and tissue engineering applications. The chapters in this book are organized into four parts: Part One introduces strategies for manipulating collagen at the molecular level, including gene targeting, mutagenesis, and functional assays. Part Two focuses on cellular and biochemical aspects of collagen biology, featuring protocols to study the effects of hypoxia, assess triple helix stability, and investigate ECM-rich environments using proteomics and mass spectrometry. Part Three covers structural and imaging techniques such as atomic force microscopy and advanced visualization of collagen suprastructures, and Part Four looks at translational and biomedical research, providing protocols for constructing 3D ECM scaffolds, engineering collagen-based biomaterials, and modeling tumor microenvironments. Written in the highly successful Methods in Molecular Biology series format, 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.
Cutting-edge and comprehensive, Collagen: Methods and Protocols, Second Edition is a valuable resource for researchers in cell biology, biochemistry, tissue engineering, and translational medicine, and will support the development of novel strategies to explore and manipulate the power of collagen.
Contents
Targeting, Mutagenesis, and Functional Testing Strategies for Large Collagens.- Next-Generation Sequencing and Variant Analysis of Fibrillar (Pro)Collagens.- Expression of Recombinant Collagen Domains and Matrikines in Eukaryotic Cells.- Recombinant Production and Characterization of Human Enzymes Responsible for Collagen Lysine Post-Translational Modifications.- Hypoxia in Cellular Behavior, Collagen Biosynthesis, and Extracellular Matrix Production.- Trypsin Digestion to Assess Triple helix Stability in Collagens as Measure of Collagen Folding.- Spatial Proteomics Analysis of Extracellular Matrix-Rich Tissues.- Labeling of Collagen in Human Musculoskeletal Connective Tissues with Stable Isotope Tracers and Preparation of Amino Acids for Mass Spectrometry.- Atomic Force Microscopy Investigations of Collagen: From Single Molecules to Fibrils.- Visualization and Organization of Collagen VI Containing Suprastructures in Articular Cartilage.- Triple Helical-Peptides: Collagen-Mimicking Tools for Tissue Engineering.- Atomic Force Microscopy to Investigate Collagen Fibrils.- Reconstructing Tumor Dynamics: CAM-ECM Scaffolds as 3D Platforms for Ex Vivo Cancer Studies and Beyond.- A Protocol for Engineering Collagen Type II Bioscaffolds to Support Stem Cell Chondrogenesis.- Construction of an In Silico Model to Study the Molecular Functions of Type I Collagen.- Targeting Collagens and LOX through Plant-Derived MicroRNAs (xenomiRs).
