TY - JOUR
T1 - An experimental toolbox for characterization of mammalian collagen type i in biological specimens
AU - Capella-Monsonís, Héctor
AU - Coentro, João Q.
AU - Graceffa, Valeria
AU - Wu, Zhuning
AU - Zeugolis, Dimitrios I.
N1 - Publisher Copyright:
© 2018 Macmillan Publishers Limited.
PY - 2018
Y1 - 2018
N2 - Collagen type I is the most abundant extracellular matrix protein, and collagen type I supramolecular assemblies (e.g., tissue grafts, biomaterials and cell-assembled systems) are used extensively in tissue engineering and regenerative medicine. Many studies, for convenience or economic reasons, do not accurately determine collagen type I purity, concentration, solubility and extent of cross-linking in biological specimens, frequently resulting in erroneous conclusions. In this protocol, we describe solubility; normal, reduced and delayed (interrupted) SDS-PAGE; hydroxyproline; Sircol collagen and Pierce BCAprotein; denaturation temperature; ninhydrin/trinitrobenzene sulfonic acid; and collagenase assays and assess them in a diverse range of biological samples (e.g., tissue samples; purified solutions or lyophilized materials; 3D scaffolds, such as sponges and hydrogels; and cell media and layers). Collectively, the described protocols provide a comprehensive, yet fast and readily implemented, toolbox for collagen type I characterization in any biological specimen.
AB - Collagen type I is the most abundant extracellular matrix protein, and collagen type I supramolecular assemblies (e.g., tissue grafts, biomaterials and cell-assembled systems) are used extensively in tissue engineering and regenerative medicine. Many studies, for convenience or economic reasons, do not accurately determine collagen type I purity, concentration, solubility and extent of cross-linking in biological specimens, frequently resulting in erroneous conclusions. In this protocol, we describe solubility; normal, reduced and delayed (interrupted) SDS-PAGE; hydroxyproline; Sircol collagen and Pierce BCAprotein; denaturation temperature; ninhydrin/trinitrobenzene sulfonic acid; and collagenase assays and assess them in a diverse range of biological samples (e.g., tissue samples; purified solutions or lyophilized materials; 3D scaffolds, such as sponges and hydrogels; and cell media and layers). Collectively, the described protocols provide a comprehensive, yet fast and readily implemented, toolbox for collagen type I characterization in any biological specimen.
UR - http://www.scopus.com/inward/record.url?scp=85048380480&partnerID=8YFLogxK
U2 - 10.1038/nprot.2017.117
DO - 10.1038/nprot.2017.117
M3 - Article
C2 - 29446773
AN - SCOPUS:85048380480
SN - 1754-2189
VL - 13
SP - 507
EP - 529
JO - Nature Protocols
JF - Nature Protocols
IS - 3
ER -