Journal
CARTILAGE
Volume 13, Issue 2_SUPPL, Pages 1742S-1754SPublisher
SAGE PUBLICATIONS INC
DOI: 10.1177/19476035211060508
Keywords
collagen; collagen subtype; mass spectrometry; collagen crosslinks; cartilage; fibrocartilage
Categories
Funding
- NIH [R01DE015038, R01AR071457, R01AR067821]
Ask authors/readers for more resources
The study developed novel assays for quantifying collagen subtypes and crosslinks using liquid chromatography-mass spectrometry, which are sensitive, cost-effective, and less time-consuming compared to traditional methods. Results showed collagen type I predominates in fibrocartilages, while collagen type II predominates in hyaline cartilages. Bottom-up proteomics revealed a spectrum of collagen ratios and quantified 42 proteins, including 24 collagen alpha-chains and 12 minor collagen types.
Introduction This study develops assays to quantify collagen subtypes and crosslinks with liquid chromatography-mass spectrometry (LC-MS) and characterizes the cartilages in the Yucatan minipig. Methods For collagen subtyping, liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis was performed on tissues digested in trypsin. For collagen crosslinks, LC-MS analysis was performed on hydrolysates. Samples were also examined histologically and with bottom-up proteomics. Ten cartilages (femoral condyle, femoral head, facet joint, floating rib, true rib, auricular cartilage, annulus fibrosus, 2 meniscus locations, and temporomandibular joint disc) were analyzed. Results The collagen subtyping assay quantified collagen types I and II. The collagen crosslinks assay quantified mature and immature crosslinks. Collagen subtyping revealed that collagen type I predominates in fibrocartilages and collagen type II in hyaline cartilages, as expected. Elastic cartilage and fibrocartilages had more mature collagen crosslink profiles than hyaline cartilages. Bottom-up proteomics revealed a spectrum of ratios between collagen types I and II, and quantified 42 proteins, including 24 collagen alpha-chains and 12 minor collagen types. Discussion The novel assays developed in this work are sensitive, inexpensive, and use a low operator time relative to other collagen analysis methods. Unlike the current collagen assays, these assays quantify collagen subtypes and crosslinks without an antibody-based approach or lengthy chromatography. They apply to any collagenous tissue, with broad applications in tissue characterization and tissue engineering. For example, a novel finding of this work was the presence of a large quantity of collagen type III in the white-white knee meniscus and a spectrum of hyaline and fibrous cartilages.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available