Histochemical quantification of collagen content in articular cartilage

Autoři: Lassi Rieppo aff001;  Lauriane Janssen aff001;  Krista Rahunen aff001;  Petri Lehenkari aff003;  Mikko A. J. Finnilä aff001;  Simo Saarakkala aff001
Působiště autorů: Research Unit of Medical Imaging, Physics and Technology, Faculty of Medicine, University of Oulu, Oulu, Finland aff001;  Microelectronics Research Unit, Faculty of Information Technology and Electrical Engineering, University of Oulu, Oulu, Finland aff002;  Department of Surgery and Intensive Care, Oulu University Hospital, Oulu, Finland aff003;  Cancer and Translational Medicine Research Unit, Faculty of Medicine, University of Oulu, Oulu, Finland aff004;  Infotech Oulu, University of Oulu, Oulu, Finland aff005;  Department of Diagnostic Radiology, Oulu University Hospital, Oulu, Finland aff006
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0224839



Articular cartilage (AC) is mainly composed of water, type II collagen, proteoglycans (PGs) and chondrocytes. The amount of PGs in AC is routinely quantified with digital densitometry (DD) from Safranin O-stained sections, but it is unclear whether similar method could be used for collagens.


The aim of this study was to clarify whether collagens can be quantified from histological AC sections using DD.

Material and methods

Sixteen human AC samples were stained with Masson’s trichrome or Picrosirius red. Optical densities of histological stains were compared to two commonly used collagen parameters (amide I and collagen CH2 side chain peak at 1338cm-1) measured using Fourier Transform Infrared (FTIR) spectroscopic imaging.


Optical density of Modified Masson’s trichrome staining, which included enzymatic removal of PGs before staining, correlated significantly with FTIR-derived collagen parameters at almost all depths of cartilage. The other studied staining protocols displayed significant correlations with the reference parameters at only few depth layers.


Based on our findings, modified Masson’s trichrome staining protocol is suitable for quantification of AC collagen content. Enzymatic removal of PGs prior to staining is critical as us allows better staining of the collagen. Further optimization of staining protocols may improve the results in the future studies.

Klíčová slova:

Amides – Cartilage – Collagens – Fourier transform infrared spectroscopy – Histology – Staining – Safranin staining – Densitometry


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