Abstract
There are several methods for determining the proteoglycan content of cartilage in biomechanics experiments. Many of these include assay-based methods and the histochemistry or spectrophotometry protocol where quantification is biochemically determined. More recently a method based on extracting data to quantify proteoglycan content has emerged using the image processing algorithms, e.g., in ImageJ, to process histological micrographs, with advantages including time saving and low cost. However, it is unknown whether or not this image analysis method produces results that are comparable to those obtained from the biochemical methodology. This paper compares the results of a well-established chemical method to those obtained using image analysis to determine the proteoglycan content of visually normal (n=33) and their progressively degraded counterparts with the protocols. The results reveal a strong linear relationship with a regression coefficient (R2) = 0.9928, leading to the conclusion that the image analysis methodology is a viable alternative to the spectrophotometry.
Highlights
Biochemical assay analyses have been commonly applied to determine the proteoglycan quantity of articular cartilage in laboratory and clinical research [1,2]
We have validated the capacity of image analysis method to accurately determine the proteoglycan content of stained histological section of articular cartilage micrographs using a well-established biochemical method of determining the protein content in solution
Spectrophotometric method to assess the proteoglycan released into the solution was adopted because of its fast, accurate and reproducible outcome
Summary
Biochemical assay analyses have been commonly applied to determine the proteoglycan quantity of articular cartilage in laboratory and clinical research [1,2] While these methods have produced reliable results over the years, they all include the destruction of the entire sample and time-consuming laboratory work. In this paper we observe that while this image analysis methodology [7] can deliver benefits over and above the biochemical or assay-based methods, there still remains the necessity to establish its accuracy and relationship to the well-established and extensively tested protocols [4,5] To this extent we propose to determine whether or not there is a relationship between the data from image analysis and a standard protein (proteoglycan) quantification method. This method was adopted because it requires very small sample volume (1 – 2 μL), has high degree of reproducibility and accuracy, and the speed of analysis is extremely fast [10,11]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
