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Archives of Orthopaedic and Trauma Surgery

, Volume 130, Issue 4, pp 557–564 | Cite as

Changes in content and synthesis of collagen types and proteoglycans in osteoarthritis of the knee joint and comparison of quantitative analysis with Photoshop-based image analysis

  • Andreas LahmEmail author
  • Eike Mrosek
  • Heiko Spank
  • Christoph Erggelet
  • Richard Kasch
  • Jan Esser
  • Harry Merk
Basic Science

Abstract

Introduction

The different cartilage layers vary in synthesis of proteoglycan and of the distinct types of collagen with the predominant collagen Type II with its associated collagens, e.g. types IX and XI, produced by normal chondrocytes. It was demonstrated that proteoglycan decreases in degenerative tissue and a switch from collagen type II to type I occurs. The aim of this study was to evaluate the correlation of real-time (RT)-PCR and Photoshop-based image analysis in detecting such lesions and find new aspects about their distribution.

Patients

We performed immunohistochemistry and histology with cartilage tissue samples from 20 patients suffering from osteoarthritis compared with 20 healthy biopsies. Furthermore, we quantified our results on the gene expression of collagen type I and II and aggrecan with the help of real-time (RT)-PCR. Proteoglycan content was measured colorimetrically. Using Adobe Photoshop the digitized images of histology and immunohistochemistry stains of collagen type I and II were stored on an external data storage device. The area occupied by any specific colour range can be specified and compared in a relative manner directly from the histogram using the “magic wand tool” in the select similar menu. In the image grow menu gray levels or luminosity (colour) of all pixels within the selected area, including mean, median and standard deviation, etc. are depicted. Statistical Analysis was performed using the t test.

Method

With the help of immunohistochemistry, RT-PCR and quantitative RT- PCR we found that not only collagen type II, but also collagen type I is synthesized by the cells of the diseased cartilage tissue, shown by increasing amounts of collagen type I mRNA especially in the later stages of osteoarthritis.

Results

A decrease of collagen type II is visible especially in the upper fibrillated area of the advanced osteoarthritic samples, which leads to an overall decrease. Analysis of proteoglycan showed a loss of the overall content and a quite uniform staining in the different zones compared to the healthy cartilage with a classical zonal formation. Correlation analysis of the proteoglycan Photoshop measurements with the RT-PCR using Spearman correlation analysis revealed strong correlation for Safranin O and collagen type I, medium for collagen type II and glycoprotein but weak correlation between PCR aggrecan results.

Conclusion

Photoshop-based image analysis might become a valuable supplement for well known histopathological grading systems of lesioned articular cartilage.

Keywords

Quantitative real-time PCR Collagen Proteoglycan Photoshop-based image analysis Osteoarthritis 

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Andreas Lahm
    • 1
    • 2
    Email author
  • Eike Mrosek
    • 3
  • Heiko Spank
    • 1
  • Christoph Erggelet
    • 4
  • Richard Kasch
    • 1
  • Jan Esser
    • 1
  • Harry Merk
    • 1
  1. 1.Section of Orthopaedic Research and Cell Biology, Department of Orthopaedics and Orthopaedic SurgeryUniversity Hospital of the Ernst-Moritz-Arndt University GreifswaldGreifswaldGermany
  2. 2.St. Anna Malteser Hospitals DuisburgDuisburgGermany
  3. 3.Cartilage and Connective Tissue Research Laboratory, Department of Orthopedic SurgeryMayo Clinic College of MedicineRochesterUSA
  4. 4.Arthrose Clinic ZürichZürichGermany

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