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Differential expressions of the lysyl oxidase family and matrix metalloproteinases-1, 2, 3 in posterior cruciate ligament fibroblasts after being co-cultured with synovial cells

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Abstract

Purpose

The adult human posterior cruciate ligament (PCL) has poor functional healing response. The synovial tissue, which surrounds the PCL ligament, might be the major regulator of the microenvironment in the joint cavity after PCL injury, thus affecting the healing process. Here we establish a novel co-culture system for PCL fibroblasts and synovial cells (SC) in vitro to explore the direct influence of paracrine on PCL cells by characterizing the different expressions of the lysyl oxidase family (LOXs) and matrix metalloproteinases (MMP-1, 2, 3), which respectively facilitate extracellular matrix (ECM) repair and degradation.

Methods

Total RNA was harvested, reverse transcribed and assessed by semi-quantitative PCR and real-time PCR for the expression of LOXs and MMP-1, 2, 3 messenger RNAs. MMP-2 activity was assayed from the collected culture media samples by using zymography.

Results

We found co-culture could promote gene expressions of the LOXs and MMP-1, 2, 3 in normal PCL fibroblasts. But in injured PCL, we found that matrix crosstalk induced an increase of the MMP-1, 2, 3 expressions and a down-regulation of the LOXs.

Conclusion

Based on these results, the crosstalk between PCL and SC strongly modified homeostatic balance of ECM and appeared to have a significant impact on PCL wound healing; decreased expression of cross-linking enzymes (LOXs) and increased expression of ECM-degrading proteinases (MMP-1, 2, 3) might be of great contribution to poor healing ability of PCL ligament.

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Acknowledgments

This study was supported by the Innovation and Attracting Talents Program for College and University (“111” Project) (B06023), NSF Projects (10672195, 30870607), CSTC2008BB5192, Sharing Fund of Chongqing University’s large-scale equipment (2009063038) and by NIH AR45635 (USA).

Declaration of interests

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. “111” Project Laboratory of Biomechanics and Tissue Repair, Bioengineering College, Chongqing University, Chongqing, 400044, China

    Chunli Wang, Jing Xie, Jiahuan Jiang, Chunming Xu, Yanjun Zhang, Li Yang & K. L. Paul Sung

  2. Department of Orthopaedics, Chongqing University of Medical Sciences, Chongqing, 400000, China

    Wei Huang, Rongfu Chen & Chunfeng Fu

  3. Departments of Bioengineering and Orthopaedics, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA, 92093-0412, USA

    Peter C. Y. Chen & K. L. Paul Sung

Authors
  1. Chunli Wang
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  2. Jing Xie
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  3. Jiahuan Jiang
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  4. Wei Huang
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  7. Yanjun Zhang
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  8. Chunfeng Fu
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  9. Li Yang
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  11. K. L. Paul Sung
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Correspondence to K. L. Paul Sung.

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Wang, C., Xie, J., Jiang, J. et al. Differential expressions of the lysyl oxidase family and matrix metalloproteinases-1, 2, 3 in posterior cruciate ligament fibroblasts after being co-cultured with synovial cells. International Orthopaedics (SICOT) 39, 183–191 (2015). https://doi.org/10.1007/s00264-014-2573-x

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  • DOI: https://doi.org/10.1007/s00264-014-2573-x

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