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Catabolic Responses of Chondrocyte-Seeded Peptide Hydrogel to Dynamic Compression

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Abstract

This study investigated the role of matrix metalloproteases and aggrecanases during dynamic compression-induced aggrecan catabolism in chondrocyte-seeded self-assembling peptide hydrogel. One- to two-week-old bovine chondrocytes were encapsulated into peptide hydrogel and cultured for 14 days prior to the application of an alternate day loading protocol. Dynamic compression-induced aggrecan catabolism was explored by evaluating GAG loss to the culture medium, zymography for matrix metalloproteases (MMPs), gene expression of MMPs and ADAMTS proteases, and Western blot analysis for aggrecan fragments. The application of loading over 4 days increased GAG loss to the medium three- to four-fold relative to free-swelling controls. Zymogram analysis detected increased concentrations of latent MMP-9 and MMP-3 in the culture medium relative to free-swelling culture. Real-time PCR showed expression levels of MMPs and ADAMTS proteases in loaded samples that ranged from 2.5- to 95-fold higher than free-swelling culture. Aggrecan fragment analysis did not detect small (50–80 kDa) molecular weight fragments in free-swelling culture; however, dynamic compression samples contained 60–80 kDa fragments that were detected by both anti-G1 and NITEGE probes, demonstrating ADAMTS but not MMP degradation. These data suggest that partially mature cartilage tissue engineering constructs may be susceptible to catabolic degradation.

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Acknowledgment

NIH-BRP grant EB003805 (MIT). NIH AR33236 (MIT).

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Authors and Affiliations

  1. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

    John D. Kisiday, Jennifer H. Lee & Alan J. Grodzinsky

  2. Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

    Alan J. Grodzinsky

  3. Department of Electrical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

    Alan J. Grodzinsky

  4. Center for Biomedical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

    Patrick N. Siparsky & Alan J. Grodzinsky

  5. Department of Clinical Science, Orthopaedic Research Center, Colorado State University, Fort Collins, CO, USA

    John D. Kisiday & David D. Frisbie

  6. Wyeth Research, Cambridge, MA, USA

    Carl R. Flannery

  7. Department of Biochemistry, Rush Medical School, Chicago, IL, USA

    John D. Sandy

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  1. John D. Kisiday
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  2. Jennifer H. Lee
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Correspondence to John D. Kisiday.

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Kisiday, J.D., Lee, J.H., Siparsky, P.N. et al. Catabolic Responses of Chondrocyte-Seeded Peptide Hydrogel to Dynamic Compression. Ann Biomed Eng 37, 1368–1375 (2009). https://doi.org/10.1007/s10439-009-9699-9

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