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High strain mechanical loading rapidly induces tendon apoptosis: an ex vivo rat tibialis anterior model
  1. A Scott1,
  2. K M Khan1,
  3. J Heer2,
  4. J L Cook3,
  5. O Lian4,
  6. V Duronio1
  1. 1Department of Family Practice & School of Human Kinetics, University of British Columbia, Vancouver, Canada
  2. 2University of Melbourne, Australia
  3. 3La Trobe University, Melbourne, Australia
  4. 4Oslo Sports Trauma Research Centre, Oslo, Norway
  1. Correspondence to:
    Assistant Professor Khan
    Department of Family Practice & School of Human Kinetics, University of British Columbia, Suite 211, 2150 Western Parkway, Vancouver V6T 1V6, Canada; kkhaninterchange.ubc.ca

Abstract

Background: The role of apoptosis, or programmed cell death, has only recently been explored in tendon.

Objective: To investigate the development of apoptosis after high strain loading of rat tendon.

Methods: The right tibialis anterior tendons of three rats were prepared for mechanical loading, and left tendons were prepared identically as non-loaded controls. Tendon was loaded with 20% strain for six hours using a 1 Hz longitudinal sine wave signal. The following were used to assess apoptosis: (a) a monoclonal mouse antibody (F7-26) to label single stranded DNA breaks; (b) a rabbit polyclonal antibody that specifically recognises the cleaved form of caspase-3.

Results: Light microscopy confirmed that the high strain protocol induced a stretch overload injury. Control tendons showed little or no staining with the F7-26 antibody, but the loaded tendons displayed numerous apoptotic cells. The percentage of apoptotic cells (20%) in the loaded tendon was significantly greater than in the control tendon (1%) (p  =  0.000). The labelled cells colocalised with abnormal nuclear morphology, including nuclear fragmentation. The staining against cleaved caspase-3 was positive in loaded tendons only, and localised both to nucleus and cytoplasm.

Conclusion: This experiment extends knowledge of human tendon apoptosis by showing that apoptosis can occur in response to short term, high strain mechanical loading. This is the first report of mechanical loading of intact tendon causing excessive apoptosis.

  • apoptosis
  • mechanical loading
  • overuse
  • rat
  • tendon

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