A Chick Embryo in-Vitro Model of Knee Morphogenesis

Document Type : RESEARCH PAPER

Authors

1 Orthopedic Trauma Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, USA

2 National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, USA

Abstract

 
 Background: In this feasibility study, a mechanically loaded in-vitro tissue culture model of joint morphogenesis using the isolated lower extremity of the 8 day old chick embryo was developed to assess the effects of mechanical loading on joint morphogenesis.
Methods: The developed in-vitro system allows controlled flexion and extension of the chick embryonic knee with a range of motion of 20 degrees from a resting position of 90-100 degrees of flexion. Joint morphogenesis at 2, 3, 4 and 7 days of culture was assessed by histology and micro MRI in 4 specimen types: undisturbed in-ovo control embryos, in-ovo paralyzed embryos, in-vitro unloaded limb cultures, and in-vitro loaded limb cultures. Relative glycosaminoglycan (GAG) concentration across the joint was assessed with an MRI technique referred to as dGEMRIC (delayed gadolinium enhanced MRI of cartilage) where T1 is proportional to glycosaminoglycan concentration.
Results: Average T1 over the entire tissue image for the normal control (IC) knee was 480 msec; for the 4 day loaded specimen average T1 was 354 msec; and for the 7 day loaded specimens T1 was 393 msec. The 4 day unloaded specimen had an average T1 of 279 msec while the 7 day unloaded specimen had an average T1 of 224 msec. The higher T1 values in loaded than unloaded specimens suggest that more glycosaminoglycan is produced in the loaded culture than in the unloaded preparation.
Conclusion: Isolated limb tissue cultures under flexion-extension load can be viable and exhibit more progression of joint differentiation and glycosaminoglycan production than similarly cultured but unloaded specimens. However, when compared with controls consisting of intact undisturbed embryos in-ovo , the isolated loaded limbs in culture do not demonstrate equivalent amounts of absolute growth or joint differentiation.

Keywords

References

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The Archives of Bone and Joint Surgery
Volume 4, Issue 2 - Serial Number 2
April 2016
Pages 109-115

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History

  • Receive Date: 20 June 2015
  • Revise Date: 17 September 2015
  • Accept Date: 21 October 2015

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