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INVESTIGATING THE EFFECTS OF HYPOTHERMIC STORAGE ON CHONDROCYTE SURVIVAL AND APOPTOSIS IN HUMAN ARTICULAR CARTILAGE.



Abstract

Due to recent advances in diagnostic technology and an increased awareness among clinicians, osteochondral damage is being detected more frequently. Thus, there is a need to preserve and store articular cartilage for the repair of joint surfaces. Chondrocytes, embedded within extracellular matrix must remain viable during storage for successful tissue transplantation. We have been able to store osteochondral tissue for over a month and maintain high chondrocyte viability. Apoptosis can be minimized in articular cartilage during hypothermic storage if biopreservation media (XVIVO-10) is used. Cadaveric osteochondral dowels are a potential source of tissue for banking and allogeneic transplantation.

The purpose of this study was to:

  • Establish a timeline and optimal conditions for storing human articular cartilage (AC).

  • Determine a suitable source of AC for banking and transplantation.

  • Distinguish apoptosis from necrosis in human AC following hypothermic storage.

Forty fresh human AC samples from femoral condyle notchplasties were used to determine a storage timeline. Each sample was divided into three portions:

  1. initial chondrocyte viability,

  2. stored in phosphate buffered saline,

  3. stored in Biowhitaker XVIVO-10.

All samples were randomly allocated to one of five time intervals (2–10 weeks). Following each time period final viability assays were conducted. Secondly, osteochondral dowels were drilled from eight cadaveric femoral condyles. Five dowels were obtained from each joint: one for initial viability/annexin V assays, the others were stored in PBS or XVIVO-10 for four and six weeks. Following storage, final viability, annexin V, and TUNEL assays were preformed.

Notchplasty samples stored in XVIVO-10 for four weeks had an average final viability of 68%, but an average loss in viability of only 6%. By one month the viability of samples stored in PBS had dropped to 5%. Osteochondral dowels stored in XVIVO-10 not only had greater chondrocyte viability, but less apoptosis.

Cadaveric dowels are a suitable source of osteochon-dral tissue for hypothermic storage and in turn allogeneic transplantation.

An osteochondral tissue bank would provide a reliable source of articular cartilage for repairing joint surfaces for patients who are not suitable candidates for total joint replacements.

Correspondence should be addressed to Cynthia Vezina, Communications Manager, COA, 4150-360 Ste. Catherine St. West, Westmount, QC H3Z 2Y5, Canada