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Chondrocyte viability is higher after prolonged storage at 37 degrees C than at 4 degrees C for osteochondral grafts
Andrea L Pallante1, Won C Bae, Albert C Chen
1Department of Bioengineering and Whitaker Institute of Biomedical Engineering, University of California-San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0412, USA.
Background:
Osteochondral allografts are currently stored at 4 degrees C for 2 to 6 weeks before implantation. At 4 degrees C, chondrocyte viability, especially in the superficial zone, deteriorates starting at 2 weeks. Alternative storage conditions could maintain chondrocyte viability beyond 2 weeks, and thereby facilitate increased graft availability and enhanced graft quality.
Purpose:
The objective of the study was to determine the effects of prolonged 37 degrees C storage compared with traditional 4 degrees C storage on chondrocyte viability and cartilage matrix content.
Study Design:
Controlled laboratory study.
Methods:
Osteochondral samples from humeral heads of adult goats were analyzed (i) fresh, or after storage in medium for (ii) 14 days at 4 degrees C including 10% fetal bovine serum, (iii) 28 days at 4 degrees C including 10% fetal bovine serum, (iv) 28 days at 37 degrees C without fetal bovine serum, (v) 28 days at 37 degrees C including 2% fetal bovine serum, or (vi) 28 days at 37 degrees C including 10% fetal bovine serum. Portions of samples were analyzed by microscopy after LIVE/DEAD staining to determine chondrocyte viability and density, both en face (to visualize the articular surface) and vertically (overall and in superficial, middle, and deep zones). The remaining cartilage was analyzed for sulfated glycosaminoglycan and collagen.
Results:
The 37 degrees C storage maintained high chondrocyte viability compared with 4 degrees C storage. Viability of samples after 28 days at 37 degrees C was approximately 80% at the cartilage surface en face, approximately 65% in the superficial zone, and approximately 70% in the middle zone, which was much higher than approximately 45%, approximately 20%, and approximately 35%, respectively, in 4 degrees C samples after 28 days, and slightly decreased from approximately 100%, approximately 85%, and approximately 95%, respectively, in fresh controls. Cartilage thickness, glycosaminoglycan content, and collagen content were maintained for 37 degrees C and 4 degrees C samples compared with fresh controls.
Conclusion:
The 37 degrees C storage of osteochondral grafts supports long-term chondrocyte viability, especially at the vulnerable surface and superficial zone of cartilage.
Clinical Relevance:
Storage of allografts at a physiologic temperature of 37 degrees C may prolong storage duration, improve graft availability, and improve treatment outcomes.
