19. EFFECTS OF TEMPERATURE AND SERUM ON STORAGE OF FRESH GOAT OSTEOCHONDRAL GRAFTS

Department: Bioengineering
Faculty Advisor(s): Robert Sah

Primary Student
Name: Andrea L Pallante
Email: apallant@ucsd.edu
Phone: 858-534-5682
Grad Year: 2011

Abstract
The objectives of this study were to determine effects of temperature (4°C, 37°C) and serum (0%, 2%, 10%) in medium on the efficacy of allograft storage for up to 28 days on chondrocyte viability, especially in the superficial zone, and cartilage matrix content. Osteochondral cores (n=65) were harvested from humeral heads of adult goats. Samples were analyzed (i) fresh, or after storage in medium for (ii) 14d at 4°C including 10% FBS, (iii) 28d at 4°C including 10% FBS, (iv) 28d at 37°C including 0% FBS, (v) 28d at 37°C including 2% FBS, or (vi) 28d at 37°C including 10% FBS. Portions of cartilage were analyzed by microscopy after LIVE/DEAD® staining to identify live and dead cells, to measure cartilage thickness, and to determine percent viability and density, both en face (to visualize the articular surface) and in the vertical profile (overall and in superficial, middle, and deep zones). The remaining cartilage was analyzed for sulfated-glycosaminoglycan and collagen. Viability varied with storage condition (p<0.001, en face and vertically). Vertically, viability also varied with zone (p<0.001), with a significant interaction between zone and storage condition (p<0.001). At 37°C, viability among 0-10% FBS samples was not significantly different in any zone (p=1.0). After 28 days of storage, viability of 0-10% FBS samples stored at 37°C was higher by ~35% at the cartilage surface en face (p<0.001), by ~45% in the superficial zone (p<0.01), and by ~35% in the middle zone (p<0.05) compared to samples stored at 4°C; in the deep zone, viability was similar following 37°C and 4°C storage (p=1.0). Cartilage thickness, glycosaminoglycan content, and collagen content were maintained for 37°C and 4°C samples compared to fresh controls. These results indicate 37°C storage of osteochondral grafts supports long-term chondrocyte viability, especially at the surface and in the superficial zone of cartilage. Such maintenance may extend acceptable storage duration and improve treatment outcomes.

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