Effects of Refrigeration and Freezing on the Electromechanical and Biomechanical Properties of Articular Cartilage

In vitro electromechanical and biomechanical testing of articular cartilage provide critical information about the structure and function of this tissue. Difficulties obtaining fresh tissue and lengthy experimental testing procedures often necessitate a storage protocol, which may adversely affect the functional properties of cartilage. The effects of storage at either $4°C$ for periods of 6 days and 12 days, or during a single freeze-thaw cycle at $−20°C$ were examined in young bovine cartilage. Non-destructive electromechanical measurements and unconfined compression testing on 3 mm diameter disks were used to assess cartilage properties, including the streaming potential integral (SPI), fibril modulus (Ef), matrix modulus (Em), and permeability $(k)$⁠. Cartilage disks were also examined histologically. Compared with controls, significant decreases in SPI (to $32.3±5.5%$ of control values, $p<0.001$⁠), Ef (to $3.1±41.3%$ of control values, $p=0.046$⁠), Em (to $6.4±8.5%$ of control values, $p<0.0001$⁠), and an increase in $k$ (to $2676.7±2562.0%$ of control values, $p=0.004$⁠) were observed at day 12 of refrigeration at $4°C$⁠, but no significant changes were detected at day 6. A trend toward detecting a decrease in SPI (to $94.2±6.2%$ of control values, $p=0.083$⁠) was identified following a single freeze-thaw cycle, but no detectable changes were observed for any biomechanical parameters. All numbers are $mean±95%$ confidence interval. These results indicate that fresh cartilage can be stored in a humid chamber at $4°C$ for a maximum of 6 days with no detrimental effects to cartilage electromechanical and biomechanical properties, while one freeze-thaw cycle produces minimal deterioration of biomechanical and electromechanical properties. A comparison to literature suggested that particular attention should be paid to the manner in which specimens are thawed after freezing, specifically by minimizing thawing time at higher temperatures.