Effects of deep-freezing and storage time on human femoral cartilage

Abstract

Surgical techniques including new, possible resources to repair injured joints and damaged cartilage are still evolving. The exact effects of cryopreservation on the collected cartilage samples require accurate determination prior to utilization. The aim of our study was to analyze the impact of cryopreservation at −80 °C on the structural properties of the human cartilage. The effects of storage time were also evaluated in conjunction with optimal utilization. The human cartilage samples were derived during operation and considered to be waste material. Samples were fresh frozen and stored at −80 °C. Cryopreservation times were: 0, 1, 3, 6, and 12 weeks. To assess the biological and structural properties of the frozen human cartilage, we performed calorimetric examinations using differential scanning calorimetry (DSC). During the first 3 weeks, the calorimetric enthalpy (ΔH cal) showed an increasing tendency compared to controls, parallel with the denaturation temperature (T m): ΔH cal (J g−1) = 1.60 versus 2.49, T m1 (°C) = 61.73 versus 63.64. After the sixth week, both the enthalpy and the transition temperature decreased, compared to the control samples. The decrease in both the calorimetric enthalpy and T m could be explained by the decrease in bound water and the time-related degeneration in the structure of the cartilage. Here we found that the duration of cryopreservation interferes with the morphology of human cartilage samples only after 6 weeks of storage time. The thermal analyzes of human cartilage by DSC could be a useful method to follow the morphological changes in the clinical practice.