The role of cryopreservation in the biomechanical properties of the intervertebral disc.

Implantation of intervertebral disc (IVD) allograft or tissue engineered disc constructs in the spine has emerged as an alternative to artificial disc replacement for the treatment of severe degenerative disc disease (DDD). Establishment of a bank of cryopreserved IVD allografts enables size matching and facilitates logistics for effective clinical management. However, the biomechanical properties of cryopreserved IVDs have not been previously reported. This study aimed to assess if cryopreservation with different concentrations of cryopreservant agents (CPA) would affect the dynamic viscoelastic properties of the IVD. Whole porcine lumbar IVDs (n = 40) were harvested and processed using various concentrations of CPA, 0 % CPA, 10 % CPA and 20 % CPA. The discs were cryopreserved using a stepwise freezing protocol and stored in liquid nitrogen. After four weeks of storage, the cryopreserved IVDs were quickly thawed at 37 °C for dynamic viscoelastic testing. The apparent modulus, elastic modulus (G'), viscous modulus (G") and loss modulus (G"/G') were calculated and compared to a fresh control group. Cryopreserved IVD without cryopreservants was significantly stiffer than the control. In the dynamic viscoelastic testing, cryopreservation with the use of CPA was able to preserve both G' and G" of an IVD. No significant differences were found between fresh IVD and IVD cryopreserved with 10 % CPA or 20 % CPA. This study demonstrated that CPAs at an optimal concentration could preserve the mechanical properties of the IVD allograft and can provide further credence for the application of long-term storage of IVD allografts for disc transplantation or tissue engineered construct applications.