Cryopreservation-Induced Stress on Long-Term Preserved Articular Cartilage

Abstract

Tissue engineered cartilage constructs have potential clinical applications in human healthcare. Their effective utilization is, however, hampered by the lack of an optimal cryopreservation procedure that ensures their availability as and when required at the patient's bedside. Cryopreservation-induced stress represents a major barrier towards the cryopreservation of such tissue constructs, and they remain a scientific challenge despite the significant progress in the long-term storage and banking of isolated chondrocytes and thin cartilage tissue slices. These stresses are caused by intra-and extracellular ice crystallization, cryoprotectant (CPA) toxicity, suboptimal rates of cooling and warming, osmotic imbalance, and altered intracellular pH that might cause cellular death and/or a disruption of extracellular matrix (ECM). This paper reviews the cryopreservation-induced stresses on tissue engineered cartilages and discusses how they influence the integrity of the tissue during its long-term preservation. We have also reported how various antioxidants, vitamins, and plant extracts have been used to inhibit and overcome the stress during cryopreservation and provide promising results. Based on the reviewed information, the paper has also proposed some novel ways which might help in increasing the postthawing cell viability of cryopreserved cartilage.

Antioxidative effect of BHA in soya bean lecithin‐based extender containing Glycerol or DMSO on freezing capacity of goat semen

“…The cryopreservation process induces various kinds of stresses to the cells mainly caused by osmotic imbalance, intra‐ and extracellular ice crystallization, lipid peroxidation and CPAs toxicity (Kaur, Pramanik, & Sarangi, ). Generally, the oxidative stress leads to the loss of total motility, progressive motility, swelling and disruption or increased permeability of the plasma membrane of spermatozoa (Tuncer et al., ).…”

The Bioactivity of Cartilage Extracellular Matrix in Articular Cartilage Regeneration

“…Most studies reporting the use of physical devitalization to process DVC have not confirmed loss of DNA or retention of GAG or collagen content. Studies exploring the freezing and thawing of tissues for cryopreservation have shown that freezing cartilage induces apoptosis and necrosis in chondrocytes . Freezing the tissue causes extracellular ice crystals to form that may cause an osmotic imbalance within the tissue creating acidic conditions, which activate degradation enzymes that degrade collagen fibers.…”

“…Protease inhibitors may also be used to increase the preservation of the ECM during freeze–thaw cycles . For a complete review of cryopreservation‐induced stresses in articular cartilage, the reader is directed to Kaur et al…”

“…Altering decellularization conditions to remove small antigenic proteins may also remove growth factors. Devitalization techniques such as freeze–thaw cycles may also increase the degradation or denaturation of latent growth factors within the ECM by activating degradation enzymes or pH changes due to physiochemical chemical stress within the tissue . Additional studies are needed to determine the mechanism of chondrogenesis seen in cartilage matrix‐based scaffolds.…”

Conditioned medium: a new alternative for cryopreservation of equine umbilical cord mesenchymal stem cells

“…Intracellular cryoprotectants, such as DMSO, are indicated during freezing at controlled rates, since they preserve the cells and their intracellular components by replacing intracellular water, reducing the formation of intracellular ice (Kaur et al, ). In our study, the 10% (v/v) DMSO concentration of associated with 20% FBS demonstrated to be effective for the cryopreservation of UCIM‐MSCs.…”