Abstract
Preserving viable articular cartilage is a promising approach to address the shortage of graft tissue and enable the clinical repair of articular cartilage defects in articulating joints, such as the knee, ankle, and hip. In this study, we developed two 2-step, dual-temperature, multicryoprotectant loading protocols to cryopreserve particulated articular cartilage (cubes ~1 mm3 in size) using a mathematical approach, and we experimentally measured chondrocyte viability, metabolic activity, cell migration, and matrix productivity after implementing the designed loading protocols, vitrification, and warming. We demonstrated that porcine and human articular cartilage cubes can be successfully vitrified and rewarmed, maintaining high cell viability and excellent cellular function. The vitrified particulated articular cartilage was stored for a period of 6 months with no significant deterioration in chondrocyte viability and functionality. Our approach enables high-quality long-term storage of viable articular cartilage that can alleviate the shortage of grafts for use in clinically repairing articular cartilage defects.
Highlights
Articular cartilage defects are difficult to self-repair due to the avascular and aneural structure of articular cartilage[1]
Particulated articular cartilage transplantation can provide viable chondrocytes in situ for articular cartilage repair, and it is regarded as a reliable treatment for articular cartilage defects in the early stage, especially for focal chondral lesions[2,3,4]
We calculated the spatial and temporal distribution of vitrifiability, and based on this calculation, we designated potential loading protocols as having sufficient cryoprotectant permeation once the minimum vitrifiability exceeded the threshold determined by Weiss et al.[25] arriving at a more optimal 7 h protocol[23] for osteochondral dowels shown experimentally to be successful at preserving chondrocyte viability and metabolism in porcine articular cartilage[32]
Summary
Articular cartilage defects are difficult to self-repair due to the avascular and aneural structure of articular cartilage[1]. Fresh articular cartilage is most commonly stored for up to 28 days at 4 °C in a tissue bank, and chondrocyte viability starts to decline after 14 days of storage at this temperature[6,7]. This short window of time for viable articular cartilage makes successful cartilage transplantation challenging in the clinical scenario, because it takes ~2 weeks to obtain regulatory clearance for disease screening and make clinical preparations such as patient matching, disease checking, operation scheduling, and sample delivery[6,8]. If viable articular cartilage could be stored for extended periods of time beyond 28 days, the shortage of articular cartilage tissue in clinics would be eased, and more widespread clinical use of articular cartilage grafts (e.g., particulated articular cartilage) for cartilage defect repair would be possible
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