Abstract
Cryopreservation of cartilage has been investigated for decades and is currently an established protocol. However, the reliability and applicability of cartilage cryopreservation for the use in electromechanical reshaping (EMR) has not been studied exclusively. A system to cryopreserve large numbers of tissue specimens provides a steady source of cartilage of similar quality for experimentation at later dates. This will reduce error that may arise from different cartilage stock, and has the potential to maximize efficiency under time constraints. Our study utilizes a unique methodology to cryopreserve septal cartilage for use in EMR studies. Rabbit septal cartilage specimens were harvested and standardized to 20 x 8 x 1 mm, and placed in one of three solutions (normal saline, PBS, 10% DMSO in PBS) for four hours in a cold storage room at 4 degrees Celsius. Then, each cartilage specimen was vacuumed and sealed in an anti-frost plastic bag and stored in a freezer at -80 degrees Celsius for 1 to 3 weeks duration. EMR was performed using 2 and 6 volts for 2 minutes application time. Bend angle measurements of the cryopreserved cartilage specimens were compared to bend angles of fresh cartilage which underwent EMR using the same parameters. Results demonstrate that normal saline, phosphate buffered saline (PBS), and PBS with DMSO were effective in cryopreservation, and indicated no significant differences in bend angle measurements when compared to no cryopreservation. Our methodology to cryopreserve cartilage specimens provides a successful approach for use in conducting large-scale EMR studies.
Highlights
Cryopreservation of cartilage has been an established protocol in the literature and has considerable importance in medical and research fields
Results comparing bend angles for fresh samples with cryopreserved samples at 2 and 6 volts for 2 minutes application time in phosphate buffered saline (PBS), normal saline, and 10% dimethyl sulfoxide (DMSO) by volume in PBS are shown in Figures 1 and 2
Fresh rabbit septal cartilage has been hydrated with either normal saline or PBS solution and has yielded promising results. 1-3 with regards to cryopreservation of cartilage, the literature has supported the use of 10% DMSO by volume in PBS to reduce freezing of the proteoglycan cellular matrix of cartilage.[5,6,7]
Summary
Cryopreservation of cartilage has been an established protocol in the literature and has considerable importance in medical and research fields. Reconstructive surgery performed in the osteochondral, chondral, meniscal, and cricoid regions currently depend on the availability of cartilage allografts Methods to preserve these grafts would naturally increase the resources available for performing these procedures.[1,2,3] Many synthetic biocompatible materials commonly used in surgical procedures require cartilage preservation.[1,2,3, 8] Likewise, cartilage reshaping modalities such as enzymatic digestion, radiofrequency reshaping, laser cartilage reshaping, and most recently, electromechanical reshaping (EMR) of cartilage are important areas that depend on a substantial amount of cartilage for data analysis.[1,2,3,4] Cryopreservation is a unique process that has been investigated for well over thirty years. Phenomena which can cause damage to cells during cryopreservation include chemical damage due to preservation in solution, extracellular and
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