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BACKGROUND Dimethyl sulfoxide (DMSO) cryoprotectant can effectively alleviate the damage to single tissue during cryopreservation and restore its physiological activity after rewarming. However, studies have not been successful for preserving large tissue. This study aimed to investigate the application conditions of DMSO in large composite-tissue by performing femoral artery perfusion and soaking in a rabbit hind-limb model. MATERIAL AND METHODS A microdialysis-freezing point osmometer was used to detect the minimum time required for effective perfusion of 10% v/v perfusion and 20% v/v perfusion group. Magnetic resonance spectroscopy (MRS) was used to detect the area under the spectrum peak of DMSO in perivascular, intramuscular, subcutaneous areas, and compare the area under the spectrum peak in the 20% vascular perfusion group and other whole immersion groups. RESULTS The minimum time required for effective perfusion of muscle in the 10% v/v group was 30 minutes, the DMSO concentration was stable at 5% v/v; In the 20% v/v group the minimum time was at 20 minutes, stable at 12% v/v. There was a statistically difference of the area under the spectrum peak in the 10% group and the 20% v/v group after vascular perfusion in different tissue locations (P&lt;0.05). The 20% vascular perfusion group and the different concentration of DMSO in the large tissue soaking group were statistically different (P&lt;0.05). There was a significant difference in the 20% v/v vascular perfusion group compared to the low concentration immersion group, but no difference compared to the 50% immersion group. CONCLUSIONS The effect of blood perfusion on cryopreservation in large tissue by using DMSO was slightly better than overall soaking, especially in preservation of skin and subcutaneous tissue.

Dimethyl Sulfoxide is Less Effective in Immersing Cryopreserved Large Pieces of Tissue: A Rabbit Hind-Limb Model