Cryoprotectant permeation through human articular cartilage.

ABSTRACT

OBJECTIVE: The cryopreservation of intact articular cartilage is constrained by minimal chondrocyte survival. It was the aim of the present study to gain an insight into the permeation kinetics of cryoprotectants through cartilage. This knowledge is essential for achieving adequate tissue permeation prior to cooling. DESIGN: The diffusion coefficients and penetration rates through human articular cartilage of dimethyl sulfoxide (Me(2)SO) and glycerol at different temperatures (4 degrees C, 17 degrees C, 27 degrees C and 37 degrees C) and at two concentrations [10% (v/v) and absolute state] were measured using diffusion nuclear magnetic imaging. Deuterated water (D(2)O) was used as a control substance. RESULTS: Glycerol penetrated faster than Me(2)SO at all temperatures and at rates that were comparable to those for D(2)O. The penetration rate of each agent increased with increasing temperature. The diffusion coefficients for glycerol and Me(2)SO increased with increasing temperature and decreased at the higher concentration, but the differences between each agent were not significant. CONCLUSIONS: The classical cryopreservation protocols expose cartilage samples to Me(2)SO at a too low temperature and/or for an insufficient time period for optimal cell survival. When considering the penetration rate, glycerol appears to be a more efficient cryoprotective agent than Me(2)SO. The present study demonstrates the power of nuclear magnetic resonance technology to elucidate key physiological factors in cryobiology.