Abstract
Cryoprotectant toxicity has become more relevant because of increased use of high concentrations of cryoprotectants for vitrification of biologic tissues. A single toxicity model that integrates cryoprotectant concentration, time and temperature is essential to optimize the cryopreservation of tissues. The Weibull probabilistic distribution has been used in environmental toxicology research. This objective of this study was to fit the Weibull model to experimental data for chondrocyte recovery from articular cartilage exposed to various concentrations of dimethyl sulfoxide at different temperatures as a function of time. This study indicated that the Weibull model is an appropriate model to describe cryoprotectant toxicity to chondrocytes in articular cartilage.
This study was designed to examine the toxicity of dimethyl sulfoxide (DMSO) on chondrocytes in porcine articular cartilage (AC) as a function of time, temperature and concentration.
The Weibull model is suitable for modeling cryoprotectant toxicity in cartilage and can be further extended to other cellular and tissue systems.
The model provides a simple method to predict toxicity and to assess the feasibility of cryopreservation protocols.
The model proved to be a good fit for the entire data set of concentration, temperature and time, yielding an R2 value of 0.87 and a maximum discrepancy of 20% between the experimental data and the model. Estimates of the model’s parameters within a confidence interval of 95% were found to be: _=30±2, _=0.67±0.05, _C=0.38±0.03, _T=−2300±300 and _CT=700±100.
Sliced porcine AC was exposed to DMSO (1, 3, 5, 6M) at different temperatures (0, 22, 37°C) for various durations. Cellular viability was determined by membrane integrity stains. Experimental data for chondrocyte recovery was fit to the global Weibull probabilistic distribution model using SPSS SigmaPlot 2000 to estimate the five parameters.
A model integrating concentration, time, and temperature of exposure is required to optimize addition and removal protocols of high concentrations of cryoprotectant for cryopreservation. The Weibull distribution is a simple and flexible model used to describe similar processes. In the current study, chondrocyte viability decreased with increased concentration, temperature and time of exposure. The model indicated a significant interaction between the toxic effects of concentration and temperature.
Correspondence should be addressed to Cynthia Vezina, Communications Manager, COA, 4150-360 Ste. Catherine St. West, Westmount, QC H3Z 2Y5, Canada
