Department of Chemical and Materials Engineering, University of Alberta, Edmonton, AlbertaT6G 1H9, Canada.
Department of Surgery, University of Alberta, Edmonton, AlbertaT6G 2B7, Canada.
J Phys Chem B. 2022 Nov 24;126(46):9566-9579. doi: 10.1021/acs.jpcb.2c05736. Epub 2022 Nov 9.
Cryopreserving articular cartilage by vitrification can increase the availability of tissue for osteochondral allograft transplantation to treat cartilage defects. Developing well-optimized vitrification protocols can be supported by mathematical modeling to reduce the amount of trial-and-error experimentation needed. Fick's law has been used to model cryoprotectant diffusion, but it assumes ideal, dilute solution behavior, neglects water movement, and assumes diffusion of each cryoprotectant is independent of the presence of other cryoprotectants. The modified triphasic model addresses some of these shortcomings by accounting for water movement and the nonideal, nondilute nature of cryoprotectant vitrification solutions. However, it currently only exists for solutions containing a single cryoprotectant. As such, we extend the modified triphasic model to include two permeating cryoprotectants so that simultaneous diffusion occurring in vitrification protocols can be more accurately modeled. Using previously published experimental data, we determine suitable values for the fitting parameters of the new model. We then model a successful vitrification protocol for particulated cartilage cubes by calculating concentration, freezing point, vitrifiability, and strain profiles at the end of each loading step. We observe that Fick's law consistently underestimates cryoprotectant concentration throughout the cartilage compared to the modified triphasic model, leading to an underestimation of tissue vitrifiability. We additionally observe that simultaneous diffusion of cryoprotectants increases the permeation rate of each individual cryoprotectant, which Fick's law fails to consider. This suggests that using the two-cryoprotectant modified triphasic model to develop vitrification protocols could reduce excess exposure to cryoprotectants and improve preserved tissue outcomes.
玻璃化冷冻保存关节软骨可以增加组织的可用性,用于治疗软骨缺损的同种异体骨软骨移植。通过数学建模来开发优化的玻璃化方案,可以减少需要进行的反复试验的数量。菲克定律已被用于模拟冷冻保护剂的扩散,但它假设理想的稀溶液行为,忽略了水的运动,并假设每种冷冻保护剂的扩散独立于其他冷冻保护剂的存在。改进的三相模型通过考虑水的运动和冷冻保护剂玻璃化溶液的非理想、非稀溶液性质来解决其中的一些缺点。然而,它目前仅适用于含有单一冷冻保护剂的溶液。因此,我们将改进的三相模型扩展到包含两种渗透冷冻保护剂,以便更准确地模拟玻璃化方案中同时发生的扩散。利用先前发表的实验数据,我们确定了新模型拟合参数的合适值。然后,我们通过计算每个加载步骤结束时的浓度、冰点、可玻璃化性和应变分布,为颗粒状软骨立方体的成功玻璃化方案建模。我们观察到,与改进的三相模型相比,菲克定律在整个软骨中始终低估了冷冻保护剂的浓度,导致对组织可玻璃化性的低估。我们还观察到,冷冻保护剂的同时扩散增加了每种冷冻保护剂的渗透速率,而菲克定律没有考虑到这一点。这表明,使用双冷冻保护剂改进的三相模型来开发玻璃化方案可以减少对冷冻保护剂的过度暴露,并改善保存组织的结果。