Minimizing ice recrystallization injury in tissues and organs has historically been sought using biological antifreeze proteins. However, the size of these compounds can limit permeation and their potential immunogenicity disqualifies them from use in several cryopreservation applications. Novel small molecule carbohydrate-derived ice recrystallization inhibitors (IRIs) are not subject to these constraints, and thus we sought to evaluate the ability of a highly active IRI to permeate liver tissue and control recrystallization. Rat liver tissue blocks (0.5 mm2) were incubated with the IRI for 6 h at 22 °C and subsequently plunged in liquid nitrogen. Ice crystals within the tissue were fixed using a formal acetic alcohol fixative as it was rewarmed from -80 °C to 22 °C over the course of 48 h. The untreated control demonstrated a gradient of increasing crystal size from the exterior to the interior region of the tissue; however, the IRI-treated condition had no such gradient and exhibited small crystals throughout. Threshold segmentation confirmed a significant reduction in the ice crystal size within the interior region of the IRI-treated condition, suggesting the IRI permeated throughout and effectively controlled recrystallization within the tissue.

中文翻译：

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使用小分子碳水化合物衍生物控制肝组织中的冰重结晶

历史上一直寻求使用生物防冻蛋白来最大限度地减少组织和器官中的冰重结晶损伤。然而，这些化合物的大小会限制渗透，并且它们潜在的免疫原性使它们无法用于多种冷冻保存应用。新型小分子碳水化合物衍生的冰重结晶抑制剂 (IRI) 不受这些限制，因此我们试图评估高活性 IRI 渗透肝组织和控制重结晶的能力。大鼠肝组织块 (0.5 mm2) 与 IRI 在 22°C 下孵育 6 小时，随后浸入液氮中。组织内的冰晶使用正式的醋酸固定剂固定，因为它在 48 小时内从 -80°C 重新升温至 22°C。未处理的对照表现出从组织的外部到内部区域增加晶体尺寸的梯度；然而，IRI 处理的条件没有这样的梯度，并且始终显示出小晶体。阈值分割证实了 IRI 处理条件内部区域内冰晶尺寸的显着减小，表明 IRI 渗透到整个组织中并有效控制了组织内的再结晶。