Biostabilization Laboratory - Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, Stadtfelddamm 34, 30625 Hannover, Germany; Unit for Reproductive Medicine - Clinic for Horses, University of Veterinary Medicine Hannover, Bünteweg 15, 30559 Hannover, Germany.
Biostabilization Laboratory - Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, Stadtfelddamm 34, 30625 Hannover, Germany.
Biochim Biophys Acta Gen Subj. 2023 Jan;1867(1):130254. doi: 10.1016/j.bbagen.2022.130254. Epub 2022 Oct 13.
In this study we investigated hydrogen bonding interactions in hydrated and frozen solutions of different cryoprotective agents (CPAs) including dimethyl sulfoxide, glycerol, ethylene glycol, propylene glycol, and trehalose. We also investigated the effect of CPAs on ice crystal growth during storage and correlated this with storage stability of liposomes.
FTIR spectroscopy was used to study hydrogen bonding interactions in CPA solutions in HO and DO, and their thermal response was analyzed using van 't Hoff analysis. The effect of CPAs on ice crystal growth during storage was investigated by microscopy and correlated with storage stability of liposomes encapsulated with a fluorescent dye.
Principal component analyses demonstrated that different CPAs can be recognized based on the shape of the OD band region only. Chemically similar molecules such as glycerol and ethylene glycol closely group together in a principal component score plot, whereas trehalose and DMSO appear as condensed separated clusters. The OH/OD band of CPA solutions exhibits an overall shift to higher wavenumbers with increasing temperature and changed fractions of weak and strong hydrogen interactions. CPAs diminish ice crystal formation in frozen samples during storage and minimize liposome leakage during freezing but cannot prevent leakage during frozen storage.
CPAs can be distinguished from one another based on the hydrogen bonding network that is formed in solution. DMSO-water mixtures behave anomalous compared to other CPAs that have OH groups. CPAs modulate ice crystal formation during frozen storage but cannot prevent liposome leakage during frozen storage.
在这项研究中，我们研究了不同冷冻保护剂（CPAs）在水合和冷冻溶液中的氢键相互作用，包括二甲基亚砜、甘油、乙二醇、丙二醇和海藻糖。我们还研究了 CPAs 对储存过程中冰晶生长的影响，并将其与脂质体的储存稳定性相关联。
傅里叶变换红外光谱（FTIR）用于研究 HO 和 DO 中 CPA 溶液中的氢键相互作用，并使用范特霍夫分析（van 't Hoff analysis）分析其热响应。通过显微镜研究 CPAs 对储存过程中冰晶生长的影响，并将其与用荧光染料包封的脂质体的储存稳定性相关联。
主成分分析（PCA）表明，仅基于 OD 带区域的形状就可以识别不同的 CPAs。化学性质相似的分子，如甘油和乙二醇，在主成分得分图中紧密聚集在一起，而海藻糖和 DMSO 则呈现出凝聚的分离簇。随着温度的升高，CPA 溶液的 OH/OD 带整体向更高的波数移动，并且弱和强氢键的分数发生变化。CPAs 可减少储存过程中冷冻样品中冰晶的形成，并在冷冻过程中最小化脂质体泄漏，但不能防止冷冻储存期间的泄漏。
可以根据溶液中形成的氢键网络来区分 CPAs。DMSO-水混合物的行为与具有 OH 基团的其他 CPAs 不同。CPAs 可调节冷冻储存过程中冰晶的形成，但不能防止冷冻储存过程中的脂质体泄漏。
