Synergistic effects of surfactants and sugars on lipoplex stability during freeze-drying and rehydration.
Synergistic effects of surfactants and sugars on lipoplex stability during freeze-drying and rehydration.
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DOI：
10.1002/jps.21564
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发表时间：
2009-09
影响因子：
3.8
通讯作者：
Anchordoquy, Thomas J.
中科院分区：
文献类型：
--
作者：
Yu, Jinxiang;Anchordoquy, Thomas J.
非病毒载体在冷冻干燥过程中的稳定性已经得到充分研究，并且已经确定糖可以在冷冻干燥过程中保护脂质复合物。然而，在冷冻干燥后经常观察到低水平的损伤，并且这种损伤在稀释的lipoplex制剂中更明显。通过研究在冷冻干燥循环中的每个步骤之后脂质复合物的稳定性（即，冷冻、一次干燥和二次干燥），我们努力了解造成损害的机制，并确定改进的稳定策略。以等摩尔的DOTAP与胆固醇的比例和3：1的DOTAP+与DNA−电荷比制备N-（1-（2，3-二油酰氧基）丙基）-N，N，N-三甲基氯化铵（DOTAP）-胆固醇/质粒DNA脂质复合物。我们的实验表明，尽管有足够水平的“稳定”糖，但当稀释的lipoplex制剂进行冷冻干燥时，仍然明显存在显著的损害。对冷冻干燥的不同阶段的分析表明，在冷冻过程中会发生显著的损伤，而糖对这种冷冻引起的损伤的保护能力有限。在蛋白质的研究中已经观察到类似的效果，并且表面活性剂已经用于蛋白质制剂中以防止表面诱导的损伤，例如，在冰晶、固体、空气或糖玻璃表面。然而，在基于脂质的制剂中使用表面活性剂固有地具有风险，这是由于改变/溶解脂质递送载体的可能性。我们的数据表明，明智地使用表面活性剂可以减少表面诱导的损伤，并导致在冷冻干燥后更好地保存lipoplex的大小和转染活性。
The stability of non-viral vectors during freeze-drying has been well-studied, and it has been established that sugars can protect lipoplexes during freeze-drying. However low levels of damage are often observed after freeze-drying, and this damage is more evident in dilute lipoplex preparations. By investigating the stability of lipoplexes after each step in the freeze-drying cycle (i.e., freezing, primary drying, and secondary drying) we strive to understand the mechanisms responsible for damage and identify improved stabilization strategies. N-(1-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP)-cholesterol/plasmid DNA lipoplexes were prepared at an equimolar DOTAP to cholesterol ratio, and a 3-to-1 DOTAP+ to DNA− charge ratio. Our experiments indicate that despite sufficient levels of “stabilizing” sugars, significant damage is still evident when dilute lipoplex preparations are subjected to freeze-drying. Analysis of the different stages of freeze-drying suggests that significant damage occurs during freezing, and that sugars have a limited capacity to protect against this freezing-induced damage. Similar effects have been observed in studies with proteins, and surfactants have been employed in protein formulations to protect against surface-induced damage, e.g., at the ice crystal, solid, air or sugar glass surfaces. However, the use of surfactants in a lipid-based formulation is inherently risky due to the potential for altering/solubilizing the lipid delivery vehicle. Our data indicate that judicious use of surfactants can reduce surface-induced damage, and result in better preservation of lipoplex size and transfection activity after freeze-drying.
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影响因子：
3.9
作者：
Kuhl, T;Guo, YQ;Hui, SW
通讯作者：
Hui, SW
影响因子：
3.3
作者：
Page, C;Dawson, P;Mire-Sluis, A
通讯作者：
Mire-Sluis, A
影响因子：
2.9
作者：
PATERNOSTRE, MT;ROUX, M;RIGAUD, JL
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RIGAUD, JL
影响因子：
3.7
作者：
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通讯作者：
PEARLMAN, R
影响因子：
3.8
作者：
Anchordoquy, TJ;Armstrong, TK;Molina, MD
通讯作者：
Molina, MD