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The influence of storage conditions on delta amino levulinic acid induced toxicity and phototoxicity in vitro

机译：贮存条件对δ氨基乙酰丙酸的体外毒性和光毒性的影响

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Delta amino levulinic acid photodynamic therapy (ALA-PDT) represents one of the most prominent advances in PDT. ALA itself or its derivatives are marketed for a variety of clinical indications. Despite the development of clinical applications, experimental ALA results are very heterogeneous and experimentally used parameters are still not standardized. This suggests that some problems remain unsolved that are likely to impair experiments to be performed but also that clinical results obtained could be greatly improved. Frequently unmentioned or imprecise data concern solvents, pH of ALA solutions, storage time, ALA degradation or ALA efficacy. In addition, diversity of experimental model is huge while capabilities of ALA transformation into PpIX are known to vary from one cell to the other. Thus, the aim of the present paper was to quantify the level of ALA degradation or changes in ALA efficacy using one single cell line without presuming of the mechanisms and determine the conditions of storage inducing the best transformation into PpIX and/or cell phototoxicity. We added ALA diluted in water, PBS or RPMI to C6 cells, a murine brain tumour cell line that can be used in vivo as an orthotopic graft. We measured in cells used as tools for final bio efficacy estimation, both the induced fluorescence and phototoxicity in various conditions of storage before use chosen to be as close as possible to the real lab conditions. Water had been found to better preserve ALA than, respectively, PBS and RPMI and this for any temperature or storage durations. The lowest temperature and the shortest duration for storage used had also been shown to better preserve ALA-induced fluorescence and phototoxicity. The fact that these properties were found to be better preserved in 7.4 buffered solvent could be in relationship with a fast ALA condensation occurring at neutral or lightly acidic pH modifying its availability for an optimal transformation into PpIX.
机译：三角洲氨基乙酰丙酸光动力疗法（ALA-PDT）代表了PDT中最突出的进展之一。 ALA本身或其衍生物可用于各种临床适应症。尽管临床应用得到了发展，但是实验ALA的结果还是非常不均一的，并且实验使用的参数仍未标准化。这表明仍然存在一些尚未解决的问题，这些问题可能会影响要进行的实验，但是所获得的临床结果也会大大改善。经常未提及或不准确的数据涉及溶剂，ALA溶液的pH值，储存时间，ALA降解或ALA功效。另外，实验模型的多样性是巨大的，而已知ALA转化为PpIX的能力会在一个单元之间变化。因此，本论文的目的是在不推测其机制的情况下，使用一种单一细胞系来量化ALA降解水平或ALA功效的变化，并确定诱导最佳转化为PpIX和/或细胞光毒性的储存条件。我们在水，PBS或RPMI中将ALA稀释后添加到C6细胞中，C6细胞是一种鼠脑肿瘤细胞系，可以在体内用作原位移植物。我们在用作最终生物功效评估工具的细胞中进行了测量，在使用前的各种储存条件下，诱导荧光和光毒性都选择为尽可能接近实际实验室条件。已经发现，在任何温度或储存时间内，水分别比PBS和RPMI更好地保存ALA。还显示了最低的温度和最短的储存时间可以更好地保留ALA诱导的荧光和光毒性。发现这些性质可以更好地保留在7.4缓冲溶剂中，这一事实可能与在中性或弱酸性pH下发生的快速ALA缩合有关，从而改变了其最佳转化为PpIX的可用性。

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