Bauhammer I, Sacha M, Haltner E
Across Barriers GmbH, Science Park 1, 66123, Saarbrücken, Germany.
Heliyon. 2019 May 8;5(5):e01618. doi: 10.1016/j.heliyon.2019.e01618. eCollection 2019 May.
In view of increasing numbers of dermatological disorders, transdermal drug delivery along with research is becoming increasingly popular. Herefore, qualified skin models are required. The objective of this study was the optimization and validation of a modified lactate dehydrogenase (LDH) release assay during the establishment of an viable human skin model, employable for a variety of skin associated disorders. Firstly, the most suitable LDH isoform for the study was determined. Subsequently, a stability study was conducted to investigate the best storage conditions of the LDH enzyme. Finally, the test system was validated in terms of linear range, range limits and system suitability. The results indicate LDH-5 as most suitable isoform due to its predominance in skin. The stability samples stored at -20 °C in the presence of polyethylene glycol (PEG) as cryoprotector displayed the targeted recovery of 100% ± 15 % until the end of the four-week study in contrast to other investigated conditions. A six-point calibration without PEG and a seven-point calibration with PEG including evaluation of system suitability and quantification limits were established with both correlation coefficients r above 0.99 and all deviations below 15%. Concluding from those results, this method can be considered valid and useful for its employment in viability determination of viable skin models.
鉴于皮肤病的数量不断增加，经皮给药及其相关研究越来越受欢迎。因此，需要合格的皮肤模型。本研究的目的是在建立一种适用于多种皮肤相关疾病的有活力的人体皮肤模型过程中，对改良的乳酸脱氢酶（LDH）释放测定法进行优化和验证。首先，确定了该研究中最合适的LDH同工酶。随后，进行了稳定性研究，以探究LDH酶的最佳储存条件。最后，对测试系统的线性范围、范围限度和系统适用性进行了验证。结果表明，LDH-5是最合适的同工酶，因为它在皮肤中占主导地位。与其他研究条件相比，在作为冷冻保护剂的聚乙二醇（PEG）存在下于-20°C储存的稳定性样品在为期四周的研究结束时显示出100%±15%的目标回收率。建立了不含PEG的六点校准和含PEG的七点校准，包括系统适用性评估和定量限，相关系数r均高于0.99，所有偏差均低于15%。从这些结果可以得出结论，该方法在有活力的皮肤模型活力测定中的应用可被认为是有效且有用的。
