Anticoagulants Influence the Performance of In Vitro Assays Intended for Characterization of Nanotechnology-Based Formulations.
Anticoagulants Influence the Performance of In Vitro Assays Intended for Characterization of Nanotechnology-Based Formulations.
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DOI：
10.3390/molecules23010012
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发表时间：
2017-12-21
期刊：
影响因子：
--
通讯作者：
Dobrovolskaia MA
中科院分区：
文献类型：
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作者：
Cedrone E;Neun BW;Rodriguez J;Vermilya A;Clogston JD;McNeil SE;Barenholz Y;Szebeni J;Dobrovolskaia MA
基于纳米技术的新型药物产品的临床前安全性评估通常依赖于体外分析，特别是在产品开发的早期阶段，因为可用于此类研究的纳米材料数量有限。大多数免疫学测试都需要捐献者的血液。为了进行这样的测试，必须防止血液凝结，这通常是通过在血液采集管中添加抗凝剂来实现的。肝素、乙二胺四乙酸(EDTA)和柠檬酸是最常用的抗凝剂。也有新的抗凝剂，如水飞蓟素，但没有得到广泛使用。尽管有观点认为某些抗凝剂可能会影响检测性能，但用于纳米技术制剂免疫学表征的体外检测中，传统抗凝剂和新型抗凝剂之间的系统比较目前尚不存在。我们在标准化免疫分析级联中比较了水飞蓟素抗凝血液和传统抗凝血液，发现抗凝剂的类型不影响溶血测试的性能。然而，水飞蓟素更适合于补体激活和白细胞增殖的检测，而传统的抗凝血剂柠檬酸盐和肝素更适合于凝血和细胞因子分泌的检测。这些结果还表明，传统的免疫控制方法，如脂多糖(LPS)，对于了解抗凝剂在检测性能中的作用并不可靠。我们观察到水飞蓟素和传统抗凝剂制备的纳米材料血液在测试结果上的差异，而传统对照组没有看到这样的效果。因此，重要的是要认识到每种抗凝剂的优点和局限性，并在个案的基础上考虑单个纳米颗粒。
The preclinical safety assessment of novel nanotechnology-based drug products frequently relies on in vitro assays, especially during the early stages of product development, due to the limited quantities of nanomaterials available for such studies. The majority of immunological tests require donor blood. To enable such tests one has to prevent the blood from coagulating, which is usually achieved by the addition of an anticoagulant into blood collection tubes. Heparin, ethylene diamine tetraacetic acid (EDTA), and citrate are the most commonly used anticoagulants. Novel anticoagulants such as hirudin are also available but are not broadly used. Despite the notion that certain anticoagulants may influence assay performance, a systematic comparison between traditional and novel anticoagulants in the in vitro assays intended for immunological characterization of nanotechnology-based formulations is currently not available. We compared hirudin-anticoagulated blood with its traditional counterparts in the standardized immunological assay cascade, and found that the type of anticoagulant did not influence the performance of the hemolysis assay. However, hirudin was more optimal for the complement activation and leukocyte proliferation assays, while traditional anticoagulants citrate and heparin were more appropriate for the coagulation and cytokine secretion assays. The results also suggest that traditional immunological controls such as lipopolysaccharide (LPS ) are not reliable for understanding the role of anticoagulant in the assay performance. We observed differences in the test results between hirudin and traditional anticoagulant-prepared blood for nanomaterials at the time when no such effects were seen with traditional controls. It is, therefore, important to recognize the advantages and limitations of each anticoagulant and consider individual nanoparticles on a case-by-case basis.
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影响因子：
6.8
作者：
Menssen, HD;Melber, K;Thiel, E
通讯作者：
Thiel, E
影响因子：
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作者：
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DOI：
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发表时间：
2018-01-01
期刊：
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影响因子：
--
作者：
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通讯作者：
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DOI：
10.1007/978-1-4939-7352-1_9
发表时间：
2018-01-01
期刊：
CHARACTERIZATION OF NANOPARTICLES INTENDED FOR DRUG DELIVERY, 2 EDITION
影响因子：
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作者：
Neun, Barry W.;Ilinskaya, Anna N.;Dobrovolskaia, Marina A.
通讯作者：
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DOI：
10.1007/978-1-4939-7352-1_10
发表时间：
2018-01-01
期刊：
CHARACTERIZATION OF NANOPARTICLES INTENDED FOR DRUG DELIVERY, 2 EDITION
影响因子：
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作者：
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通讯作者：
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