ETT S.p.A., via Sestri 37, Genoa 16154, Italy; CNR-Institute of Biophysics (IBF), Via De Marini 6, 16149 Genoa, Italy.
DiSTAV, University of Genoa, Corso Europa 26, 16132 Genoa, Italy.
Harmful Algae. 2016 May;55:230-237. doi: 10.1016/j.hal.2016.03.001. Epub 2016 Apr 1.
In the last decade, the occurrence of harmful dinoflagellate blooms of the genus Ostreopsis has increased both in frequency and in geographic distribution with adverse impacts on public health and the economy. Ostreopsis species are producers of palytoxin-like toxins (putative palytoxin and ovatoxins) which are among the most potent natural non-protein compounds known to date, exhibiting extreme toxicity in mammals, including humans. Most existing toxicological data are derived from in vivo mouse assay and are related to acute effects of pure palytoxin, without considering that the toxicity mechanism of dinoflagellates can be dependent on the varying composition of complex biotoxins mixture and on the presence of cellular components. In this study, in vitro neuronal networks coupled to microelectrode array (MEA)-based system are proposed, for the first time, as sensitive biosensors for the evaluation of marine alga toxicity on mammalian cells. Toxic effect was investigated by testing three different treatments of laboratory cultured Ostreopsis cf. ovata cells: filtered and re-suspended algal cells; filtered, re-suspended and sonicated algal cells; conditioned growth medium devoid of algal cells. The great sensitivity of this system revealed the mixture of PTLX-complex analogues naturally released in the growth medium and the different potency of the three treatments to inhibit the neuronal network spontaneous electrical activity. Moreover, by means of the multiparametric analysis of neuronal network activity, the approach revealed a different toxicity mechanism of the cellular component compared to the algal conditioned growth medium, highlighting the potential active role of the first treatment.
在过去的十年中,赤潮生物属夜光藻的有害藻华发生频率和地理分布都有所增加,对公共健康和经济造成了不利影响。夜光藻属物种是类似雪卡毒素的毒素(假定雪卡毒素和短裸甲藻毒素)的生产者,这些毒素是迄今为止已知的最有效力的天然非蛋白化合物,对包括人类在内的哺乳动物具有极强的毒性。大多数现有的毒理学数据来自体内小鼠试验,与纯雪卡毒素的急性效应有关,而没有考虑到藻类的毒性机制可能依赖于复杂生物毒素混合物的不同组成以及细胞成分的存在。在这项研究中,首次提出将体外神经元网络与基于微电极阵列(MEA)的系统相结合,作为评估海洋藻类对哺乳动物细胞毒性的敏感生物传感器。通过测试实验室培养的拟菱形藻细胞的三种不同处理方法:过滤和再悬浮藻类细胞;过滤、再悬浮和超声处理藻类细胞;不含藻类细胞的条件生长培养基,来研究毒性作用。该系统的高灵敏度揭示了自然释放到生长培养基中的 PTLX 复杂类似物混合物以及三种处理方法抑制神经元网络自发电活动的不同效力。此外,通过神经元网络活动的多参数分析,该方法揭示了细胞成分与藻类条件生长培养基相比具有不同的毒性机制,突出了第一种处理方法的潜在主动作用。