Influence of metal and alloy nanoparticles on germ cell function and embryo development - REPROTOX -

无意中暴露于越来越多的纳米颗粒（NP）可能会导致生殖细胞和胚胎的修饰。虽然这种缺陷会损害生育能力或导致先天性缺陷，NP的再生潜能已经被提出，但对其对生殖细胞的影响知之甚少。该项目旨在阐明由金属（Au，Ag）和合金（AuAg，NiTi，CrCo）制成的NP对生殖细胞的形态和遗传完整性，活力和功能以及发育胚胎的影响。颗粒将通过在液体中的激光烧蚀产生，因为这允许制造高纯度和zeta电位的胶体NP以及生产胶体合金纳米颗粒。将使用猪生殖细胞和胚胎，这是一种被广泛接受的接近人类生理学的大型动物模型。基于两个申请机构已经联合建立的AuNP试验，精子、卵母细胞和胚胎将通过共孵育暴露于颗粒，在胚胎的情况下也通过显微注射和与负载NP的精子/卵母细胞受精。将以大小、时间和剂量依赖性方式测量相关参数（精子/卵母细胞：活力、形态学、受精潜力;胚胎：发育能力、基因表达、DNA甲基化）。如果出现不利影响，将进行更详细的分析。此外，NP将被注射到睾丸中，以研究其对精子发生的影响。所获得的数据将使人们深刻了解纳米颗粒的再生潜力。

Unintended exposure to a growing variety of nanoparticles (NP) may cause modifications in germline cells and embryos. Although such defects can impair fertility or lead to congenital defects and reprotoxic potential of NP has already been suggested, only little is known about their effect on reproductive cells. The project aims to elucidate the effect of NP made of metals (Au, Ag) and alloys (AuAg, NiTi, CrCo) on morphological and genetic integrity, vitality and functionality of germ cells as well as developing embryos. Particles will be generated by laser ablation in liquid, since this allows fabrication of colloidal NP of high purity and zeta potential as well as the production of colloidal alloy nanoparticles. Porcine germ cells and embryos will be used, being a well accepted large animal model close to human physiology. On the basis of trials already established jointly by the two applicant institutions for AuNP, spermatozoa, oocytes and embryos will be exposed to the particles by coincubation, in case of embryos also by microinjection and fertilization with NP-loaded spermatozoa/oocytes. Relevant parameters (sperm/oocytes: viability, morphology, fertilising potential; embryos: developmental capacity, gene expression, DNA-methylation) will be measured in a size, time and dose-dependent manner. In case of detrimental effects, more detailed analyses will be performed. Also, NP will be injected into testicles to study their impact on spermatogenesis. The gained data will give profound insight into the reprotoxic potential of nanoparticles.