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）可能导致生殖细胞和胚胎的修饰。虽然这些缺陷可能会影响生育能力或导致先天性缺陷和NP的生殖毒性潜力，但对其对生殖细胞的影响知之甚少。本项目旨在阐明由金属（Au, Ag）和合金（AuAg, NiTi, CrCo）制成的NP对生殖细胞和发育胚胎形态和遗传完整性、活力和功能的影响。粒子将在液体中通过激光烧蚀产生，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的大型动物模型。在两家申请机构联合开展的试验基础上，精子、卵母细胞和胚胎将通过共孵育的方式暴露于颗粒中，如果是胚胎，还将通过显微注射和携带np的精子/卵母细胞受精。相关参数（精子/卵母细胞：活力、形态、受精潜力；胚胎：发育能力、基因表达、dna甲基化）将以大小、时间和剂量依赖的方式进行测量。如果有不利影响，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。获得的数据将为纳米颗粒的生殖毒性潜力提供深刻的见解。

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.