Côa Francine, Delite Fabrício de Souza, Strauss Mathias, Martinez Diego Stéfani Teodoro
Brazilian Nanotechnology National Laboratory (LNNano), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas, São Paulo, Brazil; Center for Nuclear Energy in Agriculture (CENA), University of São Paulo (USP), Piracicaba, São Paulo, Brazil.
Brazilian Nanotechnology National Laboratory (LNNano), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas, São Paulo, Brazil.
NanoImpact. 2022 Jul;27:100413. doi: 10.1016/j.impact.2022.100413. Epub 2022 Aug 6.
In this work, the toxicity and biodistribution of graphene oxide (GO) and oxidized multi-walled carbon nanotubes (MWCNT) were investigated in Caenorhabditis elegans. Bovine serum albumin (BSA) was selected as a model protein to evaluate the influence of protein corona formation on materials physicochemical properties, colloidal stability, and toxicity. Biological assays were performed to assess the effects of bare and albumin corona coated materials on survival, oxidative stress, intestinal barrier permeability, growth, reproduction, and fertility. Critical alterations in topography, surface roughness and chemistry of GO and MWCNT were observed due to albumin corona formation. These modifications were associated with changes in colloidal stability of materials and prevention of their aggregation and sedimentation in nematode testing medium. Both GO and MWCNT caused damage to nematode survival, growth, reproduction, and fertility, as well as enhanced oxidative stress and permeability of the intestinal barrier. But GO was more toxic than MWCNT to C. elegans, especially at long-term assays. Albumin corona mitigated 100% of acute and chronic effects of MWCNT. In contrast, the negative effects of GO were not completely mitigated; GO inhibited 16.2% of nematode growth, 86.5% of reproduction, and 32.0% of fertility at the highest concentration evaluated (10 mg L), while corona coated GO mitigated 50% and 100% of fertility and growth, respectively. Confocal Raman spectroscopy imaging was crucial to point out that bare and albumin corona coated GO and MWCNT crossed the C. elegans intestinal barrier reaching its reproductive organs. However, BSA corona protected the nematodes targeted organs from negative effects from MWCNT and blocked its translocation to other tissues, while coated GO was translocated inside the nematode affecting the functionality of crucial organs. In addition, coated MWCNT was excreted after 2 h of food resumption, whereas coated GO still accumulated in the nematode intestine. Our results demonstrate that the materials different translocation and excretion patterns in C. elegans had a relation to the impaired physiological functions of primary and secondary organs. This work is a contribution towards a better understanding of the impacts of protein corona on the toxicity of graphene oxide and carbon nanotubes; essential information for biological applications and nanosafety.
在本研究中,我们对线虫体内氧化石墨烯(GO)和氧化多壁碳纳米管(MWCNT)的毒性及生物分布进行了研究。选择牛血清白蛋白(BSA)作为模型蛋白,以评估蛋白质冠层的形成对材料物理化学性质、胶体稳定性和毒性的影响。进行了生物学试验,以评估裸露的和白蛋白冠层包覆的材料对生存、氧化应激、肠道屏障通透性、生长、繁殖和生育能力的影响。由于白蛋白冠层的形成,观察到GO和MWCNT在形貌、表面粗糙度和化学性质方面发生了显著变化。这些修饰与材料胶体稳定性的变化以及在线虫测试介质中防止其聚集和沉淀有关。GO和MWCNT均对线虫的生存、生长、繁殖和生育能力造成损害,并增强了氧化应激和肠道屏障的通透性。但GO对秀丽隐杆线虫的毒性比MWCNT更大,尤其是在长期试验中。白蛋白冠层减轻了MWCNT 100%的急性和慢性影响。相比之下,GO的负面影响并未完全减轻;在评估的最高浓度(10 mg/L)下,GO抑制了16.2%的线虫生长、86.5%的繁殖和32.0%的生育能力,而冠层包覆的GO分别减轻了50%和100%的生育能力和生长抑制。共聚焦拉曼光谱成像对于指出裸露的和白蛋白冠层包覆的GO和MWCNT穿过线虫肠道屏障并到达其生殖器官至关重要。然而,BSA冠层保护线虫的靶器官免受MWCNT的负面影响,并阻止其向其他组织的转运,而包覆的GO则在线虫体内转运,影响关键器官的功能。此外,在恢复喂食2小时后,包覆的MWCNT被排出,而包覆的GO仍积聚在线虫肠道中。我们的结果表明,材料在秀丽隐杆线虫体内不同的转运和排泄模式与初级和次级器官生理功能受损有关。这项工作有助于更好地理解蛋白质冠层对氧化石墨烯和碳纳米管毒性的影响;这是生物应用和纳米安全性的重要信息。