{"doi":"10.47749/t/unicamp.2024.1498854","title":"Alterações sutis desencadeadas por nanopartículas em células analisadas individualmente através de perfil multiparamétrico baseada em imagem","abstract":"The use of nanomaterials has been growing in recent decades due to their applications in industry and consumer materials, such as paints, pigments, personal care products and automotive products.These products are responsible for contaminating water, air and soil.With the widespread use of nanomaterials, the area of nanotoxicology studies should be stimulated, since many impacts on human health and the environment are still unknown.Among the most widely used nanomaterials are silver nanoparticles (AgNP).AgNPs are widely used because of their application as an antimicrobial agent, being used in cosmetics, automotive products, among others.Normally, in vitro studies to determine the toxicity of these nanoparticles are carried out with a high concentration of AgNP and a single exposure.However, taking into account exposure conditions that are closer to the relevant environmental conditions, repeated exposure at low concentrations of AgNP is more likely.The effects expected under these treatment conditions are sublethal, rather than the cell death and oxidative stress that are usually assessed in acute studies.Methods for evaluating specific signaling pathways in cells exposed to AgNP are diverse, but the use of high content analysis combined with high content imaging (HCI) is little explored in the field of nanotoxicology.HCI assays are highly sensitive and can evaluate the cellular response in an exploratory and unbiased manner.Thus, the initial objective of this work was to determine the phenotypic profiles of liver cells exposed to silver nanoparticles at sublethal concentrations, in single or repeated exposure for 15 days, and the profile was determined through non-biased imaging panels, Cell Painting and Live Cell Painting, and specific ones, Tox Painting, aimed at DNA damage, inflammation and lipidosis.We characterized the phenotypic profile of hepatoma cells exposed once or repeatedly to AgNPs, showing that 1 day of treatment alters features related to actin, Golgi and lipidosis, and 15 days of repeated exposure are related to DNA damage and inflammation.This work also explores cell exposure to nanomaterials, highlighting innovative imaging techniques and high-content analysis.We also provide detailed workflows, from image acquisition to phenotypic profile interpretation, to consolidate this research as a valuable resource for future generations.The development of Live Cell Painting, using acridine orange, demonstrated its effectiveness in revealing dynamic morphological profiles of cells.The method's application has been evidenced in drug screening and dose-response assays for silver nanoparticles, providing insights into altered pathways.Finally, we present two protocols to help interpret morphological characteristics from phenotypic profiles, where we use Morpheus for exploratory analysis and the visualization of individualized representative cells, with the help of tools such as CellProfiler.This thesis was a pioneer in establishing high content imaging and high content analysis in the research group, and it is hoped that these new approaches will be widely used in the field of nanotoxicology, to accelerate studies of the interaction between nanoparticles and cells.","journal":null,"year":2024,"id":508590,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.96,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1360588,"name":"Fernanda Garcia Fossa","orcid":null,"position":0,"is_corresponding":true}],"reference_count":0,"raw_metadata":null,"created_at":"2026-07-19T02:11:10.049781Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}