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1.
Toxins (Basel) ; 15(7)2023 06 29.
Article in English | MEDLINE | ID: mdl-37505691

ABSTRACT

Current investigations in the field of toxicology mostly rely on 2D cell cultures and animal models. Although well-accepted, the traditional 2D cell-culture approach has evident drawbacks and is distant from the in vivo microenvironment. To overcome these limitations, increasing efforts have been made in the development of alternative models that can better recapitulate the in vivo architecture of tissues and organs. Even though the use of 3D cultures is gaining popularity, there are still open questions on their robustness and standardization. In this review, we discuss the current spheroid culture and organ-on-a-chip techniques as well as the main conceptual and technical considerations for the correct establishment of such models. For each system, the toxicological functional assays are then discussed, highlighting their major advantages, disadvantages, and limitations. Finally, a focus on the applications of 3D cell culture for mycotoxin toxicity assessments is provided. Given the known difficulties in defining the safety ranges of exposure for regulatory agency policies, we are confident that the application of alternative methods may greatly improve the overall risk assessment.


Subject(s)
Cell Culture Techniques , Microphysiological Systems , Animals , Cell Culture Techniques/methods
2.
Food Chem Toxicol ; 157: 112605, 2021 Nov.
Article in English | MEDLINE | ID: mdl-34634377

ABSTRACT

Given the increasing importance of establishing better risk assessments for mycotoxins, novel in vitro tools for the evaluation of their toxicity are mandatory. In this study, an in vitro 3D spheroid model from SH-SY5Y cells, a human neuroblastoma cell line, was developed, optimized and characterized to test the cytotoxic effects caused by the mycotoxin sterigmatocystin (STE). STE induced a concentration- and time-dependent cell viability decrease in spheroids. Spheroids displayed cell disaggregation after STE exposure, increasing in a dose-dependent manner and over time. STE also induced apoptosis as confirmed by immunofluorescence staining and Western blot. Following the decreased proliferation and increased apoptosis, STE cytostasis effects were observed by migration assays both in 2D and 3D cell culture. Increased ROS generation, as well as DNA damage were also observed. Taken together, these data highlight the cytotoxic properties of STE and suggest that cell culture models play a pivotal role in the toxicological risk assessment of mycotoxins. The evaluation of cytotoxicity in spheroids (3D) rather than monolayer cultures (2D) is expected to more accurately reflect in vivo-like cell behaviour.


Subject(s)
Cell Culture Techniques, Three Dimensional/methods , Mycotoxins/toxicity , Sterigmatocystin/toxicity , Toxicity Tests/methods , Blotting, Western , Cell Line, Tumor/cytology , Cell Line, Tumor/drug effects , Cell Movement/drug effects , Comet Assay/methods , Fluorescent Antibody Technique , Humans , Neuroblastoma , Reactive Oxygen Species/metabolism , Spheroids, Cellular/drug effects
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