title: Three-dimensional microtissue assay for high-throughput cytotoxicity of nanoparticles.
abstract: Traditional in vitro nanotoxicity researches are conducted on cultured two-dimensional (2D) monolayer cells and thereby cannot reflect organism response to nanoparticle toxicities at tissue levels. This paper describes a new, high-throughput approach to test in vitro nanotoxicity in three-dimensional (3D) microtissue array, where microtissues are formed by seeding cells in nonsticky microwells, and cells are allowed to aggregate and grow into microtissues with defined size and shape. Nanoparticles attach and diffuse into microtissues gradually, causing radial cytotoxicity among cells, with more cells being killed on the outer layers of the microtissue than inside. Three classical toxicity assays [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT), glucose-6-phosphate dehydrogenase (G6DP), and calcein AM and ethidium homodimer (calcein AM/EthD-1)] have been adopted to verify the feasibility of the proposed approach. Results show that the nanotoxicities derived from this method are significantly lower than that from traditional 2D cultured monolayer cells (p < 0.05). Equipped with a microplate reader or a microscope, the nanotoxicity assay could be completed automatically without transferring the microtissue, ensuring the reliability of toxicity assay. The proposed approach provides a new strategy for high-throughput, simple, and accurate evaluation of nanoparticle toxicities by combining 3D microtissue array with a panel of classical toxicity assays.
authors: NanoScience Technology Center, University of Central Florida, Orlando, Florida 32826, USA.
authors: Yang
authors: Y
authors: Luo
authors: Chaoming
authors: C
authors: Wang
authors: Mainul
authors: M
authors: Hossain
authors: Yong
authors: Y
authors: Qiao
authors: Liyuan
authors: L
authors: Ma
authors: Jincui
authors: J
authors: An
authors: Ming
authors: M
authors: Su
mesh_terms: D001729:Bismuth / Q000737:chemistry; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004996:Ethidium / Q000031:analogs & derivatives / Q000737:chemistry / Q000633:toxicity; D005452:Fluoresceins / Q000737:chemistry / Q000633:toxicity; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism; D006367:HeLa Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry / Q000633:toxicity*; D012822:Silicon Dioxide / Q000737:chemistry; D046888:Tissue Array Analysis
chemical_list: D005452:Fluoresceins; C085925:calcein AM; C018533:ethidium homodimer; D012822:Silicon Dioxide; D005954:Glucosephosphate Dehydrogenase; D004996:Ethidium; D001729:Bismuth