Cellular Uptake and Nuclear Accumulation of Polystyrene Nanoplastics in 3T3 Fibroblasts and Hepatocytes of Rattus norvegicus
Cellular Uptake and Nuclear Accumulation of Polystyrene Nanoplastics in 3T3 Fibroblasts and Hepatocytes of Rattus norvegicus
Febriyansyah Saputra1, Alfiah Hayati1*, Adiibtia Septiani1, Manikya Pramudya1, Raden Joko Kuncoroningrat Susilo2, Vuanghao Lim3, Bayyinatul Muchtaromah4
Accumulation of PSNPs in 3T3 fibroblasts and Rattus norvegicus hepatocytes. A: Representative fluorescence images of 3T3 fibroblasts and Rattus norvegicus hepatocytes under control conditions and after exposure to PSNPs. Minimal red fluorescence is observed in the control groups, whereas PSNP-exposed cells show a marked increase in red fluorescence, indicating significant nanoparticle accumulation. White arrows indicate regions of PSNP accumulation. Images were captured at 40× magnification, and at least 100 cells were analyzed per group. B: Quantification of PSNP accumulation expressed as a percentage of the control. Data are shown as mean ± SEM from three independent experiments. Error bars represent standard error of the mean. Different letters (a, b) denote statistically significant differences between groups (p < 0.05).
Nanoplastic Distribution in 3T3 Cells. Confocal Microscopy Analysis with DAPI and Nile Red Staining, 400x Magnification. A: Control Group: Displays normal 3T3 cell morphology with round nuclei and uniformly distributed cytoplasm, indicative of healthy cellular conditions. B: PSNP-Exposed Group: Demonstrates significant nanoplastic accumulation (red asterisks) within the cytoplasm and around the nucleus, as evidenced by the intensified red fluorescence, suggesting potential cellular stress and interaction with intracellular components.
PSNP Accumulation in Rattus norvegicus Hepatocytes: Fluorescence Staining with DAPI and Nile Red. A: Control group showing normal hepatocyte morphology. B: Early PSNP exposure with mild red fluorescence indicating initial cytoplasmic accumulation. C: Progressive PSNP accumulation with increased red fluorescence in the cytoplasm and near the nucleus. D: Advanced PSNP infiltration showing intense red fluorescence within the cytoplasm and nuclei, indicating cellular stress and potential disruption. White arrows indicate hepatocyte nuclei, and yellow arrows mark the cytoplasm. Fluorescence staining with DAPI (blue) visualizes the nuclei, while Nile Red (red) highlights PSNP presence.
PSNP Distribution and Infiltration in Rattus norvegicus Hepatocytes. Confocal microscopy images showing hepatocyte nuclei (blue, DAPI) and PSNP accumulation (red, Nile Red). PSNP particles are observed in the cytoplasm (yellow arrows) and within the nucleus (red arrows), indicating successful cellular internalization and nuclear infiltration. The numbered labels (1–6) represent sequential stages of PSNP intracellular progression: (1) Extracellular localization: PSNPs are initially observed in the extracellular matrix surrounding hepatocytes. (2) Initial cytoplasmic uptake : PSNPs begin to accumulate in the cytoplasm, likely through endocytotic mechanisms. (3) Nuclear envelope interaction : PSNPs interact with and penetrate the nuclear membrane, entering the nucleus. (4) Intracellular dispersion : PSNPs are dispersed within the nucleoplasm, suggesting widespread nuclear distribution. (5) Directed movement toward nucleoli : PSNPs exhibit directed migration toward the nucleolar region. (6) Nucleolar accumulation : PSNPs accumulate near or within the nucleolus, potentially affecting nucleolar structure or function. Green arrows highlight regions of intense PSNP clustering, possibly indicating agglomeration or retention zones within or near nucleolar areas.
