Interactions of polystyrene nanoplastics with in vitro models of the human intestinal barrier

Abstract

The universal presence of micro-nanoplastics (MNPLs) and its relative unknown effects on human health is a concern demanding reliable data to evaluate their safety. As ingestion is one of the main exposure routes for humans, we have assessed their hazard using two in vitro models that simulate the human intestinal barrier and its associated lymphoid system. Two different coculture models (differentiated Caco-2/HT29 intestinal cells and Caco-2/HT29 + Raji-B cells) were exposed to polystyrene nanoparticles (PSNPs) for 24 h. Endpoints such as viability, membrane integrity, NPS localization and translocation, ROS induction, and genotoxic damage were evaluated to have a comprehensive view of their potentially harmful effects. No significant cytotoxic effects were observed in any of the analyzed systems. In addition, no adverse effects were detected in the integrity or in the permeability of the barrier model. Nevertheless, confocal microscopy analysis showed that MNPLs were highly uptaken by both of the barrier model systems, and that translocation across the membrane occurred. Thus, MNPLs were detected into Raji-B cells, placed in the basolateral compartment of the insert. The internalization followed a dose-dependent pattern, as assessed by flow cytometry. Nonetheless, no genotoxic or oxidative DNA damage induction was detected in either case. Finally, no variations in the transcription of oxidative and stress genes could be detected in any of the in vitro barrier models. Our results show that MNPLs can enter and cross the epithelial barrier of the digestive system, as demonstrated when Raji-B cells were included in the model, but without exerting apparent hazardous effects.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

References

  1. Alimba CG, Faggio C (2019) Microplastics in the marine environment: current trends in environmental pollution and mechanisms of toxicological profile. Environ Toxicol Pharmacol 68:61–74

    CAS  PubMed  Google Scholar 

  2. Araújo F, Sarmento B (2013) Towards the characterization of an in vitro triple co-culture intestine cell model for permeability studies. Int J Pharm 458:128–134

    PubMed  Google Scholar 

  3. Auta HS, Emenike CU, Fauziah SH (2017) Distribution and importance of microplastics in the marine environment: a review of the sources, fate, effects, and potential solutions. Environ Int 102:165–176

    CAS  PubMed  Google Scholar 

  4. Behrens I, Pena AI, Alonso MJ, Kissel T (2002) Comparative uptake studies of bioadhesive and non-bioadhesive nanoparticles in human intestinal cell lines and rats: the effect of mucus on particle adsorption and transport. Pharm Res 19(8):1185–1193

    CAS  PubMed  Google Scholar 

  5. Bradney L, Wijesekara H, Palansooriya KN, Obadamudalige N, Bolan NS, Ok YS, Rinklebe J, Kim KH, Kirkham MB (2019) Particulate plastics as a vector for toxic trace-element uptake by aquatic and terrestrial organisms and human health risk. Environ Int 131:104937

    CAS  PubMed  Google Scholar 

  6. Carbery M, O'Connor W, Palanisami T (2018) Trophic transfer of microplastics and mixed contaminants in the marine food web and implications for human health. Environ Int 115:400–409

    PubMed  Google Scholar 

  7. Chatterjee N, Walker GC (2017) Mechanisms of DNA damage, repair, and mutagenesis. Environ Mol Mutagen 58:235–263

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Cortés C, Domenech J, Salazar M, Pastor S, Marcos R, Hernández A (2020) Nanoplastics as potential environmental health factors. Effects of polystyrene nanoparticles on the human intestinal epithelial Caco-2 cells. Environ Sci Nano 7:272–285

    Google Scholar 

  9. Deng Y, Zhang Y, Lemos B, Ren H (2017) Tissue accumulation of microplastics in mice and biomarker responses suggest widespread health risks of exposure. Sci Rep 7:46687

    PubMed  PubMed Central  Google Scholar 

  10. des Rieux A, Fievez V, Théate I, Mast J, Préat V, Schneider YJ (2007) An improved in vitro model of human intestinal follicle-associated epithelium to study nanoparticle transport by M cells. Eur J Pharm Sci 30:380–391

    CAS  PubMed  Google Scholar 

  11. EFSA—European Food Safety Authority (2016) Presence of microplastics and nanoplastics in food, with particular focus on seafood. EFSA J 14:4501

    Google Scholar 

  12. Eriksen M, Maximenko N, Thiel M, Cummins A, Lattin G, Wilson S, Hafner J, Zellers A, Rifman S (2013) Plastic pollution in the South Pacific subtropical gyre. Mar Pollut Bull 68:71–76

    CAS  PubMed  Google Scholar 

  13. García-Rodríguez A, Vila L, Cortés C, Hernández A, Marcos R (2018a) Effects of differently shaped TiO2NPs (nanospheres, nanorods and nanowires) on the in vitro model (Caco-2/HT29) of the intestinal barrier. Part Fibre Toxicol 15:33

    PubMed  PubMed Central  Google Scholar 

  14. García-Rodríguez A, Vila L, Hernández A, Marcos R (2018b) Exploring the usefulness of the complex in vitro intestinal epithelial model Caco-2/HT29/Raji-B in nanotoxicology. Food Chem Toxicol 113:162–170

    PubMed  Google Scholar 

  15. Hesler M, Aengenheister L, Ellinger B, Drexel R, Straskraba S, Jost C, Wagner S, Meier F, von Briesen H, Büchel C, Wick P, Buerki-Thurnherr T, Kohl Y (2019) Multi-endpoint toxicological assessment of polystyrene nano- and microparticles in different biological models in vitro. Toxicol In Vitro 61:104610

    CAS  PubMed  Google Scholar 

  16. Hidalgo-Ruz V, Gutow L, Thompson RC, Thiel M (2012) Microplastics in the marine environment: a review of the methods used for identification and quantification. Environ Sci Technol 46:3060–3075

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Ioakeimidis C, Fotopoulou KN, Karapanagioti HK, Geraga M, Zeri C, Papathanassiou E, Galgani F, Papatheodorou G (2016) The degradation potential of PET bottles in the marine environment: an ATR-FTIR based approach. Sci Rep 6:23501

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Kawanishi S, Ohnishi S, Ma N, Hiraku Y, Murata M (2017) Crosstalk between DNA damage and inflammation in the multiple steps of carcinogenesis. Int J Mol Sci 18:1808

    PubMed Central  Google Scholar 

  19. Klein M, Fischer EK (2019) Microplastic abundance in atmospheric deposition within the metropolitan area of Hamburg, Germany. Sci Total Environ 685:96–103

    CAS  PubMed  Google Scholar 

  20. Kucharzik T, Lügering N, Rautenberg K, Lügering A, Schmidt MA, Stoll R, Domschke W (2000) Role of M cells in intestinal barrier function. Ann NY Acad Sci 915:171–183

    CAS  PubMed  Google Scholar 

  21. Lambert S, Wagner M (2016) Formation of microscopic particles during the degradation of different polymers. Chemosphere 161:510–517

    CAS  PubMed  Google Scholar 

  22. Li WC, Tse HF, Fok L (2016) Plastic waste in the marine environment: a review of sources, occurrence and effects. Sci Total Environ 566–567:333–349

    PubMed  Google Scholar 

  23. Lim SL, Ng CT, Zou L, Lu Y, Chen J, Bay BH, Shen HM, Ong CN (2019) Targeted metabolomics reveals differential biological effects of nanoplastics and nanoZnO in human lung cells. Nanotoxicology 13:1117–1132

    CAS  PubMed  Google Scholar 

  24. Lin W, Jiang R, Wu J, Wei S, Yin L, Xiao X, Hu S, Shen Y, Ouyang G (2019) Sorption properties of hydrophobic organic chemicals to micro-sized polystyrene particles. Sci Total Environ 690:565–572

    CAS  PubMed  Google Scholar 

  25. Liu Y, Li W, Lao F, Liu Y, Wang L, Bai R, Zhao Y, Chen C (2011) Intracellular dynamics of cationic and anionic polystyrene nanoparticles without direct interaction with mitotic spindle and chromosomes. Biomaterials 32:8291–8303

    CAS  PubMed  Google Scholar 

  26. Lozoya-Agullo I, Araújo F, González-Álvarez I, Merino-Sanjuán M, González-Álvarez M, Bermejo M, Sarmento B (2017) Usefulness of Caco-2/HT29-MTX and Caco-2/HT29-MTX/Raji B coculture models to predict intestinal and colonic permeability compared to Caco-2 monoculture. Mol Pharm 14:1264–1270

    CAS  PubMed  Google Scholar 

  27. Magrì D, Sánchez-Moreno P, Caputo G, Gatto F, Veronesi M, Bardi G, Catelani T, Guarnieri D, Athanassiou A, Pompa PP, Fragouli D (2018) Laser ablation as a versatile tool to mimic polyethylene terephthalate nanoplastic pollutants: characterization and toxicology assessment. ACS Nano 12:7690–7700

    PubMed  Google Scholar 

  28. Mahler GJ, Esch MB, Tako E, Southard TL, Archer SD, Glahn RP, Shuler ML (2012) Oral exposure to polystyrene nanoparticles affects iron absorption. Nat Nanotechnol 7:264–271

    CAS  PubMed  Google Scholar 

  29. Paget V, Dekali S, Kortulewski T, Grall R, Gamez C, Blazy K, Aguerre-Chariol O, Chevillard S, Braun A, Rat P, Lacroix G (2015) Specific uptake and genotoxicity induced by polystyrene nanobeads with distinct surface chemistry on human lung epithelial cells and macrophages. PLoS ONE 10:e0123297

    PubMed  PubMed Central  Google Scholar 

  30. Pannetier P, Cachot J, Clérandeau C, Faure F, Van Arkel K, de Alencastro LF, Levasseur C, Sciacca F, Bourgeois JP, Morin B (2019) Toxicity assessment of pollutants sorbed on environmental sample microplastics collected on beaches: part 1-adverse effects on fish cell line. Environ Pollut 248:1088–1097

    CAS  PubMed  Google Scholar 

  31. PlasticsEurope (2017) The facts 2017: analysis of European plastics production, demand and waste data. PlasticsEurope, Belgium. https://www.plasticseurope.org/application/files/5715/1717/4180/Plastics_the_facts_2017_FINAL_for_website_one_page.pdf

  32. Prata JC (2018) Airborne microplastics: consequences to human health? Environ Pollut 234:115–126

    CAS  PubMed  Google Scholar 

  33. Rist S, Carney Almroth B, Hartmann NB, Karlsson TM (2018) A critical perspective on early communications concerning human health aspects of microplastics. Sci Total Environ 626:720–726

    CAS  PubMed  Google Scholar 

  34. Rubio L, Marcos R, Hernández A (2018) Nanoceria acts as antioxidant in tumoral and transformed cells. Chem Biol Interact 291:7–15

    CAS  PubMed  Google Scholar 

  35. Rubio L, Barguilla I, Domenech J, Marcos R, Hernández A (2020) Biological effects, including oxidative stress and genotoxic damage, of polystyrene nanoparticles in different human hematopoietic cell lines. J Hazard Mater Accept. https://doi.org/10.1016/j.jhazmat.2020.122900

    Article  Google Scholar 

  36. Sambuy Y, De Angelis I, Ranaldi G, Scarino ML, Stammati A, Zucco F (2005) The Caco-2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics. Cell Biol Toxicol 21:1–26

    CAS  PubMed  Google Scholar 

  37. Santana MFM, Moreira FT, Turra A (2017) Trophic transference of microplastics under a low exposure scenario: insights on the likelihood of particle cascading along marine food-webs. Mar Pollut Bull 121:154–159

    CAS  PubMed  Google Scholar 

  38. Schirinzi GF, Pérez-Pomeda I, Sanchís J, Rossini C, Farré M, Barceló D (2017) Cytotoxic effects of commonly used nanomaterials and microplastics on cerebral and epithelial human cells. Environ Res 159:579–587

    CAS  PubMed  Google Scholar 

  39. Silva B, Bastos AS, Justino CIL, da Costa JP, Duarte AC, Rocha-Santos TAP (2018) Microplastics in the environment: challenges in analytical chemistry—a review. Anal Chim Acta 1017:1–19

    CAS  PubMed  Google Scholar 

  40. Stock V, Böhmert L, Lisicki E, Block R, Cara-Carmona J, Pack LK, Selb R, Lichtenstein D, Voss L, Henderson CJ, Zabinsky E, Sieg H, Braeuning A, Lampen A (2019) Uptake and effects of orally ingested polystyrene microplastic particles in vitro and in vivo. Arch Toxicol 993:1817–1833

    Google Scholar 

  41. Su L, Deng H, Li B, Chen Q, Pettigrove V, Wu C, Shi H (2019) The occurrence of microplastic in specific organs in commercially caught fishes from coast and estuary area of East China. J Hazard Mater 365:716–724

    CAS  PubMed  Google Scholar 

  42. Toussaint B, Raffael B, Angers-Loustau A, Gilliland D, Kestens V, Petrillo M, Rio-Echevarria IM, Van den Eede G (2019) Review of micro- and nanoplastic contamination in the food chain. Food Addit Contam Part A Chem Anal Control Expo Risk Assess 36:639–673

    CAS  PubMed  Google Scholar 

  43. Weinstein JE, Crocker BK, Gray AD (2016) From macroplastic to microplastic: degradation of high-density polyethylene, polypropylene, and polystyrene in a salt marsh habitat. Environ Toxicol Chem 35:1632–1640

    CAS  PubMed  Google Scholar 

  44. Win KY, Feng SS (2005) Effects of particle size and surface coating on cellular uptake of polymeric nanoparticles for oral delivery of anticancer drugs. Biomaterials 26:2713–2722

    CAS  PubMed  Google Scholar 

  45. Wright SL, Kelly FJ (2017) Plastic and human health: a micro issue? Environ Sci Technol 51:6634–6647

    CAS  PubMed  Google Scholar 

  46. Wu B, Wu X, Liu S, Wang Z, Chen L (2019) Size-dependent effects of polystyrene microplastics on cytotoxicity and efflux pump inhibition in human Caco-2 Cells. Chemosphere 221:333–341

    CAS  PubMed  Google Scholar 

  47. Zhu F, Zhu C, Wang C, Gu C (2019) Occurrence and ecological impacts of microplastics in soil systems: a review. Bull Environ Contam Toxicol 102:741–749

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

J. Domenech was supported by a Predoctoral Fellowship (PIF) from the Universitat Autònoma de Barcelona. We thank Dr. Victor Puentes’ group (Inorganic Nanoparticles Group, ICN2) for supplying the protocol to characterize PS nanoparticles.

Funding

This investigation has been partially supported by the Ministry of Economy and Competitiveness (SAF2015-63519-R).

Author information

Affiliations

Authors

Contributions

CC, RM, and AH planned the experiments. JD, CC, and LR carried out the experimental part. CC and JD analyzed the data, carried out the statistical analysis, and prepared tables/figures. CC, JD, AH, and RM wrote the final manuscript.

Corresponding authors

Correspondence to Ricard Marcos or Constanza Cortés.

Ethics declarations

Conflict of interest

The authors declare that there is no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Domenech, J., Hernández, A., Rubio, L. et al. Interactions of polystyrene nanoplastics with in vitro models of the human intestinal barrier. Arch Toxicol (2020). https://doi.org/10.1007/s00204-020-02805-3

Download citation

Keywords

  • Nanoplastics
  • Styrene nanoparticles
  • Intestinal barrier
  • Caco-2/HT29/Raji-B cells