Impact of Gastrointestinal Digestion In Vitro Procedure on the Characterization and Cytotoxicity of Reduced Graphene Oxide

Overview

Journal Nanomaterials (Basel)

Date 2023 Aug 26

PMID 37630872

Authors

Oscar Cebadero-Dominguez

Leticia Diez-Quijada

Sergio Lopez

Soraya Sanchez-Ballester

Maria Puerto

Ana M Camean

Angeles Jos

Affiliations

Soon will be listed here.

Abstract

The growing interest in graphene derivatives is a result of their variety of applications in many fields. Due to their use, the oral route could be a potential way of entrance for the general population. This work assesses the biotransformation of reduced graphene oxide (rGO) after an in vitro digestion procedure (mouth, gastric, intestinal, and colon digestion), and its toxic effects in different cell models (HepG2, Caco-2, and 3D intestinal model). The characterization of rGO digestas evidenced the agglomeration of samples during the in vitro gastrointestinal (g.i.) digestion. Internalization of rGO was only evident in Caco-2 cells exposed to the colonic phase and no cellular defects were observed. Digestas of rGO did not produce remarkable cytotoxicity in any of the experimental models employed at the tested concentrations (up to 200 µg/mL), neither an inflammatory response. Undigested rGO has shown cytotoxic effects in Caco-2 cells, therefore these results suggest that the digestion process could prevent the systemic toxic effects of rGO. However, additional studies are necessary to clarify the interaction of rGO with the g.i. tract and its biocompatibility profile.

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References

Barra A, Santos J, Silva M, Nunes C, Ruiz-Hitzky E, Goncalves I . Graphene Derivatives in Biopolymer-Based Composites for Food Packaging Applications. Nanomaterials (Basel). 2020; 10(10). PMC: 7589102. DOI: 10.3390/nano10102077. View

Sanchez V, Jachak A, Hurt R, Kane A . Biological interactions of graphene-family nanomaterials: an interdisciplinary review. Chem Res Toxicol. 2011; 25(1):15-34. PMC: 3259226. DOI: 10.1021/tx200339h. View

Loven K, Franzen S, Isaxon C, Messing M, Martinsson J, Gudmundsson A . Emissions and exposures of graphene nanomaterials, titanium dioxide nanofibers, and nanoparticles during down-stream industrial handling. J Expo Sci Environ Epidemiol. 2020; 31(4):736-752. PMC: 8263341. DOI: 10.1038/s41370-020-0241-3. View

Lai R, Shi P, Yi Z, Li H, Yi Y . Triple-Band Surface Plasmon Resonance Metamaterial Absorber Based on Open-Ended Prohibited Sign Type Monolayer Graphene. Micromachines (Basel). 2023; 14(5). PMC: 10221595. DOI: 10.3390/mi14050953. View

Bitounis D, Parviz D, Cao X, Amadei C, Vecitis C, Sunderland E . Synthesis and Physicochemical Transformations of Size-Sorted Graphene Oxide during Simulated Digestion and Its Toxicological Assessment against an In Vitro Model of the Human Intestinal Epithelium. Small. 2020; 16(21):e1907640. PMC: 7260083. DOI: 10.1002/smll.201907640. View

Ghulam A, Dos Santos O, Hazeem L, Pizzorno Backx B, Bououdina M, Bellucci S . Graphene Oxide (GO) Materials-Applications and Toxicity on Living Organisms and Environment. J Funct Biomater. 2022; 13(2). PMC: 9224660. DOI: 10.3390/jfb13020077. View

Diez-Quijada Jimenez L, Guzman-Guillen R, Lira A, Jos A, Camean A . In vitro assessment of cyanotoxins bioaccessibility in raw and cooked mussels. Food Chem Toxicol. 2020; 140:111391. DOI: 10.1016/j.fct.2020.111391. View

Patel T, R P, Vashi Y, Bhattacharya P . Toxic impacts and industrial potential of graphene. J Environ Sci Health C Toxicol Carcinog. 2020; 38(3):269-297. DOI: 10.1080/26896583.2020.1812335. View

Roberts M, Magnusson B, Burczynski F, Weiss M . Enterohepatic circulation: physiological, pharmacokinetic and clinical implications. Clin Pharmacokinet. 2002; 41(10):751-90. DOI: 10.2165/00003088-200241100-00005. View

10.

Fu C, Liu T, Li L, Liu H, Liang Q, Meng X . Effects of graphene oxide on the development of offspring mice in lactation period. Biomaterials. 2014; 40:23-31. DOI: 10.1016/j.biomaterials.2014.11.014. View

11.

Kejlova K, Dvorakova M, Vavrous A, Sevcik V, Kandarova H, Letasiova S . Toxicity of food contact paper evaluated by combined biological and chemical methods. Toxicol In Vitro. 2019; 59:26-34. DOI: 10.1016/j.tiv.2019.04.001. View

12.

Ahamed M, Akhtar M, Khan M, Alhadlaq H . Reduced graphene oxide mitigates cadmium-induced cytotoxicity and oxidative stress in HepG2 cells. Food Chem Toxicol. 2020; 143:111515. DOI: 10.1016/j.fct.2020.111515. View

13.

Guarnieri D, Sanchez-Moreno P, Rio Castillo A, Bonaccorso F, Gatto F, Bardi G . Biotransformation and Biological Interaction of Graphene and Graphene Oxide during Simulated Oral Ingestion. Small. 2018; 14(24):e1800227. DOI: 10.1002/smll.201800227. View

14.

Lahiani M, Gokulan K, Williams K, Khare S . Impact of Pristine Graphene on Intestinal Microbiota Assessed Using a Bioreactor-Rotary Cell Culture System. ACS Appl Mater Interfaces. 2019; 11(29):25708-25719. DOI: 10.1021/acsami.9b07635. View

15.

Kucki M, Diener L, Bohmer N, Hirsch C, Krug H, Palermo V . Uptake of label-free graphene oxide by Caco-2 cells is dependent on the cell differentiation status. J Nanobiotechnology. 2017; 15(1):46. PMC: 5480125. DOI: 10.1186/s12951-017-0280-7. View

16.

Minekus M, Alminger M, Alvito P, Ballance S, Bohn T, Bourlieu C . A standardised static in vitro digestion method suitable for food - an international consensus. Food Funct. 2014; 5(6):1113-24. DOI: 10.1039/c3fo60702j. View

17.

Novoselov K, Geim A, Morozov S, Jiang D, Zhang Y, Dubonos S . Electric field effect in atomically thin carbon films. Science. 2004; 306(5696):666-9. DOI: 10.1126/science.1102896. View

18.

Niu X, de Graaf I, van der Bij H, Groothuis G . Precision cut intestinal slices are an appropriate ex vivo model to study NSAID-induced intestinal toxicity in rats. Toxicol In Vitro. 2014; 28(7):1296-305. DOI: 10.1016/j.tiv.2014.06.010. View

19.

Gutierrez-Ruiz M, Gomez Quiroz L, Hernandez E, Bucio L, Souza V, Llorente L . Cytokine response and oxidative stress produced by ethanol, acetaldehyde and endotoxin treatment in HepG2 cells. Isr Med Assoc J. 2001; 3(2):131-6. View

20.

Patlolla A, Randolph J, Kumari S, Tchounwou P . Toxicity Evaluation of Graphene Oxide in Kidneys of Sprague-Dawley Rats. Int J Environ Res Public Health. 2016; 13(4):380. PMC: 4847042. DOI: 10.3390/ijerph13040380. View