Cyrill Buzzy from Nanomedicine Lab, University of Manchester, UK, the Lonza CytoSMART Lab for scratch assays in 2D material testing

Dr. Cyrill Bussy, School of Health Sciences, The University of Manchester, UK

In-vitro Cell Response to Graphene Oxide Exposure

Research Interest

Graphene and 2D materials are offering tremendous properties that will benefit the society in many ways. Yet, there are still concerns that these emerging materials could present novel risks to human health, due mainly to a severe knowledge-gap regarding whether they may cause harm and the level of exposure [1-4]. Graphene oxide is one of the most studied derivatives of graphene in the nanosafety field due to its inherent dispersibility in aqueous medium that do not require any chemical additives [5]. Toxicologists in nanomaterials at the University of Manchester are investigating how graphene oxide flakes interact with cells or organs and studying the impact of such interactions [6].


The Experiment

To visualize the impact of suspension of graphene oxide flakes on the mobility of cells in a wound healing test, also known as scratch assay, toxicologists at the School of Health Sciences, The University of Manchester, have used Lonza’s CytoSMART™ System. NIH/3T3 mouse embryo fibroblast cells were cultured until reaching confluence. At that point, a scratch assay was performed, creating a gap in the cell monolayer. The cell monolayer was then washed once before being exposed to either graphene oxides flakes in suspension in the normal cell culture medium for the tested cells or the cell culture medium alone. CytoSMART based time lapse microscopy was then performed and stopped when the gap between the two boundaries disappeared.

 

Video of scratch assay cell culture of NIH/3T3 cells in the presence of GO-2 recorded with CytoSMART™ Lux 10X Device 
Graph showing quantitative analysis of scratch closure in response to two different graphene oxide materials  

 

Video of scratch assay of NIH/3T3 cell line in the presence of graphene oxide material GO-1 recorded with the CytoSMART™ Lux 10X Device (Snapshot interval: 2 minutes)   Analysis of scratch closure in response to two different graphene oxide materials (GO-1 and GO-2) recorded with the CytoSMART™ System.         

Conclusion

The CytoSMART™ System can provide detailed information on the kinetic of wound healing under nanomaterial suspension exposure. Under the tested conditions, one material (GO-2, green line) was clearly delaying the wound healing process in comparison to cell not exposed to any materials (blue line). These live imaging-based results combined with additional investigations can help to understand the impact that graphene derivatives may have on cell physiology and basic functions such as division and migration. The development of biomedical applications of graphene derivatives will benefit from this knowledge [7-9].

 

References

1. Bianco A. (2013) Angew Chem Int Ed Engl 52(19):4986-97. doi: 10.1002/anie.201209099.

2. Bussy C, Ali-Boucetta H, Kostarelos K. (2013) Acc Chem Res 46(3):692-701. doi: 10.1021/ar300199e.

3. Bianco A and Prato M. (2015) 2D Mater 2 : 030201.  doi:10.1088/2053-1583/2/3/030201

4. Kostarelos K, Novoselov KS. (2014) Science 344(6181):261-3. doi:10.1126/science.1246736.

5. Bussy C, Jasim D, Lozano N, Terry D, Kostarelos K. (2015) Nanoscale  7(15):6432-5. doi: 10.1039/c5nr00236b.

6. Jasim DA, Murphy S, Newman L, Mironov A, Prestat E, McCaffrey J, Ménard-Moyon  C, Rodrigues AF, Bianco A, Haigh S, Lennon R, Kostarelos K. (2016) ACS Nano Dec 12. [Epub ahead of print]

7. Shin SR, Li YC, Jang HL, Khoshakhlagh P, Akbari M, Nasajpour A, Zhang YS, Tamayol A, Khademhosseini A. (2016) Adv Drug Deliv Rev 105(Pt B):255-274. doi: 10.1016/j.addr.2016.03.007.

8. Holt BD, Wright ZM, Arnold AM, Sydlik SA. (2016) Wiley Interdiscip Rev Nanomed Nanobiotechnol Oct 26.doi: 10.1002/wnan.1437.

9. Ding X, Liu H, Fan Y. (2015) Adv Healthc Mater 15;4(10):1451-68. doi: 10.1002/adhm.201500203.

 

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