A New Gastro-Intestinal System to Evaluate the Effect of Food Methylglyoxal

The Column

A novel modular bioreactor for dynamic in vitro studies has been set-up, connecting two-dimensional (2D) scaffolds and mimicking a multi-organ model, to study the absorption/metabolization of compounds. The effect of dietary methylglyoxal, a potentially exogenous and endogenous toxic compound, on a dynamic gastro-intestinal system has been evaluated. Bioreactors represent a powerful advance in comparison with conventional in vitro static assays and could be a potential alternative to animal testing in the future.

 

A novel modular bioreactor for dynamic in vitro studies has been set-up, connecting two-dimensional (2D) scaffolds and mimicking a multi-organ model, to study the absorption/metabolization of compounds. The effect of dietary methylglyoxal, a potentially exogenous and endogenous toxic compound, on a dynamic gastro-intestinal system has been evaluated. Bioreactors represent a powerful advance in comparison with conventional in vitro static assays and could be a potential alternative to animal testing in the future.

Q. How did the idea of this research project arise and what are the aims of this research?

A: This project aims to develop a new dynamic and multi-compartmental millifluidic bioreactor simulating a gastro-intestinal system suitable for the evaluation of the effect (absorption/metabolization and cytotoxicity) of exogenous bioactives derived from food or drug intake.

By using this new dynamic protocol, we tested methylglyoxal (MGO), which is a potentially toxic compound, produced both in food and endogenously, responsible for the formation of the advanced glycation end-products involved in many pathological chronic implications, such as diabetes, cardiovascular diseases, and ageing-associated disorders (1–4).

The experiments were also performed with the standardized Minekus’ enzymatic digestion protocol to investigate the potential of this new dynamic system compared with static tradi-tional assays (5).

Q. What are you doing in this poster that is novel?

A: The novelty aspect of this work is the setup of a new platform that gives the opportunity to simulate continuous flow conditions, reproducing different simil-physiological flows and connecting different compartments to recreate a multi-organ model. This is the first setup of a dynamic gastro-intestinal platform and such a system represents a potential advance in the study of kinetic, metabolic, and cytotoxic profiles of different substances.

 

Q. What were the main analytical challenges you had to overcome?

A: First of all, it was necessary to carefully optimize the flow conditions to treat gastric (GIST-882) and intestinal (Caco-2) cells in the dynamic system when the two compartments were connected, in order to reproduce a simil-physiological digestive process.

Then, to monitor MGO metabolic fate by using a millifluidic system, it is mandatory to collect samples at different monitoring times and to analyze them by a proper analytical technique. To detect and quantify MGO in the cell medium during gastric and intestinal phases, we used a reversed-phase high performance liquid chromatography diode-array detector (HPLC-DAD) method, slightly modified from the literature and validated by us. The choice of UV detector required a sample preparation step, consisting of a derivatization procedure to obtain a UV-detectable MGO derivative (the derivatization procedure required a setup, too).

Q. What were your main findings and why are they useful?

A: The obtained results highlight that exogenous MGO is rapidly metabolized by a safe digestive process, and this is very important for human health in relation to widespread diseases, such as diabetes and ageing-associated disorders. Furthermore, by comparing the dynamic and the static processes, it emerged that the two protocols provided complementary data, and in particular the dynamic system elucidated a new role of gastric cells, which should be further investigated in the metabolization of toxic compounds.

 

Q. How are you planning to develop this research further?

A: We are going to submit to this dynamic digestion protocol other molecules, both toxic and ac-tive compounds, also in association. Moreover, we will probably use this platform for complex matrices, such as food samples, to test the effects of the interactions among the different food components, on the absorption/metabolization of a specific molecule.

Q. Anything else you would like to add?

A: This platform could reduce in vivo experiments, in particular during preliminary investigations, and find a very promising application in high-throughput cell-based compound screening for food analysis, drug discovery, and toxicity tests.

References

  1. M. Hellwig, S. Gensberger-Reigl, T. Henle, and M. Pischetsrieder, Semin. Cancer Biol.49, 1–8 (2018).
  2. H.J. Chun, Y. Lee, A.H. Kim, and J. Lee, Neurotox. Res.29, 419–431 (2016).
  3. N.M.J. Hanssen et al., Diabetes66, 2278–2283 (2017).
  4. T.M. Jensen, D. Vistisen, T. Fleming, et al., Diabet. Med.33, 1625–1631 (2016).
  5. M. Minekus et al., Food Funct.5, 1113–1124 (2014).

Lucia Ferron, Nutraceutical and Food Chem Toxicol Analysis Laboratory, Department of Drug Sciences, University of Pavia, Italy, FlaNat Research Italia Srl, Rho (MI), Italy.

E-mail: luci.ferron01@gmail.com

This interview was first published in a special supplement from LCGC Europe: The Rising Stars of Separation Science. For more interviews with Poster Winners, go to: https://bit.ly/3aRot39