Development and assessment of an intestinal tri-cellular model to investigate the pro/anti-inflammatory potential of digested foods
IntroductionImmunonutrition, defined as the potential of foods, nutrients and dietary patterns to modulate the immune system activity, has been proposed as a strategy to enhance the immune response in both metabolic and immune-mediated diseases. However, the anti-/pro-inflammatory role of foods and...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2025-02-01
|
| Series: | Frontiers in Immunology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1545261/full |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832539892834369536 |
|---|---|
| author | Marina Ramal-Sanchez Chiara Bravo-Trippetta Veronica D’Antonio Elena Corvaglia Angela A. M. Kämpfer Roel P. F. Schins Mauro Serafini Donato Angelino |
| author_facet | Marina Ramal-Sanchez Chiara Bravo-Trippetta Veronica D’Antonio Elena Corvaglia Angela A. M. Kämpfer Roel P. F. Schins Mauro Serafini Donato Angelino |
| author_sort | Marina Ramal-Sanchez |
| collection | DOAJ |
| description | IntroductionImmunonutrition, defined as the potential of foods, nutrients and dietary patterns to modulate the immune system activity, has been proposed as a strategy to enhance the immune response in both metabolic and immune-mediated diseases. However, the anti-/pro-inflammatory role of foods and diets is far to be fully ascertained, and thus there is a continued needed for appropriate in vitro cell-culture models to investigate the role of foods in modulating cell-mediated inflammatory processes. This study aims to develop and test an in vitro tri-culture model, simulating the complexity of the intestinal tract and its multiple cell interactions.MethodsTo achieve this, the intestinal epithelial barrier was established by co-culturing human Caco-2 enterocyte-like and HT29-MTX-E12 mucus producing goblet-like colon cells, then adding human monocyte THP-1 cells to the basolateral compartment. The integrity and stability of the epithelial barrier were monitored and the inflammatory response of the model was assessed using various stressors at different concentrations, both individually and in combination (phorbol-12- myristate-13-acetate or PMA, and lipopolysaccharide or LPS), in terms of cytokines production. To test the model, different concentrations of in vitro digested broccoli (BD) were added to the apical section of the model.ResultsSupernatants from the basolateral compartment were collected and analyzed for cytokines production (IL-6, TNF-α, IL-12p70, IL-18 and IL-8) using automated ELISA (ELLA). Additionally, ZO-1 protein from the tight junctions of epithelial cells was analyzed by flow cytometry. The results indicated that 100 nM PMA added to the whole model for 20 h was the best stressor to simulate a mild-inflammatory status of the gut. Following treatment with BD, IL-6, TNF-α, IL-8 and IL-18 were significantly reduced compared to the control group, while ZO-1 expression increased at the lowest BD concentration.ConclusionsThese findings confirm the feasibility of the model for assessing the effects of food digesta on specific cytokines and permeability markers, representing a valuable strategy for investigating the role of foods in modulating the inflammatory response. The results obtained may support dietary strategies aimed at promoting wellbeing and preventing inflammatory-related metabolic diseases. |
| format | Article |
| id | doaj-art-5165c10dc55c41a3ae4f10bdb393a7c4 |
| institution | Kabale University |
| issn | 1664-3224 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj-art-5165c10dc55c41a3ae4f10bdb393a7c42025-02-05T07:32:41ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-02-011610.3389/fimmu.2025.15452611545261Development and assessment of an intestinal tri-cellular model to investigate the pro/anti-inflammatory potential of digested foodsMarina Ramal-Sanchez0Chiara Bravo-Trippetta1Veronica D’Antonio2Elena Corvaglia3Angela A. M. Kämpfer4Roel P. F. Schins5Mauro Serafini6Donato Angelino7Functional Foods and Stress Prevention Laboratory, Department of Biosciences and Technology for Food, Agriculture and Environment, University of Teramo, Teramo, ItalyFunctional Foods and Stress Prevention Laboratory, Department of Biosciences and Technology for Food, Agriculture and Environment, University of Teramo, Teramo, ItalyFunctional Foods and Stress Prevention Laboratory, Department of Biosciences and Technology for Food, Agriculture and Environment, University of Teramo, Teramo, ItalyFunctional Foods and Stress Prevention Laboratory, Department of Biosciences and Technology for Food, Agriculture and Environment, University of Teramo, Teramo, ItalyIUF – Leibniz Research Institute for Environmental Medicine, Düsseldorf, GermanyIUF – Leibniz Research Institute for Environmental Medicine, Düsseldorf, GermanyFunctional Foods and Stress Prevention Laboratory, Department of Biosciences and Technology for Food, Agriculture and Environment, University of Teramo, Teramo, ItalyFunctional Foods and Stress Prevention Laboratory, Department of Biosciences and Technology for Food, Agriculture and Environment, University of Teramo, Teramo, ItalyIntroductionImmunonutrition, defined as the potential of foods, nutrients and dietary patterns to modulate the immune system activity, has been proposed as a strategy to enhance the immune response in both metabolic and immune-mediated diseases. However, the anti-/pro-inflammatory role of foods and diets is far to be fully ascertained, and thus there is a continued needed for appropriate in vitro cell-culture models to investigate the role of foods in modulating cell-mediated inflammatory processes. This study aims to develop and test an in vitro tri-culture model, simulating the complexity of the intestinal tract and its multiple cell interactions.MethodsTo achieve this, the intestinal epithelial barrier was established by co-culturing human Caco-2 enterocyte-like and HT29-MTX-E12 mucus producing goblet-like colon cells, then adding human monocyte THP-1 cells to the basolateral compartment. The integrity and stability of the epithelial barrier were monitored and the inflammatory response of the model was assessed using various stressors at different concentrations, both individually and in combination (phorbol-12- myristate-13-acetate or PMA, and lipopolysaccharide or LPS), in terms of cytokines production. To test the model, different concentrations of in vitro digested broccoli (BD) were added to the apical section of the model.ResultsSupernatants from the basolateral compartment were collected and analyzed for cytokines production (IL-6, TNF-α, IL-12p70, IL-18 and IL-8) using automated ELISA (ELLA). Additionally, ZO-1 protein from the tight junctions of epithelial cells was analyzed by flow cytometry. The results indicated that 100 nM PMA added to the whole model for 20 h was the best stressor to simulate a mild-inflammatory status of the gut. Following treatment with BD, IL-6, TNF-α, IL-8 and IL-18 were significantly reduced compared to the control group, while ZO-1 expression increased at the lowest BD concentration.ConclusionsThese findings confirm the feasibility of the model for assessing the effects of food digesta on specific cytokines and permeability markers, representing a valuable strategy for investigating the role of foods in modulating the inflammatory response. The results obtained may support dietary strategies aimed at promoting wellbeing and preventing inflammatory-related metabolic diseases.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1545261/fullimmunonutritioncytokinesnutritiongutIBDtri-culture |
| spellingShingle | Marina Ramal-Sanchez Chiara Bravo-Trippetta Veronica D’Antonio Elena Corvaglia Angela A. M. Kämpfer Roel P. F. Schins Mauro Serafini Donato Angelino Development and assessment of an intestinal tri-cellular model to investigate the pro/anti-inflammatory potential of digested foods Frontiers in Immunology immunonutrition cytokines nutrition gut IBD tri-culture |
| title | Development and assessment of an intestinal tri-cellular model to investigate the pro/anti-inflammatory potential of digested foods |
| title_full | Development and assessment of an intestinal tri-cellular model to investigate the pro/anti-inflammatory potential of digested foods |
| title_fullStr | Development and assessment of an intestinal tri-cellular model to investigate the pro/anti-inflammatory potential of digested foods |
| title_full_unstemmed | Development and assessment of an intestinal tri-cellular model to investigate the pro/anti-inflammatory potential of digested foods |
| title_short | Development and assessment of an intestinal tri-cellular model to investigate the pro/anti-inflammatory potential of digested foods |
| title_sort | development and assessment of an intestinal tri cellular model to investigate the pro anti inflammatory potential of digested foods |
| topic | immunonutrition cytokines nutrition gut IBD tri-culture |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1545261/full |
| work_keys_str_mv | AT marinaramalsanchez developmentandassessmentofanintestinaltricellularmodeltoinvestigatetheproantiinflammatorypotentialofdigestedfoods AT chiarabravotrippetta developmentandassessmentofanintestinaltricellularmodeltoinvestigatetheproantiinflammatorypotentialofdigestedfoods AT veronicadantonio developmentandassessmentofanintestinaltricellularmodeltoinvestigatetheproantiinflammatorypotentialofdigestedfoods AT elenacorvaglia developmentandassessmentofanintestinaltricellularmodeltoinvestigatetheproantiinflammatorypotentialofdigestedfoods AT angelaamkampfer developmentandassessmentofanintestinaltricellularmodeltoinvestigatetheproantiinflammatorypotentialofdigestedfoods AT roelpfschins developmentandassessmentofanintestinaltricellularmodeltoinvestigatetheproantiinflammatorypotentialofdigestedfoods AT mauroserafini developmentandassessmentofanintestinaltricellularmodeltoinvestigatetheproantiinflammatorypotentialofdigestedfoods AT donatoangelino developmentandassessmentofanintestinaltricellularmodeltoinvestigatetheproantiinflammatorypotentialofdigestedfoods |