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Article ; Online: The application of organ-on-chip models for the prediction of human pharmacokinetic profiles during drug development.

Keuper-Navis, Marit / Walles, Markus / Poller, Birk / Myszczyszyn, Adam / van der Made, Thomas K / Donkers, Joanne / Eslami Amirabadi, Hossein / Wilmer, Martijn J / Aan, Saskia / Spee, Bart / Masereeuw, Rosalinde / van de Steeg, Evita

Pharmacological research

2023 Volume 195, Page(s) 106853

Abstract: Organ-on-chip (OoC) technology has led to in vitro models with many new possibilities compared to conventional in vitro and in vivo models. In this review, the potential of OoC models to improve the prediction of human oral bioavailability and intrinsic ... ...

Abstract	Organ-on-chip (OoC) technology has led to in vitro models with many new possibilities compared to conventional in vitro and in vivo models. In this review, the potential of OoC models to improve the prediction of human oral bioavailability and intrinsic clearance is discussed, with a focus on the functionality of the models and the application in current drug development practice. Multi-OoC models demonstrating the application for pharmacokinetic (PK) studies are summarized and existing challenges are identified. Physiological parameters for a minimal viable platform of a multi-OoC model to study PK are provided, together with PK specific read-outs and recommendations for relevant reference compounds to validate the model. Finally, the translation to in vivo PK profiles is discussed, which will be required to routinely apply OoC models during drug development.
MeSH term(s)	Humans ; Models, Biological ; Drug Development ; Biological Availability ; Microphysiological Systems
Language	English
Publishing date	2023-07-18
Publishing country	Netherlands
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	1003347-6
ISSN	1096-1186 ; 0031-6989 ; 1043-6618
ISSN (online)	1096-1186
ISSN	0031-6989 ; 1043-6618
DOI	10.1016/j.phrs.2023.106853
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Article: Recapitulating suckling-to-weaning transition in vitro using fetal intestinal organoids

Garcia, Tânia Martins / Muncan, Vanesa / Navis, Marit / van Elburg, Ruurd M / Wildenberg, Manon E

Journal of visualized experiments. 2019 Nov. 15, , no. 153

2019

Abstract: At the end of the suckling period, many mammalian species undergo major changes in the intestinal epithelium that are associated with the capability to digest solid food. This process is termed suckling-to-weaning transition and results in the ... ...

Abstract	At the end of the suckling period, many mammalian species undergo major changes in the intestinal epithelium that are associated with the capability to digest solid food. This process is termed suckling-to-weaning transition and results in the replacement of neonatal epithelium with adult epithelium which goes hand in hand with metabolic and morphological adjustments. These complex developmental changes are the result of a genetic program that is intrinsic to the intestinal epithelial cells but can, to some extent, be modulated by extrinsic factors. Prolonged culture of mouse primary intestinal epithelial cells from late fetal period, recapitulates suckling-to-weaning transition in vitro. Here, we describe a detailed protocol for mouse fetal intestinal organoid culture best suited to model this process in vitro. We describe several useful assays designed to monitor the change of intestinal functions associated with suckling-to-weaning transition over time. Additionally, we include an example of an extrinsic factor that is capable to affect suckling-to-weaning transition in vivo, as a representation of modulating the timing of suckling-to-weaning transition in vitro. This in vitro approach can be used to study molecular mechanisms of the suckling-to-weaning transition as well as modulators of this process. Importantly, with respect to animal ethics in research, replacing in vivo models by this in vitro model contributes to refinement of animal experiments and possibly to a reduction in the use of animals to study gut maturation processes.
Keywords	adults ; animal ethics ; animal experimentation ; animal use reduction ; animal use refinement ; animal use replacement ; intestinal mucosa ; mice ; suckling
Language	English
Dates of publication	2019-1115
Size	p. e60470.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/60470
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Recapitulating Suckling-to-Weaning Transition In Vitro using Fetal Intestinal Organoids.

Garcia, Tânia Martins / Navis, Marit / Wildenberg, Manon E / van Elburg, Ruurd M / Muncan, Vanesa

Journal of visualized experiments : JoVE

2019 , Issue 153

Abstract	At the end of the suckling period, many mammalian species undergo major changes in the intestinal epithelium that are associated with the capability to digest solid food. This process is termed suckling-to-weaning transition and results in the replacement of neonatal epithelium with adult epithelium which goes hand in hand with metabolic and morphological adjustments. These complex developmental changes are the result of a genetic program that is intrinsic to the intestinal epithelial cells but can, to some extent, be modulated by extrinsic factors. Prolonged culture of mouse primary intestinal epithelial cells from late fetal period, recapitulates suckling-to-weaning transition in vitro. Here, we describe a detailed protocol for mouse fetal intestinal organoid culture best suited to model this process in vitro. We describe several useful assays designed to monitor the change of intestinal functions associated with suckling-to-weaning transition over time. Additionally, we include an example of an extrinsic factor that is capable to affect suckling-to-weaning transition in vivo, as a representation of modulating the timing of suckling-to-weaning transition in vitro. This in vitro approach can be used to study molecular mechanisms of the suckling-to-weaning transition as well as modulators of this process. Importantly, with respect to animal ethics in research, replacing in vivo models by this in vitro model contributes to refinement of animal experiments and possibly to a reduction in the use of animals to study gut maturation processes.
MeSH term(s)	Animals ; Animals, Suckling/physiology ; Cells, Cultured ; Epithelial Cells/physiology ; Fetal Development/physiology ; Intestinal Mucosa/cytology ; Intestinal Mucosa/embryology ; Intestinal Mucosa/physiology ; Mice ; Organ Culture Techniques ; Organoids/cytology ; Organoids/embryology ; Organoids/physiology ; Weaning
Language	English
Publishing date	2019-11-15
Publishing country	United States
Document type	Journal Article ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/60470
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Article: Postbiotic Modulation of Retinoic Acid Imprinted Mucosal-like Dendritic Cells by Probiotic Lactobacillus reuteri 17938 In Vitro.

Haileselassie, Yeneneh / Navis, Marit / Vu, Nam / Qazi, Khaleda Rahman / Rethi, Bence / Sverremark-Ekström, Eva

Frontiers in immunology

2016 Volume 7, Page(s) 96

Abstract: Lactobacilli are widely used as probiotics with beneficial effects on infection-associated diarrhea, but also used in clinical trials of e.g., necrotizing enterocolitis and inflammatory bowel diseases. The possibility of using probiotic metabolic ... ...

Abstract	Lactobacilli are widely used as probiotics with beneficial effects on infection-associated diarrhea, but also used in clinical trials of e.g., necrotizing enterocolitis and inflammatory bowel diseases. The possibility of using probiotic metabolic products, so-called postbiotics, is desirable as it could prevent possible side effects of live bacteria in individuals with a disturbed gut epithelial barrier. Here, we studied how Lactobacillus reuteri DSM 17938 cell-free supernatant (L. reuteri-CFS) influenced retinoic acid (RA)-driven mucosal-like dendritic cells (DC) and their subsequent effect on T regulatory cells (Treg) in vitro. RA clearly imprinted a mucosal-like DC phenotype with higher IL10 production, increased CD103 and CD1d expression, and a downregulated mRNA expression of several inflammatory-associated genes (NFκB1, RELB, and TNF). Treatment with L. reuteri-CFS further influenced the tolerogenic phenotype of RA-DC by downregulating most genes involved in antigen uptake, antigen presentation, and signal transduction as well as several chemokine receptors, while upregulating IL10 production. L. reuteri-CFS also augmented CCR7 expression on RA-DC. In cocultures, RA-DC increased IL10 and FOXP3 expression in Treg, but pre-treatment with L. reuteri-CFS did not further influence the Treg phenotype. In conclusion, L. reuteri-CFS modulates the phenotype and function of mucosal-like DC, implicating its potential application as postbiotic.
Language	English
Publishing date	2016
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2606827-8
ISSN	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2016.00096
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Article: Lactobacillus reuteri and Staphylococcus aureus differentially influence the generation of monocyte-derived dendritic cells and subsequent autologous T cell responses.

Haileselassie, Yeneneh / Navis, Marit / Vu, Nam / Qazi, Khaleda Rahman / Rethi, Bence / Sverremark-Ekström, Eva

Immunity, inflammation and disease

2016 Volume 4, Issue 3, Page(s) 315–326

Abstract: Introduction: In early-life, the immature mucosal barrier allows contact between the gut microbiota and the developing immune system. Due to their strategic location and their ability to sample luminal antigen, dendritic cells (DC) play a central role ... ...

Abstract	Introduction: In early-life, the immature mucosal barrier allows contact between the gut microbiota and the developing immune system. Due to their strategic location and their ability to sample luminal antigen, dendritic cells (DC) play a central role in the interaction of microbes and immune cells in the gut. Here, we investigated how two bacteria associated with opposite immune profiles in children, that is, Lactobacillus (L.) reuteri and Staphylococcus (S.) aureus, influenced the differentiation of monocytes in vitro as well how the generated DC impacted T cell responses. Methods: We exposed monocyte cultures to cell-free supernatants (CFS) from these bacteria during their differentiation to DC. Results: The presence of L. reuteri-CFS during DC differentiation resulted in DC with a more mature phenotype, in terms of up-regulated surface markers (HLA-DR, CD86, CD83, CCR7) and enhanced cytokine production (IL6, IL10, and IL23), but had a reduced phagocytic capacity compared with non-treated monocyte-derived DC (Mo-DC). However, upon LPS activation, L. reuteri-CFS-generated DC displayed a more regulated phenotype than control Mo-DC with notable reduction of cytokine responses both at mRNA and protein levels. In contrast, S. aureus-CFS-generated DC were more similar to control Mo-DC both without and after LPS stimulation, but they were still able to induce responses in autologous T cells, in the absence of further T cell stimulation. Conclusions: We show that bacterial signals during DC differentiation have a profound impact on DC function and possibly also for shaping the T cell pool.
Language	English
Publishing date	2016-09
Publishing country	England
Document type	Journal Article
ISSN	2050-4527
ISSN	2050-4527
DOI	10.1002/iid3.115
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Article: Beneficial Effect of Mildly Pasteurized Whey Protein on Intestinal Integrity and Innate Defense in Preterm and Near-Term Piglets

Navis, Marit / Muncan, Vanesa / Sangild, Per Torp / Møller Willumsen, Line / Koelink, Pim J / Wildenberg, Manon E / Abrahamse, Evan / Thymann, Thomas / van Elburg, Ruurd M / Renes, Ingrid B

Nutrients. 2020 Apr. 17, v. 12, no. 4

2020

Abstract: Background. The human digestive tract is structurally mature at birth, yet maturation of gut functions such as digestion and mucosal barrier continues for the next 1–2 years. Human milk and infant milk formulas (IMF) seem to impact maturation of these ... ...

Abstract	Background. The human digestive tract is structurally mature at birth, yet maturation of gut functions such as digestion and mucosal barrier continues for the next 1–2 years. Human milk and infant milk formulas (IMF) seem to impact maturation of these gut functions differently, which is at least partially related to high temperature processing of IMF causing loss of bioactive proteins and formation of advanced glycation end products (AGEs). Both loss of protein bioactivity and formation of AGEs depend on heating temperature and time. The aim of this study was to investigate the impact of mildly pasteurized whey protein concentrate (MP-WPC) compared to extensively heated WPC (EH-WPC) on gut maturation in a piglet model hypersensitive to enteral nutrition. Methods. WPC was obtained by cold filtration and mildly pasteurized (73 °C, 30 s) or extensively heat treated (73 °C, 30 s + 80 °C, 6 min). Preterm (~90% gestation) and near-term piglets (~96% gestation) received enteral nutrition based on MP-WPC or EH-WPC for five days. Macroscopic and histologic lesions in the gastro-intestinal tract were evaluated and intestinal responses were further assessed by RT-qPCR, immunohistochemistry and enzyme activity analysis. Results. A diet based on MP-WPC limited epithelial intestinal damage and improved colonic integrity compared to EH-WPC. MP-WPC dampened colonic IL1-β, IL-8 and TNF-α expression and lowered T-cell influx in both preterm and near-term piglets. Anti-microbial defense as measured by neutrophil influx in the colon was only observed in near-term piglets, correlated with histological damage and was reduced by MP-WPC. Moreover, MP-WPC stimulated iALP activity in the colonic epithelium and increased differentiation into enteroendocrine cells compared to EH-WPC. Conclusions. Compared to extensively heated WPC, a formula based on mildly pasteurized WPC limits gut inflammation and stimulates gut maturation in preterm and near-term piglets and might therefore also be beneficial for preterm and (near) term infants.
Keywords	T-lymphocytes ; advanced glycation end products ; bioactive properties ; breast milk ; colon ; digestion ; digestive tract ; enteral feeding ; enzyme activity ; epithelium ; filtration ; humans ; immunohistochemistry ; infants ; inflammation ; interleukin-8 ; methodology ; milk ; models ; neutrophils ; nutrients ; pasteurization ; piglets ; pregnancy ; temperature ; whey protein concentrate
Language	English
Dates of publication	2020-0417
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
Note	NAL-light
ZDB-ID	2518386-2
ISSN	2072-6643
ISSN	2072-6643
DOI	10.3390/nu12041125
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: A Human 2D Primary Organoid-Derived Epithelial Monolayer Model to Study Host-Pathogen Interaction in the Small Intestine.

Roodsant, Thomas / Navis, Marit / Aknouch, Ikrame / Renes, Ingrid B / van Elburg, Ruurd M / Pajkrt, Dasja / Wolthers, Katja C / Schultsz, Constance / van der Ark, Kees C H / Sridhar, Adithya / Muncan, Vanesa

Frontiers in cellular and infection microbiology

2020 Volume 10, Page(s) 272

Abstract: Gut organoids are stem cell derived 3D models of the intestinal epithelium that are useful for studying interactions between enteric pathogens and their host. While the organoid model has been used for both bacterial and viral infections, this is a ... ...

Abstract	Gut organoids are stem cell derived 3D models of the intestinal epithelium that are useful for studying interactions between enteric pathogens and their host. While the organoid model has been used for both bacterial and viral infections, this is a closed system with the luminal side being inaccessible without microinjection or disruption of the organoid polarization. In order to overcome this and simplify their applicability for transepithelial studies, permeable membrane based monolayer approaches are needed. In this paper, we demonstrate a method for generating a monolayer model of the human fetal intestinal polarized epithelium that is fully characterized and validated. Proximal and distal small intestinal organoids were used to generate 2D monolayer cultures, which were characterized with respect to epithelial cell types, polarization, barrier function, and gene expression. In addition, viral replication and bacterial translocation after apical infection with enteric pathogens Enterovirus A71 and
MeSH term(s)	Animals ; Epithelial Cells ; Host-Pathogen Interactions ; Humans ; Intestinal Mucosa ; Intestine, Small ; Organoids
Language	English
Publishing date	2020-06-09
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2619676-1
ISSN	2235-2988 ; 2235-2988
ISSN (online)	2235-2988
ISSN	2235-2988
DOI	10.3389/fcimb.2020.00272
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Article ; Online: Beneficial Effect of Mildly Pasteurized Whey Protein on Intestinal Integrity and Innate Defense in Preterm and Near-Term Piglets.

Navis, Marit / Muncan, Vanesa / Sangild, Per Torp / Møller Willumsen, Line / Koelink, Pim J / Wildenberg, Manon E / Abrahamse, Evan / Thymann, Thomas / van Elburg, Ruurd M / Renes, Ingrid B

Nutrients

2020 Volume 12, Issue 4

Abstract: Background: The human digestive tract is structurally mature at birth, yet maturation of gut functions such as digestion and mucosal barrier continues for the next 1-2 years. Human milk and infant milk formulas (IMF) seem to impact maturation of these ... ...

Abstract	Background: The human digestive tract is structurally mature at birth, yet maturation of gut functions such as digestion and mucosal barrier continues for the next 1-2 years. Human milk and infant milk formulas (IMF) seem to impact maturation of these gut functions differently, which is at least partially related to high temperature processing of IMF causing loss of bioactive proteins and formation of advanced glycation end products (AGEs). Both loss of protein bioactivity and formation of AGEs depend on heating temperature and time. The aim of this study was to investigate the impact of mildly pasteurized whey protein concentrate (MP-WPC) compared to extensively heated WPC (EH-WPC) on gut maturation in a piglet model hypersensitive to enteral nutrition. Methods: WPC was obtained by cold filtration and mildly pasteurized (73 °C, 30 s) or extensively heat treated (73 °C, 30 s + 80 °C, 6 min). Preterm (~90% gestation) and near-term piglets (~96% gestation) received enteral nutrition based on MP-WPC or EH-WPC for five days. Macroscopic and histologic lesions in the gastro-intestinal tract were evaluated and intestinal responses were further assessed by RT-qPCR, immunohistochemistry and enzyme activity analysis. Results: A diet based on MP-WPC limited epithelial intestinal damage and improved colonic integrity compared to EH-WPC. MP-WPC dampened colonic IL1-β, IL-8 and TNF-α expression and lowered T-cell influx in both preterm and near-term piglets. Anti-microbial defense as measured by neutrophil influx in the colon was only observed in near-term piglets, correlated with histological damage and was reduced by MP-WPC. Moreover, MP-WPC stimulated iALP activity in the colonic epithelium and increased differentiation into enteroendocrine cells compared to EH-WPC. Conclusions: Compared to extensively heated WPC, a formula based on mildly pasteurized WPC limits gut inflammation and stimulates gut maturation in preterm and near-term piglets and might therefore also be beneficial for preterm and (near) term infants.
MeSH term(s)	Animal Nutritional Physiological Phenomena/physiology ; Animals ; Animals, Newborn ; Gastrointestinal Tract/growth & development ; Gastrointestinal Tract/immunology ; Gastrointestinal Tract/metabolism ; Gastrointestinal Tract/pathology ; Hot Temperature ; Interleukin-1beta/metabolism ; Interleukin-8/metabolism ; Neutrophil Infiltration ; Pasteurization/methods ; Premature Birth ; Swine/immunology ; Swine/metabolism ; Swine/physiology ; T-Lymphocytes/immunology ; Tumor Necrosis Factor-alpha/metabolism ; Whey Proteins/pharmacology
Chemical Substances	Interleukin-1beta ; Interleukin-8 ; Tumor Necrosis Factor-alpha ; Whey Proteins
Language	English
Publishing date	2020-04-17
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2518386-2
ISSN	2072-6643 ; 2072-6643
ISSN (online)	2072-6643
ISSN	2072-6643
DOI	10.3390/nu12041125
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Article ; Online: Mildly Pasteurized Whey Protein Promotes Gut Tolerance in Immature Piglets Compared with Extensively Heated Whey Protein.

Navis, Marit / Schwebel, Lauriane / Soendergaard Kappel, Susanne / Muncan, Vanesa / Sangild, Per Torp / Abrahamse, Evan / Aunsholt, Lise / Thymann, Thomas / van Elburg, Ruurd M / Renes, Ingrid B

Nutrients

2020 Volume 12, Issue 11

Abstract: Human milk is the optimal diet for infant development, but infant milk formula (IMF) must be available as an alternative. To develop high-quality IMF, bovine milk processing is required to ensure microbial safety and to obtain a protein composition that ... ...

Abstract	Human milk is the optimal diet for infant development, but infant milk formula (IMF) must be available as an alternative. To develop high-quality IMF, bovine milk processing is required to ensure microbial safety and to obtain a protein composition that mimics human milk. However, processing can impact the quality of milk proteins, which can influence gastro-intestinal (GI) tolerance by changing digestion, transit time and/or absorption. The aim of this study was to evaluate the impact of structural changes of proteins due to thermal processing on gastro-intestinal tolerance in the immature GI tract. Preterm and near-term piglets received enteral nutrition based on whey protein concentrate (WPC) either mildly pasteurized (MP-WPC) or extensively heated (EH-WPC). Clinical symptoms, transit time and gastric residuals were evaluated. In addition, protein coagulation and protein composition of coagulates formed during in vitro digestion were analyzed in more detail. Characterization of MP-WPC and EH-WPC revealed that mild pasteurization maintained protein nativity and reduced aggregation of β-lactoglobulin and α-lactalbumin, relative to EH-WPC. Mild pasteurization reduced the formation of coagulates during digestion, resulting in reduced gastric residual volume and increased intestinal tract content. In addition, preterm piglets receiving MP-WPC showed reduced mucosal bacterial adherence in the proximal small intestine. Finally, in vitro digestion studies revealed less protein coagulation and lower levels of β-lactoglobulin and α-lactalbumin in the coagulates of MP-WPC compared with EH-WPC. In conclusion, minimal heat treatment of WPC compared with extensive heating promoted GI tolerance in immature piglets, implying that minimal heated WPC could improve the GI tolerance of milk formulas in infants.
MeSH term(s)	Animals ; Bacterial Adhesion/drug effects ; Digestion ; Gastrointestinal Tract/drug effects ; Gastrointestinal Tract/immunology ; Gastrointestinal Tract/microbiology ; Gastrointestinal Transit/drug effects ; Gastrointestinal Transit/physiology ; Hot Temperature ; Hydrogen-Ion Concentration ; Immune Tolerance/drug effects ; Intestinal Mucosa/drug effects ; Intestinal Mucosa/microbiology ; Lysine/analogs & derivatives ; Lysine/metabolism ; Pasteurization ; Permeability ; Protein Aggregates/drug effects ; Swine ; Whey Proteins/pharmacology
Chemical Substances	Protein Aggregates ; Whey Proteins ; N(6)-carboxymethyllysine (70YDX3Z2O7) ; Lysine (K3Z4F929H6)
Language	English
Publishing date	2020-11-04
Publishing country	Switzerland
Document type	Comparative Study ; Journal Article
ZDB-ID	2518386-2
ISSN	2072-6643 ; 2072-6643
ISSN (online)	2072-6643
ISSN	2072-6643
DOI	10.3390/nu12113391
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Article: Corrigendum: Morphologically Distinct

Khan Mirzaei, Mohammadali / Haileselassie, Yeneneh / Navis, Marit / Cooper, Callum / Sverremark-Ekström, Eva / Nilsson, Anders S

Frontiers in microbiology

2017 Volume 7, Page(s) 2145

Abstract: This corrects the article on p. 437 in vol. 7, PMID: 27065990.]. ...

Abstract	[This corrects the article on p. 437 in vol. 7, PMID: 27065990.].
Language	English
Publishing date	2017-01-04
Publishing country	Switzerland
Document type	Published Erratum
ZDB-ID	2587354-4
ISSN	1664-302X
ISSN	1664-302X
DOI	10.3389/fmicb.2016.02145
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