<figure><img src="image_1.png" /></figure>

# Micellar casein and sodium caseinate supramolecular structure differently impacts subsequent food intake in pigs☆

Sylvie Guerin<sup>a</sup>, Gwénaële Henry<sup>b</sup>, Laurence Le Normand<sup>a</sup>, Armelle Cahu<sup>a</sup>, Manon Hiolle<sup>c</sup>, Alain Baniel<sup>c</sup>, Didier Dupont<sup>b</sup>, Gaëlle Boudry<sup>a, *</sup>

## ARTICLE INFO

**Keywords:**

Casein

Satiety

Protein structure

Aminoacids

## ABSTRACT

Dietary protein reduces energy intake in following meals by signaling directly or indirectly to the brain. We recently observed differences in plasma amino acid kinetics and intra-gastric behavior between micellar casein (MC) and sodium caseinate (SC) in pigs, two factors that impact food intake. Our objective was to clarify whether the supramolecular structure of casein, given as a preload to pigs, impacts on subsequent food intake.

Overnight fasted pigs were allowed to consume casein drinks differing in casein macromolecular structure (SC vs MC) within 5 min in a cross-over study. *Ad libitum* intake of their regular feed was assessed during 1 h, either 1 or 4 h after casein drink ingestion. To evaluate the potential mechanisms at play, gastric emptying of the casein drinks radiolabeled with ⁹⁹Tc-colloïd was followed using gamma-scintigraphy while plasma kinetics of ghrelin, GLP-1, insulin and free amino acids were evaluated.

The amount of feed consumed 1, but not 4 h, after SC ingestion was lower than the amount of feed consumed after MC ingestion (P = 0.03). Gastric emptying parameters, plasma ghrelin, GLP-1 and insulin kinetics after both types of casein ingestion were not significantly different (P > 0.05). However, plasma free amino acid concentrations, known to reduce food intake, increased after both SC and MC ingestion but was greater after SC than MC ingestion from 60 to 120 min (P = 0.009).

In conclusion, casein supramolecular structure in a preload drink impacts differently subsequent energy intake, likely due to difference in amino acid bioavailability. Micellar casein exhibits less anorectic effect than sodium caseinate, a property that could benefit population with high protein need but low appetite such as elderly.

## 1. Introduction

Energy intake, energy metabolism, body weight and body composition changes are influenced by the macronutrient composition of the diet on the long-term (Jean et al., 2001). On the short term, the composition of a meal also impacts eating behavior during subsequent meals (Marmonier et al., 2000). In this context, dietary protein appears to be a strong appetite inhibitor as it reduced food intake in following meals, in both pre-clinical and clinical studies (Porrini, 1997; Trigazis et al., 1997). The reduction in food intake elicited by protein snack is beyond the one that could be accounted for by the protein energy

content of the snack alone (Porrini, 1997; Trigazis et al., 1997), suggesting that protein signals might directly or indirectly affect the brain to modulate eating behavior. Indeed, several peripheral and central pathways have been unraveled that may explain the satiating effect of protein (review by (Fromentin et al., 2012)). Protein and digestive products have been shown to be detected at the intestinal epithelium level, inducing the secretion of cholecystokinin (CCK) in the proximal part of the intestine and reducing food intake by a vagus nerve-mediated effect (Raybould, 1991). Further along the gastro-intestinal tract, protein and amino acids also activate the peptide YY (PYY)-mediated ileal brake (Maljaars et al., 2008) and stimulate the secretion of glucagon-like

<footnote><sup>a</sup> Institut Numecan, INSERM, INRAE, Univ Rennes, Rennes, France</footnote>

<footnote><sup>b</sup> INRAE, Institut Agro, STLO, 35042 Rennes, France</footnote>

<footnote><sup>c</sup> Ingredia SA, Arras, France</footnote>

<footnote>Abbreviations: MC, micellar casein; SC, sodium caseinate.</footnote>

<footnote>* This article is part of a special issue entitled: 'ICFD2024' published in Food Research International.</footnote>

<footnote>* Corresponding author.</footnote>

<footnote>E-mail address: Gaëlle.Boudry@inrae.fr (G. Boudry).</footnote>
peptide-1 (GLP-1) by entero-endocrine L-cells (Bowen et al., 2006). Protein also reduces ghrelin levels compared to a high carbohydrate breakfast in lean subjects (Blom et al., 2006). Beside these gut-located effects, amino acids also signal to specific brain areas, such as the hypothalamus, reducing food intake through an AMPK-mTOR mechanisms (Morrison et al., 2007). Noteworthy, among amino acids, leucine seems to be the most potent in modulating food behavior-related brain areas (Blouet et al., 2009).

Interestingly, the type of protein in snacks or pre-loads differently influences food intake. For example, a breakfast with α-lactalbumin and gelatin had a greater energy intake reducing effect on subsequent food intake 3 h later than casein, soy protein or whey (Veldhorst et al., 2009). Similarly, casein and pea protein pre-load reduced food intake 30 min after ingestion in young healthy males but not whey protein or albumin (Abou-Samra et al., 2011). Yet, beverages with beef- vs insect-derived protein (Dai et al., 2022) or rice-puddings prepared with animal (milk) protein or plant proteins (oat, pea and potato) (Douglas & Östman, 2018) did not modify ad libitum energy intake 3.5–5 h later in young men. Interestingly, in healthy young adult females, but not males, the casein to whey ratio of milk at breakfast modified food intake 2 h later, with lower food intake after a 40: 60 casein: whey protein ratio than a 80:20, irrespective of the protein content (Kung et al., 2018). Likewise, the greater satiating effect of whey protein than that of casein observed by Hall et al. (Hall et al., 2003) is in agreement with the ‘slow and fast proteins’ concept (Boirie et al., 1997), whereby the massive influx of amino acids following ingestion of whey proteins could lead to higher satiating effect compared to the slower release of amino acids with casein.

We recently demonstrated that this slow and fast protein concept also applies to protein differing by their structure due to differences in technological processes. In a study associating *in vitro*, pre-clinical (pig) and clinical investigations, we observed striking differences in plasma amino acid kinetics as well as intra-gastric behavior between micellar casein (MC) and sodium caseinate (SC), likely due to their intra-gastric clotting properties. We thus proposed the concept of slow (MC) and rapid (SC) casein depending on protein structure (Boulier et al., 2023). Based on these observed plasma amino acids profiles, MC should have less satiating effect on subsequent food intake than SC. However, we also observed that MC distributed in the whole gastric cavity compared to SC which was mainly located in the proximal stomach. Satiation feedback signals are also the results of gastric volumetric signals produced by mechanoreceptors (Powley & Phillips, 2004). Hence, such difference in intra-gastric distribution would advocate for more satiating signals with MC than SC. The objective of the present study was therefore to clarify whether the structure of casein also impacts its preload effect on subsequent food intake in the pig model.

## 2. Material and methods

### 2.1. Casein drinks

Casein drinks were prepared fresh every day by mixing 96 g of sodium caseinate (SC, 96 g of a 100% protein (100% casein) powder) or of micellar casein (MC, Prodiet 87B Fluid, Ingredia, 120 g of an 87 % protein (92% casein) powder) with 12 g of glucose and 800 mL of water kept at 50 °C for 45 min. under constant stirring (1250 rpm). These isoproteic casein drinks were then cooled to 30 °C for 15 min. Glucose was added to ensure that pigs eat the drinks. SC and MC differed by their supramolecular structure as already described (Broyard & Gaucheron, 2015; McMahon & Oommen, 2013) and exemplified in Fig. 1.

### 2.2. Animal experiments

Animal procedures were approved by the local animal experiment committee and the Ministry of Higher Education and Research and are described in Boulier et al. (Boulier et al., 2023). Briefly, Large White × Landrace × Pietrain female pigs (initial body weight 21.5 ± 1.0 kg) were housed individually in 100 × 150 cm stainless steel cages.

In a first experiment, 6 pigs were submitted to feed intake tests. For each animal, 2 casein drinks and 2 casein drink – feed meal intervals were tested on 4 separate days. On the morning of each experiment, overnight fasted pigs were offered one of the two casein drinks. They were then offered standard diet ad libitum either 1 h or 4 h after the casein drink. During 1 h, standard feed intake was measured every 15 min.

In a second experiment, 9 pigs were trained every day during 2 weeks to consume one of the two casein drinks within 5 min. while in quadrupedal position within a Pavlov stand and to stay still for 2 h in the stand in front of the gamma camera. They were surgically implanted a catheter as already described (Boulier et al., 2023). On the morning of the experiment, overnight fasted pigs were installed in the Pavlov stand and fed one of the two casein drinks radiolabeled with 25 MBq, 99mTc-colloid sulfur CK1 (CISBio International, Saclay, France). Data acquisition was performed using high-resolution low energy collimator and with a 64 × 64 pixel matrix as already described (Boulier et al., 2023). Emptying curves (expressed as percentage of isotope vs. time) were derived and fitted to a power exponential function (Elashoff et al., 1982) to calculate gastric half-emptying time (t½) and curve shape (β). Each animal received the 2 casein drinks on separate days with a minimum of 3 days between the two sessions. During gastric emptying sessions, blood samplings (10 mL) were performed through the intra-venous catheter 5 min. before and 15, 30, 45, 60, 90 and 120 min. after casein drink consumption. Blood was distributed in several tubes (8 mM EDTA for insulin, heparin-lithium for amino acid, aprotinin 250 KIU for

A

<figure><img src="image_2.png" /><figcaption>**Fig. 1.** Casein imaging by transmission electronic microscopy. Sodium caseinate (A) and micellar (B) were imaged by transmission electron microscopy at a x80,000 magnitude.</figcaption></figure>

B

<figure><img src="image_3.png" /></figure>
ghrelin and 4 mM EDTA and 10 µL/mL of anti-Dipeptyl peptidase 4 (Millipore, Molsheim) for GLP-1 assay). All tubes were immediately centrifuged (2500g, 10 min., 4 °C) and plasma stored at -20 °C for further analyses. Plasma for ghrelin assays were acidified (10 µL HCl 1 N and 5 µLPMSF/500 µL plasma) before storage at -20 °C.

## 2.3. Plasma analyses

Total ghrelin concentration was measured in plasma samples using dedicated radio-immunoassay kits (Phoenix pharmaceuticals). Active GLP-1 concentrations in plasma samples were determined using ELISA dedicated kit (Millipore) and insulinemia was determined using a dedicated immunoassay on an automated analyzer (ST AIA-PACK IRI methodology and AIA-1800 apparatus).

Plasma amino acid concentrations were determined with the Bio-chrom 30 Automatic Amino Acid Analyzer (Biochrom Ltd., Cambridge, UK) using lithium citrate buffer as eluent, with post-column derivati-zation with ninhydrine (EZ-Nin, Biochrom).

## 2.4. Statistical analysis

Data were analyzed by paired t-test (gastric emptying parameters, satiety tests) or by two-way ANOVA for kinetics (plasma amino acid and hormones). Data are presented as mean ± SEM. Difference between means were considered significant when P < 0.05.

# 3. Results

## 3.1. Feed intake 1 h but not 4 h after micellar casein ingestion is greater than after sodium caseinate ingestion

In order to evaluate the impact of casein structure on feed intake, fasted pigs were offered one of the two casein drinks differing in casein structure and feed intake was recorded when feed was offered either 1 or

4 h after casein ingestion. When the test was performed 1 h after casein drink ingestion, the amount of feed consumed after MC drink preload was 16 % greater compared to SC (Fig. 2A). Consistently, the speed of ingestion (i.e. amount of feed eaten per min calculated on 15-min intervals) decreased with time, irrespective of the casein preload but tended to be greater (P = 0.052) after the MC preload than the SC one (Fig. 2A). On the contrary, when the test was performed 4 h after casein drink ingestion, no difference in feed intake nor speed of ingestion was observed between the two casein drinks (Fig. 2B).

## 3.2. Gastric emptying of micellar casein did not differ from that of sodium caseinate

We evaluated gastric emptying of the two casein drinks by adding radiolabeled sulfur colloid to the casein drinks and evaluating radioac-tivity emptying from the pig stomach for 2 h (Fig. 3A). No difference in gastric emptying half time (Fig. 3B) nor in the shape of the emptying curve evaluated by the β parameters of the semi-exponential fitting curve (Fig. 3C) was observed between the two casein drinks.

## 3.3. Micellar casein ingestion elicited similar eating behavior-related hormone kinetics than sodium caseinate

We next investigated eating behavior hormones kinetic after the SC and MC meals. Plasma ghrelin decreased after casein meal ingestion (time effect P = 0.01) without difference between the two casein drinks (Fig. 4A). Plasma GLP-1 increased after both casein drinks (time effect P = 0.002), without difference between the two drinks (Fig. 4B). Insuli-nemia peaked 30 min. after casein drink ingestion (time effect P < 0.0001), irrespective of the casein meal (Fig. 4C), with no further dif-ference between the two casein drinks.

<figure><img src="image_4.png" /><figcaption>**Fig. 2.** Impact of casein structure in preload drinks on subsequent feed intake. Overnight fasted pigs consumed sodium caseinate (SC) or micellar casein (MC) preload drinks in a cross-over randomized design. 1 h-feed intake and speed of ingestion over 15-min periods was recorded 1 h (A) or 4 h (B) after preload drink ingestion. Each dot represents one pig. \* P < 0.05.</figcaption></figure>

<figure><img src="image_5.png" /></figure>
<figure><img src="image_6.png" /></figure>

<figure><img src="image_7.png" /><figcaption>**Fig. 3.** Impact of casein structure on its gastric emptying. Overnight faster pigs consumed radiolabeled either sodium caseinate (SC) or micellar casein (MC) preload drinks and stayed still for 2 h in front of a γ-camera that took picture every 2 min. to monitor intra-gastric radioactivity. Emptying curves expressed as percentage of the radioactivity in the stomach versus time (A) were fitted to a power exponential function to calculate half-emptying time (t½) (B) and curve shape (β). Data are means ± SEM, n = 9 pigs in a randomized cross-over design, with a minimum of 3 days between experiments.</figcaption></figure>

**3.4. Micellar casein ingestion induced lower aminoacidemia than sodium caseinate**

Finally, we evaluated plasma amino acid levels during the 2 h
following both casein drinks. The level of free amino acids (Fig. 5A), and
more particularly of leucin (Fig. 5B), was lower after MC than SC

ingestion.

4. Discussion

Our objective was to clarify whether casein structure, comparing micelles (MC) versus smaller aggregates (SC), impacts satiety. Indeed, our previous data in pigs indicated on one hand possible enhanced stomach distension signals with MC that would reduce appetite compared to SC and on the other hand, plasma amino acid profiles indicative of greater appetite after MC than SC consumption (Boulier et al., 2023). Hence, the present study clearly indicates that MC reduces satiety on the short-term (1 h after ingestion) but not on the longer term (4 h after ingestion). Moreover, the different investigations (gastric emptying, hormone release, plasma amino acid) advocate for a role of the slower release of amino acid, including leucin, into the bloodstream as a likely mechanism inducing reduced satiety after MC compared to SC preload.

Several studies investigated the impact of the nature of dietary proteins on satiety in human trials or in animal models (Abou-Samra et al., 2011; Dai et al., 2022; Dougkas & Östman, 2018; Hall et al., 2003; Kung et al., 2018; Veldhorst et al., 2009). These different dietary proteins differed in their amino acid contents as well as structures that conferred them different intra-gastric hydrolysis properties. Here, we used the same dietary protein, caseins, that only differed in structure due to differences in extraction technology. Hence, the amino acid composition of the two casein drinks was similar. In the same way, Juvonen et al. compared the satiating effect of sodium caseinate and transglutaminase-casein (TG-casein) in healthy men (Juvonen et al., 2011). Enzymatic cross-linking with transglutaminase changes the protein network, affecting the functional characteristics of protein, resulting in a firm and high-viscosity product compared to sodium caseinate. There was almost no difference between the two types of casein for appetite rating, except for a more prolonged fullness with TG-casein that lasted until 30 min after ingestion compared to sodium caseinate whose impact on fullness was lost after 15 min. Even if no *ad libitum* meal test were performed to evaluate if this translated into reduced food intake and despite the only slight differences between the

<figure><img src="image_8.png" /></figure>

<figure><img src="image_9.png" /><figcaption>**Fig. 4.** Post-prandial plasma hormone levels. Overnight fasted pigs consumed sodium caseinate (SC) or micellar casein (MC) preload drinks. Blood samples were collected before and during 2 h after ingestion to measure ghrelin (A), GLP-1 (B) and insulin (C) levels. Data are means ± SEM, n = 9 pigs in a randomized cross-over design.</figcaption></figure>

<figure><img src="image_10.png" /></figure>
A

<figure><img src="image_11.png" /></figure>

B

<figure><img src="image_12.png" /><figcaption>**Fig. 5.** Post-prandial plasma total amino acid levels. Overnight fasted pigs consumed sodium caseinate (SC) or micellar casein (MC) preload drinks. Blood samples were collected before and during 2 h after ingestion to measure free amino acid (AA) (A) and more particularly leucine (B) levels. Data are means ± SEM, n = 9 pigs in a randomized cross-over design. \* P < 0.05 compared to SC at the same timepoint.</figcaption></figure>

two casein-based preloads, this study aligns with our results, indicating that structure is also at play in the satiety effect of dietary proteins.

One mechanism that could account for by the difference in pig feed intake after MC or SC ingestion is the difference in plasma amino acid profiles, with a lower peak of plasma amino acids after MC than SC ingestion that we attributed to difference in intra-gastric behavior and a slower hydrolysis rate (Boulier et al., 2023). In humans, several studies indicated that the level of plasma amino acid inversely correlates with the level of hunger and energy intake, especially plasma leucine level. Hence, intra-duodenal infusions of leucine at different doses in healthy men elicited different plasma leucine levels and a reduction in energy intake (−13 %) of the subsequent *ad libitum* meal served 90 min after infusion with the highest tested dose (Steinert et al., 2015). Likewise, some individual plasma amino acid (valine, lysine, leucine and isoleucine) levels correlated with hunger rating and energy intake in a clinical trial testing the effect of preloads differing in protein content but also carbohydrate/fat ratio (Korompokis et al., 2016). Aligning with the only slight differences in appetite rating (Juvonen et al., 2011), similar post-prandial plasma amino acid profiles after caseinate or TG-casein ingestion were observed in healthy men (Juvonen et al., 2012). In pigs, oral gavage with individual amino acids impacts subsequent meal pattern. In particular, oral leucin increases inter-meal intervals compared to water gavage (Müller et al., 2022), confirming that our hypothesis is plausible. However, other studies also related no difference in hunger rating despite difference in plasma amino acid kinetic (Bolster et al., 2018; Marsset-Baglieri et al., 2015; Traylor et al., 2021). Interestingly, difference in appetite rating between different types of protein were observed in healthy men with protein preloads containing 10 % but not 25 % of protein (Veldhorst et al., 2009), suggesting that differences may only appear when certain amino acids are above and below particular thresholds. Hence, we speculate that, as demonstrated in Boulier et al (Boulier et al., 2023), the structure of MC confers MC intra-gastric clotting properties that elicit a low release of amino acids into the bloodstream compared to SC.This is further supported by the plasma amino acid profiles in the present study. These difference in amino acid release may prevent the amino acid level to reach the threshold needed to induce satiety. However, this would need to be confirmed by further investigations. Besides individual amino-acids, difference in the quantity or type of peptides arising from casein hydrolysis could also be at play in the difference in satiating effect of the two types of casein (Auestad & Layman, 2021). Indeed, *in vitro* static digestion data indicate that the technical process applied to dairy proteins can modify the peptides released, both quantitatively and qualitatively (Atallah et al., 2020). Similarly, the nature of egg protein aggregate can modify the

nature of the released peptides (Nyemb et al., 2014). However, peptidomic investigation of pig intestinal contents was out of the scope of the present study.

Induction of secretion of gastro-intestinal peptides into the bloodstream by intra-intestinal amino acids upon the digestion process has also been proposed as a mechanism of dietary protein-induced satiety (Fromentin et al., 2012). This includes cysteine, which has been shown to suppress the hunger hormone ghrelin (McGavigan et al., 2015), tryptophan, which has an appetite-suppressing effect via stimulation of the satiety peptide CCK (Hajishafiee et al., 2021) and phenylalanine which is a stimulator of both CCK and PYY (Fitzgerald et al., 2020). In the present study, plasma insulin and GLP-1 levels increased, while ghrelin level decreased after both casein drink ingestion, confirming that amino acids induce gastro-intestinal peptide release in pigs (Müller et al., 2024). However, no difference between casein structures was observed. Conversely, post-prandial CCK, GLP-1 and insulin plasma profiles after TG-casein consumption were markedly different from that after sodium caseinate consumption in humans, despite no striking impact on appetite rating (Juvonen et al., 2011). Interestingly, dose-effects of amino acids on CCK and PYY release by phenylalanine (Fitzgerald et al., 2020) or ghrelin decrease by cysteine (McGavigan et al., 2015) have been described. Hence, the threshold that elicits hormone release by amino acids might have been reached with both casein drinks in our study, resulting in similar post-prandial profiles after casein ingestion.

The lower satiating effect of MC compared to SC could be interesting in population with high protein need but low appetite such as elderly. Indeed, a high-protein intake of 1-1.2 g·kg⁻¹ is recommended for older adults for the maintenance of muscle mass and strength. However, a *meta*-analysis of randomized controlled trials revealed that protein preload acutely reduces test meal energy intake in adults ≥60 years (Ben-Harchache et al., 2021). This suggests that strategies to enhance protein intake without inducing satiety are needed for this population. In the studies included in the *meta*-analysis, meal tests were performed immediately or between 1 and 3 h after the protein supplement consumption, with no effect of the timing between protein ingestion and test meal observed (Ben-Harchache et al., 2021). In our study, we tested only two time-points, 1 and 4 h. No difference between MC and SC were observed 4 h after casein drink ingestion. Yet, unlike the timepoint 1 h, no difference in plasma amino acid levels was observed 4 h after MC or SC ingestion in pigs (Boulier et al., 2023). This could explain the absence of effect of MC on appetite at 4 h.

Our study provides evidence of the impact of the dietary protein structure on protein-induced satiety. Yet we must acknowledge several
limits to our study. First, casein preloads were performed with a large quantity of protein (87 g of protein, representing 4 g.kg⁻¹ of body weight ingested in 5 min.) while protein needs are estimated around 160–170 g per day in pigs at that stage (Reeds et al., 1980). Our study was a proof-of-concept study and we made the choice of testing a large quantity of protein to maximize the chance of detecting differences between casein types. Hence, demonstration of a differential effect upon feed intake with lower protein preloads is necessary to be closer to human nutrition reality. Moreover, except for glucose addition to promote ingestion by pigs, no other ingredients than casein were incorporated in the casein drinks. Evaluation of the impact on satiety of MC and SC incorporated in real complex foods should be the next step. Indeed, while caseinate and TG-casein impact on appetite rating was barely different when tested alone (Juvonen et al., 2011), a differential impact on satiety was observed when they were used to prepare emulsions (Juvonen et al., 2015). A third limit of our study is the age of our pigs, since we performed experiments with young adolescent pigs, for which protein metabolism is different from that of older animals (Reeds et al., 1980) and mechanisms of protein-induced satiety also likely different. Finally, and although no difference in feed intake was observed 4 h after casein ingestion, we did not evaluate hormone nor amino acid plasma levels later than 2 h after casein ingestion and we cannot exclude that difference between the two types of caseins may have appeared after 2 h between the two types of casein preloads.

In conclusion, we demonstrated in pigs that the structure of casein impacts its satiety properties, with micellar casein eliciting less short-term satiety than sodium caseinate. This effect is likely due to the intra-gastric behavior of micellar casein that clotted, resulting in slow hydrolysis and a lower peak of plasma amino-acid and less central effect of amino acids.

### CRediT authorship contribution statement

**Sylvie Guerin:** Writing – review & editing, Investigation, Formal analysis. **Gwènaële Henry:** Writing – review & editing, Investigation, Formal analysis. **Laurence Le Normand:** Writing – review & editing, Formal analysis. **Armelle Cahu:** Writing – review & editing, Formal analysis. **Manon Hiolle:** Writing – review & editing, Conceptualization. **Alain Baniel:** Writing – review & editing, Conceptualization. **Didier Dupont:** Writing – review & editing, Supervision, Investigation, Funding acquisition, Conceptualization. **Gaëlle Boudry:** Writing – original draft, Supervision, Investigation, Funding acquisition, Conceptualization.

### Declaration of competing interest

The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: MH and AB are employees of Ingredia SA. SG, GH, LLN, AC, DD, GB have no conflict of interest to declare.

### Acknowledgment

The authors wish to thank Dr Charles-Henri Malbert from AniScan facility (St-Gilles, France) for his help with the gamma-scintigraphy experiments and Alain Chauvin, Julien Georges and Francis Le Gouevec (UE3P, St-Gilles, France) for taking care of the animals.

### Funding

All the experimental work was funded by Ingredia SA.

### Data sharing

Data described in the manuscript will be made available upon reasonable request to the corresponding author.

### Ethics

All animal experiments should comply with ARRIVE guidelines.

## Appendix A. Supplementary data

Supplementary data to this article can be found online at https://doi.org/10.1016/j.foodres.2024.115465.

### Data availability

Data will be made available on request.

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