Influence of product properties on mastication and simulated gastric digestion
The development of digesta viscosity for the different products will depend on the progressive disintegration of the bolus and hydration, swelling and solubilisation of different components [
31]. The rapid initial decline in viscosity and the progressive decrease in frequency and amplitude of peaks was most likely the result of disintegration of the boluses. Although this does not give a direct measure of size or amount of larger fragments the longer duration of occurring peaks could indicate a slower disintegration of the boluses of uRCB and extR (Fig.
4). This could be related to more cohesive boluses resulting from the formation a connective starch phase during mastication, as observed for uRCB and extR (Fig.
4). The thinner lamella and more disrupted and gelatinised starch granules in uRCB and extR compared with sRB and RCB (Fig.
3) may have resulted in structures which were more easily hydrated by saliva during mastication. Plasticisation of the starch phase could give more flexible structures which were compacted during mastication, rather than fractured as appeared to be the case for sRB and RCB (Fig.
4). The continuous protein phase in WB may in a similar way have contributed to a more cohesive bolus compared with sRB and RCB. Refined wheat bread has also been reported to form larger particles after mastication than wholegrain and endosperm rye sourdough bread [
32]. The viscosity curve for sRB seemed to stabilise rapidly compared with the other products, including RCB. This could be related to a weaker structure which was more easily disintegrated. Fractures were also observed in the bolus fragments (Fig.
4). The formation of fractures may have been promoted by less swollen, amylose-surrounded starch granules in sRB. sRB also appeared more disintegrated than RCB after completed gastric digestion (Fig.
4), as reflected in the particle size distribution (Fig.
5). Peaks in the viscosity curve were also observed for the semi-solid RP, indicating presence of larger bolus fragments. As RP was used in a heated state, submersion of the bolus in the colder simulated gastric fluid most likely resulted in gelling of the continuous starch phase and solidification of the bolus which then disintegrated.
In the present work only one individual was used to chew the products and expectorate prior to swallowing (and transfer into the stomach of the dynamic model), since previous investigations have reported that the inter-individual variability of food bolus particle size is very limited, as is the effect of salivary α-amylase in relation to the action of pancreatic α-amylase [
26,
33,
34]. A limitation with mastication is that smaller particles may be lost by “intermediary swallowing” following dispersion in the saliva, and thereby not included in in vitro digestion [
35]. Further studies are needed to evaluate and identify key parameters involved in cereal starch digestion and to confirm that the use of one individual to chew is representative.
The observed differences in final viscosity between the products (Fig.
5) may partially relate to fibre composition. Lower molecular weight of β-glucan (Table
1) and disrupted cell walls (Fig.
2), as observed in sRB and RCB compared with uRCB, extR and RP, is an indication of fibre degradation. β-glucan is known to be degraded by endogenous enzymes, which become active with the increased moisture content during fermentation [
36]. Comparing uRCB with extR and RP, the time needed for mixing and baking of uRCB appeared sufficient for some degradation to occur. Although not analysed, molecular weight of arabinoxylans has been shown to be degraded similarly, but not to the same extent, as β-glucan [
37,
39]. Arabinoxylan-degrading enzymes are mainly active at higher temperatures and lower pH, around 4.5, than β-glucanases [
37,
39], and in uRCB, extR and RP arabinoxylan was probably relatively unaffected. In sRB and RCB; however, some degradation likely occurred. An increase in polydispersity and slight decrease in molecular weight has also been reported for fermented rye crispbreads compared with unfermented [
38]. Furthermore, due to lower dough pH, the use of sourdough in sRB might have promoted more extensive degradation of arabinoxylan.
The digesta viscosity is also influenced by the characteristics, e.g., size and shape, of the particles present, although the relationship is not well known [
31]. This would explain the relatively low digesta viscosity of extR, despite high extractability and retained molecular weight of β-glucan and high extractability of arabinoxylan, as it contained the smallest particles of the rye products (Fig.
5).
Viscosity and the rate of disintegration during gastric digestion can influence how rapidly starch becomes available for digestion in the small intestine. Solid foods are considered to be emptied from the gastric compartment first when reaching particle size < 1–2 mm [
40]. Furthermore, in mixed meals, liquids are preferentially emptied first [
41]. As boluses of cereal foods typically form more cohesive masses than e.g., vegetables [
42], the rate of disintegration is likely to be of importance for gastric emptying and consequently the postprandial responses in humans [
43]. High viscosity can also contribute to reduced gastric emptying [
44]. Furthermore, both content of soluble fibres and viscosity can influence the diffusion rates of enzymes and glucose [
8,
9,
45]. During digestion in vivo or in dynamic in vitro models, digesta viscosity will decrease due to continuous dilution by gastric and intestinal juices. However, differences in both viscosity and diffusion rates between products resulting from variations in particle characteristics and fibre composition may still persist.
Influence of product properties on glucose release in the TIM model
The differences in glucose release between products were not clearly reflected in the progression of starch digestion as observed in the digesta from the TIM model at different time points (Fig.
7). The model has previously been used to visualise differences in starch digestion [
17], but in that study a larger difference in total release of maltose was reported and both products tested, oat and barley tempeh, were relatively similar in structure. In our study a range of products with varying structures and properties were used, so different factors may have contributed to the concentration profiles of each product.
It is possible that slower gastric disintegration of uRCB and extR (Fig.
4) contributed to a later peak in glucose concentration, but similar AUC, compared with WB and RCB by decreasing the rate at which starch became available for digestion. No large bolus fragments (>2 mm) were observed in the intestinal compartments, and gastric sieving appears to occur in the TIM model too, indicating that gastric disintegration will be a factor to consider. Furthermore, lower diffusion rates due to less degraded fibres could decrease the rate of starch hydrolysis and, in the TIM model, reduce the rate of filtration/removal of digested compounds through the dialysis filters. This might also have contributed to the shift in glucose curves observed for uRCB and extR compared with WB and RCB. The highly disrupted starch in extR could have been expected to result in faster starch hydrolysis, but the gastric disintegration and diffusion rates may be more important factors. Despite similar glucose profiles in the TIM model, the responses to uRCB and extR may differ in humans, as the lower digesta viscosity of extR may result in faster gastric emptying rate [
39]. The difference in glucose profiles between RCB and uRCB is in line with results of a recent comparison of RCB and uRCB with a refined wheat crispbread, where RCB produced an insulin response more similar to that of the wheat reference than to uRCB [
5].
Differences in the glucose profiles for RCB and WB could have been expected considering the large differences in fibre content and composition, viscosity and particle size distribution. However, at equal viscosities, lower diffusion rates have been demonstrated for solutions of arabinoxylan from wheat compared with arabinoxylan from rye [
9]. Moreover, as observed in the TIM model, particles were initially of comparable sizes for WB and RCB (Fig.
7). While starch hydrolysis rate is often reported to be higher for refined wheat breads than rye breads, measurements are usually made after simulated gastric digestion [
4,
6] and the progressive changes in particle sizes occurring in vivo are not accounted for.
For RP, the large product volume, due to its high water content, may have contributed to its high glucose AUC. Compared with the other products, RP occupied a larger fraction of the volume in the gastric compartment and less liquid could be emptied before only the product remained, which should result in faster emptying of product. In humans this may compensate for slower gastric emptying due to high digesta viscosity [
44]. Moreover, due to the high water content during preparation and the temperature of the product when initiating in vitro digestion, starch may have been more gelatinised and easily hydrolysed. The high AUC for RP compared with sRB is also in line with findings in a human study by Rosén et al. [
7], where endosperm rye porridge and whole grain rye porridge induced higher glucose and insulin responses during the first 30 min than corresponding rye breads.
Neither of the explanations above can account for the low AUC
0–90min and late peak for sRB, with degraded fibres, low digesta viscosity and what appears to be the most rapidly disintegrated bolus of the rye products. However, the results are consistent with the lower insulin or lower insulin and glucose responses compared with refined wheat bread that have been repeatedly shown for soft rye breads, both sourdough-fermented and yeast-fermented [
4,
6]. Although lactic acid produced during fermentation has been suggested to inhibit starch hydrolysis, with increased interaction between starch and gluten as the mechanism [
46], the absence of a gluten network in sRB indicates other mechanisms. Rather, sRB was the only product where the presence of an amylose layer was observed, which has been suggested to inhibit starch hydrolysis in certain rye products [
4]. The high content of resistant starch in sRB compared with the other products (Table
1) is likly related to the observed amylose layer (Fig.
3). To our knowledge, the factors contributing to the formation of an amylose layer in certain rye products are not known. However, organic acids produced during sourdough fermentation have been suggested to increase the content of resistant starch and may be a contributing factor [
47]. Presence of an amylose layer may also be related to the high water content, compared with RCB, uRCB and extR, which could promote starch retrogradation during storage [
48]. Whether the amylose layer is a result of sourdough fermentation or not warrants further investigation.