Lipid emulsion interfacial design modulates human in vivo digestion and satiation hormone response.

Lipid emulsions (LEs) with tailored digestibility have the potential to modulate satiation or act as delivery systems for lipophilic nutrients and drugs. The digestion of LEs is governed by their interfacial emulsifier layer which determines their gastric structuring and accessibility for lipases. A plethora of LEs that potentially modulate digestion have been proposed in recent years, however, in vivo validations of altered LE digestion remain scarce. Here, we report on the in vivo digestion and satiation of three novel LEs stabilized by whey protein isolate (WPI), thermo-gelling methylcellulose (MC), or cellulose nanocrystals (CNCs) in comparison to an extensively studied surfactant-stabilized LE. LE digestion and satiation were determined in terms of gastric emptying, postprandial plasma hormone and metabolite levels characteristic for lipid digestion, perceived hunger/fullness sensations, and postprandial food intake. No major variations in gastric fat emptying were observed despite distinct gastric structuring of the LEs. The plasma satiation hormone and metabolite response was fastest and highest for WPI-stabilized LEs, indicating a limited capability of proteins to prevent lipolysis due to fast hydrolysis under gastric conditions and displacement by lipases. MC-stabilized LEs show a similar gastric structuring as surfactant-stabilized LEs but slightly reduced hormone and metabolite responses, suggesting that thermo-gelling MC prevents lipase adsorption more effectively. Ultimately, CNC-stabilized LEs showed a drastic reduction (>70%) in plasma hormone and metabolite responses. This confirms the efficiency of particle (Pickering) stabilized LEs to prevent lipolysis proposed in literature based on in vitro experiments. Subjects reported more hunger and less fullness after consumption of LEs stabilized with MC and CNCs which were able to limit satiation responses. We do not find evidence for the widely postulated ileal brake, i.e. that delivery of undigested nutrients to the ileum triggers increased satiation. On the contrary, we find decreased satiation for LEs that are able to delay lipolysis. No differences in food intake were observed 5 h after LE consumption. In conclusion, LE interfacial design modulates in vivo digestion and satiation response in humans. In particular, Pickering LEs show extraordinary capability to prevent lipolysis and qualify as oral delivery systems for lipophilic nutrients and drugs.

A Rat Model of Human Lipid Emulsion Digestion.

A better understanding of how dietary lipids are processed by the human body is necessary to allow for the control of satiation and energy intake by tailored lipid systems. To examine whether rats are a valid model of human dietary lipid processing and therefore useful for further mechanistic studies in this context, we tested in rats three lipid emulsions of different stability, which alter satiety responses in humans. Different sets of 15 adult male Sprague Dawley rats, equipped with gastric catheters alone or combined with hepatic portal vein (HPV) and vena cava (VC) catheters were maintained on a medium-fat diet and adapted to an 8 h deprivation/16 h feeding schedule. Experiments were performed in a randomized cross-over study design. After gastric infusion of the lipid emulsions, we assessed gastric emptying by the paracetamol absorption test and recorded in separate experiments food intake and plasma levels of gastrointestinal hormones and metabolites in the HPV. For an acid stable emulsion, slower gastric emptying and an enhanced release of satiating gastrointestinal (GI) hormones were observed and were associated with lower short-term energy intake in rats and less hunger in humans, respectively. The magnitude of hormonal responses was related to the acid stability and redispersibility of the emulsions and thus seems to depend on the availability of lipids for digestion. Plasma metabolite levels were unaffected by the emulsion induced changes in lipolysis. The results support that structured lipid systems are digested similarly in rats and humans. Thus unstable emulsions undergo the same intragastric destabilization in both species, i.e., increased droplet size and creaming. This work establishes the rat as a viable animal model for in vivo studies on the control of satiation and energy intake by tailored lipid systems.

Adsorption and Interfacial Layer Structure of Unmodified Nanocrystalline Cellulose at Air/Water Interfaces.

Nanocrystalline cellulose (NCC) is a promising biological nanoparticle for the stabilization of fluid interfaces.  However, the adsorption and interfacial layer structure of NCC are poorly understood as it is currently unknown how to form  NCC interfacial layers. Herein, we present parameters for the adsorption of unmodified NCC at the air-water (A/W) interface. Initial NCC adsorption is limited by diffusion, followed by monolayer saturation and decrease in surface tension at the time scale of hours. These results confirm the current hypothesis of a Pickering stabilization. NCC interfacial performance can be modulated by salt-induced charge screening, enhancing adsorption kinetics, surface load, and interfacial viscoelasticity. Adsorbed NCC layers were visualized by atomic force microscopy at planar Langmuir films and curved air bubbles, whereat NCC coverage was higher at curved interfaces. Structural analysis by neutron reflectometry revealed that NCC forms a discontinuous monolayer with crystallites oriented in the interfacial plane at a contact angle < 90°, favoring NCC desorption upon area compression. This provides the fundamental framework on the formation and structure of NCC layers at the A/W interface, paving the way for exploiting NCC interfacial stabilization for tailored colloidal materials.

Tailoring Emulsions for Controlled Lipid Release: Establishing in vitro-in Vivo Correlation for Digestion of Lipids.

The use of oil-in-water emulsions for controlled lipid release is of interest to the pharmaceutical industry in the development of poorly water soluble drugs and also has gained major interest in the treatment of obesity. In this study, we focus on the relevant in vitro parameters reflecting gastric and intestinal digestion steps to reach a reliable in vitro-in vivo correlation for lipid delivery systems. We found that (i) gastric lipolysis determines early lipid release and sensing. This was mainly influenced by the emulsion stabilization mechanism. (ii) Gastric mucin influences the structure of charge-stabilized emulsion systems in the stomach, leading to destabilization or gel formation, which is supported by in vivo magnetic resonance imaging in healthy volunteers. (iii) The precursor structures of these emulsions modulate intestinal lipolysis kinetics in vitro, which is reflected in plasma triglyceride and cholecystokinin concentrations in vivo.

Microfluidic Technique for the Simultaneous Quantification of Emulsion Instabilities and Lipid Digestion Kinetics.

Quantifying the impact of environmental physicochemical changes on the microstructure of lipid delivery systems is challenging. Therefore, we have developed a methodology to quantify the coalescence of oil-in-water emulsion droplets during lipid digestion in situ on a single droplet level. This technique involves a custom-made glass microfluidic platform, in which oil droplets can be trapped as single droplets, or several droplets per trap. The physicochemical environment can be controlled, and droplet digestion, as well as coalescence, can be visualized. We show that the exchange of the physicochemical conditions in the entire reaction chamber can be reached in under 30 s. Microparticle image velocimetry allowed mapping of the flow profile and demonstrated the tuneability of the shear profile in the device. The extraction of quantitative information regarding the physical characteristics of the droplets during digestion was performed using an automated image analysis throughout the digestion process. Therefore, we were able to show that oil-in-water emulsions stabilized by proteins coalesced under human gastric conditions. This coalescence delayed the overall lipid digestion kinetics. The droplets that coalesced during digestion were hydrolyzed 1.4 times slower than individually trapped droplets. Thus, the microstructural evolution of lipid delivery systems is a crucial factor in lipid digestion kinetics. This novel technique allows the simultaneous quantification of the impact that the physicochemical environment has on both the lipid droplet microstructure and the lipid release patterns.

Blocking Gastric Lipase Adsorption and Displacement Processes with Viscoelastic Biopolymer Adsorption Layers.

Delayed fat digestion might help to fight obesity. Fat digestion begins in the stomach by adsorption of gastric lipases to oil/water interfaces. In this study we show how biopolymer covered interfaces can act as a physical barrier for recombinant dog gastric lipase (rDGL) adsorption and thus gastric lipolysis. We used ß-lactoglobulin (ß-lg) and thermosensitive methylated nanocrystalline cellulose (metNCC) as model biopolymers to investigate the role of interfacial fluid dynamics and morphology for interfacial displacement processes by rDGL and polysorbate 20 (P20) under gastric conditions. Moreover, the influence of the combination of the flexible ß-lg and the elastic metNCC was studied. The interfaces were investigated combining interfacial techniques, such as pendant drop, interfacial shear and dilatational rheology, and neutron reflectometry. Displacement of biopolymer layers depended mainly on the fluid dynamics and thickness of the layers, both of which were drastically increased by the thermal induced gelation of metNCC at body temperature. Soft, thin ß-lg interfaces were almost fully displaced from the interface, whereas the composite ß-lg-metNCC layer thermogelled to a thick interfacial layer incorporating ß-lg as filler material and therefore resisted higher shear forces than a pure metNCC layer. Hence, with metNCC alone lipolysis by rDGL was inhibited, whereas the layer performance could be increased by the combination with ß-lg.

Simultaneous control of pH and ionic strength during interfacial rheology of ß-lactoglobulin fibrils adsorbed at liquid/liquid Interfaces.

Proteins can aggregate as amyloid fibrils under denaturing and destabilizing conditions such as low pH (2) and high temperature (90 °C). Fibrils of ß-lactoglobulin are surface active and form adsorption layers at fluid-fluid interfaces. In this study, ß-lactoglobulin fibrils were adsorbed at the oil-water interface at pH 2. A shear rheometer with a bicone geometry set up was modified to allow subphase exchange without disrupting the interface, enabling the investigation of rheological properties after adsorption of the fibrils, as a function of time, different pH, and ionic strength conditions. It is shown that an increase in pH (2 to 6) leads to an increase of both the interfacial storage and loss moduli. At the isoelectric point (pH 5-6) of ß-lactoglobulin fibrils, the maximum storage and loss moduli are reached. Beyond the isoelectric point, by further increasing the pH, a decrease in viscoelastic properties can be observed. Amplitude sweeps at different pH reveal a weak strain overshoot around the isoelectric point. With increasing ionic strength, the moduli increase without a strain overshoot. The method developed in this study allows in situ subphase exchange during interfacial rheological measurements and the investigation of interfacial ordering.