Effect of Porosity on the Colonization of Digital Light-Processed 3D Hydrogel Constructs toward the Development of a Functional Intestinal Model.

With the rapid increase of the number of patients with gastrointestinal diseases in modern society, the need for the development of physiologically relevant in vitro intestinal models is key to improve the understanding of intestinal dysfunctions. This involves the development of a scaffold material exhibiting physiological stiffness and anatomical mimicry of the intestinal architecture. The current work focuses on evaluating the scaffold micromorphology of gelatin-methacryloyl-aminoethyl-methacrylate-based nonporous and porous intestinal 3D, intestine-like constructs, fabricated via digital light processing, on the cellular response. To this end, Caco-2 intestinal cells were utilized in combination with the constructs. Both porous and nonporous constructs promoted cell growth and differentiation toward enterocyte-like cells (VIL1, ALPI, SI, and OCLD expression showed via qPCR, ZO-1 via immunostaining). The porous constructs outperformed the nonporous ones regarding cell seeding efficiency and growth rate, confirmed by MTS assay, live/dead staining, and TEER measurements, due to the presence of surface roughness.

Destabilization of a model O/W/O double emulsion: From bulk to interface.

Oil-in-water-in-oil (O/W/O) double emulsions are considered an advanced oil-structuring technology that can accomplish multi-functions to improve food quality and nutrition. However, this special structure is thermodynamically unstable. This study formulated a model O/W/O double emulsion with standard surfactants, Tween 80 (4 %) and polyglycerol polyricinoleate (PGPR, 5 %), using a traditional two-step method with different homogenization parameters. Cryo-SEM and GC-FID results show that O/W/O emulsions were successfully formulated, and the release rate (RR) of medium-chain triglycerides (MCT) oil from the inner oil to the outer oil phase increased significantly with 2nd homogenization speed increasing, respectively. Interestingly, the RR of all samples reached about 75 % after 2 months of storage, suggesting that O/W/O emulsions were highly unstable. To explain the observed instability, dynamic interfacial tension and interfacial rheology were performed using a drop shape tensiometer. Results demonstrated that unadsorbed Tween 80 in the intermediate aqueous phase was a key factor in markedly decreasing the interfacial properties of the outer PGPR-assembled film by affecting the interfacial rearrangement. Additionally, it was found that the MCT release showed a positive correlation with the Tween 80 concentration, demonstrating that the formed Tween 80 micelles could transport oil molecules to strengthen the emulsion instability. Taken together, this study reveals the destabilization mechanism of model O/W/O surfactants-stabilized emulsions from bulk to interface, providing highly relevant insights for the design of stable O/W/O double emulsions.

Light-Based 3D Printing of Gelatin-Based Biomaterial Inks to Create a Physiologically Relevant In Vitro Fish Intestinal Model.

To provide prominent accessibility of fishmeal to the European population, the currently available, time- and cost-extensive feeding trials, which evaluate fish feed, should be replaced. The current paper reports on the development of a novel 3D culture platform, mimicking the microenvironment of the intestinal mucosa in vitro. The key requirements of the model include sufficient permeability for nutrients and medium-size marker molecules (equilibrium within 24 h), suitable mechanical properties (G' < 10 kPa), and close morphological similarity to the intestinal architecture. To enable processability with light-based 3D printing, a gelatin-methacryloyl-aminoethyl-methacrylate-based biomaterial ink is developed and combined with Tween 20 as porogen to ensure sufficient permeability. To assess the permeability properties of the hydrogels, a static diffusion setup is utilized, indicating that the hydrogel constructs are permeable for a medium size marker molecule (FITC-dextran 4 kg mol-1 ). Moreover, the mechanical evaluation through rheology evidence a physiologically relevant scaffold stiffness (G' = 4.83 ± 0.78 kPa). Digital light processing-based 3D printing of porogen-containing hydrogels results in the creation of constructs exhibiting a physiologically relevant microarchitecture as evidenced through cryo-scanning electron microscopy. Finally, the combination of the scaffolds with a novel rainbow trout (Oncorhynchus mykiss) intestinal epithelial cell line (RTdi-MI) evidence scaffold biocompatibility.