Aug 2026· Adolescência e Saúde· Vol 21, pp. 689-697· 0 citations· 15 references
TL;DR
Overall, the developed solid-in-oil spray-dried ethylcellulose microcapsules represent a promising oral enzyme delivery platform with potential applications in enzyme replacement therapy for exocrine pancreatic insufficiency, phenylketonuria, and related metabolic disorders.
Abstract
Exocrine pancreatic insufficiency remains a major therapeutic challenge because orally administered enzymes undergo rapid degradation in the acidic gastric environment before reaching the intestine. The present study developed a scalable solid-in-oil (s/o) spray-drying technique for encapsulating protease within semi-permeable ethylcellulose (EC) microcapsules intended for targeted intestinal enzyme delivery. Pre-formulation studies identified a critical thermal deactivation threshold above 55°C, emphasizing the need for effective thermal stabilization during spray drying. Trehalose and sucrose were investigated as thermoprotective excipients, with 1.5 M trehalose demonstrating the highest protective effect by preserving 95.7% of the native enzyme activity following thermal exposure at 75°C. Circular dichroism spectroscopy confirmed the preservation of the native secondary structure in the stabilized formulations. The optimized microcapsules exhibited a production yield of 71.8%, an encapsulation efficiency of 85.3%, a median particle size (dv50) of 5.19 μm, and a narrow particle size distribution (span = 1.80), indicating excellent manufacturing reproducibility and particle uniformity. In vitro release studies demonstrated a biphasic release profile characterized by an initial burst release followed by sustained diffusion through the ethylcellulose membrane. Mathematical modeling revealed excellent agreement with the Higuchi model (R² = 0.9974), confirming that solute transport occurred predominantly through Fickian diffusion across the hydrated polymer matrix. The semi-permeable membrane effectively retained the encapsulated enzyme while allowing diffusion of low-molecular-weight substrates and reaction products. Overall, the developed solid-in-oil spray-dried ethylcellulose microcapsules represent a promising oral enzyme delivery platform with potential applications in enzyme replacement therapy for exocrine pancreatic insufficiency, phenylketonuria, and related metabolic disorders.
Collagen peptides (CPs) possess diverse biological activities, yet their oral efficacy is limited by gastrointestinal degradation and restricted systemic exposure of intact bioactive peptide species. Herein, a stable cholesterol-free nanoliposome system was developed using soybean phospholipids and stigmasterol through high-pressure microfluidization, followed by tangential flow filtration and spray drying to obtain a stable dry formulation. The optimized nanoliposomes exhibited a particle size below 100 nm, high peptide loading, excellent redispersibility, and remarkable physicochemical stability during refrigerated storage and under different pH and thermal conditions. During simulated gastrointestinal digestion, the stigmasterol-stabilized phospholipid bilayer effectively preserved encapsulated CPs throughout the gastric phase while facilitating peptide release under intestinal conditions. Oral administration in rats significantly enhanced collagen peptide bioavailability, increasing the plasma exposure (iAUC0–8 h) of total hydroxyproline by 3.84-fold compared with free CPs. Peptide-bound hydroxyproline exposure increased 9.3-fold and accounted for approximately 94% of total absorbed hydroxyproline. UHPLC–HRMS analysis confirmed substantially enhanced systemic exposure of multiple characteristic hydroxyproline-containing dipeptides and tripeptides following nanoliposomal delivery. These findings indicate that stigmasterol-containing nanoliposomes improve the gastrointestinal stability and systemic delivery performance of collagen peptides, providing a promising strategy for enhancing the oral delivery potential of food-derived bioactive peptides.
Developing dry antimicrobial delivery systems that combine bioactive protection with biofilm-matrix disruption remains a major challenge. Building on a previously established protease-modulated pea protein isolate (PPI)-carvacrol nanoemulsion system, this study developed protease-assisted microcapsules designed to preserve carvacrol and promote the removal of preformed biofilms. Carvacrol-loaded nanoemulsions were prepared at pH 3.5, 7.0, and 10.0 and supplemented with pepsin or trypsin before spray-drying or freeze-drying with maltodextrin. Protease-specific interfacial modification markedly affected emulsion stability, powder structure, and encapsulation performance. At pH 7.0 and 10.0, trypsin reduced droplet size from 284.60 to 232.52 nm and from 149.13 to 140.38 nm, respectively. The resulting spray-dried microcapsules exhibited high encapsulation efficiency (>96%) and low surface carvacrol contents (0.95–1.24 mg/g). In contrast, pepsin caused pronounced destabilization under acidic conditions, particularly after freeze-drying, yielding porous powders with an encapsulation efficiency of 48.18% and a surface carvacrol content of 69.51 mg/g. Trypsin-assisted microcapsules achieved 90–99% removal of preformed Listeria innocua biofilm biomass within 1 h, whereas enzyme-free and pepsin-containing formulations generally remained below 40%. Microscopic observations confirmed extensive disruption and detachment of the biofilm structure. After one year at 4 °C, trypsin-loaded formulations retained high biofilm biomass removal activity (85–99% after 2 h). These findings demonstrate that protease-assisted microencapsulation can couple interfacial regulation during particle formation with enzyme-mediated biofilm-matrix disruption after rehydration, providing a sustainable carvacrol delivery platform with durable antibiofilm functionality.
Jun Ji, N. Chihib, Géraldine Agusti et al.· Current Research in Food Sci...· 0 citations
Oral enzyme replacement therapy for lactase deficiency is limited by the rapid degradation of free β-galactosidase in the gastrointestinal tract. To overcome this, β-galactosidase was encapsulated into polyelectrolyte microcapsules (PMCs) via layer-by-layer assembly using MnCO3 sacrificial templates, with CaCO3-based PMCs as a reference. MnCO3-PMCs achieved 99.4% encapsulation efficiency and retained 85.8% of initial activity, significantly outperforming CaCO3-PMCs (86.0% and 29.7%). Under simulated gastric conditions (pH 2.0, pepsin), the free enzyme and CaCO3-PMCs were completely inactivated, whereas MnCO3-PMCs preserved ~86% activity. In simulated intestinal fluid, MnCO3-PMCs exhibited a 4.3-fold activity increase within the first hour and maintained a 2.5-fold enhancement after 70 h, while the free enzyme progressively inactivated. Furthermore, MnCO3-PMCs demonstrated superior storage stability, retaining 64% of initial activity after 90 days at 4 °C, compared with 25% for CaCO3-PMCs. Although immobilization increased the Michaelis constant, the shift was smaller for MnCO3-PMCs (8.9-fold) than for CaCO3-PMCs (15.2-fold). In conclusion, MnCO3-templated PMCs effectively protect β-galactosidase from gastrointestinal degradation, sustain prolonged catalytic activity, and offer excellent storage stability, highlighting their strong potential for improving oral enzyme replacement therapy in lactose intolerance.
Yuri S. Chebykin, A. Kim, Sergey A. Tikhonenko· Gels· 0 citations
Probiotic therapy provides clinical potential for systemic health, but its therapeutic efficacy is limited by low bioavailability in the harsh gastrointestinal environment. Natural polysaccharides, such as inulin, are promising candidates for bioactive delivery; however, they easily dissolve in water and rapidly break down under acidic conditions, limiting their function as protective materials. In this study, we applied chemical modification to create a stable, acid-resistant inulin shell to protect probiotics from the gastric environment and to improve their stability during gastrointestinal transit. We prepared acetylated inulin (In–Ac) through a controlled esterification process, verifying its molecular structure by 1H nuclear magnetic resonance (NMR), Fourier transform infrared spectroscopy, and X-ray diffraction. NMR analysis confirmed successful synthesis with a high degree of acetyl substitution (79.7% ± 3.0%). Using a coaxial electrospraying system, we encapsulated Lactobacillus reuteri within these In–Ac shells. An optimized coaxial electrospraying process using a 30% (w/v) shell solution produced well-defined spherical microparticles with an average diameter of 5.62 ± 2.13 µm. The In–Ac matrix exhibited good structural integrity under acidic conditions (pH 3.0), indicating resistance against acid-induced degradation. In HuH-6 and Caco-2 cell models, the material demonstrated excellent biocompatibility, with no significant toxicity at relevant concentrations; moreover, in an in vivo mouse model, the In–Ac matrix exhibited improved probiotic stability and delivery efficiency under physiological conditions. Overall, these findings reveal that In–Ac-based microparticles fabricated via electrospraying provide a stable, biocompatible delivery platform that effectively protects probiotics during gastrointestinal transit, offering a promising platform for enhanced intestinal delivery and probiotic protection.
Goose bone paste is an underutilised poultry-processing by-product and a potential source of taste-active peptides. A nominal 1–3 kDa peptide fraction (F2), operationally designated an umami peptide fraction by analogy with comparable bone-hydrolysate fractions reported in the literature, was isolated from a neutral-protease hydrolysate by sequential ultrafiltration and encapsulated in sodium alginate (SA) microcapsules using extrusion–dripping ionic gelation. Single-factor screening identified the following formulation conditions: 2.0% (w/v) SA, 2.5% (w/v) CaCl2, 0.3% (w/v) SE-15, a core-to-wall mass ratio of 0.3, and a preparation temperature of 50 °C. A verification batch prepared under these conditions gave an encapsulation efficiency of 75.44%, with the ±1.07% denoting the SD of three technical determinations from that batch. The dried microcapsules had a moisture content of 2.98 ± 0.21% and passable flowability. The mean particle diameter was 856 ± 52 μm, with a within-batch coefficient of variation of 5.56 ± 0.28%; a complete particle-size distribution was not recorded. In pepsin-free simplified simulated gastric fluid, the apparent release from the microcapsules rose from about 6% at 1 h to about 13% at 5 h, whereas the apparent detection ratio of free F2 rose from about 56% to about 99%. The calculated concentrations fell at or below the validated limit of quantification, the microcapsule-group absorbances lay near the photometric floor of the instrument, and only three sampling times were used. These percentages and the kinetic fits are therefore qualitative to semi-quantitative. The data support only the relative statement that alginate encapsulation lowered the apparent release of F2 under the tested acidic conditions. They do not establish an exact release rate, an error estimate for the microcapsule group, or a specific release mechanism.
The development of plant protein-based delivery systems is often limited by poor stability and low retention efficiency under gastrointestinal conditions. This study investigated how pH (4 and 7) during external gelation influences the physicochemical properties, entrapment efficiency (EE), and in vitro gastrointestinal behaviour of alginate beads loaded with hemp protein concentrate (HPC), pea protein concentrate (PPC), or soy protein isolate (SPI). Zeta potential and Fourier transform infrared (FTIR) analyses suggested that at pH 4, the charge profiles of plant proteins favoured electrostatic association with anionic alginate, which was associated with higher EE values, with HPC achieving the maximum of 89.5% at pH 4. Conversely, at pH 7, electrostatic repulsion between biopolymers was associated with reduced EE. During in vitro digestion, beads formulated at pH 4 exhibited greater protein release than those prepared at pH 7, consistent with the expansion of the polymeric network under intestinal conditions. Size exclusion chromatography showed that released proteins underwent extensive proteolysis, generating low-molecular-weight fractions smaller than 300 Da. These results indicate that gelation pH is a relevant processing parameter for modulating protein retention and the extent of protein release of simulated digestion in alginate-based systems, with potential applications in protein-enriched food formulations.
Juan Cumilaf, Ever Hernández-Olivas, André Brodkorb et al.· Gels· 0 citations