Probiotic Delivery Research Articles

Optimizing the viability of microencapsulated Lactiplantibacillus plantarum using response surface methodology for dietary probiotic delivery

The study aimed to develop a microencapsulated probiotic (Lactiplantibacillus plantarum) delivery method, cryoprotected by chitosan-inulin complex microcapsules and freeze-dried to obtain maximum cell viability. Probiotic viability was assessed using simulated gastrointestinal conditions and storage at different temperatures. The L. plantarum microcapsules were developed using a Box-Behnken experimental design with varied concentrations of CaCl2, chitosan, and Inulin as an encapsulant agents. In addition, the current study used the response surface approach to explore the influence of various formulations and storage capsulate viabilities to verify an optimized L plantarum formulation. Under optimum conditions, small microbeads with the highest cell encapsulation efficiency were produced and the highest cell viability (68.33 %) was obtained. The results showed that the intelligent microcapsule of L. plantarum with SM significantly increased the viability of cells (p < 0.05) (approx. 108 log CFU/mL) and the release of the extract (85 %) in gastric simulation and intestinal simulation fluids, respectively when compared with free bacteria. In general, microencapsulated L. plantarum under previously optimized freeze-drying conditions showed promising efficiency that depends on cell viability during drying, gastrointestinal treatment, and refrigerated storage conditions and on the physicochemical properties of the powders.

Preparation and characterization of pectin-alginate-based microbeads reinforced by nano montmorillonite filler for probiotics encapsulation: Improving viability and colonic colonization

Hydrogel microbeads can be used to enhance the stability of probiotics during gastrointestinal delivery and storage. In this study, the pectin-alginate hydrogel was enhanced by adding montmorillonite filler to produce microbeads for encapsulating Lactobacillus kefiranofaciens (LK). Results showed that the viscosity of biopolymer solutions with 1 % (PAMT1) and 3 % (PAMT3) montmorillonite addition was suitable for producing regular-shaped microbeads. A layered cross-linked network was formed on the surface of PAMT3 microbeads through electrostatic interaction between pectin-alginate and montmorillonite filler, and the surrounding LK with adsorbed montmorillonite was encapsulated inside the microbeads. PAMT3 microbeads reduced the loss of viability of LK when passing through the gastric acid environment, and facilitated the slow release of LK in the intestine and colonic colonization. The maximum decrease in viability among all filler groups was 1.21 log CFU/g after two weeks of storage, while PAMT3 freeze-drying microbeads only decreased by 0.46 log CFU/g, indicating that the gel layer synergized with the adsorbed layer to provide dual protection for probiotics. Therefore, filler-reinforced microbeads are a promising bulk encapsulation carrier with great potential for the protection and delivery of probiotics and can be developed as food additives for dairy products.

Encapsulation techniques, action mechanisms, and evaluation models of probiotics: Recent advances and future prospects

AbstractThe related studies of probiotics‐based functional foods have recently attracted increasing interests in developing new types of oral administration systems. Probiotics has various biological functions, including increasing adsorption ability of nutrients, competitively rejecting pathogenic organisms, and regulating the immune system. It is well known that probiotics cannot be worked until their colonies reach the intestine alive, as well as the amount reaches 106–107 colony‐forming units/g at the end of product's shelf life. However, the delivery of probiotics to the colon through oral administration is always challenging due to weak viability under exposure to harsh conditions in the presence of ions and small molecules from the upper gastrointestinal tract. Moreover, the low viability of the probiotics due to the lack of resistance to the acidity of food matrices, high temperature during food processing, and oxygen changes during storage also limit the applications of probiotics in food products. Therefore, encapsulation of probiotics could particularly protect the probiotics from degradation and inactivation, resulting in enhanced viability during the transition from consumption to digestion in the gut. Emulsion cross‐linking, complex coacervation, microcapsules, spray drying, layer‐by‐layer self‐assembly, electrospinning, hydrogel, and other methods are used to encapsulate probiotics. Herein, this review mainly highlights the gut health functions and encapsulation techniques of probiotics, followed by the current challenges and future development prospects.

New Horizons in Probiotics: Unraveling the Potential of Edible Microbial Polysaccharides through In Vitro Digestion Models.

In vitro digestion models, as innovative assessment tools, possess advantages such as speed, high throughput, low cost, and high repeatability. They have been widely applied to the investigation of food digestion behavior and its potential impact on health. In recent years, research on edible polysaccharides in the field of intestinal health has been increasing. However, there is still a lack of systematic reviews on the application of microbial-derived edible polysaccharides in in vitro intestinal models. This review thoroughly discusses the limitations and challenges of static and dynamic in vitro digestion experiments, while providing an in-depth introduction to several typical in vitro digestion models. In light of this, we focus on the degradability of microbial polysaccharides and oligosaccharides, with a particular emphasis on edible microbial polysaccharides typically utilized in the food industry, such as xanthan gum and gellan gum, and their potential impacts on intestinal health. Through this review, a more comprehensive understanding of the latest developments in microbial polysaccharides, regarding probiotic delivery, immobilization, and probiotic potential, is expected, thus providing an expanded and deepened perspective for their application in functional foods.

Novel seamless shell-core microbead for probiotics encapsulation: Influence of gel structure on storage stability and gastrointestinal activity

Probiotics have poor stability in storage and gastrointestinal digestion, resulting in low biological activity. Therefore, we have innovatively designed a shell-core microbead that can place probiotics in an independent lipid environment as a core, with an outer layer coated with a shell film composed of gelatin and/or pectin. Addition of 3% pectin to the shell layer and cross-linking with calcium ions significantly improved the physicochemical properties of the shell film of the microbeads, including water vapor permeability (WVP), water contact angle (WCA), and gel characteristics. The microbeads have spherical structure, uniform size and low moisture content (<5%). The water solubility of gelatin microbeads (Gel-Mb, 69.62%), gelatin-pectin microbeads (Gel-Pec-Mb, 61.06%) and shell cross-linked microbeads (Cro-Pec-Mb, 52.97%) decreased gradually. Cryo-scanning electron microscopy (Cryo-SEM) showed an evident shell core structure, and an apparent gel network was observed in the surface layer of shell of Cro-Pec-Mb. SEM analysis also showed the same porous network structure on the surface of Cro-Pec-Mb. The probiotic activities of Gel-Mb, Gel-Pec-Mb and Cro-Pec-Mb were 6.87, 6.76 and 6.74 log CFU/g after 4 months storage at 4 °C, respectively. This indicates that microbead encapsulation has a high retention of probiotic activity due to the effective insulation of the lipid environment from oxygen, moisture, etc. Cro-Pec-Mb can maintain the integrity of the structure and resist the enzymatic hydrolysis of digestive enzymes, reducing probiotic inactivation during the gastrointestinal digestion, with only a decrease 1.85 log CFU/g. This study show that microbeads were a promising platform for intestinal probiotic delivery.

An Updated Comprehensive Overview of Different Food Applications of W1/O/W2 and O1/W/O2 Double Emulsions

Double emulsions (DEs) present promising applications as alternatives to conventional emulsions in the pharmaceutical, cosmetic, and food industries. However, most review articles have focused on the formulation, preparation approaches, physical stability, and release profile of encapsulants based on DEs, particularly water-in-oil-in-water (W1/O/W2), with less attention paid to specific food applications. Therefore, this review offers updated detailed research advances in potential food applications of both W1/O/W2 and oil-in-water-in-oil (O1/W/O2) DEs over the past decade. To this end, various food-relevant applications of DEs in the fortification; preservation (antioxidant and antimicrobial targets); encapsulation of enzymes; delivery and protection of probiotics; color stability; the masking of unpleasant tastes and odors; the development of healthy foods with low levels of fat, sugar, and salt; and design of novel edible packaging are discussed and their functional properties and release characteristics during storage and digestion are highlighted.

Review of the Potential of Probiotics in Disease Treatment: Mechanisms, Engineering, and Applications

Probiotics, living microorganisms with demonstrated health benefits when administered in sufficient quantities, have a rich history as dietary supplements to benefit human health. Recently, understanding of their mechanisms in the gastrointestinal tract has prompted exploration of probiotics in treating human diseases. However, the effective and precise delivery of probiotics remains a significant challenge in therapeutic applications. Here, we review the mechanisms of action of probiotics in human health and the most advanced strategies for efficient probiotic delivery. We also discuss the potential applications of engineered probiotics in disease treatment. This review contributes insights into the evolving landscape of probiotic research for therapeutic applications.

Responsively Degradable Nanoarmor-Assisted Super Resistance and Stable Colonization of Probiotics for Enhanced Inflammation-Targeted Delivery.

Manipulation of the gut microbiota using oral microecological preparations has shown great promise in treating various inflammatory disorders. However, delivering these preparations while maintaining their disease-site specificity, stability, and therapeutic efficacy is highly challenging due to the dynamic changes associated with pathological microenvironments in the gastrointestinal tract. Herein, a superior armored probiotic with an inflammation-targeting capacity is developed to enhance the efficacy and timely action of bacterial therapy against inflammatory bowel disease (IBD). The coating strategy exhibits suitability for diverse probiotic strains and has negligible influence on bacterial viability. This study demonstrates that these armored probiotics have ultraresistance to extreme intraluminal conditions and stable mucoadhesive capacity. Notably, the HA-functionalized nanoarmor equips the probiotics with inflamed-site targetability through multiple interactions, thus enhancing their efficacy in IBD therapy. Moreover, timely "awakening" of ingested probiotics through the responsive transferrin-directed degradation of the nanoarmor at the site of inflammation is highly beneficial for bacterial therapy, which requires the bacterial cells to be fully functional. Given its easy preparation and favorable biocompatibility, the developed single-cell coating approach provides an effective strategy for the advanced delivery of probiotics for biomedical applications at the cellular level.

Emerging Multiscale Biofabrication Approaches for Bacteriotherapy.

Bacteriotherapy is emerging as a strategic and effective approach to treat infections by providing putatively harmless bacteria (i.e., probiotics) as antagonists to pathogens. Proper delivery of probiotics or their metabolites (i.e., post-biotics) can facilitate their availing of biomaterial encapsulation via innovative manufacturing technologies. This review paper aims to provide the most recent biomaterial-assisted strategies proposed to treat infections or dysbiosis using bacteriotherapy. We revised the encapsulation processes across multiscale biomaterial approaches, which could be ideal for targeting different tissues and suit diverse therapeutic opportunities. Hydrogels, and specifically polysaccharides, are the focus of this review, as they have been reported to better sustain the vitality of the live cells incorporated. Specifically, the approaches used for fabricating hydrogel-based devices with increasing dimensionality (D)-namely, 0D (i.e., particles), 1D (i.e., fibers), 2D (i.e., fiber meshes), and 3D (i.e., scaffolds)-endowed with probiotics, were detailed by describing their advantages and challenges, along with a future overlook in the field. Electrospinning, electrospray, and 3D bioprinting were investigated as new biofabrication methods for probiotic encapsulation within multidimensional matrices. Finally, examples of biomaterial-based systems for cell and possibly post-biotic release were reported.

Synergistic interaction of nanoparticles and probiotic delivery: A review.

Nanotechnology is an expanding and new technology that prompts production with nanoparticle-based (1-100 nm) organic and inorganic materials. Such a tool has an imperative function in different sectors like bioengineering, pharmaceuticals, electronics, energy, nuclear energy, and fuel, and its applications are helpful for human, animal, plant, and environmental health. In exacting, the nanoparticles are synthesized by top-down and bottom-up approaches through different techniques such as chemical, physical, and biological progress. The characterization is vital and the confirmation of nanoparticle traits is done by various instrumentation analyses like UV-Vis spectrophotometry (UV-Vis), Fourier transform infrared spectroscopy, scanning electron microscope, transmission electron microscopy, X-ray diffraction, atomic force microscopy, annular dark-field imaging, and intracranial pressure. In addition, probiotics are friendly microbes which while administered in sufficient quantity confer health advantages to the host. Characterization investigation is much more significant to the identification of good probiotics. Similarly, haemolytic activity, acid and bile salt tolerance, autoaggregation, antimicrobial compound production, inhibition of pathogens, enhance the immune system, and more health-beneficial effects on the host. The synergistic effects of nanoparticles and probiotics combined delivery applications are still limited to food, feed, and biomedical applications. However, the mechanisms by which they interact with the immune system and gut microbiota in humans and animals are largely unclear. This review discusses current research advancements to fulfil research gaps and promote the successful improvement of human and animal health.

Microfluidics-Derived Microparticles with Prebiotics and Probiotics for Enhanced In Situ Colonization and Immunoregulation of Colitis.

Oral administration of probiotics orchestrates the balance between intestinal microbes and the immune response. However, effective delivery and in situ colonization are limited by the harsh environment of the gastrointestinal tract. Herein, we provide a microfluidics-derived encapsulation strategy to address this problem. A novel synergistic delivery system composed of EcN Nissle 1917 and prebiotics, including alginate sodium and inulin gel, for treating inflammatory bowel disease and colitis-associated colorectal cancer is proposed. We demonstrated that EcN@AN microparticles yielded promising gastrointestinal resistance for on-demand probiotic delivery and colon-retentive capability. EcN@AN microparticles efficiently ameliorated intestinal inflammation and modulated the gut microbiome in experimental colitis. Moreover, the prebiotic composition of EcN@AN enhanced the fermentation of relative short-chain fatty acid metabolites, a kind of postbiotics, to exert anti-inflammatory and tumor-suppressive effects in murine models. This microfluidcis-based approach for the coordinated delivery of probiotics and prebiotics may have broad implications for gastrointestinal bacteriotherapy applications.

Effect of transglutaminase on ovalbumin emulsion gels as carriers of encapsulated probiotic bacteria.

The use of emulsion gels to protect and deliver probiotics has become an important topic in the food industry. This study used transglutaminase (TGase) to regulate ovalbumin (OVA) to prepare a novel emulsion gel. The effects of OVA concentration and the addition of TGase on the microstructure, rheological properties, water-holding capacity, and stability of the emulsion gels were investigated. With the addition of TGase and the increasing OVA, the particle size of the emulsion gels decreased significantly (P < 0.05). The gels with TGase exhibited greater water holding, hardness, and chewiness to some extent by forming a more uniform and stable system. After simulated digestion, the survival rate of Bifidobacterium lactis embedded in OVA emulsion gels improved significantly in comparison with the oil-water mixture as a result of the protective effect of the emulsion gel encapsulation. By increasing the OVA content and adding TGase, the rheological characteristics, stability, and encapsulation capability of the OVA emulsion gel could be enhanced, providing a theoretical basis for the use of emulsion gels to construct probiotic delivery systems. © 2023 Society of Chemical Industry.

Fabrication of hyaluronic acid-inulin coated Enterococcus faecium for colon-targeted delivery to fight Fusobacterium nucleatum

The abundance of Fusobacterium nucleatum (F. nucleatum) is highly associated with the development and poor prognosis of colorectal cancer (CRC), which is regarded as a promising target for CRC. However, until now, the novel strategy to clear F. nucleatum in the colon and CRC has not been well proposed. Herein, a probiotic strain Enterococcus faecium (E. faecium, EF47) is verified to secrete various organic acids and bacteriocins to exert superior antimicrobial activity towards F. nucleatum. However, the oral delivery of EF47 is affected by the complex digestive tract environment, so we design the hyaluronic acid-inulin (HA-IN) coated EF47 for colon-targeted delivery to fight F. nucleatum. IN can protect EF47 from the harsh gastrointestinal tract environment and is degraded specifically in the colon, acting as prebiotics to further promote the proliferation of EF47. The exposed HA can also enhance the targeting effect to the tumor area via the interaction with the CD44 receptor on the tumor cells, which is confirmed to increase the adhesive ability in tumor tissues and inhibit the growth of F. nucleatum. Therefore, this colon-targeted delivery system provides a novel platform to realize high-activity and adhesive delivery of probiotics to assist the therapeutic efficiency of CRC.

Exploring the impact of thermal convective drying behaviours on intestinal-targeted delivery of viable probiotics encapsulated with Eudragit® L100-trehalose

Spray drying microencapsulation has emerged as a promising technique for producing highly active probiotic powders. The efficient delivery of live probiotics to their target site, particularly the intestinal tract, is crucial for their therapeutic effectiveness. This study aimed to investigate the intestinal-targeted delivery function of particles containing Lactobacillus rhamnosus GG, trehalose and Eudragit® L100 (L100) under various thermal convective drying conditions. The particle drying history, component migration within particle, and the microstructures of the final particles were thoroughly analysed. Our findings demonstrate that temperature and flow rate significantly influence the migration behaviours of trehalose and L100 within the droplets, subsequently impacting the substance distribution on the surface of the final particles. Notably, higher temperatures and flow rates promote the deposition of L100 on the particle surface, leading to enhanced intestinal-targeted delivery function. Cells encapsulated in spray-dried powders containing L100 and produced at 160 °C exhibited higher survival rate and bioaccessibility during simulated gastrointestinal digestion, compared to those obtained at 140 and 120 °C. This study provides insight into the thermal convective drying behaviours and intestinal-targeted delivery function of L100-Tre-LGG particles. It contributes to the field of probiotic delivery and holds significant potential for improving therapeutic outcomes in various applications.

Nanocarrier-mediated probiotic delivery: a systematic meta-analysis assessing the biological effects

Probiotics have gained a significant attention as a promising way to improve gut health and overall well-being. The increasing recognition of the potential health advantages associated with functional food products, leading to a specific emphasis on co-encapsulating probiotic bacteria and bioactive compounds within a unified matrix. To further explore this concept, a meta-analysis was performed to assess the effects of probiotics encapsulated in nanoparticles. A comprehensive meta-analysis was conducted, encompassing 10 papers published from 2017 to 2022, focusing on the encapsulation of probiotics within nanoparticles and their viability in various gastrointestinal conditions. The selection of these papers was based on their direct relevance to the research topic. Random-effect models were used to aggregate study-specific risk estimates. In the majority of studies, it was observed that nano-encapsulated nanoparticles showed improved viability over time compared to their free state counterparts. At various time intervals, the odds ratios (OR) with 95% confidence intervals (CI) were estimated using fixed and random effect models. At 0 min, the OR (95%CI) was 2.79 (2.79; 2.80) and 2.38 (2.14; 2.64) for. At 30 and 60 min observation was at similar rate of 2.23 (2.23; 2.24) and 2.05 (1.73; 2.43). However, at 90 min it was 1.39 (1.39; 1.39) and 1.66 (1.29; 2.14) and at 120 min 2.41 (2.41; 2.42) and 2.03 (1.63; 2.52). Overall evaluation of encapsulation revealed an improvement in probiotic bacterial viability in simulated the gastrointestinal environments.

Development of a functional Greek sheep yogurt incorporating a probiotic Lacticaseibacillus rhamnosus wild-type strain as adjunct starter culture

Greek yogurt is a fermented dairy product of high nutritional value that can be used as a matrix for the delivery of probiotics. The aim of this study was to develop a new probiotic Greek sheep yogurt with upgraded quality and functional characteristics. To do this, yogurt was manufactured by fermenting pasteurized milk with the commercial starter culture (Streptococcus thermophilus (ST), Lactobacillus bulgaricus (LB)) together with a probiotic Lacticaseibacillus rhamnosus (LR) wild-type strain (probiotic yogurt; PY). As a control, yogurt manufactured with only the starter culture (ST, LB) was used (conventional yogurt; CY) The survival of all three lactic acid bacteria (LAB) species (ST, LB, and LR) was monitored throughout the products’ shelf life (storage at 4 °C for 25 days), and also following exposure to a static in vitro digestion model (SIVDM). The population dynamics of total aerobic plate count (APC), Enterobacteriaceae, yeasts and molds grown in both yogurts were also determined. The total antioxidant activity (AA) of yogurts was comparatively determined using in parallel two different assays, whereas the Folin-Ciocalteu assay was used to determine their total phenolic content (TPC). At each sampling day, yogurts were also evaluated for their pH, titratable acidity (TA) and main sensory characteristics. The population of probiotic LR remained stable during the shelf life (and above 108 CFU/g). Yogurt starters (ST, LB) were not detected following SIVDM, whereas LR (in PY) presented a reduction of about only one log. The AA and TPC of PY were found significantly higher than that of CY (P < 0.05). At the end of storage (25th day), neither pH nor TA differed significantly between the two yogurt types, while no fungal growth was observed in the PY. Consumer sensory analysis did not reveal important differences between the two yogurt types during their shelf life. To sum up, the novel yogurt was able to deliver to consumers a high number of probiotic cells (>108 CFU/g), presented increased antioxidant power, had an expanded shelf life, and maintained its good sensory attributes.

Human in vivo assessment of the survival and germination of Heyndrickxia coagulans SNZ1969 spores delivered via gummy candies

Confectionary products hold promise as unconventional food carriers for probiotic microorganisms. This study explored the delivery of Heyndrickxia coagulans SNZ1969, a spore-forming probiotic, using gummy candies. In this study, we prepared gummy candies containing bacterial spores with a viable count that remained stable during a 24-month shelf-life period, meeting the label claim of at least one billion CFUs per serving (24 g). Then, we carried out an intervention trial involving 24 healthy adults who consumed one serving per day for two weeks followed by an additional two weeks of follow-up. Fecal samples were collected and analyzed with a protocol that allowed the viable counts of SNZ1969, both in spore and vegetative forms. The obtained results revealed that bacterial spores germinated in all volunteers. SNZ1969 persistence in the gut was monitored for two weeks after the end of gummy candy consumption, indicating its potential for prolonged colonization. These findings highlight the potential of unconventional food carriers for probiotic delivery and suggest that spore-forming probiotics can be metabolically active in the human intestine. These findings provide information for the development of food products containing spore-forming probiotics and their potential benefits in promoting gastrointestinal health.

Research Trends and Applications of Spray Drying Technology in Probiotics: Bibliometric Analysis

Spray drying is a key method for producing stable and viable probiotic powders for food and pharmaceutical applications. This study optimized spray drying parameters to maintain probiotic viability, including inlet temperature, feed rate, and protective agents. The findings revealed that microencapsulation with biopolymers, such as alginate and proteins, significantly enhances probiotic stability during spray drying. Recent innovations in encapsulating materials and combining spray drying with other preservation methods further improve probiotic efficacy. This research underscores the importance of refining spray drying techniques to enhance the effectiveness of probiotics. Future studies should optimize these methods and explore their applications in various food matrices to develop more effective functional foods and therapeutic formulations. Advancements in spray drying and encapsulation technologies offer promising opportunities for improving probiotic delivery in the food and pharmaceutical industries.

Microencapsulation and controlled release of Bacillus clausii through a novel non-digestible carbohydrate formulation as revolutionizing probiotic delivery

Probiotics have gained significant attention in recent years due to the growing awareness of physical health and well-being. However, maintaining high concentrations of probiotics throughout the product's shelf life and during the gastrointestinal tract is crucial for ensuring their health-promoting effects. After determining an optimal formulation through a fractional factorial model, this study optimizes probiotic Bacillus Clausii delivery through spray-drying microencapsulation using a novel maltodextrin-alginate-inulin (MDX-ALG-IN) formulation (optimized ratio: 7:2:1). Notably, this formulation exclusively comprises non-digestible carbohydrates, marking a novel approach in probiotic encapsulation. Achieving a high Product Yield (51.06 %) and Encapsulation Efficiency (80.53 %), the study employed SEM for morphological analysis, revealing an irregular form and extensive surface in dentations characteristic of maltodextrin involvement. With a low moisture content of 3.02 % (±0.23 %) and 90.52 % solubility, the powder displayed exceptional properties. Probiotic viability remained robust, surviving up to 60 % even after 180 days at 4 °C, 25 °C, and 37 °C. Thermal characterization unveiled microcapsule resilience, exhibiting a glass transition temperature (Tg) at 138.61 °C and a melting point of 177.28 °C. The study systematically addresses crucial aspects of microencapsulation, including formulation optimization, morphological characteristics, and powder properties. Notably, the MDX-ALG-IN microcapsules demonstrated stability in simulated gastrointestinal conditions, indicating potential application for supplements and complex food matrices. In summary, this research contributes to microencapsulation understanding, emphasizing the MDX-ALG-IN formulation's efficacy in preserving probiotic viability across production stages and simulated digestive processes.

Development of Pickering water-in-oil emulsions using a dual stabilization of candelilla wax and acylated EGCG derivatives to enhance the survival of probiotics (Lactobacillus plantarum) powder.

Probiotics have considerable interest due to their inseparable link to human health. However, probiotic products are seriously challenged during processing, preservation, and intake. Food-grade probiotic delivery systems need to be further explored as an effective way to enhance cell viability. In this study, water-in-oil (W/O) Pickering emulsions were fabricated by adding candelilla wax (CLW) as a network stabilizer based on acylated EGCG derivatives in the crystalline form as a Pickering stabilizer. The effects of acylated EGCG derivatives' concentration, CLW concentration, and oil phase volume fraction on the droplet size distribution, microstructure, and physical stability of Pickering emulsions were explored. The presence of CLW reduced the particle size and improved the physical stability of acylated EGCG-based emulsions, and the effect was more positive with increasing concentration. The protective effect of emulsions with different oil phase volume fractions on Lactobacillus plantarum during freeze-thaw cycles, storage, and gastrointestinal digestion was evaluated. The outer-phase physical barrier of W/O emulsions co-stabilized with acylated EGCG derivatives and CLW facilitated the sensitivity of probiotics to ice crystal growth, temperature changes, acidic environments, and digestive enzymes. The emulsions formulated with 40% oil phase volume fractions allowed Lactobacillus plantarum to survive up to 7.75 log CFU g-1 in the harsh gastrointestinal environment. The results offer promising strategies for applying W/O emulsion probiotic delivery systems in food processing, storage, and oral administration.