Protection Of Probiotics Research Articles

Lactiplantibacillus plantarum encapsulated by chitosan-alginate and soy protein isolate-reducing sugars conjugate for enhanced viability.

To investigate the protective effects of various wall materials on probiotics, two types of Lactiplantibacillus plantarum 90 (Lp90) microcapsules were prepared using sodium alginate and chitosan (Lp-AC), soy protein isolate (SPI) and reducing sugars conjugate (Lp -MRP) as wall materials, respectively. The physical properties, cell viability under different conditions and the application of the microcapsules were investigated. Results showed that the selected wall materials were safe to Lp90 and their simulated digestion products exhibited antioxidant activities and prebiotic properties. The encapsulation efficiencies of Lp-AC and Lp-MRP were above 80%. Both microcapsules significantly enhanced cell survival rates under various conditions including low pH, bile salts, thermal processing, mechanical force, storage, and gastrointestinal digestion, with Lp-MRP demonstrating superior protective effects. When incorporated into milk and orange juice and stored at 4°C for 28 d, the colony counts of beverages containing Lp90 microcapsules exceeded 6 Log CFU/mL, with minimal changes in total soluble solids. Lp-MRP exhibited higher cell viability and smaller viscosity changes at 25°C for 28 d. Therefore, the single-layer encapsulation using SPI and reducing sugars conjugate showed promise over traditional chitosan-alginate double-layer encapsulation concerning probiotic protection, targeted delivery, and application.

Zein and soy polysaccharide encapsulation enhances probiotic viability and modulates gut microbiota

Protein-polysaccharide systems show promise for probiotic encapsulation and delivery in food applications. This study investigated the formation of nanoparticles from zein and soluble soybean polysaccharide (SSPS) through antisolvent precipitation and their efficacy in encapsulating Bacillus subtilis. At pH 6.0, zein and SSPS formed composite aggregates (ZPS) that effectively encapsulated B. subtilis, as confirmed by scanning electron microscopy (SEM). Fourier-transform infrared spectroscopy (FTIR) analysis revealed the presence of hydrogen bonding and electrostatic interactions within the ZPS. The encapsulated probiotics exhibited significantly enhanced viability compared to free probiotics: 3.13-fold in simulated gastrointestinal digestion, 3.20-fold during pasteurization, and 1.50-fold in storage conditions. In vivo experiments in rats showed that oral administration of encapsulated probiotics increased the abundance of beneficial gut bacteria. These findings underscore the potential of zein-SSPS nanoparticles for probiotic protection and delivery, presenting a promising strategy for enhancing probiotic efficacy in food and nutraceutical applications.

Encapsulation of Lactiplantibacillus plantarum with casein-gellan gum emulsions to enhance its storage, pasteurization, and gastrointestinal survival

Probiotics serve a very important role in human health. However, probiotics have poor stability during processing, storage, and gastrointestinal digestion. The gellan gum (GG) is less susceptible to enzymatic degradation and resistant to thermal and acidic environments. This study investigated the effect of casein (CS)-GG emulsions to encapsulate Lactiplantibacillus plantarum CICC 6002 (L. plantarum CICC 6002) on its storage stability, thermal stability, and gastrointestinal digestion. L. plantarum CICC 6002 was suspended in palm oil and emulsions were prepared using CS or CS-GG complexes. We found the CS-GG emulsions improved the viability of L. plantarum CICC 6002 after storage, pasteurization, and digestion compared to the CS emulsions. In addition, we investigated the influence of the gellan gum concentration on emulsion stability, and the optimal stability was observed in the emulsion prepared by CS-0.8% GG complex. This study provided a new strategy for the protection of probiotics based on CS-GG delivery system.

Preparation of starch-based green nanofiber mats for probiotic encapsulation by electrospinning.

In this study, starch-based nanofiber mats were successfully prepared from aqueous solution by electrospinning and used for probiotic encapsulation for the first time. The physicochemical properties of the octenylsuccinated (OS) starch/poly(vinyl alcohol) (PVA) blend solutions were systematically investigated. Through Fourier transform infrared spectroscopy and X-ray diffraction spectra analysis, it was found that miscibility and hydrogen bonding interactions exist between OS starch and PVA molecules. Thermogravimetric analysis and derivative thermogravimetric analysis revealed that the produced nanofibers possess satisfactory thermal stability. Scanning electron microscopy images and diameter distribution histograms showed that continuous and defect-free nanofibers were obtained and along with the increase in the weight ratio of OS starch, the average diameter gradually decreased. In addition, it was confirmed that the probiotics were successfully encapsulated in nanofiber mats. The survival rates of Lactobacillus plantarum AB-1 and Lactobacillus rhamnosus GG encapsulated in nanofibers were as high as 94.63% and 92.42%, respectively, significantly higher than those of traditional freeze-drying. Moreover, compared to free cells, probiotics encapsulated in nanofiber mats retained better viability after 21 days of storage at 4 and 25°C, and showed remarkably higher survival rates after exposure to simulated gastric and intestinal fluid. This study showed that the developed nanofibers can be a promising encapsulation system for the protection of probiotics.

Imine-bond reinforced double interface in water-in-oil-in-water emulsions and application on Lactobacillus plantarum encapsulation

Water-in-oil-in-water (W/O/W) emulsions are kind of promising vehicles for delivering hydrophilic and hydrophobic components. In this study, the double interfaces of W/O/W emulsion was modified to improve their properties. Specifically, in situ dynamic imine chemistry was used to crosslink the interfaces in W/O/W emulsions. W/O/W emulsions were prepared that contained whey proteins in the inner and outer aqueous phases and polyglyceryl polyricinoleate (PGPR), a hydrophobic emulsifier, in the oil phase. Aldehyde oils were added to the oil phase of the emulsions to chemically react with the proteins in both the inner and outer water phases. The stability, interfacial property and permeability of W/OW emulsions were characterized using microscopy observation, tensiometer and diffusion extent determination. Then, the gastrointestinal protection and delivery of probiotics in W/O/W emulsion was determined. The results showed that the modified W/O/W emulsions possessed high storage stability. The diffusion of Rhodamine B and hydrogen ions between the internal and external aqueous phases was retarded after cinnamaldehyde interfacial modification. Atomic force microscopy images, interfacial rheology analysis and chemical composition analysis confirmed the interaction occurrence between the lipophilic aldehyde and whey protein. After the optimization of aldehyde oils, probiotics encapsulated in W/O/W emulsion exhibited enhanced cell viability during long-term storage and after going through gastric digestion site. Overall, our results provide useful information that may facilitate the design of W/O/W emulsions that can protect probiotics from harsh environments.

Double protection of probiotics in alginate hydrogel through emulsification incorporated with freeze drying and coaxial wet-electrospraying: Survivability and targeted delivery

Two probiotic strains were co-encapsulated in water in oil emulsion by two methods: (1) coaxial wet electrospraying and (2) freeze-drying methods. Optimization of the wet electrospray technique for maximum yield, minimum sphericity factor, and size resulted in an alginate concentration of 2.99% w/v, flow rates ratio of alginate to the emulsion of 4.81, and applied voltage of 10 kV. The cell viability of Lactobacillus plantarum PTCC 1896 was the same after both encapsulation techniques, while the freeze-drying method was more impressive (97.25%) than the coaxial electrospraying (86.46%) in maintaining the viability of Bifidobacterium animalis subsp. Lactis. Exposing electrospray-encapsulated probiotics to simulated gastrointestinal conditions for 4 h resulted in only a one logarithmic cycle decrease in the viability of both bacteria. The number of live cells remained at more than 108 CFU g−1. In contrast, the viability of freeze-dried probiotics was reduced to about 107 CFU g−1. The freeze-dried B. lactis was the most sensitive probiotic to simulated gastric conditions. Monitoring the viability of bacteria during storage at −18 °C for both wet electrospraying and freeze-drying methods showed that the number of living microorganisms was more than 6 Log CFU g−1 after 5 and 4 months, respectively.

Milk and Lacticaseibacillus paracasei BL23 effects on intestinal responses in a murine model of colitis.

Probiotic-containing fermented dairy foods have the potential to benefit human health, but the importance of the dairy matrix for efficacy remains unclear. We investigated the capacity of Lacticaseibacillus paracasei BL23 in phosphate-buffered saline (BL23-PBS), BL23-fermented milk (BL23-milk), and milk to modify intestinal and behavioral responses in a dextran sodium sulfate (DSS, 3% wt/vol) mouse model of colitis. Significant sex-dependent differences were found such that female mice exhibited more severe colitis, greater weight loss, and higher mortality rates. Sex differences were also found for ion transport ex vivo, colonic cytokine and tight junction gene expression, and fecal microbiota composition. Measurements of milk and BL23 effects showed BL23-PBS consumption improved weight recovery in females, whereas milk resulted in better body weight recovery in males. Occludin and Claudin-2 gene transcript levels indicated barrier function was impaired in males, but BL23-milk was still found to improve colonic ion transport in those mice. Proinflammatory and anti-inflammatory gene expression levels were increased in both male and female mice fed BL23, and to a more variable extent, milk, compared with controls. The female mouse fecal microbiota contained high proportions of Akkermansia (average of 18.1%) at baseline, and females exhibited more changes in gut microbiota composition following BL23 and milk intake. Male fecal microbiota harbored significantly more Parasutterella and less Blautia and Roseburia after DSS treatment, independent of BL23 or milk consumption. These findings show the complex interplay between dietary components and sex-dependent responses in mitigating inflammation in the digestive tract.NEW & NOTEWORTHY Sex-dependent responses to probiotic Lacticaseibacillus paracasei and milk and the potential of the dairy matrix to enhance probiotic protection against colitis in this context have not been previously explored. Female mice were more sensitive than males to colonic injury, and neither treatment effectively alleviated inflammation in both sexes. These sex-dependent responses may result from differences in the higher baseline proportions of Akkermansia in the gut microbiome of female mice.

Encapsulation of probiotic bacteria in pectin and pectin-chitosan matrices for use in confectionery products

Probiotics are live bacteria that benefit the host's health when administered in adequate quantities. However, their use may be limited due to a decrease in cell viability during production, product storage, and subsequent passage through the gastrointestinal tract. This work theoretically substantiates the use of a combined method of microcapsule formation – immobilization of probiotics into a gel and microencapsulation, which will protect microorganisms from the effects of technological and physiological factors, regulate their targeted delivery and controlled release from microcapsules at the site of deployment. Suitable carriers for coating the capsules were selected, which is crucial for ensuring adequate protection of probiotics since their properties determine the effectiveness of protecting microorganisms from harmful environmental factors and the ability to release them in the lower gastrointestinal tract. In the course of the research, microcapsules with pectin and pectin-chitosan matrices containing bifidobacteria Bifidobacterium bifidum-1 and lactobacilli Lactobacillus acidophilus Ep-317/402 were developed. The possibilities and feasibility of using the developed microcapsules with different matrices to protect cells from adverse gastrointestinal tract conditions in vitro were substantiated. The results showed that the survival rate of microencapsulated bifidobacteria in the gastric juice medium was 87 % in both matrices, in bile – in pectin 82 %, and pectin-chitosan 92 %. Survival of lactobacilli: in gastric juice – 89 % in the pectin matrix, 93 % in the pectin-chitosan matrix, in bile – 94 and 88 %, respectively. The obtained pectin and pectin-chitosan microcapsules can potentially be used as a means of delivering viable probiotic microorganisms to their location – the human large intestine and can be used in confectionery technology as ingredients to provide.

Natural Phenolic‐Metal Framework Strengthened Mesona Chinensis Polysaccharides Microgels for Improved Viability of Probiotics to Alleviate the Liver Injury and Gut Microbiota Dysbiosis

AbstractOral probiotics meet challenges during processing, storage, and gastrointestinal harsh conditions. Polysaccharide‐based hydrogels delivery system is promising in probiotic protection, but its semi‐solid and weak strength, and often requires additional cross‐links to solidify its structure. Herein a ferric ion co‐crosslinked microgel of Mesona chinensis polysaccharides and mechanically strengthened by phenolic‐metal frameworks of naturally bound brown bioactives is designed. These microgels has an intact structure in acidic condition and ruptured only at pH neutral conditions which can responsively release highly viable probiotics with high mucoadhesion and colonization. Brown bioactives are naturally bound to polysaccharides (MCPC) by hydrophobic interactions and hydrogen bonding, and they exhibited outstanding anti‐inflammatory and antioxidant properties. Moreover, these brown bioactives are the precise prebiotics which can specifically improve the abundance of Akkermansia genus in gut in vivo. Then, the probiotics Akkermansia muciniphila (AKK) are loaded into brown bioactives frameworks strengthened microgels. There is a more synergistic healthy benefit between MCPC components and loaded AKK compared with free and pasteurized AKK. MCPC microgels can deliver high viable AKK specifically to gut consequently modulating the microbiota balance, and protecting the intestinal barriers. MCPC microgels also delivered the antioxidant brown bioactives specifically into liver to alleviate the hepatic oxidative stress and inflammation.

Protection and delivery of probiotics by degraded fucoidan-chitosan nanogel-based W1/O/W2 double emulsion incorporated in self-assembled hydrogel particles

Probiotics as pharmaceutical and nutraceutical products are essential to human health, whereas maintaining their viability in adverse environments remains an obstacle. This study presents a well-designed W1/O/W2 emulsion system incorporated in calcium-alginate hydrogel particles for the targeted delivery and sustained release of Lactobacillus bulgaricus in the gastrointestinal tract. Firstly, the degraded fucoidan (DF) with the highest carboxyl content (2.99%), prepared using UV/H2O2 treatment, was more favorable for forming DF-CS nanogel through ionic cross-linking with chitosan (CS). The W1/O/W2 double emulsion fabricated by 0.4 mg/mL DF-CS nanogel possessed the highest interface stability, as concluded from visual appearance, confocal laser scanning microscope (CLSM), droplet size distribution, and ζ-potential results. The double emulsion loaded in 2% alginate hydrogel showed desirable textural properties and swelling behavior, thereby offering a high encapsulation efficiency of Lactobacillus bulgaricus (74.5%). Additionally, the hydrogel delivery system not only achieved sustained release of Lactobacillus bulgaricus in the simulated gastrointestinal condition (>107 CFU/g) but also maintained at least 71.3% and 55.1% survival of probiotics at −20 °C and 4 °C, respectively, after 28 d of long-term storage. Overall, these W1/O/W2 double emulsion-filled multilayer self-assembled hydrogel particles can be utilized for better protection and delivery of probiotics in the food and pharmaceutical industries.

Cover Image

The cover image is based on the Research Article Fabrication of dry S/O/W microcapsule and its probiotic protection against different stresses by Qianwan Guo et al., https://doi.org/10.1002/jsfa.13175.image

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.

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.

Chitosan entrapping of sodium alginate / Lycium barbarum polysaccharide gels for the encapsulation, protection and delivery of Lactiplantibacillus plantarum with enhanced viability

To preserve the viability of probiotics during digestion and storage, encapsulation techniques are necessary to withstand the challenges posed by adverse environments. A core-shell structure has been developed to provide protection for probiotics. By utilizing sodium alginate (SA) / Lycium barbarum polysaccharide (LBP) as the core material and chitosan (CS) as the shell, the probiotic load reached 9.676 log CFU/mL. This formulation not only facilitated continuous release in the gastrointestinal tract but also enhanced thermal stability and storage stability. The results obtained from Fourier transform infrared spectroscopy and thermogravimetric analysis confirmed that the addition of LBP and CS affected the microstructure of the gel by enhancing the hydrogen bond force, so as to achieve controlled release. Following the digestion of the gel within the gastrointestinal tract, the released amount was determined to be 9.657 log CFU/mL. The moisture content and storage stability tests confirmed that the encapsulated Lactiplantibacillus plantarum maintained good activity for an extended period at 4 °C, with an encapsulated count of 8.469 log CFU/mL on the 28th day. In conclusion, the newly developed core-shell gel in this study exhibits excellent probiotic protection and delivery capabilities.

The effect of aqueous extract of Arctium lappa root on the survival of Lactobacillus acidophilus La-5 and Bifidobacterium bifidum Bb-12 and sensorial and physicochemical properties ofsynbiotic yogurt.

The effect of aqueous extract of Arctium lappa root (ALE) on the survival of Lactobacillus acidophilus La-5 and Bifidobacterium bifidum Bb-12 probiotic bacteria and sensory and physicochemical properties of synbiotic yogurt was evaluated during 4 weeks storage at 4°C. According to this study, using 0.5% and 1% ALE significantly affected the survival of La-5 and Bb-12 during storage. The results showed that 1% of ALE counting of La-5 and Bb-12 has been reached from 6.96 and 8.14 Log CFU/g to 7.3 and 7.30 Log CFU/g after 28 days of storage. Moreover, adding 1% ALE to yogurt enhanced antioxidant activity and phenolic content to 1299.8 mg gallic acid/kg and 392.8 mg BHT eq./kg compared with the control (without extract) after storage, respectively. In general, in yogurt containing ALE, a decrease in Syneresis, undesirable changes in taste, texture, and appearance, and reduced overall acceptances were observed compared to the control. In conclusion, using this prebiotic compound (ALE) can improve nutritional properties and probiotic protection in yogurt during long time storage; thus, it is a good choice for application in the dairy industry.

Fabrication of dry S/O/W microcapsule and its probiotic protection against different stresses.

Encapsulation is commonly used to protect probiotics against harsh stresses. Thus, the fabrication of microcapsules with special structure is critical. In this work, microcapsules with the structure of S/O/W (solid-in-oil-in-water) emulsion were prepared for probiotics, with butterfat containing probiotics as the inner core and with whey protein isolate fibrils (WPIF) and antioxidants (epigallocatechin gallate, EGCG; glutathione, GSH) as the outer shell. Based on the high viscosity and good emulsifying ability of WPIF, dry well-dispersed microcapsules were successfully prepared via the stabilization of the butterfat emulsion during freeze-drying with 30-50 g L-1 WPIF. WPIF, WPIF + EGCG, and WPIF + GSH microcapsules with 50 g L-1 WPIF protected probiotics very well against different stresses and exhibited similar inactivation results, indicating that EGCG and GSH exerted neither harm or protection on probiotics. This significantly reduced the harmful effects of antioxidants on probiotics. Almost all the probiotics survived after pasteurization, which was critical for the use of probiotics in other foods. The inactivation values of probiotics in microcapsules were around 1 log in simulated gastric juice (SGJ), about 0.5 log in simulated intestinal juice (SIJ), and around 1 log after 40 days of ambient storage. Dry S/O/W microcapsule, with butterfat containing probiotics as the inner core and WPIF as the outer shell, significantly increased the resistance of probiotics to harsh environments. This work proposed a preparation method of dry S/O/W microcapsule with core/shell structure, which could be used in the encapsulation of probiotics and other bioactive ingredients.

Recent Advances in the Nanoshells Approach for Encapsulation of Single Probiotics.

The intestine, often referred to as the "second brain" of the human body, houses a vast microbial community that plays a crucial role in maintaining the host's balance and directly impacting overall health. Probiotics, a type of beneficial microorganism, offer various health benefits when consumed. However, probiotics face challenges such as acidic conditions in the stomach, bile acids, enzymes, and other adverse factors before they can colonize the intestinal tissues. At present, pills, dry powder, encapsulation, chemically modified bacteria, and genetically engineered bacteria have emerged as the preferred method for the stable and targeted delivery of probiotics. In particular, the use of nanoshells on the surface of single probiotics has shown promise in regulating their growth and differentiation. These nanoshells can detach from the probiotics' surface upon reaching the intestine, facilitating direct contact between the probiotics and intestinal mucosa. In this perspective, we provide an overview of the current developments in the formation of nanoshells mediated by single probiotics. We also discuss the advantages and disadvantages of different nanocoating strategies and explore future trends in probiotic protection.

Biomacromolecule based water-in-water Pickering emulsion: A fascinating artificial cell-like compartment for the encapsulation of Lactobacillus plantarum

Probiotics are considered to have the potential to be beneficial to health when consumed in adequate amounts. Maintaining excellent probiotic activity during food storage and gastrointestinal transport is a key challenge. In this study, a stable water-in-water (W/W) Pickering emulsion consisting of hydroxypropyl methylcellulose (HPMC) and dextran (DEX) was constructed. Cellulose nanocrystals (CNCs) were used as the solid stabilizers of the emulsion. The obtained emulsions had excellent long-term stability with no phase separation within 60 days. In addition, the pH and ionic strength had little influence on the stability performance of the emulsions. The W/W Pickering emulsion system was applied to the probiotic encapsulation, and the emulsion was microencapsulated by spray drying technique. The microscopy images showed that Lactobacillus plantarum tended to be enriched in the dispersed phase of W/W Pickering emulsions. The viable cell content of obtained microcapsules was 2.22 × 1010 CFU/g after spray drying. Furthermore, the viable cell content in emulsion microcapsules was 3.38 × 106 CFU/g after the simulated digestion process in artificial gastrointestinal fluid. The microcapsules containing Lactobacillus plantarum also showed excellent stability during the storage. The viable cell content of microcapsules was 1.76 × 109 CFU/g at 4 °C after 90 days of storage. This study could provide a potential method by using W/W Pickering emulsion as a system for encapsulating probiotics, which would provide protection for probiotics during spray drying and gastrointestinal digestion.

Characterization and application in yogurt of genipin-crosslinked chitosan microcapsules encapsulating with Lactiplantibacillus plantarum DMDL 9010

Microcapsules could improve the protection of probiotics in the lyophilization and gastrointestinal digestion process. The purpose of this study was to prepare Lactiplantibacillus plantarum DMDL 9010 (LP9010) microcapsules by cross-linking chitosan with genipin and to determine the encapsulation efficiency, morphological characterization, storage stability and the application of the microcapsules in fermentation. The results showed that the LP9010 microcapsules embedded in 1.00 wt% genipin cross-linked chitosan were in a uniform spherical shape with a smooth surface and satisfying agglomeration. The LP9010 microcapsules demonstrated the reasonable thermal stability and persistence of biological activity in the range of −20 °C to 25 °C. Additionally, yogurt obtained from the ST + LB + ELP9010 strain formulation with the addition of microencapsulated LP9010 had smaller particles, better taste, and better stability compared with the yogurt obtained from other strain formulations. As detected by GC–MS, the yogurt formulated with ST + LB + ELP9010 as a strain retained more flavor substances and the content of flavor substances was greater than that of the yogurt obtained from other strain formulations. Therefore, genipin cross-link chitosan could be a suitable microencapsulated material for producing yogurt fermentation strains.

A Screening Model for Probiotics Against Specific Metabolic Diseases Based on Caco-2 Monolayer Membrane

Recent studies have revealed the potency of probiotics in alleviating metabolic diseases associated with intestinal barrier dysfunction. However, an efficient model for screening probiotic strains against specific metabolic diseases has not been well developed. In the present study, a Caco-2 cell monolayer membrane model treated with tumor necrosis factor (TNF-α) or alcohol was used to evaluate the effect of 139 Lactobacillus strains on intestinal barrier function in vitro . We then selected 11 Lactobacillus strains with different regulatory abilities on the gut barrier to determine their effect against ovariectomy-induced osteoporosis or chronic alcoholic liver injury in vivo . Our results showed that the Pearson coefficient between the data of cell and animal models were 0.82 and −0.97 for the protection of probiotics against osteoporosis and alcoholic liver disease, respectively, suggesting the reliability of the cell model to simulate the in vivo protective effects of probiotics. This study established a potential in vitro approach based on a Caco-2 cell monolayer membrane model for the efficient screening of potential probiotics against specific metabolic diseases such as osteoporosis and chronic alcoholic liver disease.