Bacterial Cellulose Nanofibrils Research Articles

A 3D Cell-Culture System That Uses Nano-Fibrillated Bacterial Cellulose to Prepare a Spherical Formulation of Culture Cells

A 3-dimensional (3D) cell culture is now being actively pursued to accomplish the in vivo-like cellular morphology and biological functions in cell culture. We recently obtained nano-fibrillated bacterial cellulose (NFBC). In this study, we developed a novel NFBC-based 3D cell-culture system, the OnGel method, and the Suspension method. HepG2 human liver cancer cells were cultured via these methods and formed spherical formulations in the optimized condition, 1.0% (w/v) of NFBC in the OnGel method, and 0.06-0.10% (w/v) of NFBC in the Suspension method. Non-cancerous cells such as human-induced pluripotent stem (iPS) cells and human mesenchymal stem cells (MSCs) also formed spherical formulations. It is noteworthy that both the size and cell viability of spheroids prepared via these methods were comparable to those cultured using commercially available 3D cell-culture systems. Both OnGel and Suspension methods are less complicated than the existing 3D cell-culture systems, which is an invaluable advantage for the preparation of cancer spheroids. The NFBC-based 3D cell-culture systems introduced here show great promise as a tool to prepare cultures for cell-derived spheroids for the progress of both in vitro and in vivo studies of the biological functioning of cells.

Sustainable and Flexible Surface-Enhanced Raman Scattering Transducer: Gold Nanoparticle-Bacterial Cellulose Composite for Pesticide Monitoring in Agrifood Systems

Functionalized plasmonic nanostructure platforms are widely used for developing optical biosensors and SERS assays. In this work, we present a low-cost and scalable surface-enhanced Raman scattering (SERS) system based on an innovative optical transducer comprising gold nanoparticles (AuNPs) embedded in nano-fibrillated bacterial cellulose (BC). The AuNPs@BC composite leverages the unique nanofibrillar architecture of bacterial cellulose, which provides a high surface area, flexibility, and uniform nanoparticle distribution, enabling the formation of numerous electromagnetic “hot spots”. This structure excites localized surface plasmon resonance (LSPR), as demonstrated by a bulk sensitivity of 72 nm/RIU, and supports enhanced Raman signal amplification. The eco-friendly and disposable AuNPs@BC platform was tested for agrifood applications, focusing on the detection of thiram pesticide. The system achieved a detection limit of 0.24 ppm (1 µM), meeting the sensitivity requirements for regulatory compliance in food safety. A strong linear correlation (R2 ≈ 0.99) was observed between the SERS peak intensity at 1370 cm−1 and thiram concentrations, underscoring its potential for quantitative analysis. The combination of high sensitivity, reproducibility, and environmental sustainability makes the AuNPs@BC platform a promising solution for developing cost-effective, flexible, and portable sensors for pesticide monitoring and other biosensing applications.

Nano-Fibrillated Bacterial Cellulose Nanofiber Surface Modification with EDTA for the Effective Removal of Heavy Metal Ions in Aqueous Solutions.

Nano-fibrillated bacterial cellulose (NFBC) has very long fibers (>17 μm) with diameters of approximately 20 nm. Hence, they have a very high aspect ratio and surface area. The high specific surface area of NFBC can potentially be utilized as an adsorbent. However, NFBC has no functional groups that can bind metal ions, limiting its potential applications. In this study, the hydroxyl groups on the surface of NFBC were chemically modified with EDTA monoanhydride to convert NFBC into a metal adsorbent. The fiber morphology and crystal structures of the modified NFBC were almost identical to those of the unmodified NFBC, suggesting that the surface hydroxyl groups of NFBC were well-conjugated with the EDTA groups. Surface-modified NFBC preferentially adsorbed transition metals in aqueous solutions, such as Cu(II), Hg(II), Pb(II), and Cd(II), but hardly adsorbed Mg(II) and Cr(VI). The adsorption of heavy metal ions can be explained by the pseudo-second-order kinetics of the chemisorption process and the Langmuir isotherm model. Furthermore, the EDTA-modified NFBC is a renewable and recyclable adsorbent. The results of this study indicate that surface-modified NFBC can be utilized as a biosorbent for heavy metal removal in chemical, food, pharmaceutical, and other industrial fields.

Enhancing Starch Film Properties Using Bacterial Nanocellulose-Stabilized Pickering Emulsions.

This study aimed to address issues related to hydrophilicity, barrier properties, and mechanical performance in starch-based films by incorporating Pickering emulsions stabilized with nano-fibrillated bacterial cellulose (BC). Emulsions were added to the film-forming suspension at varying concentrations (1.0%, 2.5%, 5.0%, and 7.5% v/v) for comparison. The films were evaluated using water vapor permeability (WVP), contact angle, Fourier Transform Infrared Spectroscopy (FTIR), and tensile tests. The results showed a significant reduction in film hydrophilicity, with the contact angle increasing from 49.7° ± 1.5 to 71.0° ± 1.4, and improved water vapor barrier properties, with WVP decreasing from 0.085 ± 0.04 to 0.016 ± 0.01 g·mm/h·m2·kPa. FTIR analysis confirmed the successful incorporation of the emulsion into the starch matrix. Among the tested concentrations, 2.5% provided an optimal balance, increasing hydrophobicity while maintaining mechanical strength. These findings demonstrate that Pickering emulsions are an effective strategy for enhancing the functional properties of starch films.

Electric stimulation: a versatile manipulation technique mediated microbial applications.

Electric stimulation (ES) is a versatile technique that uses an electric field to manipulate microorganisms individually. Over the past several decades, the capabilities of ES have expanded from bioremediation to the precise motion control of cells and microorganisms. However, there is limited information on the underlying mechanisms, latest advancement and broader microbial applications of ES in various fields, such as the production of extracellular polymers with upgraded properties. This review article summarizes recent advancements in ES and discusses it as a unique external manipulation technique for microorganisms with wide applications in bioremediation, industry, biofilm deactivation, disinfection, and controlled biosynthesis. One specific application of ES discussed in this review is the extracellular biosynthesis, regulation, and organization of extracellular polymers, such as bacterial cellulose nanofibrils, curdlan, and microbial nanowires. Overall, this review aims to provide a platform for microbial biotechnologists and synthetic biologists to leverage the manipulation of microorganisms using ES for bio-based applications, including the production of extracellular polymers with enhanced properties. Researchers can engineer, manipulate, and control microorganisms for various applications by harnessing the potential of electric fields.

Facile fabrication of sulfonated bacterial cellulose-silane composite aerogels via in suit polymerization for enhanced oil/water separation performance

Cellulose-based aerogels have emerged as highly promising materials for oil-water separation because of their highly porous nature, low bulk density, cost-effectiveness, and functional performance. In this study, a novel, robust, and hydrophobic bacterial cellulose aerogel (HBCA) was reported for highly efficient oil/water separation, which was formed by incorporating methyltrimethoxysilane (MTMS) into sulfonated nano-fibrillated bacterial cellulose (SNBC) matrix through freeze-drying method. The structural integrity of the SNBC/MTMS aerogel was ensured by its three-dimensional linked network structure. The resultant aerogel demonstrated superior hydrophobicity with a contact angle of 152.4°. As an oil-absorbing material, it selectively adsorbed different types of oils and organic solvents, with a saturation adsorption capacity ranging from 42.14 to 85.37 g·g−1. Additionally, this composite aerogel demonstrated outstanding separation efficiency (98.48 %) in continuous oil-water mixture processes, suggesting promising potential applications in the treatment of industrial oil pollution and oil spill accidents. This study indicates that the proposed HBCA with sulfonated nano-fibrillated bacterial cellulose is a potential and effective material for addressing environmental challenges related to oil contamination.

Comparative In Vivo Biocompatibility of Cellulose-Derived and Synthetic Meshes in Subcutaneous Transplantation Models.

Despite the increasing interest in cellulose-derived materials in biomedical research, there remains a significant gap in comprehensive in vivo analyses of cellulosic materials obtained from various sources and processing methods. To explore durable alternatives to synthetic medical meshes, we evaluated the in vivo biocompatibility of bacterial nanocellulose, regenerated cellulose, and cellulose nanofibrils in a subcutaneous transplantation model, alongside incumbent polypropylene and polydioxanone. Notably, this study demonstrates the in vivo biocompatibility of regenerated cellulose obtained through alkali dissolution and subsequent regeneration. All cellulose-derived implants triggered the expected foreign body response in the host tissue, characterized predominantly by macrophages and foreign body giant cells. Porous materials promoted cell ingrowth and biointegration. Our results highlight the potential of bacterial nanocellulose and regenerated cellulose as safe alternatives to commercial polypropylene meshes. However, the in vivo fragmentation observed for cellulose nanofibril meshes suggests the need for measures to optimize their processing and preparation.

Application of bacterial-derived long cellulose nanofiber to suspension culture of mammalian cells as a shear protectant

Nanofibrillated bacterial cellulose (NFBC) is a bio-compatible long-fiber nanocellulose produced by cellulose-synthesizing bacteria. It forms an entangled network structure in the suspension state, thereby imparting greater viscosity than conventional media additives. In this study, we examined its application as a shear protectant in the suspension culture of mammalian cells to mitigate hydrodynamic stress imposed on the cells. The media supplemented with hydroxypropyl cellulose-adsorbed NFBC (HP-NFBC) exhibited an increase in shear viscosity according to rheometric analysis, similar to FP003, a commercially available medium additive. Suspension culture of Chinese hamster ovary cells in HP-NFBC-containing media under a high stirring rate (120 rpm) demonstrated higher cell growth and lower cell death compared to those in the medium without additives and in FP003. A 0.10 (w/v)% concentration of HP-NFBC showed the highest viable cell number among the tested concentrations. Computational fluid dynamics simulation revealed a decrease in shear rate and flow velocity within the spinner flask owing to the addition of HP-NFBC or FP003. It is suggested that the decline of these parameters in high-viscosity media suppresses the hydrodynamic stress on cells. This study highlights the potential of HP-NFBC as a shear protectant in mammalian cell suspension culture.

Detailed structural analyses and viscoelastic properties of nano-fibrillated bacterial celluloses

Nano-fibrillated bacterial cellulose (NFBC) can be prepared by cultivating a cellulose-producing bacterium in a medium containing a dispersant under agitating and aerobic conditions. Although NFBCs have various applications, their detailed structure and physical properties have not been clarified. Therefore, in this study, we performed detailed structural and physical property analyses of NFBCs to advance their potential applications. Atomic force microscopy and image analysis showed that the average fiber length of NFBCs was approximately 17 µm and fiber widths were 10–15 nm; the aspect ratios of NFBCs were > 1000, which are >10-fold higher than that of 2,2,6,6-tetramethylpioeridine-1-oxyl-oxidized cellulose nanofiber. Shear viscosity measurements showed that the NFBCs exhibited shear-thinning flow behavior even at low concentrations (0.01 wt%). Frequency sweep measurements showed that the storage modulus values were greater than the loss modulus values in the measured frequency range, indicating that the NFBCs were in a stable gel state. Thus, the NFBCs exhibited significantly longer fiber lengths, larger aspect ratios, and excellent viscoelastic properties based on these unique structural features. Our findings will help develop novel applications utilizing the ultrahigh aspect ratio unique to NFBC and its viscoelastic properties.

Bacterial cellulose nanocrystals or nanofibrils as Pickering stabilizers in low-oil emulsions: A comparative study

This investigation assessed the potential of bacterial cellulose (BC) in two distinct forms, nanocrystals (BC-NC) and oxidized nanofibrils (BC-NF), as stabilizers for low oil-in-water emulsions (1 % v/v). The research explored the impact of ionic strength and BC concentration on the physico-chemical characteristics, stability, and rheological properties of those emulsions. Nanofibrils had diameters ranging from 25 to 146 nm and lengths in the micrometer range, while nanocrystals varied in length from 133 to 870 nm and in diameter from 20 to 60 nm. Both BC-NF and BC-NC exhibited high zeta potential values (>−45 mV) and contact angles of 30-31°, indicating stability. Both nanocelluloses were effectively used as stabilizers in Pickering emulsions, namely in low-oil systems, producing small emulsion droplets with sizes between 1.42 and 4.13 μm. Further results revealed that ionic strength influenced emulsion stability, with both BC-NF and BC-NC preferentially located on the surfaces of emulsion droplets in the presence of salt, as demonstrated by microscopy images. The presence of BC at the interface contributed to creating a more robust barrier against coalescence and Ostwald ripening, influencing droplet size and rheological properties. Higher BC concentrations (1 %) increased emulsion stability in the absence of salt, while at lower BC concentrations (0.5 %), salt concentration was determinant for the long-term stability of the emulsions. These findings provide valuable insights into the production of Pickering emulsions using nanocelluloses, highlighting the advantages of bio-based nanomaterials for applications in the food industry.

Poly(butylene succinate) reinforced by small amount of grafted nanofibrillated bacterial cellulose: Toughness variability based on nanocomposites preparation method

Polybutylene succinate (PBS) is a promising biodegradable polymer; however, its low mechanical performance limits its application. To address this issue, long fiber length nanofibrillated bacterial cellulose (HP-NFBC), produced by a bottom-up process using cellulose-producing bacterium and hydroxypropyl cellulose (HPC) as a dispersing agent was used as reinforcement agent. Hydrophobic moieties were grafted on HP-NFBC surface to increase surface compatibility. Nanocomposites prepared by both solvent casting and melt-kneading were evaluated. By solvent casting, the addition of 0.25 wt% of grafted HP-NFBC increased the toughness by 212 %. Melt-kneading revealed an increase in flexural strength and young’s modulus. More evident improvement was observed when the nanocomposites were annealed, and the ultimate strength increased by 113 % at 2 wt% of grafted HP-NFBC incorporation. Great increased in toughness were achieved only by small incorporation of grafted HP-NFBC. Additionally, compost biodegradation tests were conducted to confirm that the nanocomposites had biodegradability similar to that of PBS.

In situ biomineralization reinforcing anisotropic nanocellulose scaffolds for guiding the differentiation of bone marrow-derived mesenchymal stem cells

Nanocellulose (NC) is a promising biopolymer for various biomedical applications owing to its biocompatibility and low toxicity. However, it faces challenges in tissue engineering (TE) applications due to the inconsistency of the microenvironment within the NC-based scaffolds with target tissues, including anisotropy microstructure and biomechanics. To address this challenge, a facile swelling-induced nanofiber alignment and a novel in situ biomineralization reinforcement strategies were developed for the preparation of NC-based scaffolds with tunable anisotropic structure and mechanical strength for guiding the differentiation of bone marrow-derived mesenchymal stem cells for potential TE application. The bacterial cellulose (BC) and cellulose nanofibrils (CNFs) based scaffolds with tunable swelling anisotropic index in the range of 10–100 could be prepared by controlling the swelling medium. The in situ biomineralization efficiently reinforced the scaffolds with 2–4 times and 10–20 times modulus increasement for BC and CNFs, respectively. The scaffolds with higher mechanical strength were superior in supporting cell growth and proliferation, suggesting the potential application in TE application. This work demonstrated the feasibility of the proposed strategy in the preparation of scaffolds with mechanical anisotropy to induce cells-directed differentiation for TE applications.

Bacterial cellulose nanofibrils for the physical and oxidative stability of fish oil-loaded Pickering emulsions

Fish oil, a crucial unsaturated ω-3 unsaturated fatty acid supplement, is susceptible to oxidation. To improve the stability of fish oil, this study produced a fish oil Pickering emulsion via ultrahigh-pressure homogenization pressure (UHPH) using bacterial cellulose nanofibrils (BCNFs) as the water phase. The BCNFs exhibited a high aspect ratio fibril structure with an average diameter size of 12.64 nm. The results of particle size, zeta potential, emulsion properties, and oxidative stability showed that the emulsions with a 4:6 oil-water ratio and 0.15 % BCNFs exhibited excellent oxidative stability and good physical stability. The CLSM and rheological results suggested the BCNFs acted as a stabilizer and thickener, stabilizing the oil-water medium and enhancing the oxidation stability of fish oil Pickering emulsions. The Low-field nuclear magnetic analysis results indicated the emulsion had good physical stability, and water dispersed very well in the emulsion system. Moreover, the emulsions exhibited good stability at pH 6–12 and temperature 4–75 °C. This study provided a promising method for producing fish oil Pickering emulsions and improving the stability of fish oil.

Stabilization of Pickering emulsions with bacterial cellulose nanofibrils (BCNFs) fabricated by electron beam irradiation

Bacterial cellulose (BC) had been considered as a promising Pickering stabilizer for its non-toxicity, high purity, and biocompatibility. The electron beam irradiation (EBI) technology had gained significant attention in the field of cellulose degradation due to its operational convenience and mild reaction conditions. Pickering emulsions using BC degraded by EBI as Pickering stabilizers were successfully prepared. Irradiated bacterial cellulose nanofibers (IBCNFs) exhibited a high aspect ratio(30–80 μm in length and 21–109 nm in width) and high absolute zeta potential (-50 mV). The optimal emulsification performance of IBCNFs was observed at an irradiation dosage of 100 kGy (IBC-100). The emulsions based on IBC-100 achieved an EI value of 96.15 ± 0.19%, while those based on IBC-500 obtained an EI value of 59.93 ± 1.47%. Stable Pickering emulsions were prepared when the concentration of IBC-100 stabilizer exceeded 0.3 wt% with a fixed oil phase of 30%, or when the rapeseed oil phase was below 50% while maintaining a fixed suspension of 0.5 wt% IBCNFs. Confocal laser scanning microscopy (CLSM) demonstrated that IBCNFs facilitated the formation of a dense interfacial layer at the oil-water interface and formed a three-dimensional network within the water phase. Which effectively hindered the aggregation and flocculation between individual oil droplets. Furthermore, rheological findings demonstrated that emulsions possessed primarily exhibited properties of elasticity, and their gel strength could be enhanced by adjusting the irradiation dosages of IBCNFs. Moreover, environmental stability experiments indicated that IBCNFs-based emulsions exhibited exceedingly high temperature (20–80 °C) and pH tolerance (4–12). Flocculation occurred under high acid (pH = 2) conditions and at high ionic levels (≥ 40 mM NaCl), without causing disruption to the emulsion droplet structure. This study provided an innovative approach to preparing cellulose nanofibers with high aspect ratio, which could be utilized as emulsion stabilizers. The study also demonstrated that Pickering emulsions based on IBCNFs exhibit excellent stability and rheological properties. IBCNFs have promising potential to serve as a natural Pickering emulsifier for stabilizing oil-in-water emulsions in various food applications, cosmetic and pharmaceutical formulations.

Modified bacterial nanofibril for application in superhydrophobic coating of food packaging

Superhydrophobic coatings applied to food packaging can significantly reduce unnecessary food waste. Inspired by nature, we proposed a simple method to prepare coatings with ecologically correct materials, such as beeswax and bacterial cellulose nanofibrils functionalized with nanoparticles of food silicon dioxide (BW/BCn- SiO2). The morphology of bacterial cellulose nanofibrils shows a strong reaction with SiO2 aggregated on the surface of the cellulosic material. The FTIR results confirm the efficiency of the functionalization. In addition, the functionalized material presents less thermal degradation. BW/BCn- SiO2, with a contact angle of 153° and a slip angle of 3°, features Cassie-Baxter-compatible micro/nanoscale roughness, self-cleaning properties, low-temperature storage stability, excellent adhesion and mechanical durability even after several treatments, considered nontoxic and with remarkable performance in repelling liquid foods with high viscosity, such as honey, yogurt and chocolate sauce. This work highlights the importance of innovations in food packaging that impact food waste, through superhydrophobic, ecological and safe coatings.

Tailoring the Heterogeneous Structure of Macro-Fibers Assembled by Bacterial Cellulose Nanofibrils for Tissue Engineering Scaffolds.

Bacterial cellulose/oxidized bacterial cellulose nanofibrils (BC/oxBCNFs) macro-fibers are developed as a novel scaffold for vascular tissue engineering. Utilizing a low-speed rotary coagulation spinning technique and precise solvent control, macro-fibers with a unique heterogeneous structure with dense surface and porous core are created. Enhanced by a polydopamine (PDA) coating, these macro-fibers offer robust mechanical integrity, high biocompatibility, and excellent cell adhesion. When cultured with endothelial cells (ECs) and smooth muscle cells (SMCs), the macro-fibers support healthy cell proliferation and exhibit a unique spiral SMC alignment, demonstrating their vascular suitability. This innovative strategy opens new avenues for advances in tissue engineering.

BACTERIAL CELLULOSE NANOFIBRILS FROM A BY-PRODUCT OF THE GROWING KOMBUCHA BUSINESS: OBTENTION AND USE IN THE DEVELOPMENT OF POLY(LACTIC ACID) COMPOSITES

Bacterial cellulose nanofibrils were obtained from the floating pellicle developed at the air-liquid interface during Kombucha tea production. Kombucha-derived bacterial nanocellulose (KBNC) was purified by different treatments and characterized in terms of morphology, chemical structure, thermal decomposition pattern and crystallinity. 
 Purified KBNC was then employed in the production of poly(lactic acid) (PLA)-based composites obtained by melt compounding and compression molding. The morphology and the mechanical properties of the composites with increasing contents of KBNC (i.e. 1, 3, 5, 7 wt.%) were analyzed. Results showed that KBNC was suitable for the development of PLA composites with improved stiffness.

Force-Induced Alignment of Nanofibrillated Bacterial Cellulose for the Enhancement of Cellulose Composite Macrofibers.

Bacterial cellulose, as an important renewable bioresource, exhibits excellent mechanical properties along with intrinsic biodegradability. It is expected to replace non-degradable plastics and reduce severe environmental pollution. In this study, using dry jet-wet spinning and stretching methods, we fabricate cellulose composite macrofibers using nanofibrillated bacterial cellulose (BCNFs) which were obtained by agitated fermentation. Ionic liquid (IL) was used as a solvent to perform wet spinning. In this process, force-induced alignment of BCNFs was applied to enhance the mechanical properties of the macrofibers. The results of scanning electron microscopy revealed the well-aligned structure of BCNF along the fiber axis. The fiber prepared with an extrusion rate of 30 m min-1 and a stretching ratio of 46% exhibited a strength of 174 MPa and a Young's modulus of 13.7 GPa. In addition, we investigated the co-spinning of carboxymethyl cellulose-containing BCNF with chitosan using IL as a "container", which indicated the compatibility of BCNFs with other polysaccharides. Recycling of the ionic liquid was also verified to validate the sustainability of our strategy. This study provides a scalable method to fabricate bacterial cellulose composite fibers, which can be applied in the textile or biomaterial industries with further functionalization.

Nanoemulsion-integrated gelatin/bacterial cellulose nanofibril-based multifunctional film: Fabrication, characterization, and application

The current requirements of food safety regulations and the environmental impact stemming from plastic packaging can only be addressed by developing suitable bio-nanocomposite films. Therefore, this study is dedicated to the fabrication of multifunctional film composed of gelatin, bacterial cellulose nanofibrils (BCNF), and black pepper essential oil nanoemulsion (BPEONE) and application for duck meat preservation. BCNF was prepared through ultrasonication of cellulose derived from Komagataeibacter xylinus. BPEONE observed spherical morphology with a diameter ranging from 83.7 to 118 nm. A film matrix containing a higher gelatin proportion than BCNF was more effective in trapping BPEONE. However, increasing the BPEONE fraction showed more surface abrasion and voids in the film morphology. A flexible film with good interaction, crystallinity, and greater thermal stability (421 °C) was developed. Nevertheless, film hydrophobicity (118.89°) declined, resulting in a notable effect on water solubility, swelling, and water vapor permeability. Moreover, the film had improved antibacterial and antioxidant activities, coupled with controlled release characteristics. Consequently, the developed film effectively retarded the lipid oxidation, inhibited microbial growth, and extended the shelf life of duck meat at refrigeration (4 °C) by 3 days, and made the film a promising alternative in the realm of bio-active packaging technology.

Cinnamon essential oil Pickering emulsions: Stabilization by bacterial cellulose nanofibrils and applications for active packaging films

Pickering emulsions are emulsions stabilized by solid particles with a wide range of potential applications as they typically provide more stable systems than surface-stabilized emulsions. Among the various solid stabilizers, bacterial cellulose (BC) may open up new opportunities for future Pickering emulsions due to its unique nano-size, amphiphilicity and other excellent properties. In this study, BC, TEMPO-oxidized BC (TOBC) and dialdehyde cellulose (DAC) were used as emulsifiers to stabilize cinnamon essential oil (CEO) Pickering emulsions. The microstructure, oil droplet size distribution, storage and loss moduli of the Pickering emulsions were affected by the type of emulsifier used. Compared to BC and DAC nanofibers, TOBC nanofibers exhibited better emulsifying capacity. TOBC (1%)-stabilized Pickering emulsion showed smaller droplet size, narrower size distributions, better stability and sustained release ability. Moreover, the Pickering emulsions showed high antibacterial activities against Gram-positive bacteria S. aureus and Gram-negative bacteria E. coli. Active gelatin composite films containing TOBC (1%)-stabilized CEO Pickering emulsions were successfully fabricated and showed good antibacterial and antioxidant properties, which make them suitable for food packaging application.