This study aimed to optimize a biocomposite formulation based on chitosan (CS) and periodate oxidized schizophyllan (POSPG), in order to develop a multifunctional platform suitable for biomedical applications, particularly wound-dressings. This research focuses on improving the key physicochemical and biological properties required for effective medical use. Schizophyllan (SPG) was produced by fermenting Schizophyllum commune and subsequently oxidized to yield aldehyde contents of 68

Nanoplastics and engineered nanoparticles interact in varying ways with amyloidogenic proteins, yet the mechanistic basis for the pathological effects caused by that interplay remains unclear. Here, we demonstrate how carboxylated-polystyrene nanoplastics (PsNPs) and-silica nanoparticles (SNPs) differentially modulate alpha-synuclein (alpha Syn) amyloid formation kinetics through variation in composition and binding of the protein corona, a surface layer of biological origin formed from plasma proteins. Using Taylor dispersion analysis in microfluidic capillaries under physiologically relevant flow conditions, we demonstrate that anionic PsNPs form shear-sensitive, alpha Syn-bridged clusters, which transition into stable flocs at sub-stoichiometric ratios via charge neutralization. SNPs, in contrast, maintain colloidal stability through dynamic binding equilibria. The composition of the hard corona, a tightly bound layer of protein at the NP surface with slow exchange kinetics, critically determined aggregation outcomes: PsNPs accelerated fibril formation through strong interfacial interactions, while SNPs inhibited nucleation but promoted fibril fragmentation. We identified the hard corona's slow exchange kinetics as the primary determinant of long-term aggregation, while the weakly bound "soft corona" mediated early interaction dynamics. These findings extend the current understanding of how NP surface chemistry influences corona composition and protein aggregation particularly in the context of environmentally relevant nanoplastics.

The practical application of microbial fuel cells (MFCs) is hindered by low electrochemical efficiency, primarily caused by suboptimal electron generation by electrogenic biocatalysts. In this study, a systems-based strategy was employed to rationally design a co-cultured microbial community composed of Shewanella oneidensis MR-1 and Geobacter sulfurreducens for enhanced bioelectricity production. A two-cell genome-scale metabolic model was reconstructed to capture both intracellular metabolism and interspecies metabolic interactions under MFC operating conditions. By applying a flux-based optimization framework and comparing reaction flux distributions under maximized and minimized NADH production, key bottleneck reactions governing electron generation were identified. Based on enzymatic regulation analysis using the BRENDA database, a regulated defined medium was designed to redirect intracellular metabolic fluxes toward improved NADH availability. Experimental validation in a single-chamber air-cathode MFC demonstrated that the regulated co-culture achieved a maximum power density of 3.78 mW & sdot;m-2 and an open-circuit voltage of 1.20 V, corresponding to a 1.9-fold improvement over the unregulated co-culture and up to a 46-fold increase compared with pure S. oneidensis cultures. The coulombic efficiency of the regulated system reached 131%, indicating enhanced electron recovery facilitated by optimized interspecies metabolic coupling. Overall, this study pioneers the integration of genome-scale metabolic modeling with targeted metabolic regulation as a predictive and mechanistic framework for rational microbial community design, offering a robust alternative to conventional empirical approaches for improving MFC performance.

Viability loss of probiotics during the freeze-drying process can significantly reduce their functional performance in drug and food products. In the present study, the simplex lattice design was utilized to optimize cryoprotective formulations to enhance the survival of Limosilactobacillus reuteri on freeze-drying, under simulated gastrointestinal, thermal, and storage conditions. Maltodextrin, mannitol, and saccharose were selected as cryoprotectants. The results revealed that the optimal microencapsulation efficiency (97.25%), freeze-drying yields (97.74%), and viability under in vitro gastric and intestinal conditions (85.84%) were achieved with a formulation consisting of 66.61% maltodextrin, 15.16% mannitol, and 18.23% saccharose. The absence of cryoprotective agents during freeze-drying led to a 3.88% reduction in L. reuteri viability. The highest stability was observed in the optimized composite coated with O-carboxymethyl chitosan, where only 1.06 log CFU/g of bacterial loss occurred after 42 days of storage at 4 degrees C. The presence of the maltodextrin-mannitol- saccharose composite formed a protective layer that improved the microcapsule structure, enhanced the resistance of probiotics to thermal stress, and significantly improved their stability during storage. Overall, the results indicate that the optimized composite formulation can be an effective strategy for improving the viability and stability of probiotics under various adverse conditions.

Bacterial infection is a major cause of non-healing wounds. Due to the growing problem of antibiotic resistance, there is an urgent need to develop effective treatments for chronic and acute wound infections. In this study, starch-based Ag/Fe3O4 nanocomposites (NCs) were synthesized and characterized. Antibacterial and cytotoxicity assays were subsequently performed. The therapeutic effect of NCs on infected wound healing was evaluated in normal and diabetic rats under an external static magnetic field (SMF). Ag/Fe3O4 NCs exhibited superparamagnetic behavior, with uniform spherical particles averaging 32 nm in diameter. In vitro tests revealed effective antibacterial activity and low cytotoxicity of Ag/Fe3O4 NCs compared to Ag nanoparticles. In vivo test on normal infected wounds treated with Ag/Fe3O4 NCs under a static magnetic field (NC + MF) demonstrated significant biofilm reduction by day 7 and 94 % wound closure by day 14 (P < 0.05). Hair follicles appeared only in this group by day 21. In diabetic-infected wounds, the d(NC + MF) group showed 50 % healing at day 7 and nearly complete healing by day 21, with significantly increased fibroblast proliferation and new vessels compared to control (P < 0.05). This study demonstrates the in vivo use of green-synthesized Ag/Fe3O4 NCs combined with a static magnetic field (similar to 9 mT) to enhance biofilm penetration, improve antibacterial efficacy, and reduce NCs dosage. Under SMF, wounds achieved 94 % and 83 % closure by day 14 in normal and diabetic rats, respectively. This approach is a promising non-invasive treatment option for chronic wounds by accelerating biofilm removal and enhancing the healing process.

The rational engineering of biopolymeric nanocarriers with enhanced biocompatibility and targetability remains a central goal in cancer nanomedicine. Here, we report the fabrication and comprehensive evaluation of zein-based nanoparticles (ZNp) surface-modified with polyethyleneimine (PEI) and folic acid-conjugated PEI (PEI-FA) for folate receptor (FR)-targeted delivery to breast cancer cells. Folic acid (FA) was covalently attached to branched PEI via EDC/NHS chemistry, with a conjugation efficiency of ∼45 %, confirmed by UV-Vis, 1H NMR, and FTIR spectroscopy. Functionalization with PEI-FA improved colloidal stability, reduced particle size to ∼96 nm, and increased surface charge to +32 mV in physiological media. In vitro assays revealed a higher cellular uptake of ZNp/PEI-FA in FR-overexpressing MDA-MB-231 cells compared to fibroblasts, as shown by confocal microscopy. MTT analysis demonstrated reduced cytotoxicity toward normal HFF cells while preserving selective antiproliferative effects in cancer cells (IC50 ≈ 52.5 μg/mL at 24 h). To gain deeper mechanistic insight, we combined experimental characterization with all-atom molecular dynamics simulations of zein-PEI and zein-PEI-FA complexes. This synergistic approach highlighted conformational adaptability and interfacial stability essential for targeting efficiency. Our integrated design framework underscores the potential of protein-based nanocarriers for safe, precise tumor delivery.

OBJECTIVE:Curcumin (CUR) is a natural phenolic compound with potent anticancer properties and potential as a photosensitizer (PS) for photodynamic therapy (PDT). However, its clinical application is limited by poor solubility, low bioavailability, and rapid degradation. To address these challenges, this study introduces curcumin-loaded bovine serum albumin nanoparticles (CUR-BSA NPs) as a pH-responsive drug delivery system for enhanced PDT in breast cancer treatment. METHODS:CUR-BSA NPs were synthesized using the desolvation method and characterized by using Field emission scanning electron microscopy (FESEM), Dynamic light scattering (DLS), Fourier-transform infrared (FT-IR) spectroscopy. RESULTS:The nanoparticles with size (∼170 nm), zeta potential (-36 ± 2.7 mV), and encapsulation efficiency (47.5%), demonstrated pH-responsive drug release, with higher curcumin release under acidic conditions, mimicking the tumor microenvironment. In-vitro cytotoxicity studies on MCF-7 breast cancer cells revealed that CUR-BSA NPs, in combination with blue light irradiation (420 nm, 30 J/cm2), significantly reduced cell viability to 69% after 48 h, while CUR-BSA NPs show lower cytotoxicity (45% vs. 68%) in the absence of photodynamic therapy. TUNEL assay confirmed apoptosis in 52.4% of treated cells, compared to 4.6% in the control group. Furthermore, CUR-BSA NPs displayed excellent biocompatibility in the absence of light exposure, reducing systemic toxicity. CONCLUSION:These findings establish CUR-BSA NPs as a promising nanoplatform for PDT, providing enhanced drug delivery, tumor-targeted release, and improved therapeutic efficacy in breast cancer treatment.

Carvedilol (CAR), a well-established β-blocker and a promising pharmacological agent for mitigating doxorubicin (DOX)-induced cardiotoxicity, has also demonstrated anticancer effects against breast cancer cells. Given the seemingly contradictory effects of CAR and DOX treatment on cardiomyocytes and cancer cells, particularly regarding mitochondrial function, this study investigates the synergistic potential of DOX and CAR through various delivery systems, including free form, exosomal formulations, and mitochondria-targeted delivery systems. The Chou-Talalay analysis demonstrated that combining DOX with CAR at a 1:1 ratio reduced DOX’s IC50 in MCF-7 (from 0.274 µM to 0.191 µM) and MDA-MB-231 (from 0.138 µM to 0.086 µM) cells. Drug loading in exosomal formulations was achieved with approximately 24% efficiency, maintaining the optimum ratio. While CAR exhibited minimal cytotoxicity at concentrations below 2 µM, its combination with DOX enhanced anticancer effects. Mitochondrial-targeted delivery (TPP-Exo-DOX-CAR) showed reduced cytotoxicity compared to TPP-Exo-DOX formulation. ROS production analysis and mitochondrial membrane potential assessments revealed that CAR’s protective effects become dominant in mitochondria-targeted delivery in vitro. Migration studies demonstrated significant inhibition of cell motility in CAR containing formulations. These findings suggest that while the DOX and CAR combination shows promise in cancer treatment, the delivery system significantly impacts their therapeutic efficacy, particularly in mitochondrial-targeted applications.

Paclitaxel (PTX) is recognized as one of the most potent chemotherapy agents and is widely used to treat various cancers, including ovarian, lung, breast, head, and neck cancer. Due to the limited solubility and high toxicity of PTX, its use in cancer treatment is challenging and limited. Hence, strategies have been devised to improve the solubility and bioavailability of paclitaxel. In recent years, biocompatible nanocarriers have garnered attention due to their desirable properties, including increased permeability, targeted delivery, extended circulatory half-life, and biological drug delivery for the delivery of chemotherapeutic drugs. Protein nanostructures have been widely studied for the delivery of paclitaxel due to their significant advantages, such as safety, low toxicity, availability, and relatively easy preparation. This review article reviews recent advances in the development of protein-based drug delivery systems for loading and releasing paclitaxel. These nanocarriers have great potential to improve paclitaxel's antitumor properties and efficacy. Therefore, in the future, the integration of the pharmaceutical industry and artificial intelligence techniques will provide more opportunities for research and development in the pharmaceutical field.

This study aimed to synthesize antimicrobial nanocomposite films based on periodate oxidized schizophyllan (POSPG), chitosan (CS), and green synthesized magnetic silver/iron oxide nanoparticles (Ag/Fe3O4 NPs). The average size and polydispersity index (PDI) of the Ag/Fe3O4 NPs produced by starch were 85 nm and 0.17, respectively. The schizophyllan (SPG) was first produced from Schizophyllum commune. The solution casting method was employed to prepare nanocomposite films composed of POSPG, CS and different concentrations of Ag/Fe3O4 NPs (0.66, 1.33 and 2 mu g/mL). According to the MTT results, 1.33 mu g/mL was the maximum non-toxic concentration for loading into CS/POSPG, so this film, CS/POSPG@NP1.33, was selected for further study. The appearance of a peak at 8.3 ppm in nuclear magnetic resonance (NMR) spectrum of CS/POSPG film evidenced the imine bond formation. Energy dispersive X-ray (EDX) mapping illustrated the proper distribution of Ag/ Fe3O4NPs into the matrix. The mechanical strength and water vapor permeability of the CS/POSPG@NP1.33 nanocomposite increased to 82.77 f 1.25 MPa and 0.640 f 0.02 mm g/m2.h.kPa, respectively as compared to those reported for CS/POSPG film (63.49 f 0.7 Mpa and 0.576 f 0.015 mm g/m2.h.kPa), while the swelling capacity decreased to 2.3 f 0.075. The normal human fibroblast cell survival reached 83.21 f 1.78 % after exposure to the CS/POSPG@NP1.33, indicating its biocompatibility. The inhibitory effect of the nanocomposite against S. aureus, E. coli, MRSA, P. aeruginosa, and candida was 94.6 f 3.73 %, 97.2 f 4.65 %, 88.4 f 3.87 %, 97.9 f 2.04 %, and 84.6 f 1.95 %, respectively. The eradication rate of the CS/POSPG@NP1.33 against the S. aureus biofilm was up to 47.55 f 1.72 %, which increased to 79.74 f 2.09 % after exposure to a magnetic field.

Charge heterogeneity in monoclonal antibodies (mAbs), caused by post-translational modifications, remains a substantial obstacle to ensuring consistent, stable, and effective therapeutics. Conventional optimization techniques, such as one-factor-at-a-time and design of experiments, often fail to capture the complex, nonlinear interactions between culture parameters (e.g. pH, temperature, duration) and medium components (e.g. glucose, metal ions, amino acids). This review highlights machine learning (ML) as a powerful approach for modeling these relationships and forecasting charge variant profiles in CHO cell-based mAb process development. We summarize supervised learning and regression methods used to link process conditions with charge heterogeneity and present case studies showing ML’s role in reducing acidic and basic variants. We also discuss challenges related to data quality, model interpretability, scalability, and regulatory compliance. Finally, we propose a roadmap for adaptive, ML-driven optimization strategies for bioprocess development, aligned with Quality-by-Design principles.

Background and Objective: Improving probiotics viability in digestive and storage conditions is challenging for the food and pharmaceutical industries. The present study aimed to increase viability of the microencapsulated probiotic strain of Lactobacillus reuteri ATCC 23272 in Ocarboxymethyl chitosan-coated bionanocomposite. The O-carboxymethyl chitosan was used to coat bionanocomposite containing prebiotics of pectin and inulin in presence of magnesium oxide nanoparticles. Material and Methods: Pectin and inulin were used as prebiotics with magnesium oxide nanoparticles to improve the microgel structure and O-carboxymethyl chitosan for coating the microcapsules for increasing viability and stability of the probiotics. The extrusion efficiency, viability after microwave oven drying, survival in the simulated digestive fluids, viability after heat treatment and survival rate in long-term storage at 4 and 25 degrees C after 42 d were analyzed. Optimization of inulin, pectin and O-carboxymethyl chitosan in O-carboxymethyl chitosancoated alginate-based bionanocomposite was achieved using Design-Expert software and simplex lattice mixture design. Results and Conclusion: Optimal formulation was achieved using O-carboxymethyl chitosan coating polymer (68% w/v), inulin (29.4% w/v) and pectin (2.6% w/v) with magnesium oxide nanoparticles at a constant concentration. Results showed microencapsulation efficiency (96.43%), survival after microwave oven drying (99.45%) and survival in simulated gastrointestinal conditions (88.95%). Probiotic viability entrapped in O-carboxymethyl chitosancoated microcapsules decreased by 1.46 log CFU.g-1 at 80 degrees C for 5 min. Moreover, Ocarboxymethyl chitosan-coated bionanocomposite improved the stability of probiotics by 2.93 and 3.25 log CFU.g-1 at 4 and 25 degrees C after 42 d, compared to alginate beads. Additionally, it was observed that O-carboxymethyl chitosan coating enhanced the stability of probiotics entrapped in bionanocomposite beads. Results demonstrated that O-carboxymethyl chitosan-coated bionanocomposite, as a novel microencapsulation, could significantly increase the shelf life and viability of Lactobacillus reuteri in various harsh conditions, compared to alginate beads.

Despite advances in cancer treatment, chemoresistance and toxicity remain significant challenges in breast cancer therapy. This study investigated the synergistic effects of carvedilol (CAR), a non-selective beta and alpha1 blocker with emerging anticancer properties, combined with 5-fluorouracil (5-FU), a standard chemotherapeutic agent limited by resistance development and systemic toxicity. We explored this combination as a novel drug repurposing strategy to overcome resistance while enhancing therapeutic efficacy in MCF-7 and MDA-MB231 breast cancer cell lines. The Chou-Talalay method revealed significant synergistic interactions between these drugs, with an optimal 5-FU:CAR ratio of 4:1 (CF). An exosome-mediated co-delivery system (CF-Exo) was developed for these drugs to enhance drug delivery, followed by characterization and assessment of their efficacy in vitro. Drug loading was performed through sonication, achieving about 30 % efficiency. Drug release studies demonstrated higher release rates in acidic pH (65 % for 5-FU, 36 % for CAR) compared to pH 7.4 (59 % for 5FU, 11 % for CAR). CF-Exo demonstrated enhanced cellular uptake compared to free drug combinations. The CF drug combination increased apoptosis compared to the same concentration of 5-FU or CAR in both cell lines. CFExo also maintained CF's apoptotic activity. Following studies revealed that the combination treatment primarily induced apoptosis, with increased reactive oxygen species (ROS) generation and decreased mitochondrial membrane potential (MMP). These findings suggest that the synergistic interaction between CAR and 5-FU, coupled with the exosome-mediated delivery system, may provide a promising approach for enhancing breast cancer treatment efficacy and potentially overcoming drug resistance.

Digital Twins (DTs) are digital representations of physical assets. DTs have emerged as a revolutionary concept in the field of biomanufacturing, offering virtual replicas of physical systems that enable comprehensive monitoring, analysis, and optimization of biological processes. This research provides an in-depth exploration of DTs in biomanufacturing, discussing their key components, modeling approaches, and applications. The importance of Process Analytical Technology (PAT) and sensor technology in DTs is emphasized, along with their role in real-time data acquisition and monitoring. To highlight the practical implementation and benefits of DTs, in a case study, a DT is developed for a fed-batch fermentor that produces Pichia pastoris. By adding unknown uncertainties to the fermentation process, the excellent performance of the developed DT is evaluated based on four states, namely biomass, glycerol, methanol, and protein concentration.

The production of monoclonal antibodies (mAbs) using Chinese Hamster Ovary (CHO) cells has revolutionized the treatment of numerous diseases, solidifying their position as a cornerstone of the biopharmaceutical industry. However, achieving maximum mAb production while upholding strict product quality standards remains a significant hurdle. Optimizing cell culture media emerges as a critical factor in this endeavor, requiring a nuanced understanding of the complex interplay of nutrients, growth factors, and other components that profoundly influence cellular growth, productivity, and product quality. Significant strides have been made in media optimization, including techniques such as media blending, one factor at a time, and statistical design of experiments approaches. The present review provides a comprehensive analysis of the recent advancements in culture media design strategies, focusing on the comparative application of systems biology (SB) and machine learning (ML) approaches. The applications of SB and ML in optimizing CHO cell culture medium and successful examples of their use are summarized. Finally, we highlight the immense potential of integrating SB and ML, emphasizing the development of hybrid models that leverage the strengths of both approaches for robust, efficient, and scalable optimization of mAb production in CHO cells. This review provides a roadmap for researchers and industry professionals to navigate the complex landscape of mAb production optimization, paving the way for developing next-generation CHO cell culture media that drive significant improvements in yield and productivity.

Probiotics have recently received significant attention due to their various benefits, such as the modulation of gut flora, reduction of blood sugar and insulin resistance, prevention and treatment of digestive disorders, and strengthening of the immune system. One of the major issues concerning probiotics is the maintenance of their viability in the presence of digestive conditions and extended shelf life during storage. To address this concern, numerous techniques have been explored to achieve success. Among these methods, the microencapsulation of probiotics has been proposed as the most effective way to overcome this challenge. The combination of nanomaterials with biopolymer coating is considered a novel approach to improve its viability and effective delivery. The use of polysaccharides and proteins-based bionanocomposites for microencapsulation of probiotics has emerged as an efficient and promising approach for maintaining cell viability and targeted delivery. This review article aims to investigate the use of different bionanocomposites in microencapsulation of probiotics and their effect on cell survival in long-term storage and harsh conditions in the gastrointestinal tract.

Bacterial infection, which is resistant to antibiotics, is one of the causes of non-healing wounds. This study aims to investigate the effect of increasing the penetration of silver nanoparticles into infected wounds in normal and diabetic rats by static magnetic field (SMF). After synthesis and physicochemical investigation of Ag/Fe3O4 NCs, antibacterial and cytotoxic properties were investigated. 45 normal and 27 diabetic rats with Staphylococcus aureus infected wounds were randomly divided into groups of 9 with different treatments. Ag/Fe3O4 NCs showed superparamagnetic behavior and had uniform and spherical particles with an average size of 32 nm. In normal infected wound in vivo study, a significant biofilm reduction was observed in group Ag/Fe3O4 NCs with SMF on day 7. Also, on day 14, the group's wound healing percentage was 94%. Hair follicles were observed only in the group on day 21. In diabetic rats, on day 7 wound healing in the Ag/Fe3O4 NCs with SMF group was 50%. On day 21, the high scores in epithelization, new vessel, and collagen maturation belonged to mentioned group. In vivo evaluation of Ag/Fe3O4 NCs under SMF exhibited acceleration of wound healing in diabetic rats, which can provide a noninvasive therapeutic tool for chronic wound healing of diabetic patients.

Microencapsulation is one of the most important methods to enhance the survival of bacteria when exposed to various harsh conditions. The present study evaluated the viability of L. reuteri ATCC 23272 microencapsulated in polysaccharide-based bionanocomposite. Inulin, polydextrose, and pectin were utilized as prebiotics, and magnesium oxide nanoparticles (MgO NPs) as reinforcing agent in the microgel structure. The composition of bionanocomposite was optimized using the simplex-lattice mixture method. Bionanocomposite optimal formulation was achieved by combining 91.6 % inulin and 8.4 % pectin in the presence of MgO NPs. L. reuteri prebiotic score (1.33) and E. coli (1.08), extrusion efficiency (97.57 %), viability after drying (99.37 %), and viability in simulated gastrointestinal conditions (SGI) (91.74 %) were obtained. Not using MgO NPs in the optimal composite structure caused a decrease of 2.14 log CFU/g in SGI. During 28 days of storage of bacteria at 4 and 25 °C, respectively, a reduction of 2.56 and 3.04 log CFU/g was observed for free cells compared to encapsulated cells. SEM, FTIR, and XRD analyses were performed on ingredients and microcapsules with and without bacteria. The results exhibited that the optimal bionanocomposite could be used as a beneficial encapsulation system to improve the performance of probiotics in harsh conditions.