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Lactobacillus salivarius was encapsulated by ionic gelation using high polymerization degree agave fructans and sodium alginate to obtain symbiotic microspheres stabilized by different drying methods. The microbeads were characterized by physicochemical, reconstitution, microstructure, thermal, radiographic and infrared properties. The viability of L. salivarius was evaluated for one month at 4 °C and under in vitro gastrointestinal conditions. The ionic gelation-drying technique allowed an encapsulation efficiency greater than 90 %. Oven-dried microbeads showed higher viability under in vitro gastrointestinal conditions. Freeze-drying microbeads showed higher viability during storage at 4 °C, and spray-dried microbeads showed a more notable survival at 40 °C after one month of storage. All samples showed up to 7 log CFU/g of viability after in vitro digestion, which is a benefit for consumer health. It was concluded that the ionic gelation-drying process allows obtaining dry microbeads with the potential to be added to various food products., Competing Interests: Declaration of competing interest The authors confirm that they have no conflicts of interest concerning the work described in this manuscript., (Published by Elsevier Ltd.)

Milk powder, a nutrient-rich dairy product, lacks comprehensive information summarizing its specific properties when produced by spray- and freeze-dried technologies from different sources. Therefore, this study investigated the differences in physicochemical properties, microstructure, and proteome of spray- and freeze-dried milk powders from bovine, goat, and horse sources. The results revealed that spray-dried milk powder exhibited a smaller particle size, lower air content within the powder particles, inferior reconstitution properties, and lower lactose crystallinity compared with freeze-dried milk powder. Additionally, among the studied varieties, horse milk powder showed the lowest flowability but the most effective reconstitution properties. Proteomic analysis indicated that freeze-dried milk powder exhibited higher levels of immune-related proteins, including complement C3, C7, and complement factor B, and antimicrobial enzymes such as lysozyme and lactoperoxidase compared with spray-dried milk powder. Furthermore, specific milk powders contained more immune-related proteins such as serum amyloid A, myeloid antimicrobial peptide-28, polymeric immunoglobulin receptor, and mucin-1 compared with bovine milk powder. These findings contribute to a deeper understanding of the differences in the physicochemical properties and potential biological functions of spray- and freeze-dried milk powders from various sources, which may help in further optimizing specific milk powder processing technologies., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

A bio-composite material was developed that contains chitosan, food-grade algae, and zeolite for the removal of brilliant green (BG) dye. The synthesized bio-composite was dried via two different methods (air-drying; AD, and freeze-drying; FD). The physicochemical characterization of air-dried chitosan-algae-zeolite (Cs-Alg-Zl-AD) and freeze-dried chitosan-algae-zeolite (Cs-Alg-Zl-FD) were investigated by spectroscopy (FTIR, SEM-EDX, and XPS), diffraction (XRD), surface charge via pH pzc , specific surface area (SSA) and elemental analyses. The utilization of Box-Behnken Design (BBD) was intended to optimize the three input variables, which are adsorbent dosage, pH of medium, and contact time. The adsorption optimization process yielded optimal conditions, which were verified through a desirability test and implemented in batch-mode equilibrium experiments. The Cs-Alg-Zl-FD has a higher specific surface area (SSA = 3.29 m 2 /g) compared to Cs-Alg-Zl-AD (SSA = 1.79 m 2 /g). The Cs-Alg-Zl-FD shows greater adsorptive removal of BG (98.6 %) over Cs-Alg-Zl-AD (88.6 %), in parallel agreement with differences in the SSA. Moreover, the maximum BG dye adsorption capacities of Cs-Alg-Zl-FD (119.5 mg/g) and Cs-Alg-Zl-AD (108 mg/g) at pH = 8.1 and 25 °C. The Freundlich model fits best with Cs-Alg-Zl-AD while Langmuir and Temkin models account for the Cs-Alg-Zl-FD dye adsorption. The Cs-Alg-Zl-FD shows greater dye adsorption over four adsorption cycles, as compared with the Cs-Alg-Zl-AD., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

For monoclonal antibody drug products as for other biologics, while the innovator drug products first becomes commercially available, they are often followed by one or more biosimilar products. These biosimilars often differ from the innovator product, as well as from each other, in their formulation composition. However, the impact of the formulation composition on the stability of the active pharmaceutical ingredient subjected to different 'stresses' is still not understood. We have evaluated the effect of different formulations on structural stability and aggregation behavior of a monoclonal antibody, trastuzumab (both the drug substance and the final drug product), against three most common stresses encountered during production, storage, and formulation into a lyophilized product - freeze-thaw, freeze-drying, and agitation. Irrespective of the stabilizer used, the formulations exhibited good conformational stability against all three stresses. However, the freeze-drying process caused a significant increase in the number of soluble aggregates, but only in sucrose containing formulations. On the other hand, agitation in sorbitol containing formulation led to a significant increase in insoluble aggregates. This effect could also be attributed to the absence of surfactant in this formulation composition. The stabilizing effect of trehalose appeared to be independent of its concentration. Therefore, the effect of formulation composition is more pronounced for aggregation of trastuzumab than for its conformational stability. Our findings suggest that formulation design warrants consideration of both conformational stability and aggregation behavior of the active ingredient., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [Raj Suryanarayanan reports financial support was provided by US Food and Drug Administration. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.]., (Copyright © 2025. Published by Elsevier B.V.)

Bigels in which the cationic polymer -chitosan- is combined with an anionic polymer -karaya gum, pectin or xanthan gum- were prepared. These polymers, thanks to the attraction of their charged surface groups, are liable to form a polyelectrolyte complex that would modify the characteristics of the bigel. The obtained bigels were subsequently freeze-dried and tested to study their behavior against different conditions occurring in the vaginal environment. Swelling and drug release profiles have been evaluated in a medium that simulates the ordinary vaginal conditions and simulating the conditions after ejaculation. Thanks to the formation of polyelectrolyte complexes, the bigel is able to provide a pH-independent sustained drug release. However, when one of the polymers predominates, the release of the active ingredient turns out to be pH-dependent -which can also be beneficial in some therapeutic applications-. It has been proved that the inclusion of a higher percentage of chitosan in the bigel improves its mechanical properties, while the mucoadhesivity of the system can be improved by increasing the anionic polymer. The versatility of these systems in modulating their physicochemical properties, provides a substantial advantage over the formulations currently available on the market for vaginal drug delivery., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Background: The COVID-19 pandemic has significantly affected global health, economies, and societies, and highlighted the urgent need for rapid, sensitive, affordable, and portable diagnostic devices for respiratory diseases, especially in areas with limited resources. In recent years, there has been rapid development in integrated equipments using microfluidic chips and biochemical detection technologies. However, these devices are expensive and complex to operate, showing limited feasibility for in point of care tests (PoCTs). This study aims to develop a cost-effective, portable, and practical microfluidic nucleic acid PoCT device for rapid virus diagnosis., Results: Here, we developed a device based on freeze-dried reverse transcription loop-mediated isothermal amplification (RT-LAMP) reagents for rapid nucleic acid diagnostics. Homebrew RT-LAMP reagents were optimized to eliminate non-specific amplification. The multi-step device combines nucleic acid extraction, RT-LAMP and fluorescence detection in an integrated microfluidic chip, enabling sample-in, result-out diagnosis. This device showed satisfactory sensitivity in detecting SARS-CoV-2 and Trichomonas vaginalis RNA samples, with a limit of detection (LOD) of 400 copies/μL and 80 copies/μL respectively in 45 min. The LOD of cultured Trichomonas vaginalis samples were 0.32 cells/μL in 50 min. Additionally, the freeze-dried homebrew RT-LAMP can be stored at 4 °C for up to 30 days while still maintaining high sensitivity and detection capabilities. The cost of diagnosis was reduced to as low as 0.45 $ per reaction., Significance and Novelty: Overall, by integrating freeze-dried homebrew RT-LAMP and microfluidic chip, the device achieves ready-to-use, laboratory-free, quick, resource-independent and cost-effective nucleic acid detection, and provides a feasible alternative to complex equipments. The device shows potentials for point-of-care testing of SARS-CoV-2 and other respiratory diseases in remote or resource-limited areas with proper implementation., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Mingyu Wang, Jifang Tao, Lulu Shi, Zirui Pang, Xiaohong Xie, Shiling Xie has patent pending to China National Intellectual Property Administration. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Single-cycle viruses hold great promise as fish viral vaccines due to their high protective efficacy. Although the efficacy of the vaccine in olive flounder and rainbow trout has been proven through previous research, safety must be additionally proven considering the environment of use for commercialization. This study comprehensively assesses the safety of rVHSV-GΔTM and its impact on both the host and the surrounding environment, including the coastal habitat of nearby species and seawater. We constructed an experimental tank geographically mimicking coastal conditions and evaluated its effects on co-reared olive flounder and other organisms. Results showed no detectable viral presence in co-reared organisms or concentrated culturing seawater, and the virus demonstrated limited proliferation under varying salinity conditions. The survival rate of olive flounder immunized with rVHSV-GΔTM was significantly higher (up to 90% protection against wild-type VHSV) compared to non-immunized fish. Additionally, we developed a prototype vaccine of rVHSV-GΔTM through freeze-drying. Immunization of olive flounder with the prototype vaccine conferred significantly higher protection against wild-type VHSV. These findings underscore the safety of rVHSV-GΔTM as a commercial vaccine, unaffected by the host or surrounding environment, and highlight the efficacy of freeze-drying in delivering single-cycle viruses., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025. Published by Elsevier Ltd.)

Aerogels are regarded as the next generation of thermal insulators; however, conventional aerogels suffer from issues such as brittleness, low moisture resistance, and a complex production process. Subnanowires (SNWs) are emerging materials known for their exceptional flexibility, toughness, intrinsic hydrophobicity, and gelling capabilities, making them ideal building blocks for flexible, tough, hydrophobic, and thermally insulating aerogels. Herein, we present a simple and scalable strategy to construct SNW aerogels by freeze-drying hydroxyapatite (HAP) SNW dispersions in cyclohexane. The resulting aerogels consist of three-dimensional porous flakes composed of thin layers of SNW bundles. They exhibit numerous outstanding properties, including ultralow density (12.33 mg·cm -3 ), high porosity (99.15%), remarkable flexibility and toughness, and excellent thermal insulation properties (27.53 mW·m -1 ·K -1 ). Because the HAP SNWs consist of hydroxyapatite cores capped with hydrocarbon chains, the aerogels demonstrate intrinsic hydrophobicity (138° water contact angle) and superior thermal stability compared to polymer foams. Furthermore, the HAP SNW aerogels can effectively shield against infrared radiation due to their low thermal conductivity. This work suggests that SNWs can serve as superior building blocks for flexible, tough, and intrinsically hydrophobic aerogels, paving the way for future applications in thermal insulation.

Extracellular vesicles (EVs) are widely investigated for their implications in cell-cell signaling, immune modulation, disease pathogenesis, cancer, regenerative medicine, and as a potential drug delivery vector. However, maintaining integrity and bioactivity of EVs between Good Manufacturing Practice separation/filtration and end-user application remains a consistent bottleneck towards commercialization. Milk-derived extracellular vesicles (mEVs), separated from bovine milk, could provide a relatively low-cost, scalable platform for large-scale mEV production; however, the reliance on cold supply chain for storage remains a logistical and financial burden for biologics that are unstable at room temperature. Herein, we aim to characterize and engineer a freeze-dried, mEV formulation that can be stored at room temperature without sacrificing structure/bioactivity and can be reconstituted before delivery. In addition to undertaking established mEV assays of structure and function on our preparations, we introduce a novel, efficient, high throughput assay of mEV bioactivity based on Electric Cell Substrate Impedance Sensing (ECIS) in Human dermal fibroblast monolayers. By adding appropriate excipients, such as trehalose and tryptophan, we describe a protective formulation that preserves mEV bioactivity during long-term, room temperature storage. Our identification of the efficacy of tryptophan as a novel additive to mEV lyophilization solutions could represent a significant advancement in stabilizing small extracellular vesicles outside of cold storage conditions., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: R.G.G. and S.R.M. are company officers and shareholders at the Tiny Cargo Company Inc., which has licensed technology from Virginia Tech., (© 2024. The Author(s).)

Andrographolide (AG) is a diterpene lactone with significant anti-inflammatory and antitumor activities. However, the poor water solubility limits its clinical application. An andrographolide-salicylic acid (AG-SLA) nano-cocrystal delivery system was rapidly developed using hummer acoustic resonance (HAR) technology in this research. The formulation of the AG-SLA nano-cocrystal suspension and the process parameters for HAR technology were optimized in a high-throughput manner, with SDS-Tween 80 as the optimal composite stabilizer. Nano-cocrystal suspension of AG-SLA with an average particle size of 190 nm were successfully prepared, and then the optimal formulation were tenfold scaled up. Freeze-drying was adopted to solidify the nano-cocrystal and improve its stability. Various analytical techniques were used to characterize the particle size and solid state of the nano-cocrystals. The high-energy input from the HAR instrument induced partial amorphization of the nano-cocrystals, as confirmed by PXRD and DSC analyses. Saturation solubility experiments demonstrated that the solubility in pH 1.2 hydrochloric acid buffer and pH 6.8 phosphate buffer increased by 5.74 times and 6.82 times, respectively, compared to raw AG. In vitro dissolution tests indicated that the cumulative release over 120 min in pH 1.2 hydrochloric acid buffer and pH 6.8 phosphate buffer increased by 1.60 times and 1.88 times, respectively, compared to raw AG., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Cosmetic inspection of freeze-dried products is an important part of the post-manufacturing quality control process. Traditionally done by human visual inspection, this method poses typical challenges and shortcomings that can be addressed with innovative techniques. While many cosmetic defects can occur, some are considered more critical than others as they can be harmful to the patient or affect the drug's efficacy. With the rise of artificial intelligence and computer vision technology, faster and more reproducible quality control is possible, allowing real-time monitoring on a continuous manufacturing line. In this study, several continuously freeze-dried samples were prepared using formulations and process settings that lead deliberately to specific defects faced in freeze-drying as well as defect-free samples. Two approaches (i.e. patch-based approach and multi-label classification) capable of handling high-resolution images based on Convolutional Neural Networks were developed and compared to select the optimal one. Additional visualization techniques were used to enhance model understanding further. The best approach achieved perfect precision and recall on critical defects, with a prediction time of less than 50 ms to make a decision on the acceptance or rejection of vials generated., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier B.V. All rights reserved.)

Permanent preservation of genetic resources may be indispensable for the future of humanity. This requires liquid nitrogen, as is the case for preserving animal sperm. However, this technique is expensive and poses a risk of irrecoverable sample loss on non-replenishment of liquid nitrogen in case of natural disasters. In this study, we demonstrate that lyophilization may be used as a reliable method for long-term preservation of mouse sperm at room temperature. Sperm from four mouse strains were freeze-dried and stored in a non-temperature controlled room for 5-6 years. Although the ability of the stored sperm to activate oocytes had diminished slightly, healthy offspring were obtained by artificially activating the oocytes after sperm injection. Moreover, the birth rate did not decrease even after ≤ 6 years of storage. Furthermore, owing to its low cost, safety, and ease of storage at any location, we believe that this method could be a major mode of preserving mammalian genetic resources in the future., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Herein, a mesoporous magnetic chitosan-salicylaldehyde/calcium oxide nanoparticle (CS-SL/CaO/Fe 3 O 4 ) biocomposite adsorbent that was prepared via freeze-drying. The CS-SL/CaO/Fe 3 O 4 was utilized for the adsorption of ramazol brilliant blue (RBB) dye from aqueous solution. The physicochemical properties of the CS-SL/CaO/Fe 3 O 4 were evaluated using diverse characterization techniques, including BET, XRD, FTIR, FESEM-EDX, CHNS, and pH pzc . The three main factors for adsorption included the following A: CS-SL/CaO/Fe 3 O 4 dosage (0.02-0.1 g/100 mL), B: pH (4-10), and C: Time (60-540 min). These factors were improved using statistical methods, specifically the Box-Behnken design (BBD). The optimal conditions for achieving maximum RBB removal (62.5 %) are listed: CS-SL/CaO/Fe 3 O 4 dosage of 0.1 g/100 mL, a solution pH of 7, and a contact time of 540 min. The adsorption kinetics and equilibrium isotherms were well described by the pseudo first order (PFO) kinetic and Langmuir isotherm models, respectively. Thus, the CS-SL/CaO/Fe 3 O 4 material has a maximum adsorption capacity (q max ) of 63.3 mg/g for RBB at 25 °C. The adsorption mechanism of RBB onto the CS-SL/CaO/Fe 3 O 4 surface was attributed to electrostatic forces, n-π stacking, H-bonding, and Pi-Pi interactions. Thus, CS-SL/CaO/Fe 3 O 4 represents a recoverable magnetic adsorbent with potential for capture of organic dyes from wastewater., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Antibodies and antibody mimics are extensively used in the pharmaceutical industry, where stringent safety standards are required. Implementing heat sterilization during or after the manufacturing process could help prevent contamination by viruses and bacteria. However, conventional antibodies and antibody mimics are not suitable for heat sterilization because they irreversibly denature at high temperatures. In this study, we focused on the refolding property of the cold shock protein from the hyperthermophile Thermotoga maritima (TmCSP), which denatures at elevated temperatures but regains its native structure upon re-cooling. We designed and constructed a mutant library of TmCSP in which amino acid residues in its three surface loops were diversified. From the library, mutant TmCSPs that bind to each of eight target proteins were selected by phage and yeast surface display methods. We confirmed that the secondary structure and binding affinity of all the selected mutants were restored after heat treatment followed by cooling. Additionally, freeze-drying did not impair their binding affinity. The crystal structure of a mutant TmCSP in complex with its target, the esterase from Alicyclobacillus acidocaldarius, revealed specific interactions between them. These results clearly demonstrate the feasibility of creating heat-sterilizable antibody mimics using TmCSP as a scaffold., (© 2024 The Protein Society.)

Freeze-drying is used to prolong the shelf life of pharmaceutical formulations stored in vials. To achieve this, formulations are first frozen and then dried, yielding a porous product that can in some cases be stored even at ambient conditions. In this work, the effect of different process parameters on the properties of the porous micro-structure obtained when freeze-drying dextran solutions was studied. To characterize the pore sizes, the samples were imaged with scanning electron microscopy (SEM) and the images were manually analyzed to determine the pore size distribution. To study the robustness of such manual pore characterization methodology, a reliability analysis was carried out, which showed that defining a set of guidelines leads to comparable pore size distributions among multiple participants conducting the analysis. The pore characterization methodology was then applied to products that were freeze-dried under different conditions. Higher dextran concentrations and higher cooling rates were found to lead to predominantly smaller pore sizes and longer primary drying. The conclusions of this work complement the existing literature in demonstrating the robustness of the manual pore size analysis and give valuable insight into the link between the micro-structure formed during the freezing of dextran solutions and the drying performance., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 The Authors. Published by Elsevier Inc. All rights reserved.)

Chitosan salicylaldehyde/calcium oxide nanoparticle (CS-SL/CaO) was synthesized by hydrothermal process and isolated via different drying processes, namely, air-drying (AD) and freeze-drying (FD). The physicochemical properties of freeze-dried CS-SL/CaO nanoparticle (CS-SL/CaO-FD) and air-dried CS-SL/CaO nanoparticle (CS-SL/CaO-AD) were compared. In particular, the adsorption properties reveal that the specific surface area of CS-SL/CaO-FD increased by ca. 6 times (BET SA = 7.28 m 2 /g) greater than CS-SL/CaO-FD (BET SA = 1.26 m 2 /g). Also, the adsorptive removal of acid red 88 dye (AR88) from aqueous media was optimized by employing the Box-Behnken design (BBD). The optimal adsorption conditions obtained from desirability functions test for the removal of AR88 dye employed a dosage of 0.09 g/100 mL of adsorbent dosage at a solution pH of 5.6 and 25 °C. The best AR88 dye removal was found for the adsorbents CS-SL/CaO-AD (38.2 %) and CS-SL/CaO-FD (86.1 %), which concur with differences in the adsorbent surface areas. Moreover, the adsorption kinetics and isotherm profiles for CS-SL/CaO-FD were described by the pseudo second order (PSO) and Temkin models, where the maximum adsorption capacity of AR88 by CS-SL/CaO-FD 175.4 was mg/g. These findings reveal the potential application of the CS-SL/CaO-FD towards removal of toxic cationic dye (AR88) from an aqueous environment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This article compares and explores vacuum foam-drying as an alternative drying technology to freeze-drying and spray drying for a recombinant human lipase as the model protein. Materials characteristics such as structure, surface composition and the solid-state properties of the dry materials were compared and investigated. Moreover, the technical functionality in terms of reconstitution characteristics and the lipase stability were also investigated. The stability of the lipase was evaluated through activity measurements. Sucrose and dextran D40 (40 kDa) were used as matrix former and the surfactant α-dodecyl maltoside was used as surface active additive. The study demonstrated that the drying technique greatly influenced the material structure and composition which in turn affected the reconstitution characteristics. The lipase was overrepresented at the material surface in declining order spray-dried > vacuum foam-dried > freeze-dried. The lipase activity was retained up to 10 % lipase content in solids, but at 20 % lipase a loss of activity was observed for all drying techniques. Phase separation in the solid material may be an explanation. Vacuum foam-drying shows promise as an alternative drying technique for the lipase, and potentially other proteins., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Electrostatic spray drying (ESD) of a milk protein matrix comprising whey protein isolate (WPI), skim milk powder (SMP), and lactose was compared to conventional spray drying (CSD) and freeze-drying (FD). ESD and CSD were used to produce powders at low (0.12-0.14), medium (0.16-0.17), and high (0.31-0.36) levels of water activity (a w ), while FD powders targeted low a w (0.12). Maillard reaction indicators were studied after drying and during storage for up to 28 days at 20, 40, or 60 °C by measuring free -NH 2 groups, as an indicator of available lysine, and 5-hydroxymethylfurfural (HMF). After drying, levels of residual free -NH 2 groups were ~15% higher in ESD and FD powders than in their CSD counterparts. CSD powders also had ~14% higher HMF concentrations compared to their ESD and FD counterparts. Storage led to reductions in free -NH 2 groups and increases in HMF content in all powders, the extent of which increased with increasing storage temperature. Reductions in free -NH 2 groups followed first-order reaction kinetics at 20 and 40 °C but second-order reaction kinetics at 60 °C. Lactose crystallization was detected in high-a w CSD powders after 14 d at 40 °C and in both CSD and ESD powders after 7 d at 60 °C. Overall, we found that ESD is a gentle drying technology which enables production of powders with lower Maillard reaction markers.

Here, hybrid stimuli-responsive (exhibiting pyroelectricity and piezoelectricity) porous cryogels are engineered by embedding tourmaline nanoparticles (TNs) in a cellulose nanofiber (CNF) skeleton to generate high-performance CNF-TN-based airborne particulate matter (PM) filters. First, single-layer hybrid cryogels with varying TN contents (0-5% w v -1 ) are assembled, and the design principles for multilayered filters are established based on a novel sequential pre-freezing and freeze-drying technique. As observed, the embedded TNs transformed the CNF network into a more homogeneous, isotropic, and firm structure, thus improving the structural integrity and thermal stability of the assembled cryogels while maintaining their ultrahigh porosity and low density. The TN induced piezoelectric voltage in the cryogels during filtration significantly enhanced the filtration performance. Furthermore, the patterned surface texture of the cryogels notably improved quality factor (Q f ) and the reusability of the layered filters overall. The explored hybrid cryogels, particularly those exhibiting multilayered configurations, can be deployed for high-performance airborne particulates filtration owing to the synergistic effect of their mechanical robustness, stability, and high filtration efficiency. As far as it is known that, the Q f values (>0.04) obtained by the three-layered cryogels are similar to or even higher than those of the reported best aerosol filters., (© 2024 The Author(s). Small published by Wiley‐VCH GmbH.)

This work demonstrates the preparation of fast-swelling hydrogels based on poly(vinyl alcohol) (PVA) and tamarind xyloglucan (XG), utilizing freeze-drying to achieve an interconnected macroporous structure. Although XG is non-toxic and abundant, it has poor mechanical properties. Therefore, XG was mixed with PVA and crosslinked with citric acid (CA). Without XG, the crosslinked PVA sample contained partially aligned channels several hundred microns wide. The addition of XG (25% w / w ) reduced the structural order of the hydrogels. However, the addition of XG improved the swelling ratio from 308 ± 19% in crosslinked PVA to 533.33% in crosslinked PVA/XG. XG also increased the porosity, as the porosity of the crosslinked PVA, XG, and PVA/XG samples was 56.09 ± 2.79%, 68.99 ± 2.06%, and 66.49 ± 1.62%, respectively. Resistance to compression was decreased by the incorporation of XG but was increased by CA crosslinking. The determination of the gel fraction revealed that CA crosslinking was more effective for the PVA component than the XG component. The swelling of all hydrogels was very rapid, reaching equilibrium within 10 s, due to the interconnected macroporous structure that allowed for capillary action. In conclusion, the prepared hydrogels are non-cytotoxic and well suited for biomedical applications such as drug delivery, wound dressings, and hygienic products.

Sweet potato ( Ipomoea batatas (L.) Lam.) is a nutrient-dense crop rich in fibre, minerals, and antioxidant compounds, including carotenoids and phenolic compounds, such as anthocyanins. Dehydrating sweet potato (SP) for flour production enhances its value and produces shelf-stable, health-promoting food products. This study investigated the effects of hot-air drying (HAD: 75 °C/20 h) and freeze-drying (FD: -41-30 °C/70 h) on the bioactive composition of flours from three SP varieties: Bonita (white-fleshed), Bellevue (orange-fleshed), and NP1648 (purple-fleshed). Key assessments included the total phenolic content (TPC), the total carotenoid content (TCC), and the total anthocyanin content (TAC) and the antioxidant activity (DPPH and FRAP). The results revealed distinct raw materials' bioactive profiles: Bellevue was rich in TCC (49.3 mg of β-carotene/100 g db), NP1648 showed elevated TAC (27.3 mg of cyanidin-3-glucoside/100 g db), and Bonita exhibited minimal content of bioactive compounds. Both drying methods yielded significant losses of bioactive compounds, with the TPC decreasing by over 60%, while TAC and TCC losses did not exceed 32%, revealing higher stability. Multivariate analysis indicated that the variety significantly influenced the bioactive profiles more than the drying method. The interaction between carotenoids and anthocyanins and the SP fibrous composition likely contributed to their stability during drying, indicating that FD showed no advantages over HAD. The appealing colours and high antioxidant content of Bellevue and NP1648 flours suggest their potential as ingredients for enhancing foods' bioactivity and sensory acceptance.

Foods and beverages with health benefits have become increasingly popular with consumers, and fruits and legumes are considered good sources of nutrients. In this study, sea buckthorn and fava bean were used as the main raw materials to prepare sea buckthorn-fava bean composite instant powder (S-FCP). Different drying methods (spray-drying (SD) and freeze-drying (FD)) combined with carriers (maltodextrin (MD) and inulin (INU)) were involved to investigate their effects on physicochemical properties, functional properties, and sensory attributes of instant powder. The results showed that FD better protected the color of the S-FCP and produced particles possessing more porous structures compared to SD; FD-INU (freeze-dried-inulin) had the shortest dissolution time and the largest solubility. In addition, FD-INU had the highest total phenolic and total flavonoid contents and the strongest antioxidant capacity, and FD-INU had better overall organoleptic properties and hypoglycemic potential. Therefore, FD and the use of INU as a carrier are more suitable for the production of the S-FCP. This work provides a promising approach for developing a high-valued instant powder beverage composed of sea-buckthorn/broad bean, which also contributes to the development of the functional food industry.

Haemorrhagic shock is a leading cause of death worldwide. Blood transfusions can be used to treat patients suffering severe blood loss but donated red blood cells (RBCs) have several limitations that limit their availability and use. To solve the problems associated with donated RBCs, several acellular haemoglobin-based oxygen carriers (HBOCs) have been developed to restore the most important function of blood: oxygen transport. One promising HBOC is the naturally extracellular haemoglobin (i.e. erythrocruorin) of Lumbricus terrestris (LtEc). The goal of this study was to maximise the portability of LtEc by lyophilising it and then testing its stability at elevated temperatures. To prevent oxidation, several cryoprotectants were screened to determine the optimum formulation for lyophilisation that could minimise oxidation of the haem iron and maximise recovery. Furthermore, samples were also deoxygenated prior to storage to decrease auto-oxidation, while resuspension in a solution containing ascorbic acid was shown to partially reduce LtEc that had oxidised during storage (e.g. from 42% Fe 3+ to 11% Fe 3+ ). Analysis of the oxygen equilibria and size of the resuspended LtEc showed that the lyophilisation, storage, and resuspension processes did not affect the oxygen transport properties or the structure of the LtEc, even after 6 months of storage at 40 °C. Altogether, these efforts have yielded a shelf-stable LtEc powder that can be stored for long periods at high temperatures, but future animal studies will be necessary to prove that the resuspended product is a safe and effective oxygen transporter in vivo .

A high bacterial cellulose (BC) producing Komagataeibacter intermedius (KEI6 strain) was isolated from water kefir grains in Xinjiang, China. Under optimized culture conditions, the KEI6 strain was able to produce BC (KEI6-BC) up to 7.03 g/L dry weight. In this study, the rheological properties, hydrophilicity, molar mass, and specific surface area of KEI6-BC were systematically evaluated and characterized by three different drying treatments (freeze-drying, drying at 50 °C, and high-pressure homogenization). The results showed that KEI6-BC has a storage modulus of 10 4  Pa and a weight average molecular weight of 4.19×10 5  g/mol, which exhibits a randomly curled conformational polymer structure. Interestingly, freeze-dried treated KEI6-BC exhibited a highly uniform fiber distribution as well as good functional group retention, crystallinity, and thermal stability. In addition, we used freeze-dried KEI6-BC as a carrier to load ampicillin sodium and evaluated its antibacterial activity. It was found that freeze-dried KEI6-BC was promising as a carrier for slow drug release as well as exhibited good antibacterial activity after drug loading, demonstrating its great potential as an efficient antibacterial composite film., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Freeze-drying enables delicate, heat-sensitive biomaterials to be stored in a dry form even at room temperature. However, exposure to physicochemical stress induced by freeze-drying presents challenges for maintaining material characteristics and functionality upon reconstitution, for which reason excipients are required. Although wide variety of different excipients are available for pharmaceutical applications, their protective role in the freeze-drying is not yet fully understood. In this study, our aim was to use molecular dynamics simulations to screen the properties of different sugars and amino acids, which could be combined with plant-based nanofibrillated cellulose (NFC) hydrogel to provide protective matrix system for future freeze-drying for pharmaceuticals and biologics. The changes in the NFC-based formulations before and after freeze-drying and reconstitution were evaluated using non-invasive Timegate PicoRaman spectroscopy and traditional characterization methods. We continued to the freeze-drying with the NFC hydrogel formulations including lactose with and without glycine, which showed the highest attraction preferences on NFC surface in silico. This formulation enabled successful freeze-drying and subsequent reconstitution with preserved physicochemical and rheological properties. Raman spectroscopy gave us insights of the molecular-level changes during freeze-drying, especially the mutarotation of lactose. This research showed the potential of integrating in silico screening and non-invasive spectroscopical method to design novel biomaterial-based formulations for freeze-drying. The research provided insights of the molecular-level interactions and orientational changes of the excipients, which might be crucial in future freeze-drying applications of pharmaceuticals and biologics., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper: At the time of conducting the research, authors J. Zini and J. Kekkonen were affiliated with the University of Helsinki and the University of Oulu/University of Eastern Finland, respectively. Currently, they are employed by TimeGate Instruments Ltd., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Several nanotechnology-based formulation strategies have been reported for the oral administration of biological drugs. However, a prerequisite often overlooked in developing these formulations is their adaptation to a solid dosage form. This study aimed to incorporate a freeze-drying step, using either mannitol or sucrose laurate (SLAE), into the formulation of new insulin-zinc nanocomplexes to render them resistant to intestinal fluids while maintaining a high protein loading. The resulting freeze-dried insulin-zinc nanocomplexes exhibited physicochemical properties consistent with the target product profile, including a particle size of ∼ 100 nm, a zeta potential close to neutrality (∼ -15 mV) and a high association efficiency (> 90%). Importantly, integrating the freeze-drying step in the formulation significantly improved the colloidal stability of the system and preserved the stability of the insulin molecules. Results from in vitro and in vivo studies indicated that the insulin activity remained fully retained throughout the entire formulation and freeze-drying processes. In brief, we present a novel protein formulation strategy that incorporates a critical freeze-drying step, resulting in a dry powder enabling efficient protein complexation with zinc and optimized for oral administration., (© 2024. Controlled Release Society.)

Functional microbiome development has steadily increased; with this, the viability of microbial strains must be maintained not only after the manufacturing process but also at the time of consumption. Survival is threatened by various unavoidable factors during freeze-drying and shelf storage. Here, the aim was to optimize the manufacturing process of the functional strain Lactiplantibacillus plantarum IDCC 3501 after freeze-drying and storage. Explosive growth was achieved using a medium composition with two nitrogen sources and a mineral, and growth was drastically improved by neutralizing the medium pH during the culture of L. plantarum IDCC 3501. Culture optimization involved a smaller cell size, leading to less intracellular free water. Moreover, when maltodextrin (MD) powder was directly added to the harvested cells, some intracellular free water was extracted from the bacterial cells, resulting in a dramatic increase in the viability of L. plantarum IDCC 3501 after freeze-drying and subsequent storage. Furthermore, MD enhanced survival in a dose-dependent manner. Bacterial survival was correlated with lysozyme tolerance; therefore, the positive result might have been caused by the osmotic dehydration of intracellular free water, which would potentially damage the bacterial cells via ice crystallization and/or a phase transition during freeze-drying. These critical factors of L. plantarum IDCC 3501 processing provide perspectives on survival issues for manufacturing microbiome strains. KEY POINTS: • Culture conditions for probiotic bacteria were optimized for high growth yield. • Osmotic dehydration improved bacterial survival after manufacturing and shelf storage. • Reduction in intracellular free water content is crucial for intact survival., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Probiotics have awakened a great interest in the scientific community for their potential beneficial effects on health. Although only allowed by the European Food Safety Agency as a nutrition declaration associated with the improvement of lactose digestion, recent in vitro and in vivo studies have demonstrated their varied beneficial effect for the improvement of certain pathologies. However, probiotics face stability and viability challenges, which make their delivery difficult in sufficient quantities for these effects to be observed. Thus, there is a dire need for the development and implantation of innovative technological protection procedures. In this sense, encapsulation rises as a widely applied technique, offering additional advantages. In the present study, a systematic review was conducted for the evaluation of the main encapsulation technologies applied in literature, considering operating conditions, probiotics, and encapsulation efficacy. For this purpose, several conditions are evaluated: a) the characteristics, storage conditions and viability of probiotics; b) evaluation and comparison of the probiotic stabilization for the main encapsulation methods; and c) co-encapsulation with potential bioactive molecules as a new alternative for improving cell viability. This evaluation revealed the efficacy of seven encapsulation techniques on the improvement of the stability and viability of probiotics., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Freeze-drying is a preservation method known for its effectiveness in dehydrating food products while minimizing their deterioration. However, protein denaturation and oxidation during freezing and drying can degrade the quality of meat and aquatic products. Therefore, finding the strategies to ensure the dried products' sensory, functional, and nutritional attributes is crucial. This study aimed to summarize protein denaturation mechanisms and overall quality changes in meat and aquatic products during freezing and drying, while also exploring methods for quality control. Different freeze-drying conditions result in varying levels of oxidation and functionality in meat and aquatic products, leading to changes in quality, such as altered fatty and amino acid compositions, protein digestibility, and sensory attributes. To obtain high-quality dried products by freeze-drying, several parameters should be considered, including sample type, freezing and drying temperatures, moisture content, pulverization effects, and storage conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The target of breeding red-fleshed apples is to increase their potential health benefits related to red flesh coloration and consumer acceptance. The objective of this study was to determine the usefulness of four clones (90, 120, 156, and 158) of red-fleshed apples for freeze-drying compared to the cultivar 'Trinity'. Red-fleshed apples were dried in the form of slices using a laboratory freeze-dryer. The changes in color features and image texture parameters after drying and the sensory quality of freeze-dried samples were assessed. Trends of increase in the value of the L* parameter and decrease in the a* and b* parameters after freeze-drying were observed. Furthermore, freeze-drying caused statistically significant changes in analyzed image textures named XHMean, RHMean, SHMean, VHMean, LHMean, and UHMean. Machine learning models developed based on the color parameters L* , a* , and b* distinguished raw and freeze-dried red-fleshed apples with an average accuracy of 84% for clone 90 up to 99.0% for clone 156, and models based on twenty selected image textures exhibited an accuracy of 98.5% for clone 156 to 100% for clones 90 and 158 and the cultivar 'Trinity'. The very attractive external appearance, medium-intense fruity smell, crunchiness, and intense fruity taste of all the apple slices were revealed. The innovative aspect of this study included the comparison of the drying behavior and sensory quality of the new clones and a standard cultivar of red-fleshed apples. Moreover, innovative methods and results were used to determine the effect of freeze-drying on red-fleshed apple quality, considering novel models involving thousands of image textures and machine learning algorithms.

Background/Objectives: Freeze-drying is a dehydration method that extends the shelf life and stability of drugs, vaccines, and biologics. Recently, its role has expanded beyond preservation to improve novel pharmaceuticals and their carriers, such as hydrogels, which are widely studied for both drug delivery and wound healing. The main aim of this study was to explore the multifunctional role of freeze-drying in improving the physicochemical properties of sodium alginate/poly(vinyl alcohol)-based hydrogels for medical applications. Methods: The base matrix and hydrogels containing a nanocarrier-drug system, were prepared by chemical cross-linking and then freeze-dried for 24 h at -53 °C under 0.2 mBa. Key analyses included determination of gel fraction, swelling ratio, FT-IR, SEM, TG/DTG, in vitro drug release and kinetics, and cytotoxicity assessment. Results: Freeze-drying caused an increase in the gel fraction of the hydrogel with the dual drug delivery system from 55 ± 1.6% to 72 ± 0.5%. Swelling ability was pH-dependent and remained in the same range (175-282%). Thermogravimetric analysis showed that freeze-dried hydrogels exhibited higher thermal stability than their non-freeze-dried equivalents. The temperature at 10% weight loss increased from 194.0 °C to 198.9 °C for the freeze-dried drug-loaded matrix, and from 188.4 °C to 203.1 °C for the freeze-dried drug-free matrix. The average pore size of the freeze-dried hydrogels was in the range of 1.07 µm ± 0.54 to 1.74 µm ± 0.92. In vitro drug release revealed that active substances were released in a controlled and prolonged way, according to the Korsmeyer-Peppas model. The cumulative amount of salicylic acid released at pH = 9.0 after 96 h was 63%, while that of fluocinolone acetonide reached 73%. Both hydrogels were non-toxic to human fibroblast cells, maintaining over 90% cell viability after 48 h of incubation. Conclusions: The results show a high potential for commercialisation of the obtained hydrogels as medical dressings.

New techniques for the immobilization of yeast cells have the potential for enhancement of the beer production process. Alongside conventional materials for cell immobilization, there is a rising trend toward polysaccharide-protein systems. This study focused on the immobilization of yeast cells ( Saccharomyces pastorianus ) via a freeze-drying process. The whey protein isolate, sodium alginate, maltodextrin, inulin, and their blends were used for carrier preparation. The effect of a 1.0% inulin solution as a cryoprotectant on the viability of the yeast cells after the freeze-drying process was also analyzed. The powders were assessed for cell viability, moisture content, water activity, solubility, particle size, and surface charge. According to the results, the addition of whey proteins reduced the moisture content, while solubility did not significantly decrease. Samples containing whey protein showed slight diameter variations. The negative surface charge observed in all samples, especially the control, indicates a cell's tendency to aggregate, demonstrated by optical microscopy. SEM micrographs showed successful cell immobilization in polysaccharide-protein carriers. Furthermore, inulin and whey protein addition enhanced cell protection during the immobilization of cells. The freeze-drying technique demonstrates efficacy in immobilization of yeast cells, indicating its potential for applications in the food and beverage industry.

Zirconia aerogels possess significant applications, including their use catalyst carriers, thermal insulation materials, and thermal barrier coatings. This is due to their ultrahigh temperature resistance, high porosity, and low thermal conductivity. Nonetheless, the inherent challenges associated with ZrO 2 aerogels, such as high brittleness, low compressive strength, and inadequate formability, restrict their potential applications. In this paper, with ultralow thermal conductivity and high strength zirconia aerogel composites with inorganic zirconium salt zirconium carbonate as the raw material, acetic acid as the solvent, polyvinylpyrrolidone (PVP) as the viscosity builder to stabilize the structure of the aerogel during the freeze-drying process. Additionally, yttrium nitrate hexahydrate (Y(NO 3 ) 3 ·6H 2 O) is employed as a phase stabilizer. The sol-gel method, in conjunction with the freeze-drying process, is utilized to fabricate ZrO 2 aerogel composites with an optimized microstructure. The findings indicate that optimal process parameters are achieved with a PVP solution concentration of 2.0 wt % and a zirconium carbonate concentration of 20 wt %. The mechanical properties of the resulting composites reach up to 550 kPa, while the thermal insulation performance exhibits a temperature difference of 207 °C/cm and a thermal conductivity of 0.0504 W/(m·K). This advancement addresses the mechanical stability issues commonly associated with traditional ceramic aerogels and widely used elastic insulating materials, thereby enhancing their applicability as thermal insulation and heat preservation materials.

Microneedle patch (MNP) has become a hot research topic in the field of transdermal drug delivery due to its ability to overcome the stratum corneum barrier. Among the various types of microneedles, dissolving microneedles represent one of the most promising transdermal delivery methods. However, the most used method for preparing dissolving microneedles, namely microfabrication, suffers from issues such as long drying time, susceptibility to humidity, and large batch-to-batch variability, which limit the development of dissolving microneedles. In this study, we report for the first time a method for preparing dissolving microneedles using freeze-drying technology. We screened substrates suitable for freeze-dried microneedle patch (FD-MNP) and used coating technology to enhance the mechanical strength of FD-MNP, allowing them to meet the requirements for skin penetration. We successfully prepared FD-MNP using hyaluronic acid as the substrate and insulin as the model drug. Scanning electron microscopy revealed that the microneedles had a porous structure. After coating, the mechanical strength of the microneedles was 0.61 N/Needle, and skin penetration rate was 97%, with a penetration depth of 215 μm. The tips of the FD-MNP dissolved completely within approximately 60 s after skin penetration, which is much faster than conventional MNP (180 s). In vitro transdermal experiments showed that the FD-MNP shortened the lag time for transdermal delivery of rhodamine 123 and insulin compared to conventional MNP, indicating a faster transdermal delivery rate. Pharmacological experiments showed that the FD-MNP lowered mouse blood glucose levels more effectively than conventional MNP, with a relative pharmacological availability of 96.59 ± 2.84%, higher than that of conventional MNP (84.34 ± 3.87%), P = 0.0095. After storage under 40℃ for two months, the insulin content within the FD-MNP remained high at 95.27 ± 4.46%, which was much higher than that of conventional MNP (58.73 ± 3.71%), P < 0.0001. In conclusion, freeze-drying technology is a highly valuable method for preparing dissolving microneedles with potential applications in transdermal drug delivery., (© 2024. Controlled Release Society.)

Freeze-drying (FD) and cryo-milling (CM) are common methods for preparing powder gelatinized starch samples. This study investigates the structural characterization of raw/gelatinized maize starches and digestibility after FD/CM processes to elucidate their effect on starch digestibility determination. Results showed that FD slightly increased digestibility, while higher initial glucose content in CM samples, especially for gelatinized samples. Only FD retained the granular morphology and relative crystallinity (RC), while gelatinized-FD decreased RC by 75%. CM decreased RC by 12%, while gelatinized-CM decreased it by 97%. Combined with short-range and chain structural results, FD tended to disrupt internal connected chains through volume stress, while CM cleaved glycosidic bonds in external chain. Stretched chains in gelatinized starch promoted the breakage of chains during shearing and their efficient binding with digestive enzymes. These findings would provide a basis for pre-treatment of powder samples and processes of starch- rich foods., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Messenger RNA (mRNA) vaccines have revolutionized the fight against infectious diseases and are poised to transform other therapeutic areas. Lipid nanoparticles (LNP) represent the most successful delivery system for mRNA. While the mRNA-LNP products currently in clinics are stored as frozen suspensions, there is evidence that freeze-drying mRNA-LNP into dry powders can potentially enable their storage and handling at non-freezing temperatures. Previously, we successfully applied thin-film freeze-drying (TFFD) to transform a polyadenylic acid [poly(A)]-LNP formulation from a liquid suspension to dry powders. The poly(A)-LNP were structurally multilamellar spheres without blebs, but the mRNA vaccines in clinics are comprised of mRNA-LNP that are structurally spheres surrounded by a unilamellar lipid bilayer, with some containing blebs, and it was reported that the presence of blebs increases the sensitivity of mRNA-LNP to freeze-drying-induced stress. In the present study, using an influenza A virus hemagglutinin (HA) mRNA in LNP that were structurally similar to that in the COVID-19 mRNA vaccines currently in clinic, we studied the effect of TFFD on the physical properties, internal structure, as well as immunogenicity of the HA mRNA-LNP vaccine. We concluded that TFFD can be utilized to prepare dry powders of the HA mRNA-LNP, but a sufficient amount of excipients were needed to minimize changes in the physical properties, structure, and immunogenicity of the HA mRNA-LNP vaccine., Competing Interests: Declaration of competing interest HX, CM, ROW, ZC, and DW report a relationship with TFF Pharmaceuticals, Inc., (Copyright © 2024. Published by Elsevier B.V.)

Liposomes being a promising colloidal system facilitates delivery of drugs with limited pharmacokinetic properties to achieve desirable clinical applications. However, development of a stable liposomal system is always challenging due to multiple complexities involved. Aqueous instability of liposomes and impact of various process and formulation parameters can lead to serious alteration of its therapeutic performance. In the proposed work, the authors aim to develop stable Ibrutinib-loaded liposomes using lyophilization and Quality-by-Design and assess their long-term stability. Ibrutinib-loaded liposomes were developed and optimized using Quality-by-Design technique and were further PEGylated and characterized for the same. Effect of cryoprotectants during lyophilization and other parameters are evaluated to obtain a robust formulation. The stability studies were conducted upto 6 months at various storage conditions to evaluate the effect of lyophilization. The impact of formulation, processing and lyophilization parameters on physicochemical properties of developed liposomal systems were evaluated and are critically discussed. Liquid dispersion exhibited a %degradation of 16-36% at 25 °C/60% RH which was reduced for less than 1% in lyophilized formulation for 6 months. Critical analysis and assessment of various parameters lead to identification of optimum conditions to manufacture this drug product and also opens way forward for further evaluation and translational possibilities., (© 2024. Controlled Release Society.)

Studies were conducted focusing on the drying of chili pepper fruits ( Capsicum annuum L.), cultivar Cyklon, using convective (AD), convective-microwave (AMD), vacuum (VD), and freeze-drying (FD) methods. The influence of the drying method and temperature on the kinetics of the process and selected quality attributes of the dried product were evaluated. It was demonstrated that the Midilli model best described the drying kinetics for all methods across the entire measurement range. FD and VD produced dried products with the highest brightness and the greatest value of the a* color parameter. The lowest b* color parameter was observed for the product dried using FD at 40 °C, while the highest b* value was noted for samples dried using AMD (100 W) at 60 °C. The highest carotenoid retention was achieved with the FD method at 40 °C, while the lowest carotenoid content was found in the product obtained using the AMD method (100 W) at 60 °C. The smallest losses of capsaicinoids were observed after FD drying at 40 °C, while the largest were found for AMD (100 W) at 60 °C. The analysis of chili pepper fruit extracts revealed the quantitative composition of 12 main phenolic compounds using the UHPLC-UV method. The highest polyphenol content was obtained with FD, while the lowest total polyphenol content was recorded after AD. Regardless of temperature, the total flavonoid content was highest in extracts from FD products, and the lowest flavonoid content was found after AMD at 100 W. For all drying methods analyzed, the total flavonoid content in the pepper extracts decreased with increasing temperature.

Over the past decade, continuous spin freeze-drying technology has emerged as a promising alternative to conventional batch freeze-drying, effectively addressing many of the latter's inherent disadvantages. Much of the focus during this period has been on controlling and optimizing the primary drying phase of this process. However, optimizing the secondary drying step is equally critical for the overall efficiency of the process. The primary aim of this study was to develop a comprehensive semi-mechanistic model for the secondary drying phase in continuous spin freeze-drying, accounting for the effects of process settings such as freezing rate and product temperature on desorption kinetics. Additionally, the study aimed to address discrepancies between conventional desorption models, typically applied in batch freeze-drying, and the observed data in this research. To achieve this, a residual moisture-dependent activation energy was introduced to improve the accuracy of the desorption model. Using NIR spectroscopy and IR-thermography, unknown model parameters could reliably be estimated using a simple and fast procedure. The calibrated model successfully predicted the final moisture content with an accuracy within 0.11% of the measured value under previously untested process conditions. Ultimately, the proposed semi-mechanistic model demonstrated its reliability in predicting the impact of new process conditions on both product temperature and residual moisture over time, enabling the development of a practical design space., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Thomas De Beer reports a relationship with RheaVita that includes: board membership, employment, and equity or stocks. Laurens Leys reports a relationship with RheaVita that includes: employment. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The pharmaceutical industry is progressing towards more continuous manufacturing techniques. To dry biopharmaceuticals, continuous freeze drying has several advantages on manufacturing and process analytical control compared to batch freeze-drying, including better visual inspection potential. Visual inspection of every freeze-dried product is a key quality assessment after the lyophilization process to ensure that freeze-dried products are free from foreign particles and defects. This quality assessment is labor-intensive for operators who need to assess thousands of samples for an extensive amount of time leading to certain drawbacks. Applying Artificial Intelligence, specifically computer vision, on high-resolution images from every freeze-dried product can quantitatively and qualitatively outperform human visual inspection. For this study, continuously freeze-dried samples were prepared based on a real-world pharmaceutical product using manually induced particles of different sizes and subsequently imaged using a tailor-made setup to develop an image dataset (with particle sizes from 50μm to 1 mm) used to train multiple object detection models. You Only Look Once version 7 (YOLOv7) outperforms human inspection by a large margin, obtaining particle detection precision of up to 88.9% while controlling the recall at 81.2%, thus detecting most of the object present in the images, with an inference time of less than 1 s per vial., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

New drying technologies for biologicals have recently been developed to accelerate the time-consuming batch freeze-drying (BFD) process. Among others, microwave-assisted freeze-drying (MFD) has been suggested as a faster and more effective drying technology. In this study, MFD cycles with the microwave radiation switched on and off were performed to assess the contribution of the microwave radiation to drying acceleration. It was found that thermal radiation emitted by the drying chamber walls was predominantly accelerating the drying of monodose placebos rather than microwave radiation. The combination of ultra-low chamber pressure, high thermal heat transfer and a short primary-to-secondary phase transition reduces drying times by more than 80 % compared to conventional BFD. In a second step, a design of experiment approach was used to assess the effect of thermal radiation versus microwave radiation and their combination, together with dosage properties such as fill volume and excipient concentration upon drying rate. The outcome showed the importance of high fill volume and high excipient concentration for an effective microwave contribution to the drying rate. Nevertheless, the drying acceleration for small pharmaceutical dosages with restricted solutes was mainly driven by thermal radiation rather than 2.45 GHz microwave radiation. The inability of ice to convert microwave energy into heat hampers the potential use of microwave freeze-drying for single-dose vaccines., Competing Interests: Declaration of competing interest The authors declare the following competing financial interest(s): Frédéric Mathot, Erwan Bourlès, Bernadette Scutellà and  Laurent Hansen are, or were at the time of the study employees of the GSK group of companies., (Copyright © 2024 Elsevier B.V. All rights reserved.)

To enhance the pore structure of green coffee beans (GCB) and detect the sorption capacity and extraction characteristics of flavor compounds before roasting, this study employed several methods: hot air drying (HD), freeze-drying (FD), 3-levels short-time heating with puffing (SH 1 P, SH 2 P, and SH 3 P), and 3-levels microwave with puffing (MW 45 P, MW 60 P, and MW 75 P). These methods were applied to GCBs pre-soaked in water for different times. The effects of these treatments on color change, porosity, microstructure, citric acid sorption capacity, and caffeine and chlorogenic acid extraction yield were investigated. Results indicated that, except for GCBs treated with SH 1 P, SH 2 P, SH 3 P, and MW 75 P, all other modified GCBs showed minimal color change. GCBs treated with MW 60 P exhibited favorable pore structures. MW 60 P treatments significantly improved the extraction yield of caffeine and chlorogenic acid. Furthermore, the increased porosity and improved pore size distribution of GCB after MW 60 P resulted in a significant increase in the sorption of citric acid onto modified GCB. The rate of the sorption reaction followed the pseudo-first-order kinetics. In conclusion, MW 60 P are effective treatments for enhancing the microstructure of GCB, improving sorption capacity, and improving the extraction yield of flavor compounds.

Background/objectives: Research on cyclodextrin-based metal-organic frameworks (CD-MOFs) is still in its infancy, but their potential for use in drug delivery-expressly in the lung-seems promising. We aimed to use the freeze-drying method to create a novel approach for preparing CD-MOFs. MOFs consisting of γ-cyclodextrin (γCD) and potassium cations (K + ) were employed to encapsulate the poorly water-soluble model drug Ibuprofen (IBU) for the treatment of cystic fibrosis (CF)., Methods: Using the LeanQbD ® software (v2022), we designed the experiments based on the Quality by Design (QbD) concept. According to QbD, we identified the three most critical factors, which were the molar ratio of the IBU to the γCD, incubation time, and the percentage of the organic solvent. light-, scanning electron microscope (SEM) and laser diffraction were utilized to observe the morphology and particle size of the samples. In addition, the products were characterized by Differential Scanning Calorimetry (DSC), X-ray Powder Diffraction (XRPD), Fourier Transform Infrared Spectroscopy (FT-IR) and nuclear magnetic resonance spectroscopy (NMR)., Results: Based on characterizations, we concluded that a γCD-MOF/IBU complex was also formed using the freeze-drying method. Using formulations with optimal aerodynamic properties, we achieved 38.10 ± 5.06 and 47.18 ± 4.18 Fine Particle Fraction% (FPF%) based on the Andersen Cascade Impactor measurement. With these formulations, we achieved a fast dissolution profile and increased IBU solubility., Conclusions: This research successfully demonstrates the innovative use of freeze-drying to produce γCD-MOFs for inhalable IBU delivery. The method enabled to modify the particle size, which was crucial for successful pulmonary intake, emphasizing the need for further investigation of these formulations as effective delivery systems., Competing Interests: The authors declare no conflicts of interest.

Background: Proteins are commonly used in the healthcare industry to treat various health conditions, and most proteins are sensitive to physical and chemical changes. Lyophilization, also known as freeze-drying, involves sublimating water in the form of ice from a substance at low pressure, forming a freeze-dried powder that increases its shelf life. Extreme pressure and varying temperatures in the freeze-drying process may damage the protein's structural integrity. Lyoprotectants are commonly used to protect protein conformations. It is important to choose a suitable lyoprotectant to ensure optimal effectiveness. Method: Twenty articles screened from Scopus, Web of Science, and PubMed were included in this review that discussed potential lyoprotectants and their effectiveness with different protein models. Results: Lyoprotectants were categorized into sugars, polyols, surfactants, and amino acids. Lyoprotectants can exhibit significant protective effects towards proteins, either singularly or in combination with another lyoprotectant. They exert various interactions with the protein to stabilize it, such as hydrogen bonding, hydrophobic interactions, electrostatic interactions, and osmoprotection. Conclusions: This review concludes that disaccharides are the most effective lyoprotectants, while other groups of lyoprotectants are best used in combination with other lyoprotectants., Competing Interests: The authors declare no conflicts of interest.

During pharmaceutical lyophilization processes, inter-vial drying heterogeneity remains a significant obstacle. Due to differences in heat and mass transfer based on vial position within the freeze drier, edge vials freeze differently, are typically warmer and dry faster than center vials. This vial position-dependent heterogeneity within the freeze dryer leads to tradeoffs during process development. During primary drying, process developers must be careful to avoid shelf temperatures that would result in overheating of edge vials causing the product sublimation interface temperature to rise above the critical (collapse) temperature. However, at lower shelf temperatures, center vials require longer to complete primary drying, risking collapse or melt-back due to incomplete drying. Both situations may result in poor product quality affecting drug stability, activity, and reconstitution times. We present a new approach for monitoring vial location-specific water vapor mass flow based on Tunable Diode Laser Absorption Spectroscopy (TDLAS). The single vial monitor enables measurement of the gas flow velocity, water vapor temperature, and gas concentration from the sublimating ice, enabling the calculation of the mass flow rate which can be used in combination with a heat and mass transfer model to determine vial heat transfer coefficients and product resistance to drying. These parameters can in turn be used for robust and rapid process development and control., (© 2024. The Author(s), under exclusive licence to American Association of Pharmaceutical Scientists.)

Investigating the conspicuousness of animal color patterns to different observers is crucial for understanding their function. This study examines the peculiar case of a jumping spider (Saitis barbipes) whose males display red and black ornaments during courtship despite an apparent inability to distinguish these colors. We propose that, through predator eyes, red may actually be a better match than black to the spiders' leaf litter background, and that the black fringe of hairs surrounding red ornaments may blur with red at natural predator acuities and viewing distances to produce a background-matching desaturated red. In a field experiment, we test whether red ornaments reduce predation relative to red ornaments painted black, and find that, unexpectedly, spiders with red ornaments are more heavily predated upon. Having established birds as the spiders' primary predators, we image the spiders in their natural habitat using an avian-vision camera. We find their red coloration to have similar color contrast, but lower achromatic contrast, with the background than black coloration. We also find that red and black elements blur together at typical avian acuities and viewing distances to produce lower chromatic and achromatic contrasts with the background than would be seen by animals with higher acuities and/or closer viewing distances. Interestingly, red ornaments appear orange or yellow when viewed obliquely, which reduces their achromatic, but not chromatic, contrast with the background. Our imaging results provide support for our hypothesis that red is camouflaging, whereas the results of our predation experiment do not. Any functional significance of the spiders' red coloration therefore remains unresolved., (© 2024. The Author(s).)

Debittering of pomelo juice was conducted using 3.7 g of activated resin, resulting in a 36.8% reduction in bitterness without affecting the bioactive properties of juice. The debittered juice was then encapsulated with Moringa oleifera exudate at various ratios (1-5%), yielding a powder with a slightly rough surface. Total phenol content (TPC) increased by 46-56% compared to the debittered juice. Functional yoghurt containing encapsulates at concentrations of 1% and 2% demonstrated that the 2% concentration led to longer storage duration, resulting in increased acidity and syneresis compared to the control. TPC of the yoghurt (161.89-198.22 μg Gallic acid equivalent (GAE)/g) remained significantly higher (p < 0.05) than that of the control (47.15 μg GAE/g) and acacia gum-based yoghurt (141.89-171.37 μg GAE/g), decreasing with storage duration. Addition of encapsulates significantly altered the yoghurt's texture, resulting in lower firmness (0.57 to 0.64 N) compared to the control, while adhesiveness values remained comparable (6.33 to 6.25 g.s). The highest values of G' and G" were observed in samples containing 2% encapsulates with moringa compared to those with acacia gum. This study suggests potential avenues for further exploration in functional foods with enhanced health benefits., Competing Interests: Declaration of competing interest The authors have declared no conflicts of interest for this article., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

With the increase of oily wastewater discharge and the growing demand for clean water supply, high throughput green materials for oil-water separation with anti-pollution and self-cleaning ability are urgently needed. Herein, the polysaccharide-based composite aerogels of CMC/SA@TiO 2 -MWCNTs (CSTM) with fast photo-driven self-cleaning ability have been prepared by a simple freeze-drying and ionic cross-linking strategy. The introduction of TiO 2 /MWCNTs nanocomposites effectively improves the underwater oleophobic and mechanical properties of polysaccharide aerogels and enables their photo-driven self-cleaning ability for efficient oil-water separation and purification of complex oily wastewater. For immiscible oil-water mixtures, a high separation flux of about 7650 L m -2  h -1 and a separation efficiency of up to 99.9 % was obtained. For surfactant-stabilized oil-in-water emulsion, a flux of 3952 L m -2  h -1 was achieved with a separation efficiency of up to 99.3 %. More importantly, the excellent photoluminescent self-cleaning ability and low oil adhesion contribute to the high contamination resistance, excellent reusability, and robust durability of CSTM aerogel. With the advantages of simple preparation, remarkable performance, and recyclability, this aerogel is expected to provide a green, economical, and scalable solution for the purification of oily wastewater., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: It is well known that periodontitis affects the gums and surrounding connective tissue. The chronic inflammatory response induced by bacteria in the gingival tissue leads to the loss of the collagen connection between the tooth and the bone and ultimately to bone loss., Methods: In this context, the aim of this research was the obtaining and characterization of a drug release supports in the form of sponges based on collagen, hyaluronic acid as a support and metronidazole as an antibiotic for the treatment of periodontitis. The sponges were characterized by FT-IR spectroscopy, water uptake, contact angle, SEM microscopy, in vitro metronidazole release analysis from sponges and data modeling., Results: The results showed that all the sponges had a porous structure with interconnected pores, the pore sizes being influenced by hyaluronic acid and metronidazole; the spongious structure became much more dense for samples with metronidazole content. All metronidazole-loaded sponges showed good surface wettability and an adequate swelling capacity for a suitable antimicrobial release at the periodontal pocket. The porous structures allow a controlled release, fast in the first hour, essential to control the initial microbial load at the periodontal level, which continues slowly in the following hours to ensure an effective treatment of periodontitis., Conclusions: Correlating all physical-chemical and bio-pharmaceutical results obtained, a promising solution for periodontitis treatment could be a met-ronidazole-collagen-hyaluronic system consisting of 1% collagen, 1.5% metronidazole and 0.8% hyaluronic acid, and in vitro and in vivo tests are recommended to continue studies.

Background/Objectives : This study investigates the development of 3D chitosan-x-cobalt ferrite scaffolds (x = 5, 7.5, and 10 wt%) with interconnected porosity for potential biomedical applications. The objective was to evaluate the effects of magnetic particle incorporation on the scaffolds' structural, mechanical, magnetic, and biological properties, specifically focusing on their biocompatibility and antimicrobial performance. Methods : Scaffolds were synthesized using freeze-drying, while cobalt ferrite nanoparticles were produced via a pilot-scale combustion reaction. The scaffolds were characterized for their physical and chemical properties, including porosity, swelling, and mechanical strength. Hydrophilicity was assessed through contact angle measurements. Antimicrobial efficacy was evaluated using time kill kinetics and agar diffusion assays, and biocompatibility was confirmed through cytotoxicity tests. Results : The incorporation of cobalt ferrite increased magnetic responsiveness, altered porosity profiles, and influenced swelling, biodegradation, and compressive strength, with a maximum value of 87 kPa at 7.5 wt% ferrite content. The scaffolds maintained non-toxicity and demonstrated bactericidal activity. The optimal concentration for achieving a balance between structural integrity and biological performance was found at 7.5 wt% cobalt ferrite. Conclusions : These findings suggest that magnetic chitosan-cobalt ferrite scaffolds possess significant potential for use in biomedical applications, including tissue regeneration and advanced healing therapies. The incorporation of magnetic properties enhances both the structural and biological functionalities, presenting promising opportunities for innovative therapeutic approaches in reconstructive procedures.