Ice Crystals Research Articles

Bridging the Gap in Cryopreservation Mechanism: Unraveling the Interplay between Structure, Dynamics, and Thermodynamics in Cryoprotectant Aqueous Solutions.

Cryoprotectants play a crucial role in preserving biological material, ensuring their viability during storage and facilitating crucial applications such as the conservation of medical compounds, tissues, and organs for transplantation. However, the precise mechanism by which cryoprotectants modulate the thermodynamic properties of water to impede the formation and growth of ice crystals, thus preventing long-term damage, remains elusive. This is evident in the use of empirically optimized recipes for mixtures that typically contain DMSO, glycerol, and various sugar constituents. Here, we use terahertz calorimetry, Overhauser nuclear polarization, and molecular dynamics simulations to show that DMSO exhibits a robust structuring effect on water around its methyl groups, reaching a maximum at a DMSO mole fraction of XDMSO = 0.33. In contrast, glycerol exerts a smaller water-structuring effect, even at higher concentrations (Scheme 1). These results potentially suggest that the wrapped water around DMSO's methyl group, which can be evicted upon ligand binding, may render DMSO a more surface-active cryoprotectant than glycerol, while glycerol may participate more as a viscogen that acts on the entire sample. These findings shed light on the molecular intricacies of cryoprotectant solvation behavior and have potentially significant implications for optimizing cryopreservation protocols.

Investigating the Constitutive Model of Frozen Supersulfate Saline Soil: Insights from Fractional Calculus

The study of the mechanical properties of frozen saline soil is one of the key issues in addressing the design of infrastructure in cold regions. This research focuses on the supersulfated saline soil of the Ningxia Yellow River Irrigation Area in China, conducting triaxial tests under negative temperatures (−5, −10, −15 °C, and − 20 °C) with varying water contents (12%, 16%, 20%). Based on fractional calculus theory and incorporating an exponential decay factor, this study proposes a novel fractional-order constitutive model for a unified description of the softening and hardening behaviors in frozen saline soil. The model treats frozen saline soil as a composite blend of ideal solids and ideal fluids in varying proportions, taking into account the material's inherent time-dependency and non-linear stress-strain relationships. Finally, the validity of the model is verified by the calculated values of the model and the triaxial tests. The results indicate that, based on preliminary judgment, due to the presence of salt solutes, a large amount of liquid water remains in the supersulfated saline soil at temperatures ranging from 0 to −10 °C, forming an unstable state called “warm frozen saline soil”. The mechanical properties of frozen saline soil depend on the relative content of unfrozen water, ice crystals, and salt crystals and the formation of ice and salt crystals significantly enhances the strength of frozen saline soil. The computational results of the improved fractional constitutive model align well with experimental results, effectively describing the stress-strain relationship of frozen supersulfate saline soil. In the model, the parameter κ functions analogously to an elastic modulus and exhibits a linear relationship with temperature, and the parameter α characterizes the strain hardening of saline soil, while β describes its softening behavior. The proposed fractional constitutive model, with only three parameters having clear physical significance, is convenient for practical engineering applications.

Ice-template freeze-casting synthesis of polysilsesquioxane aerogel long fibers for broadband sound absorption

Aerogel fibers inherit the merits of aerogel and fibrous materials, such as ultralow density, large specific surface area and high flexibility, demonstrating great potential as the next generation functional fibers. Herein, an ice-template freeze-casting strategy is developed to prepare polysilsesquioxane aerogel long fibers (PALFs) similar to natural fibers in structure. During the freeze-casting, the strong mutual interaction between solid and liquid phases in the hydrogel induces uniform nucleation and continuous growth of ice crystals in confined spaces. The three-dimensional monolithic hydrogel is gradually deconstructed into one-dimensional fibrous geometries by the growing columnar ice crystals. The obtained PALFs exhibit the polygonal cross-section with a uniform mesoporous structure. After simple physical assembly, the monolithic PALFs exhibit good thermal insulation performance (as low as 25.0 mW/(m·K) at 25 °C) and high-efficiency absorption of broadband sound waves (with a noise reduction coefficient up to 0.58).

A deep learning-based approach for fully automated segmentation and quantitative analysis of muscle fibers in pig skeletal muscle

Muscle fiber properties exert a significant influence on pork quality, with cross-sectional area (CSA) being a crucial parameter closely associated with various meat quality indicators, such as shear force. Effectively identifying and segmenting muscle fibers in a robust manner constitutes a vital initial step in determining CSA. This step is highly intricate and time-consuming, necessitating an accurate and automated analytical approach. One limitation of existing methods is their tendency to perform well on high signal-to-noise ratio images of intact, healthy muscle fibers but their lack of validation on more complex image datasets featuring significant morphological changes, such as the presence of ice crystals. In this study, we undertake the fully automatic segmentation of muscle fiber microscopic images stained with myosin adenosine triphosphate (mATPase) activity using a deep learning architecture known as SOLOv2. Our objective is to efficiently derive accurate measurements of muscle fiber size and distribution. Tests conducted on actual images demonstrate that our method adeptly handles the intricate task of muscle fiber segmentation, yielding quantitative results amenable to statistical analysis and displaying reliability comparable to manual analysis.

Explicit Habit‐Prediction in the Lagrangian Super‐Particle Ice Microphysics Model McSnow

AbstractThe Monte‐Carlo ice microphysics model McSnow is extended by an explicit habit prediction scheme, combined with the hydrodynamic theory of Böhm. Böhm's original cylindrical shape assumption for prolates is compared against recent lab results, showing that interpolation between cylinder and prolate yields the best agreement. For constant temperature and supersaturation, the predicted mass, size, and density of the ice crystals agree well with the laboratory results, and a comparison with real clouds using the polarizability ratio shows regimes capable of improvement. An updated form of the inherent growth function to describe the primary habit growth tendencies is proposed and combined with a habit‐dependent ventilation coefficient. The modifications contrast the results from general mass size relations and significantly impact the main ice microphysical processes. Depending on the thermodynamic regime, ice habits significantly alter depositional growth and affect aggregation and riming. The influence of primary ice behavior on precipitation formation needs to be considered in future development of microphysical parameterizations, but the consideration of secondary habits and the geometry of aggregates should be further improved in future work.

Role of Ice Mechanics on Snow Viscoplasticity

AbstractThe porous structure of snow becomes denser with time under gravity, primarily due to the creep of its ice matrix with viscoplasticity. Despite investigation of this behavior at the macroscopic scale, the driving microscopic mechanisms are still not well understood. Thanks to high‐performance computing and dedicated solvers, we modeled snow elasto‐viscoplasticity with 3D images of its microstructure and different mechanical models of ice. The comparison of our numerical experiments to oedometric compression tests measured by tomography showed that ice in snow rather behaves as a heterogeneous set of ice crystals than as homogeneous polycrystalline ice. Similarly to dense ice, the basal slip system contributed at most, in the simulations, to the total snow deformation. However, in the model, the deformation accommodation between crystals was permitted by the pore space and did not require any prismatic and pyramidal slips, whereas the latter are pre‐requisite for the simulation of dense ice.

Bidirectional anisotropic bacterial cellulose/polyvinyl alcohol/MXene aerogel phase change composites for photothermal conversion enhancement

The issues of inherent low thermal conductivity, slow heat transport rate and easy leakage are critical challenges for accelerating solar-thermal energy harvesting and storage for organic solid-liquid phase change materials (PCMs). Aerogel carriers with thermal conductivity enhancement based on ice temple methods show favorable potential for PCMs solar thermal applications. Comparing with disordered and unidirectional freezing, bidirectional freezing can assemble efficiently in multiple dimensions and result in long-range ordered structures. In this paper, bacterial cellulose/polyvinyl alcohol/MXene aerogels were constructed by bidirectional freezing. The anisotropic PCMs composites were from vacuum impregnation PEG into aerogel networks. By controlling crystallization and growth of ice crystals through the dual temperature gradients, the PCMs composites obtained enhanced heat transfer efficiency in horizontal and vertical directions. Bidirectional aligned PCMs with PDMS wedge angle of 0° and 20° exhibit high thermal conductivity of 0.716 W m−1 K−1 and 0.768 W m−1 K−1, which were 184% higher than that of PEG, and photothermal conversion efficiency of 55.17% and 76.91%. Remarkably, the PCMs composites with a 20° PDMS wedge angle also exhibited a high PCM loading capacity (96.3%) and a high enthalpy (157.7 J/g). The results show that bidirectional anisotropic PCMs composites have excellent overall performance and are expected to improve solar thermal application.

Effects of sanxan on water and ice crystal status of salt free frozen cooked noodles during freeze-thaw cycles

This study compared four biocolloids (sanxan, xanthan gum, curdlan, and guar gum) in different concentrations to replace NaCl in improving the recooking quality of salt free frozen cooked noodles (SF-FCNs). Sanxan outperformed NaCl and other biocolloids significantly improving the firmness (21.0%), chewiness (63.5%), and toughness (15.4%) of SF-FCNs after 10 freeze-thaw (FT) cycles. The results of the freezing-thawing curves showed SF-FCNs had prior FT stability when sanxan was added at 1.2%. Subsequently, the result of differential scanning calorimetry and nuclear magnetic resonance revealed sanxan reduced the content and mobility of freezable water while increasing the content of bound water. The scanning electron microscope, mercury intrusion, and optical microscopy analyses indicated that sanxan reduced the size and volume of ice crystals and the structural damage of SF-FCNs by controlling the water. The work contributes to a theoretical framework for enhancing SF-FCNs quality through precise water and ice crystal control.

High-performance Zn-ion hybrid supercapacitors based on biomass-derived hierarchical porous carbon through template-activated bifunctional induced and ice-crystal assisted strategy

As a highly desirable cathode material for Zn ion hybrid supercapacitors (ZHSCs), biomass-derived carbon precursor shrinks and collapses at elevated temperatures resulting in an impeded internal morphology, which severely hampers ion storage and transport. Herein, a unique porous carbon with multi-level pore structures is produced by a one-step carbonization-activation process. The Gelatin with rich functional groups is employed as the gel-state biomass precursor and the ZnCl2 acted as both a template and an activator. Furthermore, the growth of ice crystals during freeze-drying prevents the carbon framework from collapsing and self-polymerizing of ZnCl2, endowing the prepared porous carbon with a high specific surface area (1336 m2/g) and inheritance of biomass precursor heteroatoms (including N and O). Therefore, at 0.5 A/g, the prepared material displays a desirable specific capacitance of 337.6 F/g in 6 M KOH. In particular, the assembled ZHSC simultaneously shows excellent cycling performance (after 10000 cycles, the specific capacitance increases to 95.6% at 10 A/g) and high energy and power density (120.1 Wh/kg at 450 W/kg). This work provides important insights for the facile preparation of electrode materials for high-performance ZHSCs.

Powering aircraft with 100 % sustainable aviation fuel reduces ice crystals in contrails

Abstract. Powering aircraft by sustainable aviation fuels (SAFs) is a pathway to reduce the climate impact of aviation by lowering aviation lifecycle CO2 emissions and by reducing ice crystal numbers and radiative forcing from contrails. While the effect of SAF blends on contrails has been measured previously, here we present novel measurements on particle emission and contrails from 100 % SAF combustion. During the ECLIF3 (Emission and CLimate Impact of alternative Fuels) campaign, a collaboration between the Deutsches Zentrum für Luft- und Raumfahrt (DLR), Airbus, Rolls-Royce, and Neste, the DLR Falcon 20 research aircraft performed in situ measurements following an Airbus A350-941 source aircraft powered by Rolls-Royce Trent XWB-84 engines in 1 to 2 min old contrails at cruise altitudes. Apparent ice emission indices of 100 % HEFA-SPK (hydro-processed esters and fatty acids–synthetic paraffinic kerosene) were measured and compared to Jet A-1 fuel contrails at similar engine and ambient ice-supersaturated conditions within a single flight. A 56 % reduction in ice particle numbers per mass of burned fuel was measured for 100 % HEFA-SPK compared to Jet A-1 under engine cruise conditions. The measured 35 % reduction in soot particle numbers suggests reduced ice activation by the low-sulfur HEFA fuel. Contrail properties are consistently modeled with a contrail plume model. Global climate model simulations for the 2018 fleet conservatively estimate a 26 % decrease in contrail radiative forcing and stronger decreases for larger particle reductions. Our results indicate that higher hydrogen content fuels as well as clean engines with low particle emissions may lead to reduced climate forcing from contrails.

Effect of extruded wheat bran on the volatility and physical–chemical properties of bread during frozen storage

AbstractBackground and ObjectivesAt present, there are no sufficient data in the literature about the effect of extruded wheat bran on the volatility and physical–chemical properties of bread during frozen storage. Therefore, the current study aimed to investigate the effect of extruded wheat bran on the volatile components and physical–chemical properties of bread during frozen storage.FindingsFrozen storage could effectively extend the shelf‐life of bread. After 20 days of storage, the specific volume and cohesion of the control breads decreased significantly (2% and 21.55%, respectively). Moreover, the hardness and amylopectin retrogradation degree increased significantly (81.56%). After 30 days of storage, the amylopectin retrogradation degree of extruded wheat bran breads significantly increased (161%). After 40 days of storage, the cohesion of extruded wheat bran breads decreased significantly (7.67%). Extruded bran was conducive for improving bread stability during frozen storage. The transverse relaxation times (T2) of extruded wheat bran bread were shorter than those of the control bread. Bran inhibited water molecule migration and reduced the rate of bread quality deterioration. Further, 27 volatile components were detected in the control bread and 32 volatile components were detected in extruded wheat bran breads. The volatile components of all breads did not change significantly within the first 30 days of storage. Thereafter, the concentration of some volatile compounds decreased.ConclusionThe addition of extruded wheat bran significantly affected the volatility and physical–chemical properties of bread during frozen storage. The volatility and physical–chemical properties of extruded wheat bran bread changed gradually during frozen storage. The presence of bran inhibited water molecule migration and the formation and recrystallization of large ice crystals, and reduced the rate of bread quality deterioration during frozen storage.Significance and NoveltyThis study aimed to provide a reference for research on the storage and long‐term preservation of extruded wheat bran bread after production.

Noncontact estimation of the onset of ice accretion in turbofan stators

AbstractThe problem of crystal ice formation inside aircraft turbine engines is well‐documented, and poses a significant risk to safety. The problem is not only one of power loss in flight, but the very real possibility that a flame‐out event could occur due to ice accretion on compressor stators, with potentially catastrophic outcomes. Although many instrumentation systems have been developed for wing ice detection, incipient formation of crystal ice is somewhat more difficult to detect. This is compounded by the need for a noncontact sensor which is robust to in‐flight conditions. This paper proposes an approach to the detection of ice formation based on microwave transmission characteristics across the first and possibly the second stage of the compressor stator. It is shown that noncontact detection is feasible under realistic conditions. The contribution of this paper is twofold. First, the microwave transmission approach is motivated using wind tunnel measurements, and appropriate frequency bands are determined. Next, a signal processing approach involving higher‐order analysis of time‐frequency distribution characteristics is then put forward. Experimental results are presented to support the hypothesis that multiband detection offers a workable approach to the incipient crystal‐ice detection problem.

Effect of Moderate Static Magnetic Fields on Mice Oocyte Vitrification: Calcium-Related Genes Expression.

The ability to cryopreserve oocytes without ultrastructural injury has been a concern in the development and use of methods to preserve female reproduction. The stability of the cell membrane must be preserved to reduce the damage caused by ice crystals during vitrification. One approach that has been explored is the use of static magnetic fields (SMFs), which are believed to influence cell membrane stability. In this study, the in vitro effects of SMF that range between 20-80 mT on the vitrification of mice germinal vesicle (GV) oocytes were studied. The viability and mitochondrial (Mt) membrane potential of both vitrified and nonvitrified oocytes were assessed using Trypan blue and JC1 staining. The high in vitro maturation (IVM) rate and high Mt membrane potential in metaphase II (MII) oocytes were taken into account to determine the optimal magnetic field intensity, that is, 20 mT. None of the SMF conditions resulted in intact spindles in MII oocytes. The study also explored the expression of store-operated calcium entry (Stim1, Orai1, and Ip3r) and meiosis resumption (Ccnb, Cdk) genes in GV and MII oocytes of both vitrified and control groups. The results show that the expressions of Orai1 and Ccnb genes in Vit-MII-SMF oocytes were considerably increased. However, no significant difference in Stim1 expression was observed between the groups. The Vit-MII-SMF group exhibited a significantly higher Ccnb expression compared to other groups. In vitro fertilization (IVF) was performed to evaluate the 2 pronuclear (2PN) rates. The findings demonstrated that using 20 mT SMF improved 2PN rates compared to the nonvitrified groups. This study provides a deeper understanding of the effects of moderate SMF and vitrification on the expression of calcium channel genes in GV and MII oocytes. The results suggest that applying a 20 mT SMF can help prevent cryoinjury and enhance the characteristics of vitrified-warmed oocytes.

Vapor‐Phase Synthesis of Poly(para‐xylylene): From Coatings to Porous and Hierarchical Materials

AbstractPoly(para‐xylylene) (PPX) is a robust and biocompatible coating material that is widely used in various applications, including electronics, aerospace and defense materials, automotive materials, and biomaterials. In this progress report, recent developments in PPX technology ranging from the advancement of physical chemistry properties and structural properties for device integration to the transformation of 3D monolith materials of PPX with controls in outer and inner structures at the micro‐ and nanometer scales are highlighted. Based on emerging chemistry studies on [2.2]paracyclophanes, which are primarily used precursors to synthesize PPX via vapor deposition polymerization, functionalization, and the creation of a wide variety of functional PPX derivatives are demonstrated without resistance. Widely used as an interface coating material, PPX is processed to form integrated structural materials and has already been found to be useful in the market as part of electronic and medical implant products. Using a newly innovated transformation to fabricate PPX through templates (metal‐organic frameworks, liquid crystal, ice crystal), 3D monoliths, nanoscale particles, hierarchical and gradient interior structures, and dynamically transformable shapes at the nanoscale are demonstrated. A vast landscape of novel applications and device products is expected based on the already established R&D and market maturity of PPX.

Effects of sequential combination of cryogenic immersion freezing and ultra-cold frozen storage on the quality of Korean white kimchi

This study aimed to assess the effects of three different freezing techniques — cold room freezing (CRF; −20 °C), air-blast freezing (ABF; −70 °C), and cryogenic immersion freezing (CIF; −70 °C) — on the microstructure and physicochemical properties of Korean white kimchi (KWK) during frozen storage at −20 °C and − 45 °C for 8 weeks. CIF-treated KWK exhibited a higher freezing rate and after ultra-cold (UC; −45 °C) frozen storage, it showed a greater number and smaller diameter and area of ice crystals than CRF- and ABF-treated KWK. Drip loss was reduced and CIE color changes were delayed in KWK under UC frozen storage. The pH, titratable acidity, and reducing sugar content of frozen KWK remained unaffected by the freezing method, storage temperature, or duration. Thus, CIF combined with UC frozen storage preserves the physicochemical quality of KWK with minimal histological damage during long-term storage. Industrial relevanceIn the kimchi industry, where the shelf life of commercial products is often constrained by successive fermentation under low-temperature storage and distribution conditions, introducing frozen kimchi is a promising solution to reduce waste of overripe kimchi in the supply chain. This study highlights the effectiveness of combining CIF and UC frozen storage methods to minimize the damage caused by freezing in kimchi. This innovative technology provides a practical means for streamlining freezing processes, saving operational time, and ensuring kimchi quality during long-term storage and distribution.

Study on the coupling mechanism of water-heat-vapor-salt-mechanics in unsaturated freezing sulfate saline soil

Understanding the internal regularity of unsaturated freezing sulfate saline soil is of utmost importance concerning the durability and stability of cold region engineering in sulfate saline soil areas. One-sided freezing experiments were performed on unsaturated sulfate saline soils with varying salt and moisture contents and temperature gradients to investigate the interplay of water-vapor-salt migration, heat transfer, ice-water and water-vapor phase transformations, salt crystallization, and salt-frost heave. This study presented a novel water-heat-vapor-salt-mechanics coupling model that considers water-vapor-salt migration and its feedback on soil deformation. The results signify that the water accumulation at the top surface of unsaturated freezing sulfate saline soil is primarily due to water vapor migration. Water-vapor-salt migration, ice-water phase transition, and salt crystallization form more ice and salt crystals, filling the air-filled pores and causing significant salt-frost heave. Water vapor transfer and accumulation further exacerbate the soil deformation for unsaturated freezing sulfate saline soil. The calculating results agree with the test data and demonstrate the efficacy of the proposed five-field coupling model. These findings hold significant implications for developing practical solutions for mitigating the negative impacts of salt-frost heave induced by water-vapor-salt transfer in unsaturated freezing sulfate saline soil on construction projects.

Freezing-mediated formation of supraproteins using depletion forces

HypothesisLong-acting formulations such as microparticles, injectable depots and implantable devices can realize spatiotemporally controlled delivery of protein drugs to extend their therapeutic in vivo half-lives. To efficiently encapsulate the protein drugs into such drug delivery systems, (sub)micron-sized protein particles are needed. The formation of micronized supraproteins can be induced through the synergistic combination of attractive depletion forces and freezing. The size of the supraproteins can be fine-tuned from submicron to several microns by adjusting the ice crystallization rate through the freeze-quench depth, which is set by the target temperature.MethodsSupraprotein micron structures were prepared from protein solutions under various conditions in the presence and absence of nonadsorbing polyethylene glycol. Scanning electron microscopy and dynamic light scattering were employed to determine the sizes of the supraproteins and real-time total internal reflection fluorescent microscopy was used to follow the supraprotein formation during freezing. The protein secondary structure was measured before and after micronization by circular dichroism. A phase diagram of a protein–polyethylene glycol mixture was theoretically predicted to investigate whether the depletion interaction can elucidate the phase behavior.FindingsMicronized protein supraparticles could be prepared in a controlled manner by rapid freeze-drying of aqueous mixtures of bovine serum albumin, horseradish peroxidase and lysozyme mixed with polyethylene glycol. Upon freezing, the temperature quench initiates a phase separation process which is reminiscent of spinodal decomposition. This demixing is subsequently arrested during droplet phase separation to form protein-rich microstructures. The final size of the generated protein microparticles is determined by a competition between phase separation and cooling rate, which can be controlled by target temperature. The experimental phase diagram of the aqueous protein–polyethylene glycol dispersion aligns with predictions from depletion theory for charged colloids and nonadsorbing polymers.

Prediction of heat and mass transfer inside 3D frost microstructure

In the present study, a CFD model based on 3D frost microstructure is developed. The desublimation phase change coupled with mass transfer of water vapor to ice surface and heat conduction through the tortuous ice crystals are calculated to predict accurate heat transfer process inside the frost microstructure. Air velocity and temperature inside the frost structure after 30 min operation are significantly lower than those inside 10 min operation, which are influenced by the structural change of the ice crystals. Vertical variations in ice crystal surface temperature and ice surface mass flux are also calculated, which varied between the frost structures operated for different periods. At 70 μm height of the frost, the mass flux at the ice crystal surface shows significant decrease from 7.48 × 10−5 kg/m2s for the frost structure at 10 min to 1.82 × 10−5 kg/m2s for that at 30 min. This study provides new and fundamental information to understand the mechanism of frost growth which is very important in refrigeration, cryogenics, and aerospace industry, etc.

Effects of freeze–thaw cycles on the physicochemical and in vitro digestibility of starch in pre‐fermented frozen raw dough

SummaryIn this study, pre‐fermented frozen raw dough (PFRD) was subjected to freeze–thaw cycling (FTC) to explore the deterioration mechanism of PFRD, after which the physical quality, chemical properties and in vitro digestion characteristics of starch in the dough were analysed. The results showed that after the 5th FTC, the surface area of the dough and starch exhibited obvious depression deformation, and the damaged starch content increased by 38.21%. The random mechanical damage caused by ice crystals also had no significant regularity in the water retention and the short‐range order of starch. Moreover, the breakdown of the double helix structure of starch led to the dissolution of amylose, and the expansion and gelatinization enthalpy increased. In addition, the in vitro digestion results showed that freeze–thaw cycle twice increased the digestibility of the starch, and the hydrolysis rate reached 75.16%, Therefore, the effects of FTC twice on the starch of PFRD are controllable. This study provides a theoretical basis for the development of prefabricated frozen starch food industry technology in the future.

Effect of enzymatic hydrolysis of arabinoxylan on the quality of frozen dough during the subfreezing process.

The objective of this investigation was to examine the impact of enzymatic hydrolysis of arabinoxylan (AX) on frozen dough quality under subfreezing conditions. The dough was subjected to freezing at -40 °C for 2 h and then stored at -9, -12, and -18 °C for 15 days. The water loss, freezable water content, water migration, and microstructure of the dough were measured. The dough containing 0.8% cellulase enzymatically hydrolyzed AX (CAX) required the shortest duration when traversing the maximum ice-crystal formation zone (6.5 min). The dough with xylanase enzymatically hydrolyzed AX (XAX) demonstrated a faster freezing rate than the dough with CAX. The inclusion of both XAX and CAX in the dough resulted in the lowest freezable water loss and reduced freezable water content and free-water content levels, whereas the inclusion of xylanase-cellulase combined with enzymatically hydrolyzed AX resulted in higher free-water content levels. The textural properties of the subfreezing temperature dough were not significantly different from the dough stored at -18 °C and sometimes even approached or surpassed the quality observed in the control group rather than the dough stored at -18 °C. In addition, the gluten network structure remains well preserved in XAX- and CAX-containing doughs with minimal starch damage. The enzymatic hydrolysis of AX from wheat bran can be used as a useful additive to improve the quality of frozen dough. © 2024 Society of Chemical Industry.