Advanced Bacterial Cellulose Ionic Conductors with Gigantic Thermopower for Low-Grade Heat Harvesting.

Ionic conductors such as polymer electrolytes and ionic liquids have high thermoelectric voltages several orders of magnitude higher than electronic thermoelectric materials, while their conductivity is much lower than the latter. This work reports a novel approach to achieve high-performance ionic conductors using calcium ion (Ca2+) coordinated bacterial cellulose (CaBC) through molecular channel engineering. Through the coordination of Ca2+ with cellulose molecular chain, the distance between the cellulose molecular chains is widened, so that ions can transport along the cellulose molecular chain. Therefore, we reported ionic thermoelectric (i-TE) material based on CaBC/NaCl with a relatively high ionic Seebeck coefficient of -27.2 mV K-1 and high ionic conductivity of 204.2 mS cm-1. This ionic hydrogel is promising in the design of high-thermopower i-TE materials for low-grade heat energy harvesting.

Highly Mineralized Biomimetic Polysaccharide Nanofiber Materials Using Enzymatic Mineralization.

Many biological high-performance composites, such as bone, antler, and crustacean cuticles, are composed of densely mineralized and ordered nanofiber materials. The mimicry of even simplistic bioinspired structures, i.e., of densely and homogeneously mineralized nanofibrillar materials with controllable mechanical performance, continues to be a grand challenge. Here, using alkaline phosphatase as an enzymatic catalyst, we demonstrate the dense, homogeneous, and spatially controlled mineralization of calcium phosphate nanostructures within networks of anionically charged cellulose nanofibrils (CNFs) and cationically charged chitin nanofibrils (ChNFs)-both emerging biobased nanoscale building blocks for sustainable high-performance materials design. Our study reveals that anionic CNFs lead to a more homogeneous nanoscale mineralization with very high mineral contents up to ca. 70 wt % with a transition from amorphous to crystalline deposits, while cationic ChNFs yield rod-like crystalline morphologies. The bone-inspired CNF bulk films exhibit a significantly increased stiffness, maintain good flexibility and translucency, and have a significant gain in wet state mechanical properties. The mechanical properties can be tuned both by the enzyme concentration and the mineralization time. Moreover, we also show a spatial control of the mineralization using kinetically controlled substrate uptake in a dialysis reactor, and by spatially selectively incorporating the enzyme into 2D printed filament patterns. The strategy highlights possibilities for spatial encoding of enzymes in tailored structures and patterns and programmed mineralization processes, promoting the potential application of mineralized CNF biomaterials with complex gradients for bone substitutes and tissue regeneration in general.

Covalently grafting polycation to bacterial cellulose for antibacterial and anti-cell adhesive wound dressings.

Hydrogel-based wound dressings are becoming increasingly important for wound healing. Bacterial cellulose (BC) has been commonly used as wound dressings due to its good in vitro and in vivo biocompatibility. However, pure BC does not possess antibacterial properties. In this regard, polycation gel was grafted onto the BC using a surface-initiated activator regenerated by electron transfer atom transfer radical polymerization (SI-ARGET ATRP) with subsequent quaternization for antibacterial wound dressing. Dimethylethyl methacrylate (DMAEMA) was successfully polymerized on the BC surface which was confirmed by Fourier transform infrared spectroscopy and elemental analysis. The morphology structure, specific surface area, pore size, and mechanical properties were also characterized. The quaternized PDMAEMA grafted on the BC endowed it with excellent antibacterial activity against E. coli (Gram-negative) and S. aureus (Gram-positive) with a killing rate of 89.2 % and 93.4 %, respectively. The number of cells was significantly reduced on QPD/BC hydrogel, demonstrating its good anti-adhesion ability. In vitro cellular evaluation revealed that the antibacterial wound dressing exhibited good biocompatibility. Overall, this study provides a feasible method to develop antibacterial and anti-cell adhesive hydrogel, which has a promising potential for wound healing.

Fiber-Based Noncontact Sensor with Stretchability for Underwater Wearable Sensing and VR Applications.

The rapid development of artificial intelligent wearable devices has led to an increasing need for seamless information exchange between humans, machines, and virtual spaces, often relying on touch sensors as the primary interaction medium. Additionally, the demand for underwater detection technologies is on the rise owing to the prevalent wet and submerged environment. Here, a fiber-based capacitive sensor with superior stretchability and hydrophobicity is proposed, designed to cater to noncontact and underwater applications. The sensor is constructed using bacterial cellulose (BC)@BC/carbon nanotubes (CNTs) (BBT) helical fiber as the matrix and methyltrimethoxysilane (MTMS) as the hydrophobic modified agent, forming a hydrophobic silylated BC@BC/CNT (SBBT) helical fiber by the chemical vapor deposition (CVD) technique. These fibers exhibit an impressive contact angle of 132.8°. The SBBT helicalfiber-based capacitive sensor presents capabilities for both noncontact and underwater sensing, which exhibits a significant capacitance change of -0.27 (at a distance of 0.5 cm). We have achieved interactive control between real space and virtual space through intelligent data analysis technology with minimal interference from the presence of water. This work has laid a solid foundation of noncontact sensing with attributes such as degradability, stretchability, and hydrophobicity. Moreover, it offers promising solutions for barrier-free communication in virtual reality (VR) and underwater applications, providing avenues for smart human-machine interfaces for submerged use.

Microenvironment-regulated dual-hydrophilic coatings for glaucoma valve surface engineering.

Glaucoma valves (GVs) play an essential role in treating glaucoma. However, fibrosis after implantation has limited their long-term success in clinical applications. In this study, we aimed to develop a comprehensive surface-engineering strategy to improve the biocompatibility of GVs by constructing a microenvironment-regulated and dual-hydrophilic antifouling coating on a GV material (silicone rubber, SR). The coating was based on a superhydrophilic polydopamine (SPD) coating with good short-range superhydrophilicity and antifouling abilities. In addition, SPD coatings contain many phenolic hydroxyl groups that can effectively resist oxidative stress and the inflammatory microenvironment. Furthermore, based on its in situ photocatalytic free-radical polymerization properties, the SPD coating polymerized poly 2-methylacryloxyethylphosphocholine, providing an additional long-range hydrophilic and antifouling effect. The in vitro test results showed that the microenvironment-regulated and dual-hydrophilic coatings had anti-protein contamination, anti-oxidation, anti-inflammation, and anti-fiber proliferation capabilities. The in vivo test results indicated that this coating substantially reduced the fiber encapsulation formation of the SR material by inhibiting inflammation and fibrosis. This design strategy for dual hydrophilic coatings with microenvironmental regulation can provide a valuable reference for the surface engineering design of novel medical implantable devices. STATEMENT OF SIGNIFICANCE: Superhydrophilic polydopamine (SPD) coatings were prepared on silicone rubber (SR) by a two-electron oxidation method. Introduction of pMPC to SPD surface using photocatalytic radical polymerization to obtain a dual-hydrophilic coating. The dual-hydrophilic coating effectively modulates the oxidative and inflammatory microenvironment. This coating significantly reduced protein contamination and adhesion of inflammatory cells and fibroblasts in vitro. The coating-modified SR inhibits inflammatory and fibrosis responses in vivo, promising to serve the glaucoma valves.

Bacterial cellulose-based hydrogel with regulated rehydration and enhanced antibacterial activity for wound healing.

Bacterial cellulose (BC) hydrogels are promising medical biomaterials that have been widely used for tissue repair, wound healing and cartilage engineering. However, the high water content of BC hydrogels increases the difficulty of storage and transportation. Moreover, they will lose their original hydrogel structure after dehydration, which severely limits their practical applications. Introducing the bio-based polyelectrolytes is expected to solve this problem. Here, we modified BC and combined it with quaternized chitosan (QCS) via a chemical reaction to obtain a dehydrated dialdehyde bacterial cellulose/quaternized chitosan (DBC/QCS) hydrogel with repeated swelling behavior and good antibacterial properties. The hydrogel can recover the initial state on the macro scale with a swelling ratio over 1000 % and possesses excellent antimicrobial properties against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) with a killing rate of 80.8 % and 81.3 %, respectively. In addition, the hydrogel has excellent biocompatibility, which is conducive to the stretching of L929 cells. After 14 d of in vivo wound modeling in rats, it was found that the hydrogel loaded with pirfenidone (PFD) could promote collagen deposition and accelerate wound healing with scar prevention. This rehydratable hydrogel can be stored and transported under dry conditions, which is promising for practical applications.

Pyrolyzed bacterial cellulose: a versatile support for lithium ion battery anode materials.

A scalable, low-cost and environmentally benign strategy is developed for the facile construction of a unique kind of three-dimensional porous electrode architecture for high-performance lithium ion batteries. The methodology is based on the employment of pyrolyzed bacterial cellulose as a new three-dimensional porous scaffold to support various nanostructured active electrode materials, such as SnO2 and Ge.

Bacterial cellulose-based hydrogel with antibacterial activity and vascularization for wound healing.

Skin wounds need an appropriate wound dressing to help prevent bacterial infection and accelerate wound closure. Bacterial cellulose (BC) with a three-dimensional (3D) network structure is an important commercial dressing. However, how to effectively load antibacterial agents and balance the antibacterial activity is a lingering issue. Herein, this study aims to develop a functional BC hydrogel containing silver-loaded zeolitic imidazolate framework-8 (ZIF-8) antibacterial agent. The tensile strength of the prepared biopolymer dressing is >1 MPa, the swelling property is over 3000 %, the temperature can reach 50 °C in 5 min with near-infrared (NIR) and the release of Ag+ and Zn2+ is stable. In vitro investigation shows that the hydrogel displays enhanced antibacterial activity, and the bacteria survival ratios are only 0.85 % and 0.39 % against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). In vitro cell experiments present that BC/polydopamine/ZIF-8/Ag (BC/PDA/ZIF-8/Ag) shows satisfactory biocompatibility and promising angiogenic ability. In vivo study, the full-thickness skin defect on rats demonstrates remarkably wound healing ability and accelerated skin re-epithelialization. This work presents a competitive functional dressing with effective antibacterial properties and accelerative angiogenesis activities for wound repair.

[Preparation for and study on the property of medical bacterial cellulose].

Bacterial cellulose (BC) was prepared by Acetobacter xylinum in static culture. After purified by chemical treatment, the microstructure, chemical structure, crystal structure and mechanical property of BC were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffractometry (XRD) and tensile strength measurement respectively, and compared with those of the imported bacterial cellulose wound dressing served as control sample (XBC). The results indicated that the diameter of the BC was (22 +/- 9) nm, and the crystallinity index was 89.71%. The tensile strength and the Young's mouduls of BC were significant higher than XBC both in wet and dry states. The biocompatibility of BC and XBC were evaluated by cytotoxicity test, delayed contact sensitization study in the Guinea Pig and skin irritation test. The results showed that BC had reliable biocompatibility as well as XBC. With the unique nanostructure, high crystallinity, high mechanical strength, and reliable biocompatibility, BC produced in our country as well as XBC can be used as a safe biomaterial for the medical applications.

Self-Stretchable Fiber Liquid Sensors Made with Bacterial Cellulose/Carbon Nanotubes for Smart Diapers.

Liquid sensors for detecting water and body fluids are crucial in daily water usage and health monitoring, but it is challenging to combine sensing performance with high tensile deformation and multifunctional applications. Here, a substrate-free, self-stretchable bacterial cellulose (BC)/carbon nanotube (CNT) helical fiber liquid sensor was prepared by the solution spinning and coiling process using BC as the water-sensitive matrix and CNTs as the active sensing materials. The BC/CNT (BCT) fiber sensor has a high stretch ratio of more than 1000% and a rapid response for a current change rate of 104% within 1 s, which is almost unaffected under washing and various stretching or knotting deformations. By combination of the BCT fiber, we can design smart diapers or water level detectors, which rapidly monitor the status of smart diapers or water level, and the monitoring result can be transferred on time through an alarm device or smartphone. In short, the scalable and continuous preparation of the self-stretchable BCT helical fiber will provide a capacious platform for the development of a wearable sensor applied in daily life (such as smart diapers, water level detection, etc.).

Bacterial cellulose reinforced chitosan-based hydrogel with highly efficient self-healing and enhanced antibacterial activity for wound healing.

Biocompatible hydrogels with versatile functions are highly desired for demanding the complicated tissue issues, including irregular site and motional wound. Herein, a bio-based hydrogel with multifunctional properties is designed based on quaternized chitosan and dialdehyde bacterial cellulose. As a functional wound dressing, the hydrogel shows rapid self-healing performance and injectable behaviors due to dynamic Schiff-base interactions and presents superior antibacterial activity against E. coli (gram-negative) and S. aureus (gram-positive). The constructed 3D hydrogel also exhibits proper compressive property, desired water retention capacity. To be mentioned, the hydrogel could mimic the structure of natural extracellular matrix (ECM) in the presence of bacterial cellulose nanofibers. Thus, the biopolymer-based hydrogel shows good biocompatibility in terms of cell proliferation and cell spreading. The prepared chitosan-based hydrogel with self-healing, antibacterial, and low cost will become a promising biomaterial for wound healing.

Hydrophobic, breathable cellulose nonwoven fabrics for disposable hygiene applications.

Cellulose-based fabrics with suitably high hydrophobicity and good air-permeability are highly promising for disposable hygiene applications. Herein, a facile, one-step method is reported, which effectively converts the completely hydrophilic cellulose nonwoven substrate into highly hydrophobic fabrics (water contact angle of 130-135°) with preservation of a good air-permeability (variation ± 6% after modification compared to the original 1337 mm·s-1). Mono-isocyanates with bulky, hydrophobic moieties were adopted and 3-isocyanatopropyltrimethoxysilane (ISPTMOS) was found to be the most efficient one compared to tert-butyl isocyanate (TBIS) and m-toluene isocyanate (MTIS). The influence of the type and concentration of modifiers on the structure and key properties (hydrophobicity, air-permeability, breaking strength, flexibility) of the fabrics was systematically investigated. This approach has a great potential for industrial scale-up at the stage of post-finishing of nonwoven fabrics, which can be applied in medical, hygiene and personal care areas.

Anti-Fouling, Adhesive Polyzwitterionic Hydrogel Electrodes Toughened Using a Tannic Acid Nanoflower.

Conductive polyzwitterionic hydrogels with good adhesion properties show potential prospect in implantable electrodes and electronic devices. Adhesive property of polyzwitterionic hydrogels in humid environments can be improved by the introduction of catechol groups. However, common catechol modifiers can usually quench free radicals, resulting in a contradiction between long-term tissue adhesion and hydrogel toughness. By adding tannic acid (TA) to the dispersion of clay nanosheets and nanofibers, we designed TA-coated nanoflowers and nanofibers as the reinforcing phase to prepare polyzwitterionic hydrogels with adhesion properties. The hydrogel combines the mussel-like and zwitterionic co-adhesive mechanism to maintain long-term adhesion in underwater environments. In particular, the noncovalent cross-linking provided by the nanoflower structure effectively compensates for the defects caused by free-radical quenching so that the hydrogel obtained a high stretchability of over 2900% and a toughness of 1.16 J/m3. The hydrogel also has excellent anti-biofouling property and shows resistance to bacteria and cells. In addition, the hydrogel possesses a low modulus (<10 kPa) and ionic conductivity (0.25 S/m), making it an ideal material for the preparation of implantable electrodes.

[Clinical application of a self-developed mouth opener with a tongue base retractor in the operation of deep part of tongue base].

Objective:To test the application effect of a self-developed mouth opener with a tongue base retractor in the operation of the deep part of tongue base. Methods:The tongue base surgical field was exposed by using a self-developed mouth opener with a tongue base retractor in 8 patients who underwent deep tongue base operation via oral approach, the difficulty of operation, the effect of exposure of operation field, the tear of mucous membrane of the pharynx arch and the risk of tongue paralysis were observed. Results:The self-made mouth opener can expose the deep operative field of the tongue root by using the self-provided tongue root retractor during the operation, and the operation is conducted under the guidance of angle endoscope. The operative field of 8 patients was well exposed during the whole operation, there was no pharyngeal mucosa tearing and postoperative tongue paralysis. Conclusion:The self-made mouth opener has the advantages of simple operation, good exposure effect and less complications, but it needs rigid bending instruments in some operations.

Anisotropic bacterial cellulose hydrogels with tunable high mechanical performances, non-swelling and bionic nanofluidic ion transmission behavior.

Water-rich hydrogels with tissue-like softness, especially ion conductive hydrogels with ion signal transfer systems similar to biological areas, are promising soft electrode materials, while too poor or unstable mechanical properties that come from uncontrollable swelling and biocompatibility issues caused by introducing high concentration ions are serious obstacles in practical applications. Herein, a simple method for fabricating strong, stable, ion-conductive, anisotropic bacterial cellulose hydrogels (ABCHs) is first reported. Relying on nanofibers with high aspect ratio in bacterial cellulose (BC), a tailor-made nanofiber-network-reinforced structure is constructed by controlled dissolution, followed by aligning them well via a simple fossilizing process under stretching. Therefore, tunable high mechanical performances can be achieved and the maximum tensile strength can reach 14.3 MPa with 70% water content. It is worth noting that ABCHs will not swell in water for 30 days and maintain 93% tensile strength. Most importantly, the unique nanofluid behaviors from nanochannels in nanofibers allow effective ion transport in ABCHs relying only on low concentrations of ions in body fluids (<300 mM), avoiding sacrificing biocompatibility to achieve useful conductivity. This facile strategy might be very scalable in fabricating high-strength, non-swelling, bio-ion conductive cellulose hydrogels for application in next-generation bio-interfacing and flexible implantable devices.

TEMPO-Oxidized Bacterial Cellulose Nanofibers/Graphene Oxide Fibers for Osmotic Energy Conversion.

The large osmotic energy between river water and seawater is an inexhaustible blue energy source; however, the complicated manufacturing methods used for ion-exchange devices hinder the development of reverse electrodialysis (RED). Here, we use a wet-spinning method to continuously spin meter-scale 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)-oxidized bacterial cellulose (TOBC) nanofiber filaments, which are then used to construct nanochannels for osmotic energy conversion. These are then used to build a nacre-like structure by adding graphene oxide (GO), which provides narrow nanochannels in one-dimensional and two-dimensional nanofluid systems for rapid ion transport. With a 50-fold concentration gradient, the nanochannels in the fibers generate electricity of 0.35 W m-2, with an ionic mobility of 0.94 and an energy conversion efficiency of 38%. The assembly of GO and TOBC results in a high power density of 0.53 W m-2 using artificial seawater and river water. The RED device fabricated from TOBC/GO fibers maintains a stable power density for 15 days. This research proposes a simple method to reduce the size of nanochannels to improve the ionic conductivity, ionic selectivity, and power density of cellulose-based nanofibers to increase the possibility of their application for the conversion of osmotic energy to electrical energy.

High-Strength Superstretchable Helical Bacterial Cellulose Fibers with a "Self-Fiber-Reinforced Structure".

As a hydrogel membrane grown on the gas-liquid interface by bacterial culture that can be industrialized, bacterial cellulose (BC) cannot give full play to the advantages of its natural nanofibers. Conversion to the properties of nanofibers from high-performance to macrofibers represents a difficult material engineering challenge. Herein, we construct high-strength BC macrofibers with a "self-fiber-reinforced structure" using a dry-wet spinning method by adjusting the BC dissolution and concentration. The macrofiber with a tensile strength of 649 MPa and a strain of 17.2% can be obtained, which is one of the strongest and toughest cellulose fibers. In addition, the macrofiber can be fabricated to a superstretchable helical fiber without adding other elastomers or auxiliary materials. When the helical diameter is 1.6 mm, the ultimate stretch reaches 1240%. Meanwhile, cyclic tests show that the mechanical properties and morphology of the fiber remained stable after 100 times of 100% cyclic stretching. It is exciting that the helical fiber also owns outstanding knittability, washability, scalability, and dyeability. Furthermore, superstretchable functional helical BC fibers can be fabricated by embedding functional materials (carbon materials, conductive polymers, etc.) on BC or in the spinning dope, which can be made to wearable devices such as fiber solid-state supercapacitors. This work provides a scalable way for high-strength superstretchable and multifunctional fibers applied in wearable devices.

A 3D-printable TEMPO-oxidized bacterial cellulose/alginate hydrogel with enhanced stability via nanoclay incorporation.

Three-dimensional (3D) printing offers a novel approach to manufacture repeatable personalized structures for mass customization in medical fields. Considering the resemblance of materials in composition and microstructure to biological tissues, polysaccharide-based hydrogel is a promising printing material. However, its long-term stability of structure has always been a problem. In this work, we showed a green nanocomposite printing ink based on 2,2,6,6-tetramethylpiperidinyl-1-oxyl (TEMPO)-oxidized bacterial cellulose (TOBC), sodium alginate (SA) and laponite nanoclay (Xls). The TOBC/SA/Xls hydrogel prepared by the 3D printing ink not only exhibited structural stability, but also performed a long-term release behavior of protein which could be attractive in medical application of drug release, biomedical devices and tissue engineering.

Urethra-inspired biomimetic scaffold: A therapeutic strategy to promote angiogenesis for urethral regeneration in a rabbit model.

Limited angiogenesis and epithelialization make urethral regeneration using conventional tissue-engineered grafts a great challenge. Consequently, inspired from the native urethra, bacterial cellulose (BC) and bladder acellular matrix (BAM) were combined to design a three dimensional (3D) biomimetic scaffold. The developed BC/BAM scaffold was engineered for accelerating urethral regeneration by enhancing angiogenesis and epithelialization. The BC/BAM scaffold reveals the closest mimic of native urethra in terms of the 3D porous nanofibrous structure and component including collagen, glycosaminoglycans, and intrinsic vascular endothelial growth factor (VEGF). In vitro studies showed that the bioinspired BC/BAM scaffold promoted in vitro angiogenesis by facilitating human umbilical vein endothelial cells (HUVECs) growth, expression of endothelial function related proteins and capillary-like tube formation. Effect of the BC/BAM scaffold on angiogenesis and epithelialization was studied by its implantation in a rabbit urethral defect model for 1 and 3 months. Results demonstrated that the improved blood vessels formation in the urethra-inspired BC/BAM scaffold significantly promoted epithelialization and accelerated urethral regeneration. The urethra-inspired BC/BAM scaffold provides us a new design approach to construct grafts for urethral regeneration. STATEMENT OF SIGNIFICANCE: Findings in urethral regeneration demonstrate that an ideal tissue-engineered urethra should have adequate angiogenesis to support epithelialization for urethral regeneration in vivo. In this study, inspired from the native urethra, a bioinspired bacterial cellulose/bladder acellular matrix (BC/BAM) scaffold was developed to promote angiogenesis and epithelialization. The designed scaffold showed the closest physical structure and component to natural urethra, which is beneficial to angiogenesis and regeneration of urethral epithelium. This is the first time to utilize BC and dissolved BAM to develop biomimetic scaffold in urethral tissue engineering. Our biomimetic strategy on urethra graft design provided enhanced angiogenesis and epithelialization to achieve an accelerated and successful rabbit urethral repair. We believe that our urethra-inspired biomimetic scaffold would provide new insights into the design of urethral tissue engineering grafts.

Silver Nanowire-Bacterial Cellulose Composite Fiber-Based Sensor for Highly Sensitive Detection of Pressure and Proximity.

Fiber-based sensors are desirable to provide an immersive experience for users in the human-computer interface. We report a hierarchically porous silver nanowire-bacterial cellulose fiber that can be utilized for sensitive detection of both pressure and proximity of human fingers. The conductive fiber was synthesized via continuous wet-spinning at a speed of 20 m/min, with a diameter of 53 µm, the electrical conductivity of 1.3 × 104 S/cm, a tensile strength of 198 MPa, and elongation strain of 3.0% at break. The fibers were coaxially coated with a 10 µm thick poly(dimethylsiloxane) dielectric elastomer to form the fiber sensor element which is thinner than a human hair. Two of the sensor fibers were laid diagonally, and the capacitance changes between the conductive cores were measured in response to pressure and proximity. In the touch mode, a fiber-based sensor experienced monotonic capacitance increase in the pressure range from 0 to 460 kPa, and a linear response with a high sensitivity of 5.49 kPa-1 was obtained in the low-pressure regime (<0.5 kPa). In touchless mode, the sensor is highly sensitive to objects at a distance of up to 30 cm. Also, the fiber can be easily stitched into garments as comfortable and fashionable sensors to detect heartbeat and vocal pulses. A fiber sensor array is able to serve as a touchless piano to play music and accurately determine the proximity of an object. A 2 × 2 array was further shown for two- and three-dimensional location detection of remote objects.