Nature-Inspired Nano Cellulose Materials, Advancements in Nano Cellulose Preparation and Versatile Applications

*Hanumanthu Jeevan Rao, Sanjay Singh, Perumalla Janaki Ramulu, Narender Singh, Thiago F. Santos, Caroliny M. Santos, Nandini Robin Nadar and Gara Dheeraj Kumar*

#### **Abstract**

A promising ecofriendly, cost-effective biomaterial derived from natural sources, i.e., lignocellulose natural fibers from trees, plants, agri waste, fruits, vegetables, seeds, and leaves. It clicked the attention of the researchers due to promising properties and versatility. The aim of the study is to examine the recent developments and preparation methods and extraction techniques for nanolignocellulose materials from nature. It also discusses the wide range of applications that use nano cellulose's remarkable properties for a variety of research fields. Current work discussed advancements in nano cellulose preparation techniques, innovative methods, and cutting-edge applications. The ease of nano cellulose excels as a material for tissue engineering scaffolds, wound dressings, flexible and sustainable electronics, and drug delivery systems in the biomedical industry. It is desirable component in composites due to its remarkable reinforcing abilities in polymers. The use of environmental applications such as water purification, oil spill cleanup, and biodegradable packaging is also highlighted in this research. The studies emphasize the need for more investigation and optimization of extraction processes, characterization, and applications. This multidisciplinary study intends to motivate academics and scientists to fully utilize nano cellulose and contribute to the creation of environmentally friendly and sustainable solutions across a range of industries.

**Keywords:** nano cellulose, nature-inspired materials, nano cellulose applications, natural fibers, nanocomposites, sustainable materials

#### **1. Introduction**

Nanocellulose has received significant interest due to its mechanical and optical properties [1], biodegradability [2], availability [3], recyclability [4], renewability [5], and low coefficient of thermal expansion (CTE) [6] as exhibited in **Figure 1**. This

#### **Figure 1.**

*Availability, extraction, and applications of nano cellulose.*

nanomaterial has been reported as an intriguing sustainable and tunable platform for the manufacturing of a variety of high-value products. It can take the shape of nanofibrils or nanocrystals [7]. Microcellulose materials derived from nature constitute a promising class of biomaterials distinguished by remarkable qualities and a broad range of possible applications. These naturally occurring materials have attracted a lot of interest because of their exceptional mechanical strength, large surface area, biocompatibility, and environmentally favorable characteristics [8, 9]. An increasingly popular substitute for synthetic materials in a variety of industries is nanocellulose, a derivative made from abundant and sustainable plant sources [10]. With a focus on cutting-edge extraction methods, this study provides a thorough review of the most recent developments in nanocellulose synthesis and its many applications. Cellulose, the most common biopolymer on Earth, serves as the fundamental building element of plant cell walls. Microfibrils make up its hierarchical structure, which can be further broken down into nanoscale dimensions [11]. Numerous extraction techniques are used to produce nanocellulose, each of which has unique benefits and features. Techniques for mechanical disintegration that efficiently shrink cellulose fibers to nanoscale dimensions include high-pressure homogenization and ultrasonication [12]. Meanwhile, amorphous portions are selectively removed using chemical methods such acid hydrolysis and oxidation, producing extremely crystalline nanocellulose [13]. Furthermore, environmentally conscious extraction methods involve enzymatic processes that harness cellulase enzymes to derive nanocellulose from cellulose-rich sources [14].

In recent trends, the efficiency and sustainability of nanocellulose extraction processes have taken center stage. The quest for greener alternatives has led to the exploration of eco-friendly solvents, aiming to replace traditional chemicals and minimize environmental impact. Enzymatic techniques have gained prominence due to their exceptional selectivity, low energy requirements, and minimal waste generation. Researchers have also investigated synergistic combinations of multiple extraction methods to maximize nanocellulose yield and enhance its properties. The overarching objective of ongoing research endeavors is to establish scalable and cost-effective extraction methodologies that enable large-scale production of nanocellulose for industrial applications [15]. In essence, the remarkable attributes of nature-inspired nanocellulose materials, coupled with advancements in extraction

#### *Nature-Inspired Nano Cellulose Materials, Advancements in Nano Cellulose Preparation… DOI: http://dx.doi.org/10.5772/intechopen.114222*

techniques, hold the promise of revolutionizing various industries through sustainable and innovative solutions [16]. This article provides a comprehensive exploration of these breakthroughs, shedding light on the potential of nanocellulose as a versatile and environmentally conscious biomaterial as shown in **Figure 2**.

Its characteristics have been further improved and its prospective uses have been increased because of improvements in nanocellulose preparation procedures [17]. Functional additives have been added to nano cellulose to give it certain functions, enabling customized applications in a variety of industries. These additions include nanoparticles, polymers, and biomolecules. To fine-tune the surface features and increase interfacial adhesion in composite materials, surface modifications through grafting or chemical treatments have been used. These preparation methods have led the way for the creation of hydrogels, films, coatings, and composites based on nanocellulose that exhibit extraordinary properties and have a wide range of applications, from electronics to healthcare [18]. Nano cellulose's adaptability is demonstrated in a wide range of study fields. Due to their biocompatibility and biodegradability, nano cellulose-based materials are used in biomedicine as diagnostic platforms, scaffolds for tissue engineering, wound dressings, and drug delivery systems [19]. Taking use of its superior mechanical qualities and electrical conductivity, nano cellulose has demonstrated promise as a sustainable option for flexible electronic devices and sensors in nanoelectronics. Additionally, due to its remarkable adsorption capability and biodegradability, nano cellulose has attracted interest in environmental applications such as water filtration, oil spill repair, and sustainable packaging options. A paradigm change in materials research and engineering has been sparked by natureinspired nano cellulose materials, which provide a green and sustainable alternative to traditional materials. The goal of this study work is to present a thorough grasp of the developments in nano cellulose preparation methods, flexible applications, and the most recent extraction method trends. It is believed that by putting light on the most recent advancements in this area, researchers and scientists would be motivated to investigate the enormous potential of nano cellulose, advancing the development of sustainable technologies and applications in a variety of industries.

**Figure 2.** *Extraction process of nano cellulose.*

#### **2. Work carried by the researchers in the last decade**

The number of papers published annually varied from 2012 to 2022, but it witnessed a rise as of 2017 with 38 articles and reached a peak in 2022 with 72 published articles. **Figure 3** depicts the annual trends in publications on this topic (nano cellulose; preparation; synthesis; applications) based on a sample of 436 articles gathered on August 2022. As shown in **Figure 3**, the publication on nanocellulose for versatile applications increases every year, especially for packaging, capacitors, batteries, drug delivery, water filtration, and sensing devices applications [20]. The rise in papers discussing the use of nanocellulose for batteries, packaging, capacitors, medicine delivery, water filtering, and sensing devices is a clear sign of the material's possible future uses. Because of its exceptional physiochemical and biological properties, nanocellulose has emerged as a material with potential for a variety of applications thanks to the rapid growth of nanotechnology and materials science. According to **Figure 3**, the number of articles has been increasing exponentially while the number of citations has decreased during the past 5 years in the field of nanocellulose. This was due to a balance between new discoveries and existing advances. This may result in a temporary decrease in citations as the scientific community works to incorporate and evaluate the newly available information. Considering the aforementioned properties of nanocellulose, transforming this renewable material, which reduces carbon footprint into advanced products is a crucial step toward sustainable development [21].

#### **2.1 2010–2012**

The researchers aimed to identify and develop the various natural fibers and micro cellulose and nano cellulose extraction techniques for lignocellulose materials. Various mechanical techniques such as high-pressure homogenization and ultrasonication were

*Nature-Inspired Nano Cellulose Materials, Advancements in Nano Cellulose Preparation… DOI: http://dx.doi.org/10.5772/intechopen.114222*

identified to extract the desired efficient micro and nano fibrils and, they break down lignocellulose biomass into nano scale [22, 23]. Different chemical treatments, such as acid hydrolysis and oxidative methods, were studied, implemented, and investigated to convert the material phases and generation of crystalline nano lignocellulose [24]. In addition to that, enzymatic approaches received attention due to sustainable and ecofriendly nature and particularly in producing cellulose to nano fibrils and particles.

#### **2.2 2013–2015**

Scientific community focused on multiplying the productivity and adaptability for suitable applications of nano cellulose development methods. New and eco-friendly solutions, chemical combinations were developed to substitute older chemicals in extraction methods, intended to minimize the environmental effects. Also, there is development in combination of different techniques to improve the quality of the materials extractions and these combinations minimize the energy consumption and time for the processes [25]. Also, researchers focused on characterization of the Nano lignocellulose and investigations carrying on its structural, mechanical, chemical, thermal and Morphological properties for suitable sustainable applications.

#### **2.3 2016–2018**

The enhancement in the nano cellulose excitation continued with various sources, also the functionalization of the materials is very important in the applications like energy storage, water filtration, drug delivery and bio textiles during the extraction process [26, 27]. Also, the scientific community started adding some nano materials to improve the specific properties nano cellulose, making it more adaptive for desired applications. By using various chemical treatments, the surface modifications of nano cellulose are improved and investigated various combinations, it is enhancing the different parameter of the nano cellulose thermal stability, crystallinity, surface roughness, morphology is very useful for development of advanced composite materials [28].

#### **2.4 2019–2022**

Researchers are currently concentrating more on the difficulties associated with extracting a significant amount of nanocellulose from different types of biomasses [29–31] and their byproducts as shown in **Figure4**. Numerous adjustments were made to extraction procedures to improve production effectiveness and lower costs, making nanocellulose economically feasible for a variety of sectors. To fulfill industrial objectives, scaling-up enzymatic procedures and continuous flow extraction techniques became popular. To further increase the accessibility and sustainability of nanocellulose, researchers looked into other sources of cellulose-rich materials [32–37]. Interdisciplinary research partnerships also developed, bringing together specialists from diverse sectors to create cutting-edge, environmentally friendly extraction methods. Research efforts to comprehend the potential negative effects of nanocellulose extraction on human health and the environment increased, resulting in the creation of safer and more environmentally friendly extraction techniques. Overall, the field of nano cellulose extraction made tremendous strides between 2010 and 2022 as researchers worked to increase the effectiveness, scalability, and sustainability of extraction techniques. These developments pave the way for the widespread use of nanocellulose materials inspired by nature in a variety of fields, such as biomedicine, electronics, packaging, and environmental remediation [38].

**Figure 4.** *Anotomy and stages of Nano cellulose.*

### **3. Materials and methods**

#### **3.1 Fourier transform infrared (FTIR) analysis**

FTIR identified characteristic peaks for cellulose, hemicellulose, and lignin in the biomass-based nano cellulose was carried out using a Thermos Scientific Nicolet iS50 FTIR Spectrometer, and the characterization revealed the real functional groups connected to the with nano cellulose sample. Crystallinity degree of nano cellulose was determined, indicating its potential for structural applications, FTIR analysis provided insights into the thermal stability of the nano cellulose, relevant for its processing and applications to view the FTIR spectra on any nano cellulose particles, it mixed with potassium bromide in the appropriate ratio, and samples are subsequently pelletized using an XLR pellet press. With a scan rate resolution, the FTIR spectra identified the range between 4000 and 400 cm− 1 .

#### **3.2 X-ray diffraction (XRD) analysis**

XRD helps quantify the crystallinity index (CI), a measure of the ordered crystalline portion of the nano cellulose. High CI indicates strong cellulosic chains and potentially superior mechanical properties. XRD peak broadening can be used to estimate the average size of crystalline domains (crystallites) in the nano cellulose. Larger crystallites often improve strength and stiffness, while smaller ones enhance surface area and reactivity. XRD patterns can also provide information about crystal morphology, such as the presence of preferred orientations. Besides cellulose, natural fibers may contain other components like hemicellulose, lignin, and minerals. XRD can identify these impurities by their characteristic peaks, allowing assessment of the purity of the extracted nano cellulose. This knowledge is crucial for understanding the properties and potential applications of the material. Different XRD peak intensities and profiles reveal the arrangement of cellulose chains within the orientations. The Bruker D8 Advance X-Ray diffraction instrument was used to compute the nano cellulose's crystallography and identify its crystalline characteristics. The mechanical, thermal, and tribological properties of the fiber were affected by these crystalline

*Nature-Inspired Nano Cellulose Materials, Advancements in Nano Cellulose Preparation… DOI: http://dx.doi.org/10.5772/intechopen.114222*

effects on a small number of the nanocellulose particles that were characterized. With a spectrum magnification mode of 2 and a step size of 0.02° at room temperature, the instrument scanning range ranged from 10° to 80°.

#### **3.3 Thermogravimetry analysis (TGA)**

TGA measures the mass loss of a material as its temperature increases. For nano cellulose, TGA identifies the decomposition temperature, marking the onset of significant mass loss. A higher decomposition temperature indicates greater thermal stability, making the material suitable for high-temperature applications. TGA also reveals the onset temperature, at which initial mass loss starts, providing insights into the initial thermal behavior. TGA curves help elucidate the mechanisms of thermal degradation in nano cellulose. Multiple mass loss steps often indicate the breakdown of different components like cellulose, hemicellulose, and lignin at different temperatures. Analyzing the weight loss percentages and temperature ranges can reveal the dominant degradation mechanisms, such as dehydration, depolymerization, and charring, TGA can be used to assess the impact of processing methods and chemical modifications on the thermal stability of nano cellulose. Treatments like mercerization or acetylation can alter the thermal degradation profile by influencing crystallinity, chain structure, and functional groups. Understanding these effects helps optimize processing and tailor nano cellulose for specific thermal performance requirements Thermo-gravimetric analysis (TGA), which was carried out with the TGA-DTA Perkin Elmer STA6000, Perkin Elmer Diamond, and an instrument with an integrated Pure Platinum Pan Furnace Holder with Ring for Sample Handling with Temperature Rise and Appropriate Balancing, revealed the separation of functional groups of the nano cellulose regarding weight loss. The loaded fibril specimen's thermal conditions steadily increase up to 20 C to 1000 C with a temperature ramp of 20.00 C/min. However, pure Nitrogen is delivered at a 20 ml/min effective flow rate, maintaining the camber conditions.

#### **3.4 Differential scanning calorimetry (DSC)**

By using a Netzsch DSC 204f1 Phoenix Instruments, researchers were able to measure the thermal characteristics of nanocellulose in relation to the material's transition temperature as well as the amount of heat emitted by the sample. The thermal transition rate was 20–400°C. In conjunction with nitrogen gas, the sample was placed in an aluminum tray with a pierced cap, which escalated the temperature until the fiber took the form of ash.

#### **3.5 Transmission electron microscopy (TEM)**

TEM offers high-resolution images of NFC, revealing its individual fibrils, their diameter, length, and surface features. This level of detail is crucial for understanding the influence of processing methods (e.g., acid hydrolysis, mechanical fibrillation) on the morphology of NFC, which impacts its properties and applications. TEM analysis can be coupled with selected area electron diffraction (SAED) to examine the crystal structure of NFC.SAED patterns reveal the arrangement of cellulose chains within the crystallites, providing information about the crystallinity index (CI) and crystal orientation. High CI indicates a well-ordered crystalline structure, potentially leading to superior mechanical properties in NFC-based materials. TEM can also detect

the presence of aggregates, bundles, or entangled regions within the NFC structure, providing insights into its dispersion behavior. Transmission electron microscopy is typically employed by Meditec, Inc., Oberkochen, Germany, to determine the micro anatomy of nanocellulose. Prior to bath sonication, samples were prepared as 200–300 nm-sized particles in water and ethanol. To hasten the volatilization of fluids from the grid, ethanol was added. Also, TEM can be used to investigate specific features of NFC, such as the presence of pores, internal defects, or surface modifications by chemical treatments. It allows visualization of functional groups attached to the surface, which are crucial for further modifications and interactions with other materials. This information is valuable for tailoring NFC for specific applications like composites, biofilms, or drug delivery systems.

#### **3.6 Scanning electron microscopy (SEM)**

SEM offers high-magnification images, revealing the surface texture, roughness, and presence of features like pores, cracks, or fibril bundles in NFC.

This information is crucial for understanding the adhesion and interaction of NFC with other materials, impacting its performance in composites, membranes, or biobased materials. SEM can also visualize the distribution of chemical modifications or coatings applied to the NFC surface, providing insights into its surface functionality. SEM allows observation of the arrangement and connectivity of individual NFC fibrils within the network. It can reveal the presence of entanglement, aggregation, or bridging between fibrils, influencing the overall porosity, strength, and stability of the NFC scaffold. Understanding the network structure helps tailor processing methods to achieve desired properties in NFC-based materials like gels, films, or scaffolds for tissue engineering. SEM can identify and characterize the presence of any remaining impurities or contaminants in the NFC, such as residual lignin, hemicellulose, or mineral particles. These impurities can affect the mechanical properties, surface chemistry, and functionality of NFC and therefore need to be minimized for specific applications. By providing visual evidence of their presence and distribution, SEM guides optimization of purification processes for obtaining high-quality NFCWith the JEOL - JSM-6390 - Scanning Electron Microscope (SEM) instrument, the scanning of the Nano lignocellulose is observed with a focused beam of electrons. The electrons interact with Nano lignocellulose atoms to produce the topography of the fiber surface with quality images seen in high convention scanning electron microscopy mode with 4.0 nm, and the electron beam's accelerating voltage is 0.5 to 30 kV at room temperature.

#### **3.7 Particle size analyzer**

A particle size analyzer (Nano-ZS, Malvern Instruments Ltd., UK) was used to measure the physical parameters of nanocellulose, including diameter, size distribution, and zeta potential. Just before analysis, the samples were diluted by deionized water five times to measure zeta potential.

#### **3.8 Atomic force microscopy (AFM)**

Unlike SEM, AFM operates at the atomic level, providing incredibly detailed images of the NFC surface. It can visualize individual cellulose chains, their

*Nature-Inspired Nano Cellulose Materials, Advancements in Nano Cellulose Preparation… DOI: http://dx.doi.org/10.5772/intechopen.114222*

orientation, and surface features like pores, roughness, and defects with unparalleled resolution. This information is crucial for understanding the surface chemistry, reactivity, and interaction of NFC with other materials at the molecular level. AFM goes beyond imaging by measuring the surface forces and mechanical properties of NFC at the nanoscale. It can determine the elasticity, stiffness, adhesion, and friction of individual fibrils within the NFC network. This knowledge contributes to tailoring NFC for specific applications requiring high strength, flexibility, or controlled interactions with other materials. AFM can be used to directly observe the distribution and morphology of chemical modifications or coatings applied to the NFC surface. It can visualize grafted functional groups, polymer attachments, or nanoparticles deposited on the cellulose chains. This information is vital for understanding the effectiveness of surface modifications in enhancing specific properties of NFC for applications like filtration, drug delivery, or biocatalysis. The potential information of Nano cellulose surface roughness, average surface roughness-(Ra), Root mean square (RMS)roughness-(Rq), Different point average surface roughness-(Rz), Skewness-(Rsk), Kurtosis-(RKU), and it exhibits the highest peak-to-valley height (Rt), was observed using the scanning probe microscopy of AFM technique. Asylum Research, MFP-3D BIO AFM equipment (USA) was used to perform the scanning.

#### **3.9 Energy dispersive X-ray analysis (EDAX)**

To determine the primary elements and chemical combinations on the surface of the nanocellulose particles, this analytical technique focuses on elemental analysis using X-ray excitation. The summary of several compounds and elements, including Ni, Be, Ti, Zn, C, O, Na, Mg, Al, Si, Ag, P, Cl, K, Ca, and S, is distributed. The model Jeol 6390LA scanning electron microscope was characterized (**Table 1**).




*Nature-Inspired Nano Cellulose Materials, Advancements in Nano Cellulose Preparation… DOI: http://dx.doi.org/10.5772/intechopen.114222*

#### **Table 1.**

*Nano cellulose source and extraction techniques.*

### **4. Versatile applications of nature-inspired nano cellulose**

While nature-inspired nano cellulose materials hold great promise in these applications, challenges remain in terms of scalability, cost-effectiveness, and standardization of production methods. Additionally, ensuring compatibility with existing recycling and composting systems is crucial for widespread adoption. The integration of nature-inspired nano cellulose materials in barrier coatings and biodegradable packaging solutions presents a transformative pathway for the packaging industry [62, 63]. These applications not only enhance product preservation and consumer experience but also contribute to a more sustainable and environmentally conscious future by reducing plastic waste and promoting circular economy principles. Continued research and development in this field are essential to fully unlock the potential of nature-inspired nano cellulose in addressing the evolving needs of modern packaging, innovative capacitors, and batteries, advanced drug delivery systems, efficient water filtration materials, and new sensing devices as showed in **Figure 5** [64, 65].

#### **4.1 Barrier coatings and biodegradable packaging**

Nature-inspired nano cellulose materials have demonstrated exceptional potential in revolutionizing packaging technologies by offering both barrier coatings for extending shelf life and innovative solutions for biodegradable and compostable packaging. These applications address critical challenges in the packaging industry, such as food preservation, environmental sustainability, and reduced plastic waste.

#### *4.1.1 Barrier coatings to enhance packaging shelf life*

A natural contender for extending the shelf life of many packaged items is nano cellulose, which has outstanding barrier characteristics against gases, moisture, and

**Figure 5.** *Applications for nature-inspired nano cellulose materials.*

*Nature-Inspired Nano Cellulose Materials, Advancements in Nano Cellulose Preparation… DOI: http://dx.doi.org/10.5772/intechopen.114222*

other external pollutants. Food waste can be decreased, and sustainable consumption encouraged by using nano cellulose as a barrier layer to dramatically extend the freshness and quality of perishable goods.

#### *4.1.1.1 Benefits*


#### *4.1.1.2 Applications*


Lightweight, strength and mimicking nature's strength, nature inspired nano cellulose provides high strength-to-weight ratio, ideal for aircraft parts and bio-based composites. Derived from biomass like wood or plant waste, it promotes a circular economy and reduces environmental impact. It is safe for use in medical applications like wound dressings and drug delivery systems. Blocks moisture, oil, and gases, making it valuable for food packaging and protective coatings. Versatile modification allows tuning for specific applications like filtration, biosensing, and catalytic supports. Scalable production from various sources ensures cost-effectiveness and widespread accessibility. Enhances energy efficiency in buildings and materials due to its low thermal conductivity. Certain compositions offer inherent flame resistance, improving safety in construction and transportation. Adjustable pore size facilitates selective filtration, purification, and controlled release applications. Also, Paves the way for replacing petroleum-based materials with sustainable and eco-friendly alternatives.

#### *4.1.2 Biodegradable and compostable packaging solutions*

Biodegradable and compostable packaging options have drawn a lot of attention in response to the growing worries about plastic pollution and its effects on the environment. Given that it can be produced from renewable resources, exhibits biodegradability, and has a smaller environmental impact than traditional plastic packaging materials, nature-inspired nano cellulose presents an attractive option.

#### *4.1.2.1 Benefits*


#### *4.1.2.2 Applications*


#### **4.2 Functional nanocomposites applications of nature-inspired nano cellulose**

Nature-inspired nano cellulose has emerged as a remarkable candidate for enhancing the performance of polymer composites, offering a wide array of mechanical, thermal, and electrical enhancements [66]. The integration of nano cellulose into polymer matrices imparts unique properties that open up innovative applications in various industries. This section explores the multifaceted applications of nano cellulose as a reinforcement in polymer composites, highlighting its contributions to mechanical strength, thermal stability, and electrical conductivity [67, 68]. While significant advancements have been made in the development of nano cellulose-based functional nanocomposites, ongoing research aims to further optimize dispersion techniques, tailor surface modifications, and explore novel polymer/nano cellulose combinations. These efforts will likely lead to even more diverse and advanced applications in fields such as smart materials, 3D printing, and multifunctional coatings [69]. The incorporation of nature-inspired nano cellulose into polymer composites for mechanical, thermal, and electrical enhancements holds immense promise for a wide range of applications. By harnessing the remarkable properties of nano cellulose, researchers and industries are driving innovation toward sustainable and high-performance materials that address contemporary challenges and pave the way for a more efficient and environmentally conscious future.

#### *4.2.1 Nano cellulose reinforcement in polymer composites*

Nano cellulose, due to its exceptional mechanical properties and abundant surface functionalities, has gained attention as an effective reinforcement material in polymer composites. By dispersing nano cellulose within a polymer matrix, these composites exhibit enhanced mechanical strength, stiffness, and durability compared to their neat polymer counterparts [70]. Nano cellulose's complex network structure has a synergistic effect that distributes and absorbs stress, enhancing the composite's overall mechanical performance [71].

*Nature-Inspired Nano Cellulose Materials, Advancements in Nano Cellulose Preparation… DOI: http://dx.doi.org/10.5772/intechopen.114222*

#### *4.2.2 Mechanical enhancements*

The mechanical characteristics of polymer matrices are significantly enhanced by adding nanocellulose. Enhancing the strength and toughness of composite materials are nanocellulose fibers, which have a high aspect ratio and exceptional tensile strength [72, 73]. This reinforcement is especially useful in sectors that demand materials that are low weight but nevertheless durable, such as athletic products, aircraft constructions, and automotive components [74–76].

#### *4.2.3 Thermal enhancements*

Nano cellulose's low thermal expansion coefficient and high thermal stability make it a valuable component for enhancing the thermal performance of polymer composites. By integrating nanocellulose, it is possible to lessen the thermal expansion mismatch between the polymer and other materials, potentially reducing warping or cracking due to temperature changes. This is helpful for applications like electronic packaging, thermal insulation, and flame-resistant materials where thermal stability is important.

#### *4.2.4 Electrical enhancements*

Nano cellulose can be used into polymer composites to add electrical conductivity because of its special electrical characteristics, including its high surface area and capacity to carry charges. By adjusting the nanocellulose quantity and dispersion within the polymer matrix, this conductivity can be adjusted. These conductive composites are used in wearable technology, flexible electronics, sensors, and electromagnetic interference shielding.

#### *4.2.5 Environmental benefits*

The use of nature-inspired nano cellulose as a reinforcement in polymer composites aligns with sustainability goals. Nano cellulose is derived from renewable sources and is biodegradable, making it an eco-friendly alternative to synthetic reinforcements. The reduced reliance on fossil-based materials contributes to a more environmentally responsible approach to composite manufacturing.

#### **4.3 Biomedical applications of nature-inspired nano cellulose**

Nanocellulose materials that are inspired by nature have shown incredible promise in the field of biomedicine, providing ground-breaking approaches to drug delivery, wound healing, and tissue engineering [77]. Nanocellulose-based materials have distinctive features that make them ideal candidates for many biomedical applications, drawing inspiration from the intricate architectures present in natural systems [78]. There are obstacles to overcome despite the apparent potential of nature-inspired nanocellulose materials in biomedical applications. In order to move these materials from the laboratory to clinical practice, it is essential to ensure batch-to-batch consistency, maximize scalability, and obtain regulatory approvals. To ensure patient safety, a thorough assessment of the long-term biocompatibility and degrading behavior of nanocellulose-based materials is required [79]. Nano cellulose materials with natural inspirations have opened up new avenues for biomedical uses. Nano cellulose-based materials provide inventive solutions that take advantage of their special qualities and biomimetic traits, including adaptable drug delivery carriers, wound healing assistance, and tissue engineering [80]. These substances have the potential to revolutionize how we approach healthcare and regenerative medicine as research develops, leading to better patient outcomes and quality of life.

#### *4.3.1 Drug delivery carriers*

Materials made of nanocellulose have shown promise as regulated and precise medication delivery systems. Their large surface area, adaptable surface chemistry, and biocompatibility make it possible for medicinal substances to be loaded and released quickly. Numerous medications, including hydrophilic and hydrophobic substances, proteins, and nucleic acids, can be enclosed in nanocellulose. Nano cellulose's porosity structure permits continuous and regulated medication release, reducing adverse effects and improving therapeutic results. Precision medicine techniques are made possible by the surface modification of nanocellulose with targeting ligands, which increases its specificity to cell types or regions.

#### *4.3.2 Wound healing materials*

Wound dressings made of nano cellulose have drawn attention because of their outstanding abilities to speed up the healing process. These substances have a great capacity to store water and can keep a wound wet, which promotes cell migration, proliferation, and tissue regeneration. Nano cellulose dressings also possess antibacterial qualities, which can aid in the healing process and help prevent infections. Nanocellulose wound dressings resemble the extracellular matrix of tissues to give the wound mechanical support and can be customized for various wound types and stages.

#### *4.3.3 Tissue engineering scaffolds for regenerative medicine*

Nano cellulose scaffolds are essential for tissue engineering because they provide a three-dimensional environment for cell adhesion, growth, and differentiation. These scaffolds offer a favorable environment for tissue regeneration because they closely mirror the structural design of tissues and organs. The mechanical properties of nano cellulose can be modified to correspond to those of particular tissues, ensuring optimal mechanical support throughout the healing process. Nanocellulose scaffolds are excellent candidates for applications like bone, cartilage, and nerve tissue engineering because they can direct cell behavior and encourage tissue creation through surface functionalization and the insertion of bioactive chemicals.

#### **4.4 Water purification and filtration of nature-inspired nano cellulose**

Nature-inspired nano cellulose materials have shown promise in tackling water filtration and purification issues [81]. They are successful in eliminating impurities and raising the quality of the water due to their special characteristics.

#### *4.4.1 Adsorbent materials for heavy metals and pollutant removal*

Nano cellulose-based adsorbents are ideal for removing heavy metals and other contaminants from water sources because they have a large surface area and a lot

#### *Nature-Inspired Nano Cellulose Materials, Advancements in Nano Cellulose Preparation… DOI: http://dx.doi.org/10.5772/intechopen.114222*

of binding sites. Lead, mercury, cadmium, and arsenic can be effectively removed thanks to their natural affinity for metal ions. This application helps to protect public health and lessen the negative effects of industrial activity on the environment.

#### *4.4.2 Microfiltration and ultrafiltration membranes*

To precisely separate particles, bacteria, and macromolecules from water, nanocellulose materials can be designed into microfiltration and ultrafiltration membranes. These membranes can be functionalized to increase selectivity and permeability and have adjustable pore diameters. Their use in water treatment processes aids in the removal of suspended solids, bacteria, viruses, and other impurities, yielding cleaner and safer water resources.

#### *4.4.3 Enhanced adsorption and filtration performance*

The hierarchical structure of nano cellulose materials, reminiscent of natural systems, allows for tailored modification and functionalization. This flexibility enables the design of materials with enhanced adsorption capacities and filtration efficiencies. By optimizing surface chemistry and morphology, researchers can fine-tune the materials' performance for specific water treatment needs.

#### *4.4.4 Sustainable water management*

Incorporating nano cellulose-based materials into water purification and filtration technologies aligns with sustainability goals. These materials are derived from renewable resources and can be produced using eco-friendly processes, reducing the overall environmental impact of water treatment systems. The application of nature-inspired nano cellulose materials in water purification and filtration highlights their potential to contribute to cleaner and safer water sources. As ongoing research continues to advance these materials, their role in addressing global water challenges becomes increasingly significant. Whether through adsorption, membrane filtration, or tailored modifications, nano cellulose materials offer innovative solutions for sustainable water management and environmental protection [82]. Materials made of nanocellulose that are inspired by nature are useful for environmental cleanup following disasters or industrial accidents that taint water sources. Their effective adsorption and filtering abilities facilitate the quick removal of contaminants, assisting in ecosystem restoration and aquatic life preservation.

#### **4.5 Sustainable energy solutions of nature-inspired nano cellulose**

The performance of flexible electronics, energy storage devices, batteries, and capacitors has been significantly improved by nature-inspired nano cellulose, which has emerged as a promising contender for different sustainable energy solutions [83]. Nano cellulose's special qualities, such as its large surface area, mechanical toughness, and ion conductivity, make it a desirable material for developing energy innovations [84].

#### *4.5.1 Nano cellulose in flexible electronics*

• Materials that are not only lightweight and flexible but also ecologically benign are needed for flexible and wearable electronics. Nano cellulose, which is made from renewable resources, completely satisfies these requirements.


#### *4.5.2 Energy storage devices*


#### *4.5.3 Enhancing battery performance*


#### *4.5.4 Capacitors with nano cellulose*


### **5. Future directions and challenges**

• The search for nature-inspired nanocellulose materials, improvements in manufacturing processes, and a wide range of uses is an ongoing adventure that offers *Nature-Inspired Nano Cellulose Materials, Advancements in Nano Cellulose Preparation… DOI: http://dx.doi.org/10.5772/intechopen.114222*

both thrilling possibilities and difficult obstacles. The trajectory of research and innovation in this discipline is likely to be shaped by several future objectives and obstacles as the field matures.


#### **5.1 Emerging trends in nano cellulose research**

Several new trends that are emerging are influencing the direction that nanocellulose research will take as it continues to develop. Researchers are concentrating on creating unique nano cellulose variants with specialized features for certain applications, such as cellulose nanocrystals and cellulose nanofibrils. Techniques for surface functionalization and modification are becoming more popular to increase compatibility with various matrices and boost performance. Additionally, multidisciplinary partnerships between engineers, biotechnologists, and material scientists are increasingly widespread, resulting in ground-breaking discoveries and multifunctional materials. By combining nanocellulose with other cutting-edge materials like graphene and nanoparticles, new opportunities for synergistic effects and improved functionality are becoming possible. To develop materials with superior mechanical characteristics and self-healing capacities, efforts are also focused on comprehending and imitating nature's hierarchical structures. Another intriguing area that shows promise for the future is investigating the possibilities of nanocellulose in the realm of nanomedicine, including targeted medication delivery and tissue engineering.

#### **5.2 Addressing scalability and cost-effectiveness**

Although nano cellulose has great potential, scaling and cost-effectiveness issues make it difficult to use on a large scale. Large-scale manufacturing of nano cellulose is economically difficult since it frequently includes energy-intensive processes and specialized equipment. To increase sustainability and lower costs, researchers are actively striving to improve production techniques, create more energy-efficient processes, and investigate the use of waste streams as feedstock. Production expansion is still a top aim as long as constant quality and minimal environmental impact are maintained. To create affordable and scalable processes for producing nanocellulose, collaboration between government, business, and academia is essential. Additionally, investigating bioengineering techniques to increase cellulose yield in plants or microorganisms may provide creative ideas to get around scalability obstacles.

#### **5.3 Environmental considerations and life cycle analysis**

Addressing the environmental impact of nano cellulose production and use is of utmost importance as the demand for sustainable materials rises. To examine the entire environmental impact of nano cellulose materials and pinpoint any potential problem

areas in their manufacturing, use, and disposal, thorough life cycle assessments (LCAs) must be carried out. To lessen the usage of harsh chemicals and energy-intensive stages, researchers are actively investigating greener and more environmentally friendly extraction technologies, such as enzyme-assisted processes. For nanocellulose-based goods, recycling and circular economy strategies are being researched to reduce waste generation and advance resource efficiency. For responsible development, it is also essential to comprehend the potential environmental dangers connected to nanocellulose materials, such as their behavior in aquatic systems and potential ecotoxicity. To ensure the safe and sustainable usage of nanocellulose across a range of applications, regulatory frameworks and guidelines may need to be established.

#### **5.4 Holistic approaches and collaborative initiatives**

A comprehensive strategy that incorporates scientific research, engineering innovation, commercial viability, and environmental stewardship is needed to address the issues and realize the full potential of nano cellulose materials. Knowledge exchange, technology transfer, and the creation of standardized protocols can be facilitated by collaborative projects involving academia, business, governmental organizations, and non-governmental organizations. Openly exchanging data, processes, and best practices can hasten development and reduce effort duplication. International partnerships can promote a global perspective on research into nanocellulose by allowing the sharing of various perspectives and experiences from various places. The multidisciplinary nature of nano cellulose research necessitates global collaboration and knowledge exchange among scientists, engineers, and stakeholders from different regions. Open-access platforms, conferences, and research networks will play a crucial role in facilitating collaboration and accelerating advancements. In conclusion, the future of nano cellulose materials holds great promise, with emerging trends pushing the boundaries of possibilities. Addressing challenges related to scalability, cost-effectiveness, and environmental sustainability is essential for unlocking the full potential of these nature-inspired materials and realizing their transformative impact across various industries. By adopting a multidisciplinary and collaborative approach, researchers and stakeholders can drive innovation and shape a more sustainable future.

#### **6. Conclusions**

From the nature perspective, the current study summarizes the available resources for extraction of nano cellulose, and their extraction methods with the potential applications. Based on the current research trend, the scientific community identified the nano cellulose and existed from forest biomass, agro waste, and ocean biomass, for many applications in Biomedical, aerospace, energy, pharmaceutical and other versatile applications in emerging areas. The economical, thermal, mechanical, morphological, and chemical properties of the nano cellulose broadened the applications where it fit. Since there is huge demand of nano cellulose materials for sustainable applications is indeed requirement for large production. Also, nano cellulose materials are increasing the economic aspects of productivity, and it is playing a very important role in environmental safety and protection.

The quest for efficient and eco-friendly nano cellulose preparation methods has been guided by nature's ingenious processes. Biomimetic extraction techniques, mechanical disintegration, and self-assembly have all contributed to unlocking the potential of nano cellulose. Understanding the structural, mechanical, and thermal

#### *Nature-Inspired Nano Cellulose Materials, Advancements in Nano Cellulose Preparation… DOI: http://dx.doi.org/10.5772/intechopen.114222*

characteristics of nanocellulose materials has benefited from thorough characterization. Applications of nature-inspired nano cellulose materials include biomedical devices, textiles, water purification, and energy solutions in addition to sustainable packaging. A more sustainable and technologically advanced future is now possible thanks to the extraordinary qualities of nano cellulose, such as its mechanical strength, thermal stability, and biocompatibility.

Nanocellulose materials with natural inspiration have been instrumental in the development of sustainable energy systems. Their use in flexible electronics, energy storage systems, batteries, and capacitors has improved performance, increased the efficiency of energy storage, and made it possible to develop more ecologically friendly solutions for energy storage and utilization. In conclusion, the development of nanocellulose materials that are inspired by nature, improvements in the methods used to prepare them, and their wide range of applications highlight the incredible promise of biomimetic methods in materials research. This journey of discovery stands as a testament to the ingenuity of researchers and their commitment to harnessing nature's wisdom to address contemporary challenges. As this field continues to evolve, we anticipate even greater strides in sustainable innovation, offering a brighter and more sustainable future for generations to come.

#### **Conflict of interest**

The authors declare no conflict of interest.

### **Author details**

Hanumanthu Jeevan Rao1 \*, Sanjay Singh1 , Perumalla Janaki Ramulu2 , Narender Singh1 , Thiago F. Santos3 , Caroliny M. Santos3 , Nandini Robin Nadar4 and Gara Dheeraj Kumar4

1 Amity Institute of Aerospace Engineering, Amity University Uttar Pradesh, Noida, Uttar Pradesh, India

2 Department of Mechanical Engineering, School of Mechanical, Chemical and Materials Engineering, ASTU, Adama, Ethiopia

3 Postgraduate Program in Chemical Engineering, Technology Center, Federal University of Rio Grande do Norte, Brazil

4 Department of Aerospace, Jain (Deemed-to-be University), Bengaluru, Karnataka, India

\*Address all correspondence to: jeevan.rao14@gmail.com

© 2024 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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[83] Xiao J, Li H, Zhang H, He S, Zhang Q, Liu K, et al. Nanocellulose and its derived composite electrodes toward supercapacitors: Fabrication, properties, and challenges. Journal of Bioresources and Bioproducts. 2022;**7**:245-269. DOI: 10.1016/j.jobab.2022.05.003

[84] Li R, Tian D, Chen L, Zhuang B, Feng H, Li Q, et al. The application of cellulose nanofibrils in energy systems. Batteries. 2023;**9**:399. DOI: 10.3390/ batteries9080399

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[88] Chen C, Hu L. Nanocellulose toward advanced energy storage devices: Structure and electrochemistry. Accounts of Chemical Research. 2018;**51**:3154- 3165. DOI: 10.1021/acs.accounts.8b00391

#### **Chapter 4**

## Fascinating Properties and Applications of Nanocellulose in the Food Industry

*Elham Asghari-Varzaneh and Hajar Shekarchizadeh*

#### **Abstract**

Nanocellulose, a material derived from cellulose fibers, has gained significant attention in various industries due to its unique properties and potential applications. From an economic perspective, using nanocellulose in industry offers several advantages, such as cost-effectiveness, enhanced product performance, environmental benefits, and diversified applications. Also, it is utilized in the food industry because of its distinct properties, including high surface area, rheological behavior, water absorption ability, crystallinity, and no cytotoxicity. A significant application of nanocellulose is its potential to replace fats, carbohydrates, and proteins and serve as stabilizing agents in high-calorie foods. Moreover, nanocellulose has demonstrated exceptional efficacy as a delivery system, making it an ideal choice for preserving nutrients and active ingredients in food products. A primary objective in the packaging industry is to maintain food quality, extend its shelf life, and minimize waste. Since nanocellulose is both renewable and natural and offers oxygen and water vapor barrier properties, it emerges as a suitable candidate for the packaging industry. However, despite its promising features and applications, there are uncertainties around its non-toxicity and the potential impact on human health, issues that are currently being examined by scientists in the food industry.

**Keywords:** nanocellulose, food stabilizer, dietary fiber, encapsulating agent, packaging

#### **1. Introduction**

Cellulose (C6H10O5)n, one of the most abundant natural polymers in the world, was first discovered by Anselme Payene. Cellulose is the main component of the cell walls of most plants. The main sources of cellulose are wood pulp, cotton stalks, straw, bacteria, sea animals, and agricultural waste (fruits and vegetables) [1]. Renewability, non-toxicity, environmentally-friendly, wide availability, excellent mechanical properties, low weight, reinforcing capabilities, and biodegradability, with no adverse effects on health and environment, are among the important and practical features of cellulose used in various industries such as food, paper, bio-material, and pharmaceutical industries [2, 3]. Cellulose is a homopolysaccharide compound formed by joining large numbers of β-D-glucose monomers with β (1,4) glycosidic bonds [4]. Cellulose forms intra- and intermolecular hydrogen bonds due to hydroxyl groups in its structure. In general, these hydroxyl groups and their ability to create hydrogen bonds affect the biodegradability, chirality, and hydrophilic properties of cellulose and also create important functions, such as microfibrillated structure, hierarchical organization (crystalline and amorphous fractions), and cohesive nature [5, 6].

Cellulose derivatives, including cellulose acetate, cellulose sulfate, cellulose nitrate, carboxymethyl cellulose, ethyl cellulose, methylcellulose, and nanocellulose*,* exhibit distinct properties (sizes and shapes) and characteristics based on the production method and materials employed [1].

Nanocellulose consists of cellulose particles with dimensions less than 100 nm. The nanocellulose fibers can be obtained through chemical modification, such as acid hydrolysis, or physical modification, such as ultrasonication [7]. Nanocellulose possesses several practical and significant properties that make it an ideal choice for preparing various polymer composites, including low density, non-abrasiveness, combustibility, excellent potential for chemical modification, crystalline structure, increased specific surface area, advantageous rheological properties, outstanding resistance to water vapor transmission and oxygen permeation, effective barrier against aqueous liquids and oil grease, non-toxicity, and inexpensiveness compared to other synthetic polymers [8, 9].

Nanocellulose, an environmentally friendly and biodegradable biomaterial, has emerged as a promising alternative to synthetic materials. In recent years, cellulose nanocomposites have found applications in various industries, including food, medical, biomedical, pharmaceutical, civil construction, automotive, electronics, packaging, construction, and wastewater treatment [8, 10].

For example, nanocellulose-based biomaterials have properties similar to natural tissue, making them proper for cell attachment and growth. Nanocrystalline suspensions can be used as a cell culture environment, and cellulose nanofibers have been studied to grow membranes containing healing and bacteriostatic agents for treating skin burns [11]. Also, in the field of application in the construction industry, it is possible to mention the efforts of researchers to produce products compatible with the environment. For example, in 2019, a study was conducted to investigate the mechanical performance of integrating cellulose fibers into cement systems. The findings showed that the material had improved properties, including compressive strength, modulus of elasticity, and toughness, indicating its potential as a promising material with ductile and more resistant behavior [12].

#### **2. Economic aspects of using nanocellulose in industry**

Nanocellulose can be produced from different raw materials, which depend on the availability of the raw material, its cost, and environmental conditions, and these factors can directly affect the cost of nanocellulose production. This is observed, even though the properties of the produced nanocellulose and its application in the industry are influenced by the raw material and production method. As research and innovation in nanocellulose-based technologies continue, new sources and production methods are expected to be recognized and developed, further expanding its potential applications over the years [13]. In general, the economic advantages of the use of nanocellulose in industry can be mentioned as follows:

1. Cost-effectiveness: nanocellulose can be produced from renewable sources such as wood pulp, agricultural residues, or waste paper. This makes it a cost-effective and supportable alternative to customary materials. Additionally, the production

#### *Fascinating Properties and Applications of Nanocellulose in the Food Industry DOI: http://dx.doi.org/10.5772/intechopen.114085*

processes for nanocellulose are becoming more efficient, further reducing production costs; 2. Lightweight and strong: nanocellulose is lightweight yet incredibly strong, making it an attractive material for industries such as automotive, aerospace, and construction. Its high strength-to-weight ratio can lead to cost savings in manufacturing and transportation; 3. Enhanced product performance: the interpolation of nanocellulose into various products can improve their mechanical strength, barrier properties, and persistence. This can result in longer-lasting products with reduced maintenance and replacement costs; 4. Environmental benefits: nanocellulose is biodegradable and non-toxic, making it an environmentally friendly alternative to synthetic materials. As sustainability becomes increasingly important to consumers and regulatory bodies, the use of nanocellulose can provide a competitive advantage for companies looking to reduce their environmental impact; 5. Variegated applications: nanocellulose has a wide range of potential applications across industries such as packaging, textiles, electronics, healthcare, and energy storage. This versatility opens up new market opportunities for companies utilizing nanocellulose in their products.

Apart from the economic benefits of using nanocellulose in the industry, there are several disadvantages and barriers facing nanocellulose and its wide application in various industries, which can be mentioned as 1. Initial investment: implementing and developing efficient and valuable methods of nanocellulose production can be expensive, which is the biggest challenge for manufacturing companies; 2. Competition: nanocellulose faces competition from other materials, and companies may need to invest in marketing and education efforts to promote its advantages and gain market share; 3. Uncertain demand: due to the novelty of using nanocellulose in the industry, this material may face uncertainty in the market and price; 4. Regulatory challenges: regulatory requirements and standards for nanocellulose products may impact production costs and market access, requiring companies to navigate complex regulatory landscapes [13, 14].

#### **3. Types of nanocellulose**

In general, depending on the source of cellulose, the production method, their size, and function, there are three types of nanocellulose with similar chemical composition and different morphology, namely nano-fibrillated cellulose (NFC), cellulose nanocrystals (CNCs), and bacterial nanocellulose (BNC) [15].

Nano-fibrillated celluloses are 1–100 nm in diameter and 500–2000 nm in length (depending on the type of raw material), and all are long, entangled, and flexible. Moreover, they consist of 100% cellulose in both amorphous and crystalline forms, and in comparison to CNCs, NFCs possess a greater surface area, higher aspect ratio, and longer length [16]. Also, among various types of nanocelluloses, NFCs have the simplest production method, owing to their avoidance of harsh chemical processes that degrade the molecular structure of cellulose. Generally, they are prepared by the physical separation of cellulose fibers, such as grinding, homogenization, and ultrasonication, or by chemical methods, including oxidation of wood raw material by 2,6,6-Tetramethylpiperidine-1-oxyl radical (TEMPO) under gentle stirring, and finally by a combination of two chemical and physical methods that include carboxymethylation and high-pressure homogenization [17].

Mostly, CNCs are obtained through acid hydrolysis of cellulose nanofibrils. Their diameter and length are 2–20 μm and 100–500 nm, respectively. Their structure is

rod- or whisker-shaped and consists entirely of crystalline cellulose (around 54–88% crystallinity). CNCs exhibit an exceptionally high crystallinity [18]. The production of nanocellulose involves two fundamental steps. First, impurities of the raw material are eliminated using an alkaline (NaOH) solution or by a bleaching method. Subsequently, the raw material is heated in an acidic environment to eliminate the amorphous portion of cellulose, and after centrifugation and dialysis, CNC is obtained. It is worth noting that the type of raw material, type of acid, and hydrolysis temperature and time affect the dimensions of CNCs [19, 20].

Bacterial nanocelluloses have a twisted ribbon-like shape and length of 20–100 nm, which are chiefly obtained from *Gluconacetobacter xylinus, Acetobacter*, *Rhizobium*, *Agrobacterium*, *Aerobacter*, *Achromobacter*, *Azotobacter*, *Salmonella*, *Escherichia*, and *Sarcina* systems or cell-free systems [21, 22]. BNCs form a hydrogel at the water-air interface. Their synthesis involves the production of β-1,4-glucan chains within the bacterial cell excreted through the cell membrane as protofibrils. These protofibrils then crystallize to create ribbon-shaped microfibrils, leading to the formation of a pellicle. The physical and mechanical properties of BNCs are different depending on the type of bacterial strain, synthesis method (cell and cell-free systems), cultural conditions (carbon and nitrogen source), structural features, and arrangement of fibers [23].

#### **4. Application of nanocellulose in the food industry**

Nanocelluloses possess exceptional utility and distinctiveness across various industries, particularly in the food sector, attributed to their remarkable capacity for forming hydrogen bonds, expansive surface area, and aspect ratio compared to cellulose fibers. Additionally, their inexhaustible primary sources further enhance their significance. Different types of nanocellulose are used in the food industry as emulsifiers, fat substitutes, thickeners, and in food packaging. Nanocellulose also acts as a carrier agent in biological systems to carry nutrients like curcumin, vitamins, omega-3-fatty acids, minerals, amino acids, nutraceuticals, probiotics, polyphenols, antioxidants, micronutrients, enzymes, and essential oils [23].

#### **4.1 Stabilizer and emulsifier**

In the food industry, many natural substances such as gums (like xanthan, carrageenan, gellan, and locust bean gum) [24] and hydrophilic polysaccharides extracted from seaweed and microorganisms are used as natural stabilizers and emulsifiers [25]. The use of stabilizers in products like gravies, creams, sauces, frostings, additives, icings, salad dressings, foams, soups, puddings, dips, desserts, texture enhancers, thickeners, and frozen dairy products is highly prevalent [26]. However, subjects such as thinning, phase separation, and lack of phase stability over time are the main problems of the food industry. Hence, it is crucial to employ a cost-effective and easily extractable natural material that not only preserves the quality but also prevents any physical changes in the products [27]. Nanocellulose (NC), as a natural emulsifying and stabilizing ingredient, is of great interest in food products due to its unique rheological properties. Also, it can extend the shelf life and preserve the shape of frozen desserts. While increasing the amount of other stabilizers can improve shape retention, it may negatively impact the texture and flavor of the dessert. However, NC can prolong the structure retention time without compromising other desirable

#### *Fascinating Properties and Applications of Nanocellulose in the Food Industry DOI: http://dx.doi.org/10.5772/intechopen.114085*

characteristics of the dessert [28]. In 2022, researchers successfully created medium internal phase oil-in-water (O/W) Pickering emulsions using bamboo shoots nanocellulose (BSNC) as a stabilizer. The nanocellulose extracted from bamboo shoots demonstrated promising potential as an emulsifier in the Pickering emulsions. Emulsions with a BSNC content of 0.5 wt% at a 5:5 oil-to-water volume ratio had smaller particle sizes of around 25. However, increasing the BSNC content resulted in larger droplet sizes and even demulsification. Subjecting the emulsions to twice shearing improved their physicochemical properties significantly by reducing droplet size. As the BSNC content increased, the apparent viscosity initially increased and then decreased, with all emulsions exhibiting elastic behaviors [29].

Another application of NC is the development of Pickering emulsions in various industries such as dairy, meat, and bakery. For example, in the dairy industry, it is a suitable option for replacing butter in baking cakes and replacing cream in preparing frozen yogurt and ice cream. In 2019, research was conducted to investigate the addition of NC to ice cream and its effect on the structure and performance of the frozen product. The results showed that NC has a direct effect on fat absorption. So, the presence of NC caused enhanced sensory attributes in samples containing less fat, and crystal growth, ice cream hardness, and rheology remained unchanged at subzero temperatures [30]. The surface tension between oil and water decreases by adding NC to different emulsions, which improves the stability of the emulsion system [31]. In this regard, the effect of adding NC as a stabilizer on meat sausage was investigated. The findings revealed that the addition of NC enhanced the binding capacity of fat and moisture, the sausage's hardness, elasticity, and chewability, resulting in reduced cooking loss and improved emulsion stability [32]. So, in general, Pickering emulsions stabilized with NCs have more stability and better compatibility than traditional surfactants.

#### **4.2 Fat replacer**

Today, with the progress of science, the world has moved toward consuming healthy and smart foods with low fat and sugar content and, as a result, suitable calories. Obesity and overweight are some of the basic problems among people of different societies, which have forced scientists to research the replacement of fattening factors, especially fat, with appropriate and natural substances. The use of NC in many foods, especially high-consumption and high-demand foods such as ice cream and processed meats, is one of the effective options for reducing the amount of fat and, as a result, reducing calories, maintaining the quality of the product's texture, and improving nutritional and health aspects. In this regard, Velásquez-Cock et al. [30] added NC to the ice cream formulation to produce ice cream with low fat and calories. The results showed that the presence of NC in low-fat ice creams caused a significant decrease in the melting rate of reduced-fat ice cream. A study conducted in 2022 aimed to develop reduced-fat mayonnaise formulations with 5, 15, and 30% fat content using varying concentrations of nanocellulose synthesized from palm-pressed fiber. The study also included a 20-day stability assessment. The results showed that the reduced-fat mayonnaise had smaller oil droplets closely packed together when viewed under a light microscope. However, significant oil droplet coalescence was observed during storage, which could lead to viscosity loss. These findings suggest that nanocellulose was effective as a fat mimetic during mayonnaise formulation, but its effectiveness during long-term storage is uncertain [33].

#### **4.3 Dietary fiber**

Nanocellulose, as a dietary fiber, has a significant impact on the general health of the body through three mechanisms: promoting satiety (effect on protein), regulating blood sugar (effect on glucose), and regulating blood lipids (impact on lipid and fat). Foods containing high protein are mainly used to control obesity and weight due to their high satiety effect; therefore, a more complex protein has a slower digestion rate, and as a result, the time to induce satiety in the body increases. In general, the speed of food digestion determines the availability of nutrients in the body, and the delay in emptying the stomach may evoke the satiety effect [34, 35]. The results of recent years have shown that NC, like anionic polysaccharides (pectin or carrageenan), effectively enhances digesta viscosity, impedes proteolysis, and decelerates gastric emptying through their interaction with protein solutions. NCs with a negative charge can decrease protein proteolysis in the stomach by attracting proteins with opposite charges through electrostatic forces. Consequently, this leads to stable satiety and reduced consumption of other foods (**Figure 1**) [36].

Another benefit of NC as a dietary fiber is its role in the treatment of obesity and diabetes. NC impedes glucose absorption in the body by slowing down the digestion and absorption of glucose. NC spontaneously combines with glucosedecomposing enzymes to form a hydrophobic complex, which changes the protein structure of the enzyme and disrupts the activity of α-amylase and amyl glucosidase (**Figure 2**) [37]. In 2019, scientists also investigated the effect of NC, extracted from bleached softwood pulp, on digestive viscosity, glucose digestion, and absorption. The findings indicated that concentrations exceeding 0.5 wt% substantially enhanced digesta viscosity, slowed hydrolysis, minimized glucose release, and delayed glucose diffusion [39]. Also, in 2023, to increase the value of waste chili stems and promote the recycling of green resources, cellulose was extracted from chili stems using a nitric acid-ethanol method, and nanocellulose was then prepared using a sulfuric acid hydrolysis method. The study demonstrated that biscuits with satisfactory overall quality could be made using 7% nanocellulose, resulting in a regular appearance and a relatively smooth surface. Mice experiments revealed that consuming biscuits containing nanocellulose reduced food intake significantly and inhibited weight gain in mice. Therefore, the research suggests that whole wheat biscuits with nanocellulose could be considered as a high-fiber food option [40].

**Figure 1.** *Schematic of the effect of NC on protein digestion [36].*

*Fascinating Properties and Applications of Nanocellulose in the Food Industry DOI: http://dx.doi.org/10.5772/intechopen.114085*

**Figure 2.** *Schematic of NC effect on glucose digestion [37, 38].*

Another application of NC in the food industry is its use as an agent that disrupts the digestion and absorption of lipids in the body. The presence of lipids and fats in daily meals, including milk, ice cream, hamburgers, bread, and dairy products, plays a significant role in weight gain and blood pressure increase. Therefore, the production of targeted and intelligent food holds a key role in the future of the food industry. In general, NCs, with their absorption power and high water content, increase food's viscosity, which in turn reduces the food's caloric content and causes disorders in fat digestion. So, NCs effectively hinder the digestion and full absorption of lipids in the body through various fundamental mechanisms. These include (1) thickening or gelling gastrointestinal fluids to prevent digestive enzymes from reaching the fat; (2) combining with gastrointestinal constituents such as bile salt to form a precipitate; (3) reducing the surface area exposed to digestive enzymes by promoting flocculation of fat; and (4) forming a thin coating around the fat to reduce its exposure to digestive enzymes (**Figure 3**) [41]. To investigate the effect of NC on lipids, scientists simulated the conditions and environment of the digestive system. They found that NC reduces the activity of the lipase enzyme, increases the viscosity of intestinal digesta, and is more effective than cellulose in blocking the effect of bile salts and absorbing cholesterol in the body [42].

**Figure 3.** *Schematic of NC effect on fat digestion [41].*

#### **4.4 Retrogradation inhibitor in starchy food**

Another nutritional application of NC is its use in the starchy food industry. Scientists have found that nanocellulose (especially CNC) can be used to produce healthy starchy foods and support weight loss efforts. A major issue in the starchy food industry is "retrogradation," which typically occurs during the cooling and storage stage of gelatinized starch, reducing the quality of the product [43]. With a large specific surface area and abundant hydroxyl groups, CNC effectively interacts with amylose through hydrogen bonding, inhibiting short-term retrogradation. Furthermore, the interaction between CNC and amylopectin inhibits long-term retrogradation (**Figure 4**) [44]. There have been extensive studies on the effect of NC on starch. For example, a 2017 study investigating the impact of NC on oxidized potato starch found that NC influenced enzyme activity and the amount of resistant starch. Notably, the addition of NC to gelatinized starch reduced the amount of quickly digestible and slowly digestible starch but increased the amount of resistant starch. Resistant starch is a type of starch that resists enzymatic digestion and cannot be digested in the small intestine. However, it can be fermented with volatile fatty acids in the human intestinal tract, specifically in the colon [45].

#### **4.5 Food additive**

Today, nanocelluloses, especially BNCs, due to small diameter, hydrophobicity, and water-holding capacity, are added to food and are used to reduce calories, increase and improve their absorption in the body, and improve nutritional properties. Among the applications of nanocellulose as a food additive, it can be mentioned to replace BNCs instead of flavonoids, cara gum, and other stabilizers to the ice cream formula to obtain a new type of ice cream. For example, in 2018, Guo et al. [46] showed that adding bacteria to ice cream increased its resistance to melting and could maintain its shape for at least an hour after being taken out of the freezer. Since BNCs are easily digested and absorbed by the digestive system, they are used as a quality improver based on meat. For example, in 2019, Guo et al. [47] demonstrated that BNC can effectively reduce moisture loss in chicken cake without altering the chemical

**Figure 4.** *Schematic of NC effect on starch digestion [37, 38].*

structure or protein groups in the chicken. Another application of nanocellulose is the enrichment of beverages and dairy products in the market. For example, nanocelluloses can produce quality yogurt with a soft texture and higher stability [48].

#### **4.6 Encapsulating agent**

The food industry faces a significant challenge in preserving micronutrients, vitamins, minerals, and active components (such as probiotics) during production, preparation, transportation, and storage. Moreover, digestive enzymes often compromise these nutrients in the stomach and bile salts before reaching their intended destination in the body, primarily the intestine [49]. Therefore, food industry scientists are striving to protect nutrients from adverse factors and conditions and improve the performance of the delivery system using encapsulation techniques. Encapsulation is an active substance transfer system that uses natural polymer materials to safeguard nutrients, thereby enhancing the bioavailability and biological activity of substances. The polymer materials used in encapsulation should ideally be natural, readily available, inexpensive, non-toxic, free of pollution, easily modifiable, and have good biodegradability and biocompatibility [34]. Among the materials used for encapsulation, NC has drawn the attention of numerous researchers due to its distinctive biological properties, such as high safety, excellent biodegradability, and non-toxicity. A noteworthy study in this area has explored the use of alginate and NC in the microencapsulation of probiotics. The results of the study indicated that the inclusion of NC increases the compression strength of the produced hydrogels by more than 30% and diminishes the porosity induced by freeze-drying. Additionally, the introduction of 13% NC significantly extended the survival time of the probiotic strain (*Lactobacillus rhamnosus*) within the alginate microbeads [50]. In 2020, researchers encapsulated the probiotic *Lactobacillus rhamnosus* using a mixture of whey protein isolate, inulin, and NC (CNC). They found that NC significantly improved the resistance and survivability of the microcapsules under simulated digestive system conditions [51]. Also, Wang et al. [52] developed a soy protein isolate combined with CNC composite nanoparticles for the delivery of curcumin. The resulting complex composite nanoparticles demonstrated improved encapsulation efficiency and controlled release in simulated gastrointestinal conditions. CNC was found to be essential in the delivery system for the active compounds. In 2021, a compelling study was conducted on the encapsulation of ascorbic acid (Vitamin C) using NC. In this experiment, Vitamin C was encapsulated through an electrostatic interface with glycidyltrimethylammonium chloride-modified chitosan (GCS) and then cross-linking with phosphorylated CNC (PCNC) to produce Vitamin C-GCS-PCNC nanocapsules. The results of the investigation revealed that the inclusion of NC in the encapsulation formulation enhanced the efficiency, provided better control of the release action, and increased the antioxidant and antibacterial capabilities of vitamin C [53].

#### **4.7 Food packaging materials**

Packaging plays an essential role in protecting food from spoilage factors, including light, oxygen, heat, and bacteria. This makes it a vital component within the comprehensive system of the food industry. The use of natural and renewable materials in efficient packaging not only preserves food and extends its shelf life but also maintains the quality and nutritional value of the food until its expiration date, thereby helping to minimize waste in the sector. The prevalent use of plastic and

its derivatives in the packaging industry, combined with increasing concern about environmental pollution and public health, has promoted scientists to explore natural and biodegradable materials for the production of suitable packaging. Among these materials, cellulose and its derivatives, especially NCs, have emerged as significant and essential candidates for the creation of biodegradable and intelligent packaging. One of the prominent attributes of NC that inspires scientists to employ it in the packaging industry includes its natural origin, high biodegradability, strength, resistance to oxygen and moisture penetration, and antibacterial properties. Generally, the exceptional oxygen barrier properties of NCs result from a combination of their high crystallinity and a network structure sustained by strong inter- and intramolecular hydrogen bonds [54]. Extensive research is being conducted on the application of NC in the packaging industry, particularly in intelligent packaging. For instance, in 2021, researchers manufactured nanocomposite films to regulate D-limonene permeability by utilizing nanocrystals and starch. Their study results indicated that the amount of NC, along with its appropriate aspect ratio, could autonomously manage D-limonene permeability [55]. In another study, Perumal, et al. [56] discovered that incorporating NC into the formulation of a biocomposite film (comprising clay and polyvinyl alcohol (PVA)) not only prolonged the storage life of mangoes but also enhanced the thermal and tensile properties of the films by increasing the amount of CNC filler up to 6 wt %. Currently, researchers are exploring the use of NC in intelligent packaging systems, leveraging its antibacterial and antioxidant properties. In 2020, researchers produced a probiotic nanocomposite film utilizing *Lactobacillus plantarum*, NC, and inulin. Their findings indicated that the fabricated probiotic packaging films exhibited potent antibacterial properties and effectively extended the shelf life of chicken fillets [57]. Another study produced composite films using polyhydroxybutyrate (PHB) and nanocellulose (CNC) via the solvent casting method. This study showed that, besides enhancing the barrier performance, mechanical strength, transparency, and other attributes of the composite film, CNC did not impact the inherent PHB [58].

Another beneficial effect of incorporating nanocellulose into packaging systems is enhancing the mechanical properties, such as high tensile strength, elongation, and flexural modulus [59]. For instance, a 2020 study observed a sevenfold increase in tensile strength and a sixfold increase in elongation at break when CNC was added to a PVA film [60]. Moreover, in research conducted by Chen et al. [61], it was found that integrating 0.5% stearic acid-modified microcrystalline cellulose/nanocellulose into cassava starch film significantly increased tensile strength by 484.5%.

#### **5. Safety and regulatory aspects**

Wood pulp and powdered cellulose are generally recognized as safe (GRAS) and are commonly used as raw materials in food-contact materials or food additives. As NC is one of the most widely used cellulose derivatives in different industries, evaluating its biological effects on the ecosystem and the human body solely based on its chemical characteristics is not sufficient. The interactions of NC with cells and other living organisms can be influenced by various factors, including its size, shape, aggregation characteristics, and other unknown factors [62]. One of the primary routes for NC to enter the body is through the skin and respiratory system. If the amount of NC in the skin and respiratory system exceeds permissible limits, it can cause skin toxicity and pulmonary inflammation [34, 63]. The use of commercial

#### *Fascinating Properties and Applications of Nanocellulose in the Food Industry DOI: http://dx.doi.org/10.5772/intechopen.114085*

foodborne nanoparticles, such as titanium dioxide (TiO2) and zinc oxide (ZnO), in the food industry can lead to complications like colitis [64], and this raises significant concerns among consumers and researchers about the safety of NC following ingestion through the gastrointestinal tract. Therefore, extensive studies have been conducted to investigate the effect of NC and related problems on food and the human body, using both *in vivo* and *in vitro* methods.

In 2010, the toxicity effect, including the *in vitro* cytotoxic and genotoxic properties of NC, obtained from birch pulp, with dimensions ranging between 20 and 60 nm to 100 and 350 nm, on human and animal cells was investigated. Additionally, the study examined the lethal effect of NC on human cells. Also, the effect of NC damage on DNA using a bacteriological assay (the Ames test) was also studied [65]. In 2011, another *in vitro* study focused on the toxicity, health, and environmental safety of NC with dimensions of 20–30 nm. The study evaluated its effect on human and mouse macrophages, its impact on cell survival, as well as the cytokinetic characteristics of cells [66]. Also, in recent years, extensive studies have been conducted on the oral toxicity of NCs using both *in vitro* cellular models and/or *in vivo* animal models. For instance, in 2019, the toxicological effects of ingested NC were investigated using both an *in vitro* intestinal epithelium model and an *in vivo* rat model [67]. In 2020, the impact of CNF on gut microbial health was studied by examining its cytotoxicity and *in vivo* toxicity [68], and in 2020, the hepatotoxicity of NC modified with oxalate ester in rats was investigated [69]. Furthermore, in 2021, the oral toxicity of carboxymethylated CNF was evaluated using *in vitro* cellular studies and an *in vivo* mice repeated gavage model [70]. All these studies consistently concluded that both original and functionalized NC showed no cytotoxicity. Additionally, it was observed that NC was non-toxic in an *in vivo* animal model when used at a reasonable concentration, with an upper limit of 3.5 wt%. Therefore, to gather clearer information regarding the safety effects of nanomaterials like NC, it is crucial to establish internationally standardized methods that guarantee their safety. Moreover, prior to the commercialization of food products utilizing new nanotechnologies, it is essential to proactively anticipate, understand, and effectively manage both the potential positive and negative consequences that may arise from consuming nanomaterials [62].

#### **6. Conclusion and prospects**

Cellulose, the most abundant polymer in nature, is the primary resource for the production of NC. The unique properties of NC, including its biocompatibility, biodegradability, non-toxicity, inexhaustible nature, nanoscale size, high aspect ratios, large surface area, liquid crystalline, and rheological properties, have made it an appealing choice for use in different industries such as food industry. Generally, NC has a greater affinity for oil than water. It can be used as a barrier to mix water and oil emulsions, making it a valuable stabilizing agent for emulsions and food products. This characteristic explains the use of NC in the food industry, particularly in foods such as sauces, soups, puddings, etc. Moreover, due to its ability to create stable emulsions with low-oil content, NC can contribute to the production of low-oil, low-calorie food products. NC can also serve as a suitable coating and carrier for bioactive compounds, such as probiotics, protecting them from the effects of digestive enzymes. Consequently, NC can play a crucial role in food enrichment. When utilized in packaging systems, NC enhances the mechanical and barrier properties of packaging films. As a result, food products are less exposed to spoilage factors,

and their shelf life is increased. From the economic point of view, the use of nanocellulose in the industry due to its cheap source, low weight, and high strength, as well as enhanced product performance, can have a bright future, provided that it can overcome the upcoming obstacles, such as the expensive production method and the instability of the market. Also, there are still significant challenges faced during the application of NC in the food industry. These challenges include improving technology and reducing production costs, developing new methods for modifying the hydrophobicity of NC, further investigating its nutritional properties and impact on consumer health, and conducting more comprehensive research on the safety aspects of NC. As research and development continue to advance the production and utilization of nanocellulose, its economic viability is expected to further improve in the coming years.

#### **Author details**

Elham Asghari-Varzaneh and Hajar Shekarchizadeh\* Department of Food Science and Technology, College of Agriculture, Isfahan University of Technology, Isfahan, Iran

\*Address all correspondence to: shekarchizadeh@iut.ac.ir

© 2023 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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#### **Chapter 5**

## Bacterial Nanocellulose: Methods, Properties, and Biomedical Applications

*Haiyong Ao and Xiaowei Xun*

#### **Abstract**

Unlike plant and wood-origin cellulose, bacterial nanocellulose (BNC) produced by bacteria exhibits the highest purity and natural nanofiber morphology, attracting increasing interest from many researchers and industrial sectors. It has numerous unique features including the biomimetic nanoscale three-dimensional (3D) network, high water holding capacity, and moldability in different shapes, accepted wet strength, outstanding gas permeability, and good biocompatibility, which makes the BNC show great potential in a wide variety of biomedical applications. Extensive research has verified the feasibility of application in wound dressing, bone/cartilage tissue regeneration, vascular tissue engineering, and so on. This chapter focuses on the production and properties of BNC, the fabrication of BNC-based biomaterials, and the biomedical applications of BNC.

**Keywords:** bacterial nanocellulose, fabrication techniques, properties, biomaterials, biomedical applications

#### **1. Introduction**

Bacterial nanocellulose (BNC) as a type of extracellular polysaccharide polymer synthesized by bacteria has unique physical and chemical properties, including high purity (free from lignin and hemicellulose) and crystallinity (84–89%), high degree of polymerization, good mechanical properties, high water retention, and threedimensional (3D) nanofibrous structure [1, 2]. All these features make BNC attractive for material scientists and engineers.

Importantly, BNC is environment-friendly due to its nontoxic, nonimmunogenic, biocompatible, biodegradable, and renewable nature [3]. The nanofibrillar network of BNC is similar to the structure of collagen in the native extracellular matrix (ECM), and the 3D uniform and interconnected pores facilitate cell infiltration and nutrient and waste exchange [4, 5]. As a promising natural biomaterial, BC has been extensively utilized in various biomedical fields for wound healing, bone, cartilage, and blood vessel engineering [6, 7].

During the past two decades, many studies have been dedicated to developing various BC-based biomaterials. The 3D porous structure and easily modified surface (abundant OH groups) of BNC are beneficial for introducing into BC varieties of reinforcement substances including biomolecules, nanoparticles, and polymers to acquire new materials with highly desirable properties, which can lead to the formation of BC-based functional materials for biomedical applications [3]. Moreover, BC-based scaffolds have been specifically designed to mimic the 3D structures of native tissues to support and provide the microenvironments required for cell adhesion, proliferation, migration, and differentiation [8]. These extensive efforts have accelerated the development of BNC-based biomaterial.

In this chapter, we briefly introduced the biosynthesis and properties of BNC. Furthermore, the strategies for fabricating the BNC biomaterial and applications in various biomedical fields were summarized.

#### **2. Biosynthesis and properties of BNC**

#### **2.1 Biosynthesis and production of BNC**

The bacterial synthesis of cellulose was first reported by Brown as early as 1886 [9]. Brown found that a white gel-like thin layer was formed on the surface of the culture medium after incubating *Bacterium xylinum'* in a static state. Chemical analysis determined that the main component of the white gel is cellulose and does not contain hemicellulose and lignin, and further research found that the diameter of this cellulose is at the nanoscale.

*Gluconacetobacter genus*, as one of the most efficient BNC producers, is widely used in the biosynthesis of BNC [10, 11]. The general process to produce BNC is schematically described in **Figure 1A**. The biosynthesis process of BNC is mainly divided into four steps (**Figure 1B**): (1) glucose is converted into glucose-6-phosphate, (2) glucose-6-phosphate is converted into glucose-1-phosphate by phosphoglucomutase, (3) uridine diphosphate glucose (UDP-glucose) pyrophosphorylase converts glucose-1-phosphate to UDP-glucose, (4) the UDP-glucose is converted to cellulose by the action of cellulose synthase (**Figure 1C**) [13, 14]. The self-assembly and crystallization of BNC nanofibrils are regulated by the cell itself [15]. The synthesized chains are sequentially gathered in the culture medium at a rate of up to 200,000 glucose molecules per second and form protofibrils or sub-elementary fibrils, which are 2–4 nm in diameter. Each sub-elementary protofibril is composed of 12–16 glucan chains, where the glucan chains are aligned and stacked into ordered nanostructures called the microfibrils. The micro- and macrofibrils and loose bundles finally form 30–100 nm ribbon-like structures. The ribbons are highly porous and have 3D fibrous network.

For many years, many researches have been focused on BNC production. Various culture methods have been used by controlling the fermentation conditions to increase the yield of BNC or obtain BNC with different characteristics [16]. Under static culture conditions, bacteria need to float on the surface of the medium to obtain enough oxygen, and cellulose microfibrils are extruded from the bacteria and synthesize a tight BNC pellicle. The BNC formed under static conditions usually has high crystallinity and tensile strength. In contrast, a fluffy, spherical, or irregular lump BNC was obtained *via* the agitating culture method. Although the agitating culture has faster cell growth rate than static conditions because of more oxygen filling in the

*Bacterial Nanocellulose: Methods, Properties, and Biomedical Applications DOI: http://dx.doi.org/10.5772/intechopen.114223*

#### **Figure 1.**

*(A) A schematic illustration of BNC production. (B) Schematic illustration of biochemistry of cellulose synthesis in a bacterial cell and the extracellular transport of cellulose chains and the formation of highly ordered structures. (C) Chemical structure of BNC. (D) Schematic illustration of the aerosol-assisted biosynthesis of BNC [12].*

culture medium, the production of BNC decreased, which perhaps due to the uniform aeration of cultures induced cells to grow intensively instead of the polymer synthesis. Based on this, the bioreactors have been developed that can produce BNC pellicles at higher yields under static conditions (**Figure 1D**) [12].

#### **2.2 Properties of BNC**

Compared to vegetal cellulose, its unique chemical composition and physical structure endows BNC with numerous unparalleled physical, chemical, and biological properties (**Figure 2**). BCN is an attractive candidate for widespread applications in various fields, especially in applications related to biomedical.

**Figure 2.** *A schematic representation of properties of BNC.*

#### *2.2.1 Physical properties*

BNC has natural 3D porous network structure, which is composed of nanofibers and exhibits enormous mechanical properties. The Young's modulus of BNC sheets up to 15 GPa, which is much greater than plant cellulose and several synthetic fibers [17]. The excellent mechanical properties make BNC to be used for blood vessels and bone tissue engineering. The water holding capacity of BNC is about 100 times its dry weight, which is attributed to the high surface area and pore volume of BNC that can intercept more water [18]. In addition, the average diameter of BNC nanofibers is 1.5 nm, showing higher surface area and flexibility. These fascinating physical properties make BNC become desirable wound dressing material.

#### *2.2.2 Chemical properties*

BNC is composed of linear homopolysaccharides conjugated by β-D-glucose units linked by 1,4-β-glycosidic linkages and has considerably higher crystallinity (80–90%) and degree of polymerization (up to 8000), which means that pure BNC can be obtained with simple processing. The abundant hydroxyl groups and highlevel hydrogen bonds on the surface of BNC nanofibers allow for manipulation in

their loading of functional molecules. Although BNC exhibits significant intrinsic characteristics, it is still necessary to develop more modified properties of BNC to meet the requirements of the required biomedical applications.

#### *2.2.3 Biological properties*

BNC has wide applications in biomedical engineering because of its excellent biological properties. Héctor and coworkers confirmed the nontoxicity and extremely low bacterial endotoxin of BNC nanofibers by using *in vitro* and *in vivo* tests [19]. In addition, the good biocompatibility of BNC is due to its peculiar 3D nanofibrous network structure that supports cell penetration and proliferation. Helenius et al. subcutaneously implanted the BNC in rats to systematically evaluate the biocompatibility of BNC [20]. The results showed no inflammation around the implants and the fibroblasts infiltrated BNC, which indicates the good biocompatibility of BNC and has the potential to be used as a scaffold in tissue engineering.

#### **3. Fabrication of BNC biomaterials**

Although the above excellent inherent characteristics make BNC have broad application prospects in the field of biomedical, pure BC possesses certain restrictions that limit its application due to the lack of unique biological functions. In recent years, several approaches have been developed to improve the physicochemical properties and function of BNC, including producing BNC composites, surface medication, and changes in porosity. These efforts have significantly improved the surface properties and biological functions of BNC and tremendously expanded its application in various tissue engineering fields.

#### **3.1 Surface modifications**

#### *3.1.1 Physical modifications*

The BNC nanofibers were physical modifications by using bioactive materials, which not only preserves the unique properties but also enhances the biological functions to meet the requirements of biomedical applications. To improve the biocompatibility of BNC, Luo et al. selected a new type of coating, chondroitin sulfate (CS) modified with gelatin (Gel), to modify BNC scaffold [21]. The coating on the surface of the BNC nanofibers is linked by hydrogen bonding. The results of *in vitro* cell studies indicated that the CS/Gel coatings significantly promote cell proliferation, adhesion, differentiation, and ingrowth into scaffolds. Nanomaterials can also be used for surface modification of BNC. He et al. developed a Cu2+ loaded phase-transited lysozyme (PTL) nanocoating for surface modification of BNC. The coating endowed the BNC with inhibiting bacterial growth and inflammation and simultaneously induced vascularization, collagen deposition, and reepithelialization of wounds promising dressings for healing infected wound, which was considered a promising dressing for treating infected wounds. In addition, polymer composite nanoparticles are also used for surface modification of fibers. Ma et al. developed a homogeneous AgNP-loaded polydopamine (PDA)/polyethyleneimine (PEI) coating on the surface of BNC nanofibers, which exhibited excellent antibacterial activities and cytocompatibility [22].

#### *3.1.2 Chemical modifications*

The only surface functional group on the surface of BNC nanofibers is the hydroxyl group, which is not suitable for cell proliferation [23]. Nevertheless, the hydroxyl groups on the surface of BNC nanofibers are beneficial for surface chemical modifications; they can be treated with various chemical reagents tomodify their chemical structure and incorporate additional functionalities. Therefore, the surface chemical modifications of BNC are required for biomedical applications. Oxidation is an important reaction for adding new functional groups to BNC, which creates infinite possibilities for the functionalization of BNC nanofibers. TEMPO (2,2,6,6-Tetramethylpiperidinyloxy or 2,2,6,6-Tetramethylpiperidine-1 oxyl radical)-mediated oxidization is a simple and effective chemical reaction, in which C6 of a hydroglucose units selectively gain an anionic carboxyl group [24]. A new bone repair composite scaffold (CS/OBC/nHAP) was constructed by evenly dispersing the *in situ* crystalline nano-hydroxyapatite (nHAP) in oxidized bacterial cellulose (OBC) and chitosan (CS) scaffolds [7]. Shahriari-Khalaji et al. achieved high carboxylate content by optimizing the TEMPO-mediated oxidation of BNC and then covalently bonded the ε-poly-L-lysine (PLL) with oxidized BNC to develop an O-BNC-based functional wound dressing [25]. Furthermore, Xie et al. prepared biofunctional group-modified bacterial cellulose (DCBC) by carboxymethylation and selective oxidation to achieve the perfect compound of cellulose and chitosan [26]. The obtained BNC-based dressing can effectively inhibit bacterial proliferation in wounds and kill the bacteria.

#### **3.2 BNC-based composites**

#### *3.2.1 BNC nanocomposites*

To improve the mechanical and biological properties of BNC biomaterials, various BNC-based nanocomposites were prepared by incorporating different kinds of nanomaterials, including carbon-based nanoparticles (NPs) [27], metal/metal oxide NPs [28], and other inorganic NPs [29]. The simplest and most convenient method for fabricated BNC nanocomposites is the mechanical mixing method. Yang et al. prepared the BNC/Ti3C2Tx nanocomposites using mechanical mixing. Although this strategy allows for sufficient mixing of nanomaterials with BNC nanofibers, it damages the intrinsically continuous 3D structure of BNC. To maintain the intrinsically continuous 3D structure of BC, Luo et al. developed a cost-effective, scalable, and efficient approach, membrane-liquid interface (MLI) culture, to prepare BNCbased nanocomposite [30]. The MLI culture technology is schematically illustrated in **Figure 3A** [31]. First, a layer of BNC film (around 3 mm in thickness) was grown as a substrate using a conventional static culture method. Then, a nanomaterialsdispersed medium was sprayed onto the BNC substrate followed by the culturing of BNC on the substrate–medium interface. After the sprayed medium was completely consumed off by the BNC growth, another layer of the medium was sprayed. This process was repeated until the designed thickness was achieved. It is noted that the thickness of nanocomposites prepared using the MLI method can be accurately controlled by the spray cycles of the medium. In addition, the sample shape and dimension can be facially designed by the geometry of the containers used for the preparation. This method has been widely used to prepare various nanocomposites (**Figure 3B**) [12].

*Bacterial Nanocellulose: Methods, Properties, and Biomedical Applications DOI: http://dx.doi.org/10.5772/intechopen.114223*

#### **Figure 3.**

*(A) Schematic illustration of the preparation of BNC nanocomposites by using membrane-liquid interface culture technology [31]. (B) SEM and photographs of BC-based nanocomposites were prepared by using membraneliquid interface culture technology [12].*

#### *3.2.2 BNC-based biocomposites*

Compared with other natural biopolymers, pristine BNC lacks cytocompatibility and important biological functions. The abundant ∙OH groups on the surface of BNC provide binding stable sites for biopolymers in biocomposites. Therefore, various techniques including *in situ* addition, solution impregnation, and chemical cross-linking were used to combine various bioactive substances to obtain the BNC-based biocomposites with enhanced physicomechanical, antimicrobial, and biocompatible properties. In order to enhance the antibacterial properties and cytocompatibility of BNC, Zhou et al. composited the collagen I (Col-I) and the antibacterial agent hydroxypropyltrimethyl ammonium chloride chitosan (HACC) into the BNC 3D network structure by a novel membrane-liquid interface (MLI) culture (**Figure 4A** and **B**) [32]. The introduction of HACC and Col-I makes BNC have outstanding antibacterial properties and improved cytocompatibility to promote NIH3T3 cell and HUVEC proliferation and

#### **Figure 4.**

*(A) Schematic illustration of the preparation of BNC-based biocomposites. (B) SEM images of BNC-based biocomposites. (C) The characteristice of BNC-based biocomposites [32].*

spread (**Figure 4C**). The BNC-(polypyrrole) Ppy composites were prepared to mimic the natural myocardial microenvironment by *in situ* polymerization [33]. The composites were flexible and still maintained 3D network structure and displayed electrical conductivities in the range of native cardiac tissue. Wan et al. reported extracellular matrix (ECM)-mimetic scaffolds by conjugating electrospun cellulose acetate (CA) submicrofibers with BNC nanofibers *via* a facile and scalable dispersion freeze-drying process [34]. It is found that the composites have a 3D porous network structure and improved cell behavior.

#### **3.3 3D porous BNC scaffold**

As a promising biomaterial, BNC has some intrinsic disadvantages when applied in tissue engineering; its dense nanofibrous network (the pore sizes of pristine BNC are only approximately 0.02–10 μm) markedly limits cell migration and 3D tissue regeneration. To date, different methods have been reported for the fabrication of 3D porous BNC scaffolds. The paraffin microparticles, potato starch, agarose microparticles, and gelatin microspheres as porogens have also been used to fabricate 3D porous BNC scaffolds by using the *in situ* porogen technique [35–38]. Cui et al. described the utilization of gelatin microspheres in BNC production for porosity enhancement, and the observed porosity depended on the diameter of microspheres (**Figure 5A**) [36]. However, a nonuniform pore structure would be formed due to the bacteria moving to the air/medium interface during the culture process of the *in situ* porogen impregnation technique.

Laser patterning as an efficient method is used to prepare 3D porous BNC scaffold. Laser treatment has universality, and the obtained scaffolds do not contain pollutants and chemical cross-linking agents, thereby having better biocompatibility. The pore structure prepared by laser is parallel microchannels. Yang et al. fabricated the nanosubmicrofibrous cellulose scaffolds with microchannels by laser-aided punching (**Figure 5B**) [39]. The cell study found that the presence of microchannels favors cell proliferation and migration at an optimum microchannel size. However, laser-aided punching prepared microchannels can lead to cell leakage from the scaffold, making it difficult to achieve 3D culture.

Freeze-drying is the most common technique for preparing a 3D porous BNC scaffold, which can obtain the BNC sponges with higher porosity and specific surface. However, the pore size of freeze-dried BNC sheets is unable to precisely regulate, and the pore structure is instability. Therefore, improvements need to be made to address these issues. Xun et al. first reported an improved strategy to fabricating 3D macroporous BNC scaffolds with controllable pore size by freeze-drying and cross-linking the mechanically disintegrating shortcut BNC nanofiber suspensions (**Figure 5C**) [40]. The pore sizes of the MP-BNC scaffolds were controlled by adjusting the concentration of BNC in the suspensions. The fabrication process was facile, scalable, and effective in controlling the pore structure. The cross-linked BNC scaffolds exhibited excellent compression properties and shape recovery ability compared to the original BNC. Moreover, the results of *in vitro* and *in vivo* studies demonstrated that the scaffolds had excellent biocompatibility and were effective in regenerating cartilage tissue.

Additive manufacturing or 3D printing is an emerging technology to prepare 3D porous scaffolds through rapid prototyping. In recent years, it has been widely used in personalized customization of tissue engineering scaffolds. Li et al. used the gelatin methacrylate (GelMA)/BNC bioink formulations to develop heterogeneous tissue-engineered skin (HTS) containing layers of fibroblast networks with larger

*Bacterial Nanocellulose: Methods, Properties, and Biomedical Applications DOI: http://dx.doi.org/10.5772/intechopen.114223*

#### **Figure 5.**

*Different 3D porous BNC scaffolds were fabricated by using* in situ *porogen [34], laser patterning [39], freeze-drying and cross-linking [40], and 3D bioprinting [41].*

pores, basal layers with smaller pores, and multilayered keratinocytes (**Figure 5D**) [41]. The results revealed that the 10%GelMA/0.3%BNC bioink was better to bioprint dermis due to its high printability and cell-friendly sparse microenvironment. The approaches developed in the above-described studies and their findings suggest that BNC has great potential to be printed into 3D microstructures for the development of scaffolds and medical devices for various biomedical applications.

#### **4. Biomedical applications of BNC**

In the past two decades, BNC has aroused great attention to fundamental and scientific research for biomedical applications due to its unique properties [42]. Functionalized BNC and its composites have been applied in several medical applications, such as wound dressing, bone and cartilage tissue regeneration, and the

development of artificial organs and blood vessels' substitutes (**Figure 6**) [23, 43]. This section focuses on the research progress of the main biomedical applications of BNC in recent years.

#### **4.1 Wound healing**

The skin is the main protective barrier of the human body and has many functions, such as controlling body temperature and maintaining a balance of electrolytes and water [44]. However, large-scale skin damage caused by trauma, ulcers, and burns is difficult to heal on its own and requires human intervention to promote wound healing [45, 46]. Wound dressings have been developed to prevent infection and dehydration of wound, reduce inflammatory responses, and promote wound healing. BNC is one of the most promising wound dressings due to its ultrafine 3D network structure, excellent gas permeability, high water absorbency, and favorable biocompatibility [47]. Furthermore, BNC-based dressings can also prevent bacterial invasion, absorb excess exudates, and retain moisture in wounds to promote the growth of granulation.

In the healing process, it is important to prevent bacterial infection on the wound site. However, BNC does not have intrinsic bactericidal properties to eliminate the colonized bacteria. Therefore, the initial research focus was on antibacterial BNCbased dressings. Hence, various promising antibacterial agents are introduced into BNC, such as metals and their oxides, antimicrobial peptides, and biological and synthetic polymers (chitosan) [48]. Ao et al. fabricated the quaternized chitosan (HACC)/BNC antibacterial wound dressing by using *in situ* synthesis possessed, and

#### *Bacterial Nanocellulose: Methods, Properties, and Biomedical Applications DOI: http://dx.doi.org/10.5772/intechopen.114223*

the obtained dressing had favorable antibacterial properties [49]. To further improve the antibacterial properties of BNC dressings, silver nanoparticles (AgNPs) were introduced into the BNC structure [22]. The results of the standard plate count assay indicated that the antibacterial BNC dressing showed antibacterial rates of over 99.9% against *Escherichia coli* and *Staphylococcus aureus*. The *in vivo* assay demonstrated that the antibacterial BNC dressing could inhibit infection and inflammation and accelerate wound healing within 12 days compared with BNC. To reduce the cytotoxicity of sliver-based dressing and promote wound healing, a novel multifunctional sliver nanowires (AgNWs)/collagen I (Col I)/BNC dressing was constructed *via* compositing AgNWs into BNC and adsorbing Col I [50]. The antibacterial assay demonstrated that the multifunctional AgNWs/Col I/BNC dressing could kill *S. ausreus* and *E. coli* colonizing on the surface of material due to the silver ions released. Importantly, this dressing had lower cytotoxicity and promoted collagen deposition, hair follicle growth, neovascularization, and wound healing. A novel copper ion (Cu2+)-loaded BNC-based antibacterial wound dressing was prepared *via* codeposition of polydopamine (PDA) and Cu2+ [51]. The *in vivo* study revealed that the dressing can eliminate *S. aureus* infections and inflammatory response and promote collagen deposition, capillary angiogenesis, and wound healing.

In addition, BNC composites containing nanomaterials, biopolymers, antioxidants, antibiotics, and blood clotting agents have been fabricated to improve the wound healing of BNC-based dressings. Cai et al. fabricated a composite adhesive organo hydrogel by introducing BNC and platelet-rich plasma (PRP) into a poly-N- (tris[hydroxymethyl]methyl)acrylamide (THMA)/N-acryloyl aspartic acid (AASP) hybrid gel network infiltrated with glycerol/water binary solvent [52]. The PRPloaded organo hydrogel has good tissue adhesion properties and releases a variety of growth factors to accelerate the wound healing process through collagen deposition and angiogenesis. Shen et al. developed an aggregation-induced emission (AIE) molecule BITT-composited BNC for wound healing [53]. The BNC-BITT composites retained the advantages of biocompatible of BNC and displayed photodynamic and photothermal synergistic antibacterial effects under irradiation of a 660 nm laser, which endowed the dressings with excellent wound healing performance in a mouse full-thickness skin wound model infected by multidrug-resistant bacteria. In response to hemostasis and repair of irregular and deep skin wounds, an injectable aldehyde BNC/polydopamine (DBNC/PDA) photothermal cryogel was prepared by oxidation polymerization method [6]. The PDA enhances the photothermal properties of DBNC/PDA cryogel to kill most bacteria and provides wound protection under near-infrared light. Otherwise, the DBNC/PDA low temperature gel has rapid hemostatic effect in the face of irregular and deep skin wounds. It is worth noting that the nanoenzyme with excellent peroxidase (POD) activity has been used to prepare BNC-based wound dressing. Zhang et al. introduced the metal-organic frameworks (MOF)-based nanocatalysts loaded with glucose oxidase (GOx) into the BNC-reinforced hydrogel for the treatment of diabetic foot ulcers [54]. The designed nanoenzyme could effectively catalyze the decomposition of glucose and *in situ* generate •OH for bacteria killing. In addition, this nanoenzyme-based hydrogel exhibited excellent hemostatic properties owing to the enhanced absorption capacity.

#### **4.2 Bone/cartilage tissue regeneration**

Another area of potential exciting application for BNC biomaterial is bone/cartilage tissue engineering due to its biomimetic ECM properties, excellent mechanical

properties, and highly porous structure [7, 55, 56]. Biosynthetic BNC has some intrinsic disadvantages (the dense nanofibrous network) that markedly limit its applications in tissue engineering. However, the *in situ* biosynthetic BNC could serve as a surface coating for Ti implants to improve their biological function [57, 58]. Liu et al. reported a metal ions-containing BNC coating for functional Ti implant by *in situ* biosynthesis on the surface of Ti with complex shapes [59]. The results of *in vitro* and *in vivo* experiments confirmed that the functional BNC coating on the Ti can integrate the operative crevices and promote osteogenesis. To optimize the pore structure of BNC to meet the requirements of bone and cartilage tissue, Xun et al. developed a 3D macroporous BNC scaffold by freeze-drying and cross-linking the BNC shortcut nanofibers [40]. After the 3D macroporous BNC scaffolds were implanted into nude mice subcutaneously for 8 weeks, the neocartilage tissue with native cartilage appearance and abundant cartilage-specific extracellular matrix deposition was successfully regenerated. To construct the BNC scaffolds with structurally and biochemically biomimetic cartilage tissue microenvironment, the 3D hierarchical porous BNC/ decellularized cartilage extracellular matrix (DCECM) scaffold was fabricated by freeze-drying technique after EDC/NHS chemical crosslinking [60]. The *in vitro* and *in vivo* tests indicated that the BC/DCECM scaffolds achieved satisfactory neocartilage tissue regeneration with superior original shape fidelity, exterior natural cartilage-like appearance, and histologically cartilage-specific lacuna formation and ECM deposition. Ling et al. fabricated a hierarchically porous SF/BC/MXene (FSCM) scaffold with ~20.0 μm macropore and nanofibrillar wall, which has excellent bone defect repair ability [61]. In addition, the BNC can also be used for the osteochondral repair. Lou et al. designed a bilayer structure osteochondral scaffold with a dense γ-Polyglutamic acid/carboxymethyl chitosan/BNC (PGA/CMCS/BNC) hydrogel cartilage layer and a porous nano HA-containing PGA/CMCS/BNC hydrogel osteogenic layer [62]. The *in vivo* experiments indicated that the scaffold with bioactive ions had a much better effect on the repair of osteochondral defects.

#### **4.3 Blood vessels**

BNC is a possible material to use for artificial blood vessels for small- or largesized vascular grafts due to its good mechanical strength (a burst pressure of up to 880 mmHg), blood biocompatibility, and moldability [63]. The bioreactor to produce tubular BNC was developed to prepared BNC-based vascular grafts with excellent cytocompatibility and hemocompatibility [64, 65]. A tubular BNC graft with greater mechanical strength and thinner walls was obtained by mercerization; this technology made fewer platelets adhere to the luminal surface and promoted the proliferation of endothelial cells [63]. Mimicking the morphological structure of native blood vessels is critical for the development of vascular grafts. Vascular grafts with BNC nanofibers and submicrofibrous cellulose acetate (CA) were fabricated to mimic the morphological structure of native blood vessels. Regulating the content of BNC can reduce the thrombosis potential of stents and enhance endothelialization.

#### **4.4 Other biomedical application**

In addition to the above biomedical applications, BNC has been applied in other biomedical fields, such as nerve repair [66, 67] and muscle [33], corneal [68, 69], urethral [70], and intervertebral disc [71]. These researches will be sure of certain theoretical value and practical significance to the biomedical application of BNC.

*Bacterial Nanocellulose: Methods, Properties, and Biomedical Applications DOI: http://dx.doi.org/10.5772/intechopen.114223*

#### **5. Conclusion**

In this chapter, we provide an overview of the fabrication and biomedical application of BNC-based biomaterials. First, the biosynthesis of BNC in biology, chemistry, and physics is introduced, and the properties of BNC are summarized. Furthermore, we introduce and discuss the various techniques to fabricate BNC-based composites and 3D porous BNC scaffolds to enhance the mechanical and biological properties. Due to the rapid development of the abovementioned technologies, BNC has a broad range of applications in biomedicine, including wound dressing, bone/cartilage tissue regeneration, vascular tissue engineering, and so on.

#### **Acknowledgements**

This work was supported by National Natural Science Foundation of China (grant nos. 82160355), the Science and Technology Research Project of Jiangxi Education Department (grant nos. GJJ2200657), and Natural Science Foundation of Jiangxi Province (grant nos. 20212ACB214002).

#### **Conflict of interest**

The authors declare no conflict of interest.

#### **Author details**

Haiyong Ao\* and Xiaowei Xun School of Materials Science and Engineering, East China Jiaotong University, Nanchang, PR China

\*Address all correspondence to: aohyong@126.com

© 2024 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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### Section 3
