Advantages and disadvantages of bacterial cellulose and solutions to improve its properties.
The new in situ properties interfere with the nanofibers crosslinking. The main objective of such modification are new characteristics in the matrix, changing its biophysical properties. The additives become part of the nanofibers, interacting with the –OH portions present in the bacterial cellulose chains and forming new hydrogen bonds. Chitosan, a polysaccharide derived from chitin, has biocompatibility, antibacterial and antifungal properties. The combination of bacterial cellulose, in a dressing, exhibited favorable antibacterial activities and no cytotoxicity
[61]. Zhou et al.
[62] demonstrated that their bacterial cellulose bandage associated with collagen I and hydroxypropyltrimethyl ammonium chloride chitosan exhibited excellent antibacterial activity, cytocompatibility and promoted the growth and proliferation of NIH3T3 cells and HUVECs cells. Silver nanoparticles and polydopamine incorporated into bacterial cellulose demonstrated antibacterial activity, increased cell viability, showed no cytotoxicity to fibroblast cells, granulation tissue formation, angiogenesis and re-epithelialization upon histopathological examination
[63]. Several nanotubes, nanosheets were also incorporated into cellulose culture media. Park et al.
[64] produced hybrid compounds of bacterial cellulose and carbon nanotubes that showed osteoconductivity and osteoinductivity. Likewise, Khalid et al.
[65] demonstrated that the bandage, composed of bacterial cellulose and carbon nanotubes, acted as a mechanical and antibacterial barrier to fragile healing tissue, aided in moisture retention, reduced inflammation, and resulted in efficient wound healing. Graphene nanosheets were incorporated into the bacterial cellulose matrix, resulting in decreased crystallinity, improved mechanical and electrical properties. Luo et al.
[66] produced a compound that exhibited high tensile strength with 93% improvement compared to pure bacterial cellulose film. In addition, the film also showed excellent flexibility with good conductivity. The association between nano zinc oxide and bacterial cellulose increased porosity and pore sizes, which increased water vapor permeability (an important factor for a bandage), it also showed antibacterial activity, good physical properties, non-cytotoxicity and good biocompatibility
[67]. Although in situ modifications allow a uniform material distribution, the fermentation conditions of the biosynthesis process limit the incorporation of other materials.
Ex-situ modifications seek to alter the physicochemical and functional properties of the matrix after biosynthesis and purification of bacterial cellulose. The nanometric materials can be aggregated through diffusion to pass through the network pores. This type of modification can be divided into the chemical modification and composites development
[68].
In the chemical modification process, bacterial cellulose is treated with several chemical reagents to modify its chemical structure and incorporate additional functionalities. The most common chemical modification is oxidation but there are also modifications by acetylation
[69], benzoylation
[70], succinylation
[71] and phosphorylation
[72].
Oxidation seeks to add new functional groups to cellulose. Oxidized cellulose is the most precious by-product of cellulose, and several chemical and physical properties of oxidized cellulose can be obtained under various oxidizing conditions (nature, temperature, pH and reaction duration)
[73]. Many agents can be used, such as hydrogen peroxide, persulfates, permanganates, nitrogen dioxide, chlorine dioxide and phosphoric acids
[74]. However, water-soluble 2, 2, 6, 6-tetramethylpiperidine-1-oxyl (TEMPO) is widely used to oxidize cellulose. Oxidized BC has been investigated for different applications such as adsorption of heavy metals, oil removal and various biomedical applications
[75][76]. In etherification, the reaction is carried out in two steps; in the first step, cellulose is activated by treatment with an alkaline solution, followed by an etherification reaction with monochloroacetic acid or its sodium salt. Carboxymethylcellulose is one of the most important cellulose derivatives and can be used in pharmaceutical, cosmetic, food and biomedical areas
[77]. Sulfation synthesizes cellulose in sulfuric acid in isopropyl alcohol or with SO3-pyridine complex in ionic liquids
[78][79]. Cellulose sulfate has as its main characteristic, anticoagulating, antivirus and antibacterial properties
[80]. Benzoylation treats bacterial cellulose with benzoyl citrate, adding to the material the potential for sensors, piezoelectric materials and optical properties
[81]. Phosphorylation is developed for textiles and flame retardant materials, as it can induce the formation of calcium phosphate making the material suitable for biomedical applications
[16].
Despite its advantages, cellulose has no antibacterial capacity and moderate mechanical properties. The development of composites aims to improve some properties that limit the application of bacterial cellulose in biomedical and tissue engineering. To improve mechanical and biological properties, researchers have incorporated different types of materials into bacterial cellulose, including polymers, carbon-based nanoparticles, metal/metal oxide nanoparticles, and other inorganic nanoparticles
[82].
Bacterial cellulose fragments were immersed in the chitosan solution followed by lyophilization to produce a scaffold to aid in ovarian cancer diagnosis. The scaffold obtained showed better interaction with the cells compared to pure BC
[83]. JU et al.
[84] produced a bacterial cellulose film, in which the cellulose suspension and the polyvinyl alcohol solution were mixed, followed by the incorporation of chitosan in bulk form or nanoparticle form. The bulk form of chitosan increased the mechanical and elastic properties of the film, while the nanoparticle form showed higher antibacterial properties. The gelatin and hydroxyapatite incorporation in bacterial cellulose showed a composite with high mechanical properties, positive cell adhesion, proliferation and differentiation
[85]. Yan et al.
[86] achieved a scaffold with reduced porosity, high mechanical properties and great in vitro biocompatibility, by incorporating bacterial nano-cellulose into alginate and collagen. The incorporation of graphene, a carbon nanomaterial with a 2D structure, and carbon nanotubes add to bacterial cellulose better mechanical, electrical and thermal properties
[16][87].
In summary, several biopolymers and biomaterials can be incorporated into bacterial cellulose to improve its properties, reducing its applicability limitations. The in situ and ex-situ modifications are methods that work on the incorporation of these materials homogeneously. Although the in situ modifications present several advantages of materials aggregation, the method is limited because some materials do not support the biosynthetic process. On the other hand, ex-situ modification expands the range of materials that can be incorporated; however, scientists still seek completely homogeneous incorporation in this process.
5. Bacterial Cellulose for Cardiac Tissue Regeneration
Understanding the environment nanoscale is essential to produce biomaterials that mimic the cellular microenvironment. The environment properties employ a total influence on cell adhesion, proliferation, maturation and differentiation, and consequently generate impacts on the function of a tissue. Cellulose is a very versatile material with its adaptable properties that allow its application in systems with different chemical and biophysical environments. Cellulose-based biomaterials provide important advantages over conventional synthetic materials, which demonstrates their promise of advancing scientific knowledge. The role of the extracellular matrix is established, and we know that it not only allows cellular attachment but also sends biochemical and biophysical clues to the nascent cells and tissues. Such data support studies on the application of scaffolds of decellularized tissues and organs in tissue engineering and regenerative medicine. The mimicry of natural conditions both in the tissue and in the ECM requires adequate adhesion and growth properties that maintain the tissue’s normal structure, and the results of biopolymers’ application involving celluloses mentioned above reveal successful results.
The bacterial cellulose use in cardiac tissue regeneration still needs more studies. In the literature, only one study was found that tested the cellulose membrane viability, acting as an adhesive, loaded with co-cultured cells. Simeoni et al.
[88] produced a patch loaded with skeletal myoblasts and mesenchymal stem cells that was surgically inserted into the epicardial region of the left ventricle, where they found that the cellulose patch can protect the myocardium against the deleterious effects and pathological remodeling of the ischemic heart; this beneficial result was not obtained only with cell therapy. Other studies demonstrated the applicability of cellulose, modified cellulose and its composites. Only Simoeni et al.
[88] describe the bacterial cellulose use itself. Chen et al.
[89] developed a polyurethane/cellulose scaffold that presented greater mechanical strength and essential characteristics for the survival and function of cardiac cells with native anisotropy. As such, Entcheva et al.
[90] tested the potential of cellulose acetate and reduced cellulose scaffolds for the growth of cardiomyocytes in vitro. They attested that the surface of these materials promoted cell growth, while increasing gap junctions, and electrical functionality. Such studies open doors to new possibilities for applications of bacterial cellulose, at the same time highlighting the potential of this biomaterial in cardiac tissue regeneration.