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Polysaccharide-Protein Nanoassemblies: Novel Soft Materials for Biomedical and Biotechnological Applications
Abstract and Figures
Polysaccharide and proteins are the major constituent building blocks of biological systems and often occur as highly organized macromolecular architectures (e.g. the capsid of viruses). Both can occur in the same or in different biological physiological environment interacting in specific or non-specific ways. When isolated and purified, these macromolecules can harness self-assembled (SA) soft nanomaterials by non-covalent electrostatic complexation. Although polysaccharide-protein electrostatic SA systems of this type have been studied for more than two decades, the possibility to design materials with enhanced biological function and improved technological advantages over those based on synthetic or inorganic components, has only started to be recognized and is yet to be fully realized. In this review we address two main type of SA polysaccharideprotein systems, namely, those based on chitosan-protein and those based on polyanionic polysaccharide (pectin, hyaluronic acid or alginate) - protein ones. The physical properties of chitosan- and polyanion-based SA nanocomplexes with oppositely charged proteins depend on the composition and conditions as reviewed here with reference to some specific systems.
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... Hyaluronan exhibits outstanding hydrodynamic qualities, particularly in terms of its viscosity and water-retentiveness. It used to be thought of as crucial for organ structural support and joint lubricant (Fuenzalida and Goycoolea 2015). Furthermore, it was discovered that HA was necessary for healthy cellular proliferation, embryo development, repairing procedures, and inflammatory, and tumor growth. ...
Polysaccharides that are naturally sourced have enormous promise as wound dressings, due to their wider availability and reasonable cost and good biocompatibility. Furthermore, nanosilver extensively applied in wound treatment is attributed to its broad spectrum of antimicrobial effects and lesser drug resistance. Consequently, wound dressings in corporating nanosilver have attracted wide-scale interest in wound healing, and nanosilver-functionalized polysaccharide-based wound dressings present an affordable option for healing of chronic wounds. This review encompasses preparation methods, classification, and antibacterial performances of nanosilver wound dressings. The prospective research arenas of nanosilver-based wound polysaccharide dressings are also elaborated. The review attempts to include a summary of the most recent advancements in silver nanotechnology as well as guidance for the investigation of nanosilver-functionalized polysaccharide-based wound dressings.
... A pre-culture of B. licheniformis was grown in LB medium (1% (w/v) NaCl, 1% (w/v) peptone, 0.5% yeast extract, pH 7) at 37°C under agitation at 250 rpm overnight. Antibacterial assay was performed in a 24-well flat bottom deep well plate by mixing 50 µL of bacterial suspension or medium as a blank, with 1 mL of chitosan solution and 4 mL of MM1 P100 medium (3% (w/v) glucose, 0.13% (w/v) MgSO 4 107 , yielding final chitosan concentrations of 0, 10, 20, 30, 40, 50, 75, or 100 µg mL −1 . After overnight incubation at 37°C without shaking, 20 mL per well were diluted with 180 µL of MM1 P100 medium in a separate 96-well plate, and the OD was measured at λ = 600 nm (OD 600 ) using a UV/Vis microplate reader (Epoch 2 Microplate Spectrophotometer, Agilent Technologies, Santa Clara, CA, USA). ...
Chitosans are versatile biopolymers with multiple biological activities and potential applications. They are linear copolymers of glucosamine and N-acetylglucosamine defined by their degree of polymerisation (DP), fraction of acetylation (FA), and pattern of acetylation (PA). Technical chitosans produced chemically from chitin possess defined DP and FA but random PA, while enzymatically produced natural chitosans probably have non-random PA. This natural process has not been replicated using biotechnology because chitin de-N-acetylases do not efficiently deacetylate crystalline chitin. Here, we show that such enzymes can partially N-acetylate fully deacetylated chitosan in the presence of excess acetate, yielding chitosans with FA up to 0.7 and an enzyme-dependent non-random PA. The biotech chitosans differ from technical chitosans both in terms of physicochemical and nanoscale solution properties and biological activities. As with synthetic block co-polymers, controlling the distribution of building blocks within the biopolymer chain will open a new dimension of chitosan research and exploitation. Degree of polymerisation, fraction and pattern of acetylation change the material and biological properties of chitosan. Here, the authors show that enzymes can N-acetylate fully deacetylated chitosan to replicate the natural control over acetylation pattern not found in chemically produced chitosan allowing more control over properties.
... In addition, the use of self-assembly of proteins and polysaccharides to embed and control the release of bioactive substances is attracting in recent years (Cornelus, Kruif, Fanny, Weinbreck, & Renko, 2004;Chen, Fu, Niu, Hao, & Xin, 2018;Espinosa-Andrews, Báez-González, Cruz-Sosa, & Vernon-Carter, 2007). The opposite charges of protein and polysaccharide are used to coat each other, and the polyelectrolyte complex is formed through electrostatic interaction (Goycoolea & Fuenzalida, 2015). Composite nanoparticles have more excellent rationalization and functionality (Chuah, Roberts, Billa, Abdullah, & Rosli, 2014;Luo, Teng, Li, & Wang, 2015). ...
Two quercetin loading zein nanoparticles with shell of caseinate (SC-ZNP) and caseinate-chitosan double layer (CHC-ZNP) were fabricated by anti-solvent method. The formulas of the two nanoparticles were optimized and their physicochemical properties and bioavailability enhancement effect were compared in both in vitro and in vivo. The particle size, zeta potential, encapsulation efficiency and loading efficiency of SC-ZNP were 188 nm, -29.33 mV, 82.78% and 1.58%, while these values for CHC-ZNP were 551 nm, +57.47 mV, 78.26% and 2.91%, respectively. Physicochemical characterization showed that quercetin lost its crystalline structure and existed in amorphous form in both nanoparticles. After freeze-drying, SC-ZNP showed better redispersibility and smaller size change than CHC-ZNP. Both nanoparticles significantly improved the solubility and passive diffusion velocity of quercetin. The coating of caseinate shell provided better colloidal stability (pH, salt and storage) than caseinate-chitosan double layers. Both SC-ZNP and CHC-ZNP exhibited mucoadhesive properties and significantly decreased the feces excretion of quercetin in rat. Because of smaller particles size, SC-ZNP penetrated into the mucous layer more easily. After oral administration, few free quercetin was found in rat plasma, while many metabolites were detected. Through semi-quantification of its most dominant metabolite, the bioavailability of quercetin in SC-ZNP and CHC-ZNP groups were 2.34 and 1.89 times enhanced.
... Such events reduce dosing frequency in a chemotherapy regimen. Among various types of nanocarriers, naturally occurring compound-based nanocomplexes have been receiving extensive attention in drug delivery applications [9][10][11]. These molecules can produce a spontaneous self-assembly or complexation with therapeutic agents. ...
Pancreatic cancer (PanCa) is a lethal disease. Conventional chemotherapies for PanCa offer severe systemic toxicities. Thus, the development of a successful nanomedicine-based therapeutic regimen with augmented therapeutic efficacy is highly sought. Naturally occurring pectin and modified pectin-based drug delivery systems exhibit remarkable self-targeting ability via galactose residues to various cancer cells. Herein, we developed and used an innovative approach of highly stable nanocomplexes based on modified pectin and tannic acid (MPT-NCs). The nanocomplex formation was enabled by strong intermolecular interactions between pectin and tannic acid under very mild conditions. These nanocomplexes were characterized by particle size and morphology (DLS, TEM, and SEM), FT-IR spectroscopy, and zeta potential measurements. Additionally, MPT-NCs were capable of encapsulating anticancer drugs (5-fluorouracil, gemcitabine, and irinotecan) through tannic acid binding. The in vitro bioactivity of these drug MPT-NCs were evaluated in pancreatic cancer adenocarcinoma (PDAC) cell lines (HPAF-II and PANC-1). A dose-dependent internalization of nanocomplexes was evident from microscopy and flow cytometry analysis. Both proliferation and colony formation assays indicated the anticancer potential of pectin drug nanocomplexes against PDAC cells compared to that of free drug treatments. Together, the pectin-based nanocomplexes could be a reliable and efficient drug delivery strategy for cancer therapy.
Significant advancements have been noticed in cancer therapy for decades. Despite this, there are still many critical challenges ahead, including multidrug resistance, drug instability, and side effects. To overcome obstacles of these problems, various types of materials in biomedical research have been explored. Chief among them, the applications of natural compounds have grown rapidly due to their superb biological activities. Natural compounds, especially polyphenolic compounds, play a positive and great role in cancer therapy. Tannic acid (TA), one of the most famous polyphenols, has attracted widespread attention in the field of cancer treatment with unique structural, physicochemical, pharmaceutical, anticancer, antiviral, antioxidant and other strong biological features. This review concentrated on the basic structure along with the important role of TA in tuning oncological signal pathways firstly, and then focused on the use of TA in chemotherapy and preparation of delivery systems including nanoparticles and hydrogels for cancer therapy. Besides, the application of TA/Fe3+ complex coating in photothermal therapy, chemodynamic therapy, combined therapy and theranostics is discussed.
Statistical methods were used to provide insight into a polymer complex system composed of lysozyme and alginate to quantify the effects of such parameters as pH, and ionic composition of the mixing solution on the properties of the complexes including composition, particle diameter, and zeta potential. Various crosslinkers (calcium, barium, iron[III], and bovine serum albumin), were used with lysozyme for complex formation to investigate the effect of crosslinker charge density on protein release kinetics, modelled using ktⁿ. Multivariate statistical analysis showed that the kinetic parameters associated with the release were, not surprisingly highly dependent on the ionic strength of the release media, with higher ionic strength leading to faster release. The release parameter k was also shown to depend on the protein properties (size, ionic strength) while n was slightly, but not statistically dependent on the charge density of the crosslinker demonstrating that the nature of the crosslinker had minimal impact on drug release. The multivariate statistical has the potential to be used for optimization of the complexes and prediction of physical properties and degradation rates.
Nanoencapsulation of hydrophobic nutraceuticals with food ingredients has become one of topical research subjects in food science and pharmaceutical fields. To fabricate food protein-based nano-architectures as nanovehicles is one of effective strategies or approaches to improve water solubility, stability, bioavailability and bioactivities of poorly soluble or hydrophobic nutraceuticals. Milk proteins or their components exhibit a great potential to assemble or co-assemble with other components into a variety of nano-architectures (e.g., nano-micelles, nanocomplexes, nanogels, or nanoparticles) as potential nanovehicles for encapsulation and delivery of nutraceuticals. This article provides a comprehensive review about the state-of-art knowledge in utilizing milk proteins to assemble or co-assemble into a variety of nano-architectures as promising encapsulation and delivery nano-systems for hydrophobic nutraceuticals. First, a brief summary about composition, structure and physicochemical properties of milk proteins, especially caseins (or casein micelles) and whey proteins, is presented. Then, the disassembly and reassembly behavior of caseins or whey proteins into nano-architectures is critically reviewed. For caseins, casein micelles can be dissociated and further re-associated into novel micelles, through pH- or high hydrostatic pressure-mediated disassembly and reassembly strategy, or can be directly formed from caseinates through a reassembly process. In contrast, the assembly of whey protein into nano-architectures usually needs a structural unfolding and subsequent aggregation process, which can be induced by heating, enzymatic hydrolysis, high hydrostatic pressure and ethanol treatments. Third, the co-assembly of milk proteins with other components into nano-architectures is also summarized. Last, the potential and effectiveness of assembled milk protein nano-architectures, including reassembled casein micelles, thermally induced whey protein nano-aggregates, α-lactalbumin nanotubes or nanospheres, co-assembled milk protein-polysaccharide nanocomplexes or nanoparticles, as nanovehicles for nutraceuticals (especially those hydrophobic) are comprehensively reviewed. Due to the fact that milk proteins are an important part of diets for human nutrition and health, the review is of crucial importance not only for the development of novel milk protein-based functional foods enriched with hydrophobic nutraceuticals, but also for providing the newest knowledge in the utilization of food protein assembly behavior in the nanoencapsulation of nutraceuticals.
An emerging understanding of natural biolubricants such as saliva and synovial fluid reveals that they rely on synergistic interactions between biomacromolecules in aqueous media, wherein one polymeric component promotes adsorption to biological substrates while another facilitates hydration lubrication. We hypothesise that this phenomenon can be achieved by combining the strong adsorption characteristics of proteins with the hydration of polysaccharides. To confirm this hypothesis, hydrated soluble complexes were formed through electrostatic interactions between positively-charged lysozyme molecules and anionic residues on pectic polysaccharides extracted from Plantago ovata seed mucilage. Boundary friction was measured between smooth hydrophobic polydimethylsiloxane (PDMS) surfaces using a ball-and-disk tribometer, while adsorption was assessed using a combination of the quartz-crystal microbalance with dissipation monitoring technique and ellipsometry. The results indicate that complexation between polysaccharides and proteins leads to improved adsorption on hydrophobic PDMS surfaces, which in turn decreases boundary friction in sliding contact. By comparing the adsorption behaviour of the complexes and individual components, we conclude that pectic polymers maintain the film’s hydration while lysozyme improves surface adsorption, thereby providing new evidence that physical complexes can be utilised to design bi-functional surface layers. Our findings open up a range of opportunities for application in food and oral healthcare products, particularly in the use of food-compatible ingredients to control aqueous lubrication.
Food-derived biomaterials have attracted increasing attention due to their renewability, excellent biocompatible and biodegradable properties, tunable solubility, and multi-active binding sites for introducing novel functionalities. As the three major food biomaterials, polysaccharides, lipids, and proteins from diverse natural sources are environmentally friendly and possess potential functions for emerging applications. Food-derived nanomaterials can be obtained from one or more types of food biomaterials or the combination of the various biopolymers by many manufacturing methods. Applications of food-derived nanomaterials or bionanocomposites to biosensing are current research trends owing to their new optical, catalytic, and structural functionalities. This review provides a broad overview of the functional properties of food-derived nanomaterials, the preparation techniques for creating food-derived nanomaterials, and their potential applications for biosensing. Moreover, the major challenges and future perspectives of this fast-growing field are also highlighted.
The present invention provides colloidally stable dispersions of nanoparticles comprising beta-lactoglobulin and a polysaccharide which are transparent when diluted in aqueous media. In particular these colloidally stable dispersions of nanoparticles are useful as delivery vehicles of hydrophobic nutraceuticals and fat-soluble vitamins, for enrichment of food products, especially of transparent beverages and other non-fat or low fat foods and drinks. The present invention further provides methods for the preparation of said colloidally stable dispersions.
Several polymers of both natural and synthetic origin have been used for a variety of biomedical applications including pharmaceutical preparations, drug targeting, imaging, drug delivery, prosthetics, and tissue engineering scaffolds. Due to their reproducible characteristics in terms of their molecular weight, degradation and mechanical properties, synthetic polymers are attractive for a variety of the aforementioned applications. However, synthetic polymers from the biological standpoint, synthetic polymers often lack much-desired bioactivity and biocompatibility, which may translate into adverse side effects. Natural polymers on the other hand are abundant and resemble the components present in biological extracellular matrices. Thus, natural polymers are readily accepted by the body and possess high bioactivity and biocompatibility. Natural polymers can be divided into three major classes according to their chemical structure: (i) polysaccharides, (ii) proteins, and (iii) polyesters. This chapter presents an overview of the polysaccharide-based biomaterials, their structure property, and applications in the area of drug delivery and tissue engineering. Particular emphasis is given to polysaccharides such as (i) hyaluronic acid (HA), (ii) chondroitin sulfate, (iii) chitin and chitosan, (iv) alginates, and (v) cellulose. These polymers and their popular derivatives are also discussed in the context of their chemical and biological properties. Polysaccharides in their native form may not be able to provide all the desired properties for a particular biomedical application. Thus, the chapter also focuses on the polysaccharide derivatives and their blend with other polymers for a variety of biomedical applications.
This book provides a critical overview of the advances being made towards overcoming biological barriers through the contribution of nanosciences and nanotechnologies. Overcoming these barriers is of primary importance for solving the problems of many current drugs and vaccines and it is also especially relevant for the commercial exploitation of new therapeutic strategies i.e. gene and cellular therapies. Despite important information that has been covered in recently published books related to nanomedicine (mainly oriented to technologies and applications), there is still an important gap related to the relationship between the chemical properties of nanobiomaterials and their interaction with the biological environment. Thus, this comprehensive new book that is presented with a clear and organised structure dedicated to the different biological barriers relevant for drug delivery applications will focus on (i) mechanistic issues related to the interaction between drug delivery systems and biological barriers and (ii) the analysis of the critical factors determining the efficacy of nanobiomaterials for specific applications. Moreover, this field has become highly multidisciplinary over the last decade, leading to the development of new products, as well as to significant advances from the regulatory point of view. Moreover, this book will also provide an up-to-date view on the clinical relevance of nanomedicine and on its possible impact on global health.
The polyelectrolyte titration is widely used to monitor the cationic and anionic demand of papermaking suspensions. Most laboratory and online instrumentation is based upon the streaming current detector. In most cases users simply use the end-point generated by the instrumentation. This paper describes the factors controlling the shape of the titration curve and gives examples of systems where viewing the endpoint alone gives misleading results.
In the present work we investigated the electrostatic self-assembly of nanocomplexes (NCXs) made of lysozyme (Lyz) and different alginates (Alg A Mw ~4kD and M/G ~1.42 or Alg B Mw ~7kD and M/G~5.00) to establish a relationship between the alginate structure and both the physical (size and zeta potential, optical density) and the thermodynamic properties of the systems. It was found that the alginate rich in mannuronic unit (Alg B) had a greater binding stoichiometry and lower affinity than Alg A, a fact that correlated with a more extended structure that in turn affected the capacity to form hydrogen bonds associated with the differences in charge distribution. Both alginates show higher affinity and temperature stability than poly-acrylic acid (PAA), showing the importance of the polyasaccharide structure in the NCXs formation. For the first time we showed that the M/G ratio plays a role on the affinity of interaction of alginate and lysozyme. Finally, these NCXs seem promising systems for biomedical application due to their in vitro biocompatibility and macromolecules retention capacity.
Self-assembly is the autonomous organization of components into patterns or structures without human intervention. Self-assembling processes are common throughout nature and technology. They involve components from the molecular (crystals) to the planetary (weather systems) scale and many different kinds of interactions. The concept of self-assembly is used increasingly in many disciplines, with a different flavor and emphasis in each.
Pectins belong to the group of hetero-polysaccharides and are present in all plant primary cell walls. Depending on the molecular composition there are different types of pectin characterising themselves particularly by different gelling mechanisms. In addition to the manufacturing process of pectin, the chapter describes the relation between pectin structure and gelling mechanism. Furthermore, health aspects of pectins are discussed as well as diverse applications in the food sector. Various formulation examples show the different application areas in the food range.
Mucoadhesion is defined as the ability of a material to adhere to the mucosal surface of tissues for an extended period of time [1]. A mucoadhesive drug delivery system can improve the controlled delivery and optimize the bioavailability of drugs for a prolonged period of time. Polysaccharides are among the most versatile families of macromolecules and their potential use in transmucosal drug delivery application is fully recognized [2,3]. The mucoadhesion of various polysaccharide families of varying structural characteristics and natural sources was explored by a sensitive micorviscosimetric method in order to contribute to a more rational design of mucoadhesive polysaccharide-based nanoparticles as potential carriers of compounds that interfere either with the adhesion or with the avaibility of essential nutrients of the H. pylori in the stomach mucosa. Here we show how deviation in viscosity of mixed solutions of polysaccharides and soluble fraction of mucin with respect to the viscosity of the original stock solutions can be considered as diagnostic of molecular associative interactions [4]. In the light of the results obtained we could observe a strong correlation between interaction and the ability of polysaccharide coils to contract in the presence of salt (i.e. chain flexibility). Moreover, the “more neutral” polysaccharides such as dextran, mesquite gum and a bacterial exopolysaccharide from Streptococcus thermophilus, showed no interaction with mucin. The adopted approach is perhaps an oversimplification of the mucoadhesion behavior in vivo during the interaction of polysaccharides with the mucous epithelia. However, it has allowed gaining further insight into the possible mechanisms underlying the mucoadhesion phenomena.