Preparation, properties and drug controlled release of chitin-based hydrogels: An updated review.

Chitin, as the second abundant biopolymers on earth after cellulose, has been used for synthesis of hydrogels in a wide range of applications due to its biocompatibility, biodegradability and nontoxicity. This review aims to provide an overview of the latest developments in the preparation, properties and drug controlled release of chitin-based hydrogels. In the first part, the preparation of hydrogels from native chitin, nano chitin and chitin derivatives via physical or chemical procedures was discussed in detail. The properties of chitin-based hydrogels, including gel strength, swelling degree and smart response characteristics, were described in the second part. This review also introduced how chitin-based hydrogel as a drug controlled release carrier is affected by composition, pH, temperature, magnetic field, electric field and other factors. We hope this review can provide guidelines for the rational design of chitin-based hydrogels in drug delivery systems.

Fabrication of chitin monoliths with controllable morphology by thermally induced phase separation of chemically modified chitin.

As a natural polymer, chitin has excellent biological properties such as biodegradability and immunological, antibacterial, and wound-healing activities and has numerous applications in cosmetics, drug delivery, and pharmaceuticals. Organic polymer monoliths have also drawn significant attention, owing to their high permeability, large surface area, and high mechanical strength. They are usually applied to separation, ion exchange, catalysis, and chromatography. We have previously prepared cellulose monoliths using biopolymers; however, because chitin possesses amide groups on its side chain, it is superior to cellulose for further chemical modification and applications. However, the utilization of chitin is restricted by its insolubility in water and common organic solvents. In this study, for the first time, a monolith was prepared by chemical modification of chitin using a thermally induced phase separation (TIPS) method. First, we prepared dibutyrylchitin (DBC) as a starting polymer that is soluble in organic solvents. To prepare the monolith, DBC was dissolved completely in dimethyl sulfoxide (DMSO) while heating, and deionized water was added to the solution. It was then cooled at 20 °C to form a monolith via phase separation. The porous morphology of the DBC monolith was altered by regulating the DBC concentration, DMSO/H2O ratio, and aging temperature. The DBC monolith was converted to a chitin monolith by the alkaline hydrolysis of butyryl ester. The successful hydrolysis of butyryl ester was confirmed by the disappearance of the peak at 1735 cm-1 in the FT-IR spectra, which is related to the ester moiety of DBC. The chitin monolith has the potential to be utilized under water flow for catalysis, metal capture from wastewater, dye sorption, and drug delivery systems.

Versatile antibacterial surface with amphiphilic quaternized chitin-based derivatives for catheter associated infection prevention.

Microbial colonization of catheter surfaces is responsible for most healthcare-associated infections. Quaternized chitin and chitosan have excellent antimicrobial and biocompatible properties and can be used to provide safe and prolonged protection for biomedical catheters. Herein, we prepared quaternized ß-chitin derivative (QC)- and quaternized chitosan derivative (QCS)-based antimicrobial surfaces. The quaternized polysaccharides modified TPU surfaces exhibited hydrophilicity, good biocompatibility. Among these, QCS2-modified TPU exhibited excellent antibacterial properties against Gram-positive and Gram-negative bacteria, and prevented the adherence of bacteria compared with pristine TPU. The antibacterial activity of QCS2-modified surfaces maintained for 8 weeks under the condition of immersion in serum. An in vivo subcutaneous implantation experiment revealed 99.87% reduction of bacteria and reduced expression of inflammation-related factors in the surrounding tissue five days after implantation with QCS2-modified TPU. Therefore, quaternized polysaccharide-modified surfaces have promising potential in preventing medical catheter-associated infections.

Identification of a Chitooligosaccharide Mechanism against Bacterial Leaf Blight on Rice by In Vitro and In Silico Studies.

This study focuses on a commercial plant elicitor based on chitooligosaccharides (BIG®), which aids in rice plant growth and disease resistance to bacterial leaf blight (BLB). When the pathogen (Xoo) vigorously attacks rice that has suffered yield losses, it can cause damage in up to 20% of the plant. Furthermore, Xoo is a seed-borne pathogen that can survive in rice seeds for an extended period. In this study, when rice seeds were soaked and sprayed with BIG®, there was a significant increase in shoot and root length, as well as plant biomass. Furthermore, BIG®-treated rice plants showed a significant reduction in BLB severity of more than 33%. Synchrotron radiation-based Fourier transform infrared (SR-FTIR) analysis was used to characterize BIG®'s mechanism in the chemical structure of rice leaves. The SR-FTIR results at 1650, 1735, and 1114 cm-1 indicated changes in biochemical components such as pectins, lignins, proteins, and celluloses. These findings demonstrated that commercial BIG® not only increased rice growth but also induced resistance to BLB. The drug's target enzyme, Xoo 1075 from Xanthomonas oryzae (PDB ID: 5CY8), was analyzed for its interactions with polymer ingredients, specifically chitooligosaccharides, to gain molecular insights down to the atomic level. The results are intriguing, with a strong binding of the chitooligosaccharide polymer with the drug target, revealing 10 hydrogen bonds between the protein and polymer. Overall, the computational analysis supported the experimentally demonstrated strong binding of chitooligosaccharides to the drug target.

Modified porous carboxymethyl chitin microspheres by an organic solvent-free process for rapid hemostasis.

Rapid and effective hemorrhage control is essential to reduce mortality following traumatic injuries. Herein we developed an organic solvent-free process to prepare carboxymethyl chitin microsphere (CMCHm) in an aqueous two-phase system through heating and freeze-drying. To further enhance the hemostatic performance of CMCHm, we loaded calcium ions and in-situ polymerized dopamine to get modified hemostatic microspheres CMCHm-Ca2+ and CMCHm-PDA, respectively. The size of these microspheres was mainly distributed between 50 µm and 150 µm, and the porous microstructure was observed by SEM. The data of in vitro degradation, cell cytotoxicity, and hemolysis test indicated good biocompatibility of these microspheres. Importantly, CMCHm-Ca2+ and CMCHm-PDA displayed better hemostatic performance compared with CMCHm and the positive controls Yunnan baiyao® and Quickclean®. Especially, the bleeding time was reduced to 59 s (CMCHm-Ca2+) and 45 s (CMCHm-PDA) in the femoral artery/vein cut model, respectively. All these demonstrate CMCHm-Ca2+ and CMCHm-PDA hold great potential for rapid hemostasis.

Efficacy of a Chitin-Based Water-Soluble Derivative in Inducing Purpureocillium lilacinum against Nematode Disease (Meloidogyne incognita).

Plant-parasitic nematodes cause severe economic losses annually which has been a persistent problem worldwide. As current nematicides are highly toxic, prone to drug resistance, and have poor stability, there is an urgent need to develop safe, efficient, and green strategies. Natural active polysaccharides such as chitin and chitosan with good biocompatibility and biodegradability and inducing plant disease resistance have attracted much attention, but their application is limited due to their poor solubility. Here, we prepared 6-oxychitin with good water solubility by introducing carboxylic acid groups based on retaining the original skeleton of chitin and evaluated its potential for nematode control. The results showed that 6-oxychitin is a better promoter of the nematicidal potential of Purpureocillium lilacinum than other water-soluble chitin derivatives. After treatment, the movement of J2s and egg hatching were obviously inhibited. Further plant experiments found that it can destroy the accumulation and invasion of nematodes, and has a growth-promoting effect. Therefore, 6-oxychitin has great application potential in the nematode control area.

Transglutaminase-Mediated Caseinate Oligochitosan Glycation Enhances the Effect of Caseinate Hydrolysate to Ameliorate the LPS-Induced Damage on the Intestinal Barrier Function in IEC-6 Cells.

Some food components can regulate the intestinal barrier function. Herein, the effect of transglutaminase-type oligochitosan glycation on caseinate hydrolysate for its ability to maintain intestinal epithelial integrity and the tight junction (TJ) structure was investigated by assessing and comparing the bioactivities of glycated caseinate hydrolysate and caseinate hydrolysate against the lipopolysaccharide-induced barrier damage in the model cells (rat intestinal epithelial IEC-6 cells). The results from liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis demonstrated that oligochitosan glycation occurred at the Gln residues of α-S1-casein and α-S2-casein. The two hydrolysates retarded the lipopolysaccharide cytotoxicity toward IEC-6 cells and enhanced the barrier integrity by increasing the transepithelial electrical resistance or decreasing the paracellular permeability. In addition, these two hydrolysates could upregulate both mRNA and protein expression of three TJ proteins in IEC-6 cells. More importantly, the glycated caseinate hydrolysate had higher potential than caseinate hydrolysate to protect IEC-6 cells against the lipopolysaccharide-induced barrier damage, suggesting that the transglutaminase-mediated oligochitosan glycation of proteins is a useful approach to enforce protein biofunctions in the intestine.

Chemoenzymatic production of chitooligosaccharides employing ionic liquids and Thermomyces lanuginosus chitinase.

The aim of this work was to study a two-step chemoenzymatic method for production of short chain chitooligosaccharides. Chitin was chemically pretreated using sulphuric acid, sodium hydroxide and two different ionic liquids, 1-Ethyl-3-methylimidazolium bromide and Trihexyltetradecylphosphonium bis(2,4,4-trimethylpentyl)phosphinate under mild processing conditions. Pretreated chitin was further hydrolyzed employing purified chitinase from Thermomyces lanuginosus ITCC 8895. Trihexyltetradecylphosphonium bis(2,4,4-trimethylpentyl)phosphinate treated chitin appeared amorphous and resulted in generation of 1.10 ± 0.89 mg ml-1 of (GlcNAc)2 and 1.07 ± 0.92 mg ml-1 of (GlcNAc)3. Further derivation of optimum conditions through two-factor-9 run experiments resulted in to 1.5 and 1.3 fold increments in (GlcNAc)2 and (GlcNAc)3 production, respectively. 0.1 g of both (GlcNAc)2 and (GlcNAc)3 has been purified from the Trihexyltetradecylphosphonium bis(2,4,4-trimethylpentyl)phosphinate pretreated chitin (1 g) employing cation exchange chromatography. The present study will lay the foundation for development of a green sustainable solution for cost effective upcycling of coastal residual resources to chito-bioactives.

Multi-functional carboxymethyl chitin-based nanoparticles for modulation of tumor-associated macrophage polarity.

Current challenge of using cytokines is its poor distribution and systemic side effects. To avoid this issue, we prepared the tumor-targeted and microenvironment-responsive nanocarriers (TRN), which were consisted of α-tocopheryl succinate (α-TOS) loaded mesoporous silica nanoparticles as cores, and surface-modified by thioketal-linkage, electrostatically coated with carboxymethyl chitin, and further anchored glucose-regulated protein 78-binding peptide as shells for encapsulating IL-12. TRN showed a size of 260 nm after encapsulated IL-12 and α-TOS with loading content of 0.0206% and 7.21%, respectively, and exhibited good biocompatibility to 4 T1 cells and macrophages. Moreover, IL-12/α-TOS loaded TRN displayed obvious anti-tumor efficacy on BALB/c nude mice bearing 4 T1 tumors, which was derived from promoted targeting to tumor tissue, endocytosed by macrophages and locally release IL-12 to subsequently repolarize tumor-associated macrophages into tumoricidal M1 phenotype with reduced side effects. The nanosystem exhibited as a promising strategy with functional conversion of macrophages in tumor microenvironment for anti-tumor therapy.

Biochemical characterization of a novel acidic chitinase with antifungal activity from Paenibacillus xylanexedens Z2-4.

A chitinase gene (PxChi52) from Paenibacillus xylanexedens Z2-4 was cloned and heterologously expressed in Escherichia coli BL21 (DE3). PxChi52 shared the highest identity of 91% with a glycoside hydrolase family 18 chitinase (ChiD) from Bacillus circulans. The recombinant enzyme (PxChi52) was purified and biochemically characterized. PxChi52 had a molecular mass of 52.8 kDa. It was most active at pH 4.5 and 65 °C, respectively, and stable in a wide pH range of 4.0-13.0 and up to 50 °C. The enzyme exhibited the highest specific activity of 16.0 U/mg towards colloidal chitin, followed by ethylene glycol chitin (5.4 U/mg) and ball milled chitin (0.4 U/mg). The Km and Vmax values of PxChi52 towards colloidal chitin were determined to be 3.06 mg/mL and 71.38 U/mg, respectively, PxChi52 hydrolyzed colloidal chitin and chitooligosaccharides with degree of polymerization 2-5 to release mainly N-acetyl chitobiose. In addition, PxChi52 displayed inhibition effects on the growth of some phytopathogenic fungi, including Alternaria alstroemeriae, Botrytis cinerea, Rhizoctonia solani, Sclerotinia sclerotiorum and Valsa mali. The unique properties of PxChi52 may enable it potential application in agriculture field as a biocontrol agent.

Alginate-chitosan oligosaccharide-ZnO composite hydrogel for accelerating wound healing.

Moist, breathable and antibacterial microenvironment can promote cell proliferation and migration, which is beneficial to wound healing. Here, we fabricated a novel sodium alginate-chitosan oligosaccharide­zinc oxide (SA-COS-ZnO) composite hydrogel by spontaneous Schiff base reaction, using aldehydated sodium alginate (SA), chitosan oligosaccharide (COS), and zinc oxide (ZnO) nanoparticles, which can provide a moist and antibacterial environment for wound healing. The porosity and swelling degree of SA-COS-ZnO hydrogel are 80% and 150%, respectively, and its water vapor permeability is 682 g/m2/24h. The composite hydrogel showed good biocompatibility to blood cells, 3T3 cells, and 293T cells, and significant antibacterial activity against Escherichia coli, Staphylococcus aureus, Candida albicans, and Bacillus subtilis. Moreover, the hydrogel showed a promoting effect on wound healing in a rat scald model. The present study suggests that marine carbohydrates composite hydrogels are promising in wound care management.

The impact of chitooligosaccharides and their derivatives on the in vitro and in vivo antitumor activity: A comprehensive review.

Chitooligosaccharides (COS) are the degraded products of chitin or chitosan. COS is water-soluble, non-cytotoxic to organisms, readily absorbed through the intestine, and eliminated primarily through the kidneys. COS possess a wide range of biological activities, including immunomodulation, cholesterol-lowering, and antitumor activity. Although work on COS goes back at least forty years, several aspects remain unclear. This review narrates the recent developments in COS antitumor activities, while paying considerable attention to the impacts of physicochemical properties (such as molecular weight and degrees of deacetylation) and chemical modifications both in vitro and in vivo. COS derivatives not only improve some physicochemical properties, but also expand the range of applications in drug and gene delivery. COS (itself or as a drug carrier) can inhibit tumor cell proliferation and metastasis, which might be attributed to its ability to stimulate the immune response along with its anti-angiogenic activity. Further, an attempt has been made to report limitations and future research. The potential health benefits of COS and its derivatives against cancer may offer a new insight on their applications in food and medical fields.

Interaction between bovine serum albumin and chitooligosaccharides: I. Molecular mechanism.

The interaction between chitooligosaccharides (COS2-6) and bovine serum albumin (BSA) is worthy of investigation, which provides support for improving the physical properties (gelling, foaming, and emulsifying) of food proteins via COS addition and in vivo research on COS bioactivity. Component analysis indicated that COS2 and COS3 were enriched in the COS2-6-BSA precipitate. The fluorescence binding constant (1.73 × 103 M-1), ΔG of isothermal titration calorimetry (-6.7 kJ/mol), and the predicted ΔG of molecular docking (-10 to -5 kJ/mol) confirmed the weak interaction of COS2-6-BSA. Quartz crystal microbalance dissipation and molecular docking indicated that electrostatic and hydrophobic interactions were the main stabilization forces. Molecular docking showed that the predicted ΔG of COS2-6 to BSA decreased with the increasing degree of polymerization. This work clarified the weak and selective interaction between COS2-6 and BSA via various methods, which is useful for the food application of COS.

Characterization of Different Salt Forms of Chitooligosaccharides and Their Effects on Nitric Oxide Secretion by Macrophages.

In this paper, chitooligosaccharides in different salt forms, such as chitooligosaccharide lactate, citrate, adipate, etc., were prepared by the microwave method. They were characterized by SEM, FTIR, NMR, etc., and the nitric oxide (NO) expression was determined in RAW 264.7 cells. The results showed that pure chitooligosaccharide was an irregular spherical shape with rough surface, and its different salt type products are amorphous solid with different honeycomb sizes. In addition to the characteristic absorption peaks of chitooligosaccharides, in FTIR, the characteristic absorption of carboxyl group, methylene group, and aromatic group in corresponding acid appeared. The characteristic absorption peaks of carbon in carboxyl group, hydrogen and carbon in methyl, methylene group, and aromatic group in corresponding acid also appeared in NMR. Therefore, the sugar ring structure and linking mode of chitooligosaccharides did not change after salt formation of chitooligosaccharides. Different salt chitooligosaccharides are completely different in promoting NO secretion by macrophages, and pure chitooligosaccharides are the best.

Biochemical characterization of a novel bifunctional chitosanase from Paenibacillus barengoltzii for chitooligosaccharide production.

A novel chitosanase gene, designated as PbCsn8, was cloned from Paenibacillus barengoltzii. It shared the highest identity of 73% with the glycoside hydrolase (GH) family 8 chitosanase from Bacillus thuringiensis JAM-GG01. The gene was heterologously expressed in Bacillus subtilis as an extracellular protein, and the highest chitosanase yield of 1, 108 U/mL was obtained by high-cell density fermentation in a 5-L fermentor. The recombinant chitosanase (PbCsn8) was purified to homogeneity and biochemically characterized. PbCsn8 was most active at pH 5.5 and 70 °C, respectively. It was stable in a wide pH range of 5.0-11.0 and up to 55 °C. PbCsn8 was a bifunctional enzyme, exhibiting both chitosanase and glucanase activities, with the highest specificity towards chitosan (360 U/mg), followed by barley ß-glucan (72 U/mg) and lichenan (13 U/mg). It hydrolyzed chitosan to release mainly chitooligosaccharides (COSs) with degree of polymerization (DP) 2-3, while hydrolyzed barley ß-glucan to yield mainly glucooligosaccharides with DP > 5. PbCsn8 was further applied in COS production, and the highest COS yield of 79.3% (w/w) was obtained. This is the first report on a GH family 8 chitosanase from P. barengoltzii. The high yield and remarkable hydrolysis properties may make PbCsn8 a good candidate in industrial application.

Composite of MOF and chitin as an efficient catalyst for photodegradation of organic dyes.

A novel composite has been fabricated by using MOF and chitin as a natural and biocompatible compound. To this purpose, MOF was synthesized by using 2-aminoterephthalic acid and iron (III) chloride hexahydrate and then reacted with Cl-functionalized chitin. The resulting composite was characterized and utilized as a catalyst for degradation of methylene blue both in dark condition and under visible light irradiation. The results indicated superior catalytic activity under visible light irradiation. Furthermore, study of the reaction variables, including basicity, dye concentration and catalyst loading showed that the highest catalytic activity was achieved at basic condition. It was also found that both initial dye concentration and catalyst loading can affect the catalytic activity. To disclose the merits of the composite compared to its individual components, kinetic studies of the photo-degradation process in the presence of the composite, chitin and MOF have been performed. The results confirmed superior activity the composite compared to its components. The study of the mechanism of the reaction using scavengers confirmed that the created holes (h+) are the most effective species in the process of photocatalytic degradation of MB. Notably, the catalyst was recyclable and could be used for degradation of other dyes.

Construction of hydrogels based on the homogeneous carboxymethylated chitin from Hericium erinaceus residue: Role of carboxymethylation degree.

Carboxymethyl chitin hydrogels with different degree of substitution (DS) were prepared by the homogeneous carboxymethylation of chitin extracted from Hericium erinaceus residue. The effect of DS on gel structure and property were studied. Results showed that the DS of carboxymethyl chitin hydrogels can be increased by increasing the amount of sodium chloroacetate. The equilibrium swelling degree and pH swelling sensitivity of the hydrogels were enhanced as the increase of DS. Zeta potential, low-field nuclear magnetic resonance, contact angle and molecular dynamics simulation results suggested that the introduction of carboxymethyl functional group enhanced the negative charge, water mobility, surface hydrophilicity and the ability to form hydrogen bonds with water of the hydrogels, resulting in an increased swelling degree of the hydrogels. Moreover, the prepared hydrogels showed different adsorption capability to various dyes, and the adsorption performance of the prepared hydrogels for cationic dyes could be enhanced as the increase of DS.

Antimicrobial Activity of Chitosan Oligosaccharides with Special Attention to Antiparasitic Potential.

The global rise of infectious disease outbreaks and the progression of microbial resistance reinforce the importance of researching new biomolecules. Obtained from the hydrolysis of chitosan, chitooligosaccharides (COSs) have demonstrated several biological properties, including antimicrobial, and greater advantage over chitosan due to their higher solubility and lower viscosity. Despite the evidence of the biotechnological potential of COSs, their effects on trypanosomatids are still scarce. The objectives of this study were the enzymatic production, characterization, and in vitro evaluation of the cytotoxic, antibacterial, antifungal, and antiparasitic effects of COSs. NMR and mass spectrometry analyses indicated the presence of a mixture with 81% deacetylated COS and acetylated hexamers. COSs demonstrated no evidence of cytotoxicity upon 2 mg/mL. In addition, COSs showed interesting activity against bacteria and yeasts and a time-dependent parasitic inhibition. Scanning electron microscopy images indicated a parasite aggregation ability of COSs. Thus, the broad biological effect of COSs makes them a promising molecule for the biomedical industry.

Computational Assessment of Chito-Oligosaccharides Interactions with Plasma Proteins.

It is widely recognized that chitin and chitosan are potential sources of bioactive materials and that their oligosaccharides reveal various biological activities (including antimicrobial) that are correlated with their structures and physicochemical properties. This study uses the molecular docking approach to assess the interactions of small chito-oligosaccharides (MW< 1500 Da) with plasma proteins in order to obtain information regarding their fate of distribution in the human organism. There are favorable interactions of small chito-oligomers with plasma proteins, the interactions with human serum albumin being stronger than those with α-1-acid glycoprotein. The interaction energies increase with increasing the molecular weight, decrease with increasing deacetylation degrees and are reliant on the deacetylation pattern. This study could inform the application of chito-oligosaccharides with varying molecular weights, degrees, and patterns of deacetylation in human health.

Distinct genetic basis for root responses to lipo-chitooligosaccharide signal molecules from different microbial origins.

Lipo-chitooligosaccharides (LCOs) were originally found as symbiotic signals called Nod Factors (Nod-LCOs) controlling the nodulation of legumes by rhizobia. More recently, LCOs were also found in symbiotic fungi and, more surprisingly, very widely in the kingdom Fungi, including in saprophytic and pathogenic fungi. The LCO-V(C18:1, fucosylated/methyl fucosylated), hereafter called Fung-LCOs, are the LCO structures most commonly found in fungi. This raises the question of how legume plants such as Medicago truncatula can discriminate between Nod-LCOs and Fung-LCOs. To address this question, we performed a genome-wide association study on 173 natural accessions of M. truncatula, using a root branching phenotype and a newly developed local score approach. Both Nod-LCOs and Fung-LCOs stimulated root branching in most accessions, but the root responses to these two types of LCO molecules were not correlated. In addition, the heritability of the root response was higher for Nod-LCOs than for Fung-LCOs. We identified 123 loci for Nod-LCO and 71 for Fung-LCO responses, of which only one was common. This suggests that Nod-LCOs and Fung-LCOs both control root branching but use different molecular mechanisms. The tighter genetic constraint of the root response to Fung-LCOs possibly reflects the ancestral origin of the biological activity of these molecules.