Development of chitosan based microencapsulated spray dried powder of tuna fish oil: oil load impact and oxidative stability / Desenvolvimento de pó microencapsulado por pulverização de óleo de atum à base de quitosana: impacto da carga de óleo e estabilidade oxidativa

Abstract The impact of fish oil concentration on the oxidative stability of microcapsules through the spray drying process using chitosan and maltodextrin as wall material was studied. Emulsions were prepared with different Tuna fish oil (TFO) content (TFO-10%, TFO20%, TF030% TF0-40%) while wall material concentration was kept constant. Microencapsulated powder resulting from emulsion prepared with high fish oil load have high moisture content, wettability, total oil and low encapsulation efficiency, hygroscopicity and bulk tapped density. Oxidative stability was evaluated periodically by placing microcapsules at room temperature. Microcapsules prepared with TFO-10% presented high oxidative stability in terms of peroxide value (2.94±0.04) and anisidine value (1.54±0.02) after 30 days of storage. It was concluded that optimal amounts of fish oil for microencapsulation are 10% and 20% using chitosan and maltodextrin that extended its shelf life during study period.

Resumo Foi estudado o impacto da concentração de óleo de peixe na estabilidade oxidativa de microcápsulas por meio do processo de secagem por atomização, utilizando quitosana e maltodextrina como material de parede. As emulsões foram preparadas com diferentes teores de óleo de atum (TFO) (TFO-10%, TFO20%, TF030% TF0-40%), enquanto a concentração de material de parede foi mantida constante. O pó microencapsulado resultante da emulsão preparada com alta carga de óleo de peixe tem alto teor de umidade, molhabilidade e óleo total e baixa eficiência de encapsulação, higroscopicidade e densidade extraída a granel. A estabilidade oxidativa foi avaliada periodicamente colocando microcápsulas à temperatura ambiente. As microcápsulas preparadas com TFO-10% apresentaram alta estabilidade oxidativa em termos de valor de peróxido (2,94 ± 0,04) e valor de anisidina (1,54 ± 0,02) após 30 dias de armazenamento. Concluiu-se que as quantidades ideais de óleo de peixe para microencapsulação são de 10% e 20% usando quitosana e maltodextrina que prolongaram sua vida útil durante o período de estudo.

pH-responsive chitosan/acrylamide/gold/nanocomposite supported with silver nanoparticles for controlled release of anticancer drug.

In this study, we prepared a pH-responsive nanocomposite hydrogel based on chitosan grafted with acrylamide monomer and gold nanoparticles using gamma irradiation method (Cs-g-PAAm/AuNPs). The nanocomposite was enhanced with a layer coating of silver nanoparticles to improve the controlled release of the anticancer drug fluorouracil while increasing antimicrobial activity and decreasing the cytotoxicity of silver nanoparticles in nanocomposite hydrogel by combining with gold nanoparticles to enhance the ability to kill a high number of liver cancer cells. The structure of the nanocomposite materials was studied using FTIR spectroscopy and XRD patterns, which demonstrated the entrapment of gold and silver nanoparticles within the prepared polymer matrix. Dynamic light scattering data revealed the presence of gold and silver in the nanoscale with the polydispersity indexes in the mid-range values, indicating that distribution systems work best. Swelling experiments at various pH levels revealed that the prepared Cs-g-PAAm/Au-Ag-NPs nanocomposite hydrogels were highly responsive to pH changes. Bimetallic pH-responsive Cs-g-PAAm/Au-Ag-NPs nanocomposites exhibit strong antimicrobial activity. The presence of AuNPs reduced the cytotoxicity of AgNPs while increasing their ability to kill a high number of liver cancer cells.Cs-g-PAAm/Au-Ag-NPs has a high amount of fluorouracil drug loaded at pH 7.4 reaching 95 mg/g with a maximum drug release of 97% within 300 min. Cs-g-PAAm/Au-Ag-NPs have been recommended to use as oral delivery of anticancer drugs because they secure the encapsulated drug in the acidic medium of the stomach and release it in the intestinal pH.

Antimicrobial and Osteogenic Effects of Collagen Membrane Decorated with Chitosan-Nano-Hydroxyapatite.

Collagen membranes are routinely used in oral surgery for bone regeneration. Despite their numerous advantages, such as stimulating bone growth, bacterial contamination still remains one of the disadvantages of membrane use. Thus, we assessed the biocompatibility and osteogenic and antibacterial properties of a collagen membrane (OsteoBiol) modified with chitosan (CHI) and hydroxyapatite nanoparticles (HApNPs). Attenuated total reflectance-Fourier transform infrared spectroscopy (ATR FT-IR), X-ray powder diffraction (XRD), and field emission scanning electron microscopy (FE-SEM) were performed for membrane characterization. Biocompatibility was assessed on dental pulp stem cells (DPSCs) by an MTT assay, while the osteogenic effect was assessed by an ALP activity assay and qPCR analysis of osteogenic markers (BMP4, ALP, RUNX2, and OCN). Antimicrobial properties were investigated by counting colony-forming units (CFUs) of Streptococcus mitis, Porphyromonas gingivalis, and Fusobaterium nucleatum on membranes and in the surrounding medium. Membranes showed no cytotoxicity. ALP activity was higher and ALP, BMP4, and OCN genes were up-regulated in DPSCs on modified membranes compared to unmodified membranes. The CFUs were reduced on modified membranes and in the medium. Modified membranes showed great biocompatibility and a high osteoinductive effect. Additionally, they showed antimicrobial and antibiofilm effects against periopathogens. It can be concluded that the incorporation of CHI and hydroxyapatite nanoparticles in collagen membranes may be advantageous to promote osteogenesis and reduce bacterial adhesion.

Tyrosinase Magnetic Cross-Linked Enzyme Aggregates: Biocatalytic Study in Deep Eutectic Solvent Aqueous Solutions.

In the field of biocatalysis, the implementation of sustainable processes such as enzyme immobilization or employment of environmentally friendly solvents, like Deep Eutectic Solvents (DESs) are of paramount importance. In this work, tyrosinase was extracted from fresh mushrooms and used in a carrier-free immobilization towards the preparation of both non-magnetic and magnetic cross-linked enzyme aggregates (CLEAs). The prepared biocatalyst was characterized and the biocatalytic and structural traits of free tyrosinase and tyrosinase magnetic CLEAs (mCLEAs) were evaluated in numerous DES aqueous solutions. The results showed that the nature and the concentration of the DESs used as co-solvents significantly affected the catalytic activity and stability of tyrosinase, while the immobilization enhanced the activity of the enzyme in comparison with the non-immobilized enzyme up to 3.6-fold. The biocatalyst retained the 100% of its initial activity after storage at -20 °C for 1 year and the 90% of its activity after 5 repeated cycles. Tyrosinase mCLEAs were further applied in the homogeneous modification of chitosan with caffeic acid in the presence of DES. The biocatalyst demonstrated great ability in the functionalization of chitosan with caffeic acid in the presence of 10% v/v DES [Bet:Gly (1:3)], enhancing the antioxidant activity of the films.

Quaternization of high molecular weight chitosan for increasing intestinal drug absorption using Caco-2 cells as an in vitro intestinal model.

Potential use of a quaternized chitosan (MW 600 kDa) with 65% of 3-chloro-2-hydroxypropyltrimethylammonium (600-HPTChC65) as an absorptive enhancer was investigated in Caco-2 monolayers. 600-HPTChC65 (0.005% w/v) quickly reduced transepithelial electrical resistance (TEER) to the maximum level in 40 min with full recovery within 6 h after removal. Its TEER reduction was corresponded to increased FD4 transport across the monolayers and disrupted localization of tight junction proteins ZO-1 and occludin at the cell borders. 600-HPTChC65 was densely localized at the membrane surface and intercellular junctions. This chitosan (0.08-0.32% w/v) reduced the efflux ratio of [3H]-digoxin by 1.7- 2 folds, suggesting an increased [3H]-digoxin transport across the monolayers. Its binding with P-gp on Caco-2 monolayer increased the signal of fluorescence-labeled anti-P-gp (UIC2) reactivity due to conformational change. 600-HPTChC65 (0.32% w/v) had no effect on P-gp expression in the Caco-2 monolayers. These results suggest that 600-HPTChC65 could enhance drug absorption through tight junction opening and decreased P-gp function. Its interaction with the absorptive barrier mainly resulted in disrupting ZO-1 and occludin organization as well as changing in P-gp conformation.

Synthesis of L-methionine-loaded chitosan nanoparticles for controlled release and their in vitro and in vivo evaluation.

Therapeutically popular controlled release-enabling technology has forayed into the nutrition sector. Polymer coated forms of L-methionine used in soy protein diets, and its intermediate metabolite, S-adenosyl-L-methionine, used in myriad of medical conditions have proved more efficacious over (highly catabolized) free forms. In this premier study, L-methionine-loaded chitosan nanoparticles (M-NPs) were synthesized using ionic gelation method and their efficacy was evaluated. Biophysical characterization of the NPs was done using a Nanopartica SZ 100 analyser, transmission electron microscopy, and Fourier transform infrared spectroscopy. The M-NPs were spherical and smooth and 218.9 ± 7.4 nm in size and in vitro testing confirmed the controlled release of methionine. A 60-days feeding trial in L. rohita fish fingerlings was conducted. A basal diet suboptimal (0.85%) in methionine was provided with one of the supplements as under: none (control), 0.8% chitosan NPs (0.8% NPs), 1.2% L-methionine (1.2% M) (crystalline free form), 0.6% M-NPs and 1.2% M-NPs. While the addition of 0.6% M-NPs to the basal diet complemented towards meeting the established dietary requirement and resulted in significantly highest (P < 0.05) growth and protein efficiency and sero-immunological test scores (serum total protein, serum globulin, serum albumin: globulin ratio, phagocytic respiratory burst/NBT reduction and lysozyme activity), 1.2% supplementation in either form (free or nano), for being 0.85% excess, was counterproductive. Liver transaminases and dehydrogenases corroborated enhanced growth. It was inferred that part of the methionine requirement in nano form (M-NPs) can confer intended performance and health benefits in animals relying on plant proteins-based diets limiting in this essential amino acid. The study also paves the way for exploring chitosan NPs-based sustained delivery of amino acids in human medical conditions.

[Effects of chitosan oligosaccharide on bone metabolism and IKK/NF-κB pathway in rats with osteoporosis and periodontitis].

PURPOSE: To investigate the effects of chitosan oligosaccharide on bone metabolism and IKK/NF-κB pathway in mice with osteoporosis and periodontitis. METHODS: Thirty rats were randomly divided into 3 groups, with 10 rats in each group. They were divided into control group, ovariectomized periodontitis group and chitosan oligosaccharide treatment group. Except for the control group, the other two groups were ovariectomized and smeared with Porphyromonas gingivalis fluid to establish the model of osteoporosis with periodontitis. Four weeks after ligation, the rats in chitosan oligosaccharide treatment group were gavaged with 200 mg/kg chitosan oligosaccharide, and the other two groups were gavaged with equal volume of normal saline once a day for 90 days. The periodontal tissues of each group were observed before administration, and the bone mineral density of rats was detected by dual energy X-ray animal bone mineral density and body composition analysis system. After 90 days of administration, the bone mineral density was detected again. After administration, blood was collected from tail vein, and the contents of serum alkaline phosphatase (ALP), bone Gla protein (BGP) and tartrate resistant acid phosphatase 5b (TRACP5b) were measured by enzyme-linked immunodeficient assay. The gingival index and periodontal attachment loss of rats in each group were obtained by visual examination and exploratory examination. The maxilla was removed, and the distance from the enamel cementum boundary to the alveolar crest was measured to obtain alveolar bone absorption value. H-E staining was used to observe the pathology of maxilla in each group. RT-PCR and Western blot were used to detect the nuclear factors in periodontal tissue of rats in each group. SPSS 22.0 software package was used for statistical analysis. RESULTS: Before administration, the gums of the control group were pink without bleeding, and the gums of the other two groups were red and swollen with slight bleeding. After administration, compared with the control group, the bone mineral density, serum ALP, BGP of ovariectomized periodontitis group decreased significantly(P＜0.05); while TRACP5b, gingival index, loss of periodontal attachment and alveolar bone resorption, NF-κB and IKK mRNA and protein expression in periodontal tissue increased significantly(P＜0.05). Compared with the ovariectomized periodontitis group, the bone mineral density, serum ALP, BGP were significantly increased(P＜0.05); while TRACP5b, gingival index, periodontal attachment loss and alveolar bone resorption, NF-κB and IKK mRNA and protein expression in periodontal tissue were significantly decreased (P＜0.05). In the ovariectomized periodontitis group, the periodontal tissue combined with epithelium was separated from the tooth surface, the dental pocket was obvious and deep, and the height of alveolar bone decreased. Although dental pocket could be observed in the periodontal tissue of rats treated with chitosan oligosaccharide, it was not obvious, and new bone appeared around the alveolar bone. CONCLUSIONS: Chitosan oligosaccharide can induce biochemical indexes of bone metabolism to become normal, alleviate the symptoms of periodontitis, this may be related to the inhibition of IKK/NF-κB pathway by chitosan oligosaccharide.

Facile synthesis of hydroxypropyl chitosan-egg white hydrogel dressing with antibacterial and antioxidative activities for accelerating the healing of burn wounds.

Burn injury is the fourth most common injury worldwide. Deep partial-thickness burns are susceptible to bacterial infections due to the absence of a skin shield, which can lead to severe pain, scarring, and even death. Therefore, developing a wound dressing that can promote wound repair accompanied by excellent antibacterial effects is crucial for clinical application. Herein, a facile self-healing hydroxypropyl chitosan-egg white hydrogel (HPCS-EWH) with excellent biocompatibility, antioxidant activity, anti-inflammation and antibacterial properties was prepared. This physical crosslinking hydrogel was endowed with the intrinsic merits of its parental components, such as reactive oxygen species (ROS) scavenging, antibiosis, and thriving cell growth in vitro. In an in vivo model of Staphylococcus aureus-infected burn wounds, HPCS-EWH could accelerate wound healing due to its anti-inflammatory and antibacterial activities, and promote cell proliferation and angiogenesis. Therefore, HPCS-EWH could be used to heal deep partial-thickness skin burn wounds.

Development of an Enzymatic Biosensor Using Glutamate Oxidase on Organic-Inorganic-Structured, Electrospun Nanofiber-Modified Electrodes for Monosodium Glutamate Detection.

Herein, dendrimer-modified montmorillonite (Mt)-decorated poly-Ɛ-caprolactone (PCL) and chitosan (CHIT)-based nanofibers were prepared. Mt was modified with a poly(amidoamine) generation 1 (PAMAMG1) dendrimer, and the obtained PAMAMG1-Mt was incorporated into the PCL-CHIT nanofiber's structure. The PCL-CHIT/PAMAMG1-Mt nanofibers were conjugated with glutamate oxidase (GluOx) to design a bio-based detection system for monosodium glutamate (MSG). PAMAMG1-Mt was added to the PCL-CHIT backbone to provide a multipoint binding side to immobilize GluOx via covalent bonds. After the characterization of PCL-CHIT/PAMAMG1-Mt/GluOx, it was calibrated for MSG. The linear ranges were determined from 0.025 to 0.25 mM MSG using PCL-CHIT/Mt/GluOx and from 0.0025 to 0.175 mM MSG using PCL-CHIT/PAMAMG1-Mt/GluOx (with a detection limit of 7.019 µM for PCL-CHIT/Mt/GluOx and 1.045 µM for PCL-CHIT/PAMAMG1-Mt/GluOx). Finally, PCL-CHIT/PAMAMG1-Mt/GluOx was applied to analyze MSG content in tomato soup without interfering with the sample matrix, giving a recovery percentage of 103.125%. Hence, the nanofiber modification with dendrimer-intercalated Mt and GluOx conjugation onto the formed nanocomposite structures was performed, and the PCL-CHIT/PAMAMG1-Mt/GluOx system was successfully developed for MSG detection.

Wettability of mg-ha/Chitosan-based membrane surfaces: blood vs. autologous platelet liquid (APL).

OBJECTIVE: The physical and physical chemistry is able to influence the interaction of the scaffolds and bone substitutes with the body fluid and blood. The aim of the present investigation was to evaluate the wettability properties of an Mg-HA Chitosan-based Gel with blood vs. autologous platelet gel. MATERIALS AND METHODS: A total of 6 study groups were evaluated according to the Mg-HA Chitosan-based Gel thickness (1, 2 and 3 mm) and the fluids (blood vs. autologous platelet gel). The biomaterial wettability was conducted through the sessile drop technique. RESULTS: The study findings showed a significant difference in contact angle between the APL and blood groups (p<0.05). The MG-Ha Chitosan-based membrane thicknesses seem to produce no significant effects on contact angles measurement for all groups (p>0.05). CONCLUSIONS: In the present investigation, a similar MG/Ha gel membranes wettability was reported between APL and blood groups. In addition, a high hydrophilicity of MG/Ha gel membranes was reported with a potential advantage in terms of a more effective osteogenic capability in the clinical practice.

Modifications in steroid and triterpenoid metabolism in Calendula officinalis plants and hairy root culture in response to chitosan treatment.

BACKGROUND: Chitosan, a deacetylated derivative of chitin, is one of the most preferred biopolymers for use as biostimulants and biofertilizers in organic agriculture and as elicitors to enhance the productivity of plant in vitro cultures. Valued as a non-toxic, biodegradable, and environment-friendly agent, it is widely applied to improve plant growth and yield, the content of bioactive specialized metabolites, and resistance to stress conditions and pathogens. However, the influence of chitosan on the growth-defense trade-off, particularly the interplay between steroid and triterpenoid metabolism, has not been extensively investigated. RESULTS: In this study, Calendula officinalis pot plants and hairy root cultures exposed to chitosan treatment displayed reduced biomass and altered steroid and triterpenoid metabolism. Biosynthesis and accumulation of free forms of sterols (particularly stigmasterol) were inhibited, while the content of sterol esters increased remarkably. The content of some triterpenoids (mainly free triterpenoid acids) was slightly enhanced; however, the biosynthesis of triterpenoid saponins was negatively affected. CONCLUSIONS: These results indicate that in certain plants, chitosan treatment might not positively influence the growth and metabolite production. Therefore, to avoid unexpected effects, initial studies of the conditions of chitosan treatment are recommended, including the dose and the number of chitosan applications, the type of treatment (e.g., foliar or soil), and the vegetative stage of the treated plants.

Chitosan-grafted-caffeic acid combined with ultrasound inhibits the oxidation and degradation of myofibrillar proteins in pompano (Trachinotus ovatus) during ice storage.

This study aimed to investigate the impact of chitosan-grafted-caffeic acid (CS-g-CA) and ultrasound (US) on myofibrillar proteins (MPs) in pompano (Trachinotus ovatus) during 24 days of ice storage. Fresh fish slices were treated with US (20 kHz, 600 W), CS-g-CA (G), and US combined with CS-g-CA (USG) for 10 min, respectively. Samples treated with sterile water served as study controls (CK). All samples were then stored in ice at 4 °C. The oxidation and degradation of MPs were evaluated at 4-day intervals. The results showed that US slightly accelerated the fragmentation of myofibrils, as confirmed by the increased myofibril fragmentation index (MFI). However, on day 24, the surface hydrophobicity (SH) of USG samples was 4.09 µg BPB bound/mg protein lower than that of G samples, and the total sulfhydryl content of USG samples was 0.50 µmol g-1 higher than that of G samples, suggesting that US could reinforce the antioxidant capacity of CS-g-CA. Regarding degradation of MPs, USG treatment maintained the secondary and tertiary structure of MPs by reducing the transition from ordered to disordered structures and by reducing the exposure of tryptophan residues. Sodium dodecyl sulphate- polyacrylamide gel electrophoresis (SDS-PAGE) showed that the inhibitory effect of USG on protein degradation may be related to the binding of CS-g-CA to MPs. The results of scanning electron microscopy (SEM) further clarified the fact that the USG treatment can protect the myofibril microstructure by maintaining the compact arrangement of muscle fibers. Additionally, USG treatment could improve the sensory properties of pompano. Overall, the synergistic effects of US and CS-g-CA can effectively delay the protein oxidation and degradation. The results provided in this study are valuable for the quality maintenance of marine fish.

Controlled drug delivery from chitosan-coated heparin-loaded nanopores anodically grown on nitinol shape-memory alloy.

Nitinol (NiTi shape-memory alloy) is an interesting candidate in various medical applications like dental, orthopedic, and cardiovascular devices, owing to its unique mechanical behaviors and proper biocompatibility. The aim of this work is the local controlled delivery of a cardiovascular drug, heparin, loaded onto nitinol treated by electrochemical anodizing and chitosan coating. In this regard, the structure, wettability, drug release kinetics, and cell cytocompatibility of the specimens were analyzed in vitro. The two-stage anodizing process successfully developed a regular nanoporous layer of Ni-Ti-O on nitinol, which considerably decreased the sessile water contact angle and induced hydrophilicity. The application of the chitosan coatings controlled the release of heparin mainly by a diffusional mechanism, where the drug release mechanisms were evaluated by the Higuchi, first-order, zero-order, and Korsmeyer-Pepass models. Human umbilical cord endothelial cells (HUVECs) viability assay also showed the non-cytotoxicity of the samples, so that the best performance was found for the chitosan-coated samples. It is concluded that the designed drug delivery systems are promising for cardiovascular, particularly stent applications.

Magnetic chitosan hydrogel induces neuronal differentiation of neural stem cells by activating RAS-dependent signal cascade.

Our aim was to modulate magnetic cues to influence the differentiation of neural stem cell (NSC) into neuron during nerve repair and to explore corresponding mechanisms. Here, a magnetic hydrogel composed of chitosan matrices and magnetic nanoparticles (MNPs) with different content was prepared as the magnetic-stimulation platform to apply intrinsically-present magnetic cue and externally-applied magnetic field to NSC grown on the hydrogel. The MNP content had regulatory effects on neuronal differentiation and the MNPs-50 samples exhibited the best neuronal potential and appropriate biocompatibility in vitro, as well as accelerated the subsequent neuronal regeneration in vivo. Remarkably, the use of proteomics analysis parsed the underlying mechanism of magnetic cue-mediated neuronal differentiation form the perspective of protein corona and intracellular signal transduction. The intrinsically-present magnetic cues in hydrogel contributed to the activation of intracellular RAS-dependent signal cascades, thus facilitating neuronal differentiation. Magnetic cue-dependent changes in NSCs benefited from the upregulation of adsorbed proteins related to "neuronal differentiation", "cell-cell interaction", "receptor", "protein activation cascade", and "protein kinase activity" in the protein corona. Additionally, magnetic hydrogel acted cooperatively with the exterior magnetic field, showing further improving neurogenesis. The findings clarified the mechanism for magnetic cue-mediated neuronal differentiation, coupling protein corona and intracellular signal transduction.

Drug-free and non-crosslinked chitosan/hyaluronic acid hybrid hydrogel for synergistic healing of infected diabetic wounds.

The management of infected diabetic wounds remains a major challenge in clinical practice. Recently, multifunctional hydrogels have attracted much attention in the area of wound healing. Herein, we developed the drug-free and non-crosslinked chitosan (CS)/hyaluronic acid (HA) hybrid hydrogel, so as to combine the multiple functions of CS and HA for synergistic healing of the methicillin-resistant Staphylococcus aureus (MRSA)-infected diabetic wound. As a result, CS/HA hydrogel showed the broad-spectrum antibacterial activity, the great capacity for promoting fibroblasts proliferation and migration, the excellent reactive oxygen species (ROS) scavenging ability, and the great cell-protection effects under oxidative stress. In the MRSA-infected diabetic mouse wounds, CS/HA hydrogel significantly promoted the wound healing via eliminating MRSA infection and enhancing epidermal regeneration, collagen deposition and angiogenesis. Considering the drug-free feature, the ready availability, the great biocompatibility and the excellent wound healing efficacy, CS/HA hydrogel may have great potentials in clinical use for the management of chronic diabetic wounds.

Guanidinylated/PEGylated chitosan in the bioink promotes the formation of multi-layered keratinocytes in a human skin equivalent.

The biological differences of skin between rodent and human beings and the strong appeal to replace the experimental animals have led to the development of alternative models with structures similar to the real human skin. Keratinocytes cultured in vitro on conventional dermal scaffolds tend to form monolayer rather than multi-layer epithelial tissue architectures. How to construct human skin or epidermal equivalents with multi-layered keratinocytes similar to real human epidermis remains one of the greatest challenges. Herein, a human skin equivalent with multi-layered keratinocytes was constructed by 3D bioprinting fibroblasts and subsequent culturing epidermal keratinocytes. Biocompatible guanidinylated/PEGylated chitosan (GPCS) was used as the main component of bioink to 3D bioprint tissue-engineered dermis. The function of GPCS to promote HaCat cell proliferation and connection was confirmed at the genetic, cellular, and histological levels. Compared with the skin tissues with mono-layered keratinocytes engineered with collagen and gelatin, adding GPCS in the bioink generated tissue-engineered human skin equivalents with multi-layered keratinocytes. Such human skin equivalents could be alternative models for biomedical, toxicological, and pharmaceutical research.

Application of chitosan with different molecular weights in cartilage tissue engineering.

Cartilage tissue engineering involves the invention of novel implantable cartilage replacement materials to help heal cartilage injuries that do not heal themselves, aiming to overcome the shortcomings of current clinical cartilage treatments. Chitosan has been widely used in cartilage tissue engineering because of its similar structure to glycine aminoglycan, which is widely distributed in connective tissues. The molecular weight, as an important structural parameter of chitosan, affects not only the method of chitosan composite scaffold preparation but also the effect on cartilage tissue healing. Thus, this review identifies methods for the preparation of chitosan composite scaffolds with low, medium and high molecular weights, as well as a range of chitosan molecular weights appropriate for cartilage tissue repair, by summarizing the application of different molecular weights of chitosan in cartilage repair in recent years.

UV cross-linked injectable non-swelling dihydrocaffeic acid grafted chitosan hydrogel for promoting wound healing.

Hydrogels are widely used as wound dressings for wound healing, but when hydrogels absorb wound exudate, swelling occurs and compresses the surrounding tissue, affecting healing. A chitosan injectable (CS/4-PA/CAT) hydrogel based on catechol and 4-glutenoic acid was prepared to avoid swelling and promote wound healing. After cross-linking by UV light, pentenyl groups formed hydrophobic alkyl chains which give the hydrogel a hydrophobic network and thus control its swelling. CS/4-PA/CAT hydrogels retained their non-swelling for a long time in PBS solution at 37 °C. CS/4-PA/CAT hydrogels had good injectable and adhesive properties, and had a good killing effect on E. coli and S. aureus and could remove the bacterial biofilms of E. coli and S. aureus. CS/4-PA/CAT hydrogels had good in vitro coagulation function by absorbing red blood cells and platelets. When used in a whole skin injury model, CS/4-PA/CAT-1 hydrogel stimulated fibroblast migration, promoted epithelialization and accelerated collagen deposition to promote defect healing, and showed good hemostatic effects in liver and femoral artery defects in mice. In summary, the non-swelling injectable hydrogel with free radical scavenging, rapid hemostasis, and antibacterial effects would be a promising treatment for defect repair.

Chitosan-grafted phenolic acids as an efficient biopolymer for food packaging films/coatings.

Chitosan (CS), a bio-renewable natural material, has the potential to be utilized as a biopolymer for food packaging films (PFs)/coatings. However, its low solubility in dilute acid solutions and poor antioxidant and antimicrobial activities limit its application in PFs/coatings. To address these restrictions, chemical modification of CS has garnered increasing interest, with graft copolymerization being the most extensively used method. Phenolic acids (PAs) as natural small molecules are used as excellent candidates for CS grafting. This work focuses on the progress of CS grafted PA (CS-g-PA) based films, introducing the chemistry and methods of preparing CS-g-PA, particularly the effects of different PAs grafting on the properties of CS films. In addition, this work discusses the application of different CS-g-PA functionalized PFs/coatings for food preservation. It is concluded that the food preservation capability of CS-based films/coatings can be improved by modifying the properties of CS-based films through PA grafting.

Photodynamic antibacterial chitosan/nitrogen-doped carbon dots composite packaging film for food preservation applications.

In this study, we synthesized nitrogen-doped carbon dots (N-CDs) with remarkable photodynamic antibacterial properties by a hydrothermal method. The composite film was prepared by solvent casting method, compounding N-CDs with chitosan (CS). The morphology and structure of the films were analyzed by Fourier-transformed infrared spectroscopy (FTIR), scanning electron microscope (SEM), atomic force microscope (AFM), and transmission electron microscope (TEM) techniques. The films' mechanical, barrier, thermal stability, and antibacterial properties were analyzed. A preservation test of the films was studied on the samples of pork, volatile base nitrogen (TVB-N), total viable count (TVC), and pH were determined. Besides, the effect of film on the preservation of blueberries was observed. The study found that, compared with the CS film, the CS/N-CDs composite film is strong and flexible, with good UV light barrier performance. The prepared CS/7 % N-CDs composites showed high photodynamic antibacterial rates of 91.2 % and 99.9 % for E. coli and S. aureus, respectively. In the preservation of pork, it was found that its pH, TVB-N, and TVC indicators were significantly lower. The extent of mold contamination and anthocyanin loss was less in the CS/3 % N-CDs composite film-coated group, which could greatly extend the shelf life of food.