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1.
FEMS Microbiol Lett ; 2024 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-39113276

RESUMO

The first steps in chitin degradation in marine bacteria involve chitinase, which produces N,N'-diacetylchitobiose (GlcNAc)2 from chitin. Moreover, in Vibrio bacteria, chitinase activity is enhanced by heterodisaccharide ß-N-acetyl-D-glucosaminyl-(1,4)-D-glucosamine (GlcNAc-GlcN) produced from (GlcNAc)2 by chitin oligosaccharide deacetylase (COD). However, the role of COD in other marine bacteria, such as Shewanella, remains unexplored. This study investigates GlcNAc-GlcN's impact on chitinase gene expression and enzyme production in S. baltica ATCC BAA-1091, drawing parallels with Vibrio parahaemolyticus RIMD2210633. Using real-time quantitative PCR, the study assesses the up-regulation of chitinase gene expression in S. baltica in response to GlcNAc-GlcN, informed by COD's known ability to produce GlcNAc-GlcN from (GlcNAc)2. In Vibrio, GlcNAc-GlcN considerably up-regulates chitinase gene expression. This study posits a similar regulatory mechanism in S. baltica, with preliminary investigations indicating COD's capacity to produce GlcNAc-GlcN. This study highlights the importance of exploring GlcNAc-GlcN's regulatory role in chitin metabolism across diverse marine bacteria. The potential induction of chitinase production in S. baltica suggests broader ecological implications. Further research is crucial for a comprehensive understanding of chitin utilization and regulatory pathways in marine bacterial genera.

2.
Front Microbiol ; 15: 1427143, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39113839

RESUMO

Introduction: Chitin, abundant in marine environments, presents significant challenges in terms of transformation and utilization. A strain, T22.7.1T, with notable chitin deacetylation capabilities, was isolated from the rhizosphere of Acanthus ebracteatus in the North Sea of China. Comparative 16S rDNA sequence analysis showed that the new isolate had the highest sequence similarity (99.79%) with Rhodococcus indonesiensis CSLK01-03T, followed by R. ruber DSM 43338T, R. electrodiphilus JC435T, and R. aetherivorans 10bc312T (98.97%, 98.81%, and 98.83%, respectively). Subsequent genome sequencing and phylogenetic analysis confirmed that strain T22.7.1T belongs to the R. indonesiensis species. However, additional taxonomic characterization identified strain T22.7.1T as a novel type strain of R. indonesiensis distinct from CSLK01-03T. Methods: This study refines the taxonomic description of R. indonesiensis and investigates its application in converting chitin into chitosan. The chitin deacetylase (RiCDA) activity of strain T22.7.1T was optimized, and the enzyme was isolated and purified from the fermentation products. Results: Through optimization, the RiCDA activity of strain T22.7.1T reached 287.02 U/mL, which is 34.88 times greater than the original enzyme's activity (8.0 U/mL). The natural CDA enzyme was purified with a purification factor of 31.83, and the specific activity of the enzyme solution reached 1200.33 U/mg. RiCDA exhibited good pH and temperature adaptability and stability, along with a wide range of substrate adaptabilities, effectively deacetylating chitin, chitooligosaccharides, N-acetylglucosamine, and other substrates. Discussion: Product analysis revealed that RiCDA treatment increased the deacetylation degree (DD) of natural chitin to 83%, surpassing that of commercial chitosan. Therefore, RiCDA demonstrates significant potential as an efficient deacetylation tool for natural chitin and chitooligosaccharides, highlighting its applicability in the biorefining of natural polysaccharides.

3.
Artigo em Inglês | MEDLINE | ID: mdl-39115691

RESUMO

The natural polymer chitin is an abundant source for valuable N-acetylchitooligosaccharides and N-acetylglucosamine applicable in several industries. The endochitinase Chit36-TA from Trichoderma asperellum was recombinantly expressed in Komagataella phaffii for the enzymatic degradation of chitin from unused insect exuviae into N-acetylchitooligosaccharides. Chit36-TA was purified by Ni-NTA affinity chromatography and subsequently biochemically characterized. After deglycosylation, the endochitinase had a molecular weight of 36 kDa. The optimum pH for Chit36-TA was 4.5. The temperature maximum of Chit36-TA was determined to be 50 °C, while it maintained > 93% activity up to 60 °C. The chitinase was thermostable up to 45 °C and exhibited ~ 50% activity after a 15 min incubation at 57 °C. Chit36-TA had a maximum specific enzyme activity of 50 nkat/mg with a Km value of 289 µM with 4-methylumbelliferyl-N,N',N″-triacetyl-ß-chitotrioside as substrate. Most tested cations, organic solvents and reagents were well-tolerated by the endochitinase, except for SDS (1 mM), Cu2+ (10 mM) and Mn2+ (10 mM), which had stronger inhibitory effects with residual activities of 3, 41 and 28%, respectively. With a degree of hydrolysis of 32% applying colloidal shrimp chitin (1% (w/v)) and 12% on insect larvae (1% (w/v)) after 24 h, the endochitinase was found to be suitable for the conversion of colloidal chitin as well as chitin from black soldier fly larvae into water-soluble N-acetylchitooligosaccharides. To prove scalability, a bioreactor process was developed in which a 55-fold higher enzyme activity of 49 µkat/l and a tenfold higher protein expression of 1258 mg/l were achieved.

4.
Environ Technol ; : 1-12, 2024 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-39092595

RESUMO

This is the first record on literature to use biochar as support for CoFe2O4 to applicate and evaluate it as photocatalyst for degradation of organic pollutants. The support was verified by XRD, FT-IR, SEM, EDS and band gap. Composites CFO1BQ3, CFO1BQ1, and CFO3BQ1 showed 100% degradation in 60 min. This outstanding performance can be related to the drop in band gap energy and recombination rate of e¯/h + . The composites showed better efficiency when compared to pure CoFe2O4 (∼78%). This might be associate to the fact that biochar has a high concentration of phenolic, hydroxyl and carboxylic functional groups on its surface. In this reaction h+, O2•-, and •OH were the reactive species involved in the degradation. The toxicity of ponceau was tested before and after the treatment, through biochemical biomarkers in Danio rerio fish. In general, the treatment proved to be efficient in reducing ponceau toxicity in D. rerio fish.

5.
Int J Biol Macromol ; 277(Pt 3): 134425, 2024 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-39097065

RESUMO

In this study, deep eutectic solvent (DES) prepared from choline chloride, lactic acid, and one of the four polyols (ethylene glycol, glycerol, xylitol, and sorbitol) were compared and assessed for their effectiveness in extracting chitin from lobster shells. Our results revealed that as the number of hydroxyl groups in polyols increased, the hydrogen bond network within the DESs became denser. However, this led to a corresponding increase in viscosity, which impacted the efficiency of chitin extraction. Among all prepared DESs, choline chloride-lactic acid/glycerol (CCLaGly) exhibited superior extractive ability, resulting in the extraction of pure chitin from lobster shells. The purity, crystallinity, and molecular weight of the extracted chitin using CCLaGly DES were comparable to those of chemically-isolated chitin, with purity reaching 94.76 ± 0.33 %, crystallinity at 78.78 %, and a molecular weight of 655 kDa. Additionally, the physicochemical properties of the DES-extracted chitins were characterized using Fourier-transform infrared spectroscopy, X-ray diffraction, thermogravimetric analysis, and scanning electron microscopy. This study conducted a comparative analysis of polyol effects on chitin extraction from lobster shells, thereby opening a promising avenue for the utilization of various crustacean shells in sustainable biomaterial production.

6.
J Biol Chem ; : 107622, 2024 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-39098522

RESUMO

The primary distinction between insect and bacterial chitin degradation systems lies in the presence of a multi-modular endo-acting chitinase ChtII, in contrast to a processive exo-acting chitinase. Although the essential role of ChtII during insect development and its synergistic action with processive chitinase during chitin degradation have been established, the mechanistic understanding of how it deconstructs chitin remains largely elusive. Here OfChtII from the insect Ostrinia furnacalis was investigated employing comprehensive approaches encompassing biochemical and microscopic analyses. The results demonstrated that OfChtII truncations with more carbohydrate-binding modules (CBMs) exhibited enhanced hydrolysis activity, effectively yielding a greater proportion of fibrillary fractions from the compacted chitin substrate. At the single-molecule level, the CBMs in these OfChtII truncations have been shown to primarily facilitate chitin substrate association rather than dissociation. Furthermore, a greater number of CBMs was demonstrated to be essential for the enzyme to effectively bind to chitin substrates with high crystallinity. Through real-time imaging by high-speed atomic force microscopy, the OfChtII-B4C1 truncation with three CBMs was observed to shear chitin fibers, thereby generating fibrillary fragments and deconstructing the compacted chitin structure. This work pioneers in revealing the nanoscale mechanism of endo-acting multi-modular chitinase involved in chitin degradation, which provides an important reference for the rational design of chitinases or other glycoside hydrolases.

7.
Int J Biol Macromol ; 277(Pt 3): 134475, 2024 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-39102917

RESUMO

This work valorizes rejects from Tenebrio Molitor TM breeding through the production of chitin and chitosan. Two processes are proposed for extracting chitin from larval exuviae and adult. The first process P1 provides chitin with high contents compared to literature data but the characterization shows the presence of impurities in the exuviae chitin responsible for the shifts in the values of the physicochemical characteristics towards those presented by γ chitin. These impurities are removed by delipidation and pure α chitin is obtained. The effective delipidation of this chitin would be linked to its fibrous surface structure. The analysis of the results of P1 led us to develop a second extraction process P2 which provides pure chitin with improved yields using delipidation followed by deproteinization. The N-deacetylation of chitin according to Kurita or Broussignac process makes possible the preparation of pure, highly deacetylated chitosan samples (2 % < DA < 12 %) with high yields and controlled molar masses (Mv). A kinetic study of molecular degradation during deacetylation is carried out. A comparison with Hermetia illucens allows to extend the use of insects as a potential source of chitin and chitosan and confirms the role of the source and the processes in the determination of their characteristics.

8.
Mar Pollut Bull ; 206: 116709, 2024 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-38991607

RESUMO

The reliable quantification of microplastic contamination in chitinous organisms requires validated methods to remove interfering complex organic and inorganic material. This study trialled KOH, H2O2 and HNO3 digestion methods on the digestive tracts of two large decapods (Panulirus cygnus and Portunus armatus) to validate a protocol that facilitates reliable microplastic extraction. KOH digestion provided the best recovery (>95 %) of all polymers (e.g. polyamide, polyethylene, polyethylene terephthalate, polypropylene, polystyrene and polyvinyl chloride), with the lowest impact to their physical morphology and chemical spectra. While HNO3, and HNO3 + H2O2 treatments were more effective at digesting chitin, they destroyed polyamide, and altered several other polymers. High digestion efficiency did not result in high matrix clarification or high microplastic recovery for large decapods. This study emphasises the importance of validating species-specific microplastic extraction methods, whilst proposing additional post-digestion protocols, such as density separation, for complex samples, that can be applied in future research investigating plastic contamination in large decapods.

9.
Int J Biol Macromol ; 277(Pt 1): 134124, 2024 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-39067733

RESUMO

Chitin has garnered significant attention due to its renewable, biocompatibility and biodegradability, while its practical application seriously hindered as the functionality of chitin itself can no longer meet people's increasing requirements for materials. Here, an effective method is successfully built for high-performance chitin fibers fabrication through a multi-step strategy that involved chemical pre-crosslinking, followed by wet-twisting and wet-stretching techniques, combined with physical cross-linking. The as-prepared chitin fiber exhibited a smooth surface, adjustable diameter, and mechanical strong properties (144.6 MPa). More importantly, functional chitin fiber with magnetic or conductive abilities can be easily obtained by spraying Fe3O4 particles or Ag nanowire on the chemical pre-crosslinking chitin gel film before stretching and twisting. The doped functional inorganic particles exist in a continuous ribbon structure in the fiber reduced the decrease in material strength caused by uneven particles dispersion, resulting 88.4 % of stress and 91.6 % of strain retention. This work not only bestow invaluable insights into the fabrication of functional chitin fibers but also provide a novel approach to solve the problem of poor compatibility between organic and inorganic composite materials.

10.
Int J Biol Macromol ; : 134235, 2024 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-39079565

RESUMO

Petrochemical resources are non-renewable, which has impeded the development of synthetic polymers. The poor degradability of synthetic polymers poses substantial environmental pressure. Additionally, the high cost of synthetic biopolymers with excellent degradation performance limits their widespread application. Thus, it is crucial to seek green, sustainable, low-cost polymers as alternatives to petrochemical-based synthetic polymers and synthetic biopolymers. Chitin is a natural and renewable biopolymer discovered in crustacean shells, insect exoskeletons, and fungal cell walls. Chitin chains consist of crystalline and amorphous regions. Note that various treatments can be employed to remove the amorphous region, enhancing the crystallinity of chitin. Chitin nanowhiskers are a high crystallinity nanoscale chitin product with a high aspect ratio, a large surface area, adjustable surface morphology, and biocompatibility. They discover widespread applications in biomedicine, environmental treatment, food packaging, and biomaterials. Various methods can be utilized for preparing chitin nanowhiskers, including chemical, ionic liquids, deacetylation, and mechanical methods. However, developing an environmentally friendly preparation process remains a big challenge for expanding their applications in different materials and large-scale production. This article comprehensively analyzes chitin nanowhiskers' preparation strategies and their drawbacks. It also highlights the extensive application in different materials and various fields, besides the potential for commercial application.

11.
Microorganisms ; 12(7)2024 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-39065035

RESUMO

Planctomycetes of the genus Singulisphaera are common inhabitants of soils and peatlands. Although described members of this genus are characterized as possessing hydrolytic capabilities, the ability to degrade chitin has not yet been reported for these bacteria. In this study, a novel Singulisphaera representative, strain Ch08, was isolated from a chitinolytic enrichment culture obtained from a boreal fen in Northern European Russia. The 16S rRNA gene sequence of this isolate displayed 98.2% similarity to that of Singulisphaera acidiphila MOB10T. Substrate utilization tests confirmed that strain Ch08 is capable of growth on amorphous chitin. The complete genome of strain Ch08 determined in this study was 10.85 Mb in size and encoded two predicted chitinases, which were only distantly related to each other and affiliated with the glycoside hydrolase family GH18. One of these chitinases had a close homologue in the genome of S. acidiphila MOB10T. The experimental verification of S. acidiphila MOB10T growth on amorphous chitin was also positive. Transcriptome analysis performed with glucose- and chitin-growth cells of strain Ch08 showed upregulation of the predicted chitinase shared by strain Ch08 and S. acidiphila MOB10T. The gene encoding this protein was expressed in Escherichia coli, and the endochitinase activity of the recombinant enzyme was confirmed. The ability to utilize chitin, a major constituent of fungal cell walls and arthropod exoskeletons, appears to be one of the previously unrecognized ecological functions of Singulisphaera-like planctomycetes.

12.
Microorganisms ; 12(7)2024 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-39065092

RESUMO

Magnaporthe oryzae is one of the most important fungal pathogens of rice. Chitin and avirulent strains can induce two layers of immunity response, pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI), in rice with cognate R genes. However, little is known about the assembly of the rice microbiome induced by PTI and ETI in rice. In this study, we investigate the impact of continuous treatment of the avirulent M. oryzae strain with AvrPi9 and chitin on the bacterial endophytic community of rice varieties harboring resistant gene Pi9 and their antagonistic activity against rice blast fungus. Analysis of the 16S rRNA showed a significant increase in the diversity and microbial co-occurrence network complexity and the number of beneficial taxa-Bacillus, Pseudomonas, Microbacterium, and Stenotrophomonas spp.-following the chitin and avirulent strain treatments. The antifungal assay with bacterial endophytes recovered from the leaves showed few bacteria with antagonistic potential in rice treated with avirulent strains, suggesting that the sequential treatment of the avirulent strain decreased the antagonistic bacteria against M. oryzae. Moreover, we identified Bacillus safensis Ch_66 and Bacillus altitudinis Nc_68 with overall antagonistic activities in vivo and in vitro. Our findings provide a novel insight into rice microbiome assembly in response to different innate immunity reactions.

13.
Polymers (Basel) ; 16(14)2024 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-39065272

RESUMO

Nanocomposites made via electrospinning were constructed of polyvinyl alcohol (PVA) and chitin. Chitin was extracted from a natural source (Fomes fomentarius), which allowed for precise control of the chemical properties of the resulting material. Chitin was chosen as a filler due to its low cost and widespread availability. Increasing the degree of acetylation of the chitin increased the Young's Modulus of the resulting fiber mats but only at relatively high levels. While composites at lower acetylation levels were stable, no increase in the Young's Modulus was observed, presumably due to decreased intermolecular bonding among fibers. The results suggest that precise control of the degree of acetylation of chitin, more than the loading amount and dispersibility, significantly impacts composite formation.

14.
Sci Total Environ ; 947: 174560, 2024 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-38972425

RESUMO

Over the last decade, commercialization of insects for food and feed has been exponentially increasing. Insect protein is emerging as a sustainable livestock feed and human food alternative due to its low land and carbon footprint. The principles of insect industry are deeply embedded in the core values of sustainability and circular economy. Black soldier fly (BSF) is the crown jewel of insect industry and is one of the most commercially farmed insects. However, this steadfast growth is accompanied by generation of insect based biowaste such as dead flies and pupae exuviae. This will be a major waste fraction from this industry. This study discusses the valorization potential of this waste into chitin (which finds application in cosmetics, bioplastics, and pesticides, among other industries), biogas, fertilizer, and biochar. There is need to conduct more explorative research on value proposition of insect based biowaste to ensure that this industry can comply fully with circular economy and sustainability principles.


Assuntos
Insetos , Animais , Agricultura/métodos
15.
Microbiol Resour Announc ; : e0030824, 2024 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-39037313

RESUMO

We present the draft genome sequence of Collimonas sp. strain H4R21, isolated from the rhizosphere of Fagus sylvatica in the forest experimental site of Montiers (France). This genome features coding capacity for plant growth promotion, such as the ability to solubilize minerals, to produce siderophores and antifungal secondary metabolites.

16.
Animal ; 18(8): 101239, 2024 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-39053154

RESUMO

The demand for sustainable and ethically farmed animal products is on the rise as consumers become more environmentally and animal welfare conscious. The need to diminish the consumption of soybean meal is urgent, and companies are looking for ways to respond to this necessity by looking for alternatives to soybean meal. This study assessed the impact of introducing whole dehydrated and live black soldier fly larvae (BSFL) into the diet of an indigenous chicken breed as environmental enrichment. A total of 144 39-day-old male Bianca di Saluzzo chickens were distributed among 18 pens and assigned to three different experimental groups. The control group received a diet where soybean meal was entirely replaced by alternative ingredients. The two experimental groups were given the same diet supplemented with 5% of the expected daily feed intake of whole dehydrated BSFL or whole live BSFL. Throughout the trial period (from the bird age of 39-174 days of age), live weight was recorded every 21 days, and the average daily gain, daily feed intake, and feed conversion ratio were calculated. The time required for the birds to consume the larvae was recorded three times a week. At age 147 and 174 days, 12 birds per treatment were selected based on mean live weight and slaughtered. Measurements included hot and chilled carcass weights, organ weights (spleen, liver, heart, stomach), breast and thigh muscle weights, and the corresponding yields were calculated. Acid protease activity was measured in proventriculus extract, and chitinase and chitosanase activity was calculated based on the release of reducing sugars from chitin or chitosan. The results showed little improvement in final live weights and daily feed intakes of the animals fed the insect larvae compared with control birds. Larva supplementation had no negative impact on the overall well-being of the animals assessed by blood analysis and histopathological assessment of the intestinal tract, spleen, and liver. No differences were found between the dehydrated vs live insect larvae consumption times, with all larvae being eaten up very rapidly (< 3 min). The birds fed BSFL showed an increase in chitinase activity. These findings support the potential use of whole BSFL as a form of environmental enrichment, particularly in their dehydrated form, being more convenient to use and store, which would also encourage the uptake of this practice by farmers.

17.
Int J Biol Macromol ; 276(Pt 2): 133980, 2024 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-39032901

RESUMO

N-acetyl-oligosaccharides exhibit antioxidant and antibacterial activities. However, the low catalytic efficiency of chitinase on crystalline chitin hinders the eco-friendly production of N-acetyl-oligosaccharides. A marine-derived chitinase-producing strain Chitiniphilus eburneus YS-30 was screened in this study. The genome of C. eburneus YS-30 spans 4,522,240 bp, with a G + C content of 63.96 % and 4244 coding genes. Among the chitinases secreted by C. eburneus YS-30, Ce0303 showed the highest content at 19.10 %, with a molecular weight of 73.5 kDa. Recombinant Ce0303 exhibited optimal activity at 50 °C and pH 5.0, maintaining stability across pH 4.0-10.0. Ce0303 demonstrated strict substrate specificity, with a specific activity toward colloidal chitin of 6.41 U mg-1, Km of 2.34 mg mL-1, and kcat of 3.27 s-1. The specific activity of Ce0303 toward α-chitin was 18.87 % of its activity on colloidal chitin. Ce0303 displayed both exo- and endo-hydrolytic properties, primarily producing (GlcNAc)1-3 from colloidal chitin. The structure of Ce0303 includes one catalytic domain and two chitin-binding domains. Docking results revealed that the GlcNAc at -1 subsite formed two hydrogen bonds with conserved Trp380. The hydrolytic properties of Ce0303 will provide technical support for the comprehensive utilization of crustacean raw materials.

18.
Int J Biol Macromol ; 276(Pt 2): 133954, 2024 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-39029834

RESUMO

Mycelium-based leather substitutes with a three-dimensional reticulated structure have attracted attention owing to the negative environmental impacts of natural and synthetic leather. This study utilised Ganoderma lucidum mycelium to prepare a mycelium-based leather substitute with zinc cross-linking (MF-Zn) and evaluated its physicochemical properties and sensory performance; the conventional Cr3+ tanning method was used as reference. Results demonstrated that Zn2+ and Cr3+ formed cross-links with the -OH and -NHOCH3 groups in the polysaccharides of chitin, while Zn2+ selectively bonded to a fraction of -NH2 groups in cystine and phenylalanine. The mycelium-based leather substitute with Zn cross-linking exhibited impressive tensile strength and tear strength of 7.0 MPa and 16.4 kN/m, respectively, while demonstrating desirable organoleptic properties. The free radical-scavenging capacity of MF-Zn was assessed, revealing a DPPH radical and hydroxyl radical scavenging rates of 39.4% and 52.7%, respectively. By successfully investigating the cross-linking mechanism of mycelial fibres with Zn2+ and obtaining the stabilised mycelium-based leather substitute, this study establishes a fundamental basis for the development of sustainable leather substitutes, meeting the requirements and facilitating significant advancements in low-carbon leather substitute production.

19.
Nat Prod Res ; : 1-2, 2024 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-38963904

RESUMO

This letter emphasizes the potential of chemically modified chitin and chitosan in natural product research. Extracted from crustacean shells, these biopolymers are known for their biocompatibility, biodegradability and non-toxicity. Chemical modifications improve their solubility, adsorption capacity and antimicrobial properties, making them ideal for applications in drug delivery, wound healing and pollutant removal. Furthermore, combining natural products with modified chitosan creates novel therapeutic agents with increased efficacy and fewer side effects. This research highlights the significance of exploring the various applications of chitin and chitosan, aligning with the journal's focus on innovative natural product solutions.

20.
J Biomed Mater Res A ; 2024 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-39007419

RESUMO

As the cornerstone of tissue engineering and regeneration medicine research, developing a cost-effective and bionic extracellular matrix (ECM) that can precisely modulate cellular behavior and form functional tissue remains challenging. An artificial ECM combining polysaccharides and fibrillar proteins to mimic the structure and composition of natural ECM provides a promising solution for cardiac tissue regeneration. In this study, we developed a bionic hydrogel scaffold by combining a quaternized ß-chitin derivative (QC) and fibrin-matrigel (FM) in different ratios to mimic a natural ECM. We evaluated the stiffness of those composite hydrogels with different mixing ratios and their effects on the growth of human umbilical vein endothelial cells (HUVECs). The optimal hydrogels, QCFM1 hydrogels were further applied to load HUVECs into nude mice for in vivo angiogenesis. Besides, we encapsulated human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) into QCFM hydrogels and employed 3D bioprinting to achieve batch fabrication of human-engineered heart tissue (hEHT). Finally, the myocardial structure and electrophysiological function of hEHT were evaluated by immunofluorescence and optical mapping. Designed artificial ECM has a tunable modulus (220-1380 Pa), which determines the different cellular behavior of HUVECs when encapsulated in these. QCFM1 composite hydrogels with optimal stiffness (800 Pa) and porous architecture were finally identified, which could adapt for in vitro cell spreading and in vivo angiogenesis of HUVECs. Moreover, QCFM1 hydrogels were applied in 3D bioprinting successfully to achieve batch fabrication of both ring-shaped and patch-shaped hEHT. These QCFM1 hydrogels-based hEHTs possess organized sarcomeres and advanced function characteristics comparable to reported hEHTs. The chitin-derived hydrogels are first used for cardiac tissue engineering and achieve the batch fabrication of functionalized artificial myocardium. Specifically, these novel QCFM1 hydrogels provided a reliable and economical choice serving as ideal ECM for application in tissue engineering and regeneration medicine.

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