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1.
Nat Commun ; 15(1): 3289, 2024 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-38632231

RESUMO

Endowing textiles with perceptual function, similar to human skin, is crucial for the development of next-generation smart wearables. To date, the creation of perceptual textiles capable of sensing potential dangers and accurately pinpointing finger touch remains elusive. In this study, we present the design and fabrication of intelligent perceptual textiles capable of electrically responding to external dangers and precisely detecting human touch, based on conductive silk fibroin-based ionic hydrogel (SIH) fibers. These fibers possess excellent fracture strength (55 MPa), extensibility (530%), stable and good conductivity (0.45 S·m-1) due to oriented structures and ionic incorporation. We fabricated SIH fiber-based protective textiles that can respond to fire, water, and sharp objects, protecting robots from potential injuries. Additionally, we designed perceptual textiles that can specifically pinpoint finger touch, serving as convenient human-machine interfaces. Our work sheds new light on the design of next-generation smart wearables and the reshaping of human-machine interfaces.


Assuntos
Fibroínas , Seda , Humanos , Seda/química , Têxteis , Condutividade Elétrica , Fibroínas/química , Tato
2.
Int J Mol Sci ; 25(6)2024 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-38542528

RESUMO

Spider silk has extraordinary mechanical properties, displaying high tensile strength, elasticity, and toughness. Given the high performance of natural fibers, one of the long-term goals of the silk community is to manufacture large-scale synthetic spider silk. This process requires vast quantities of recombinant proteins for wet-spinning applications. Attempts to synthesize large amounts of native size recombinant spidroins in diverse cell types have been unsuccessful. In these studies, we design and express recombinant miniature black widow MaSp1 spidroins in bacteria that incorporate the N-terminal and C-terminal domain (NTD and CTD), along with varying numbers of codon-optimized internal block repeats. Following spidroin overexpression, we perform quantitative analysis of the bacterial proteome to identify proteins associated with spidroin synthesis. Liquid chromatography with tandem mass spectrometry (LC MS/MS) reveals a list of molecular targets that are differentially expressed after enforced mini-spidroin production. This list included proteins involved in energy management, proteostasis, translation, cell wall biosynthesis, and oxidative stress. Taken together, the purpose of this study was to identify genes within the genome of Escherichia coli for molecular targeting to overcome bottlenecks that throttle spidroin overexpression in microorganisms.


Assuntos
Fibroínas , Aranhas , Animais , Fibroínas/química , Proteômica , Espectrometria de Massas em Tandem , Seda/química , Proteínas Recombinantes/química , Bactérias , Aranhas/genética
3.
Sheng Wu Gong Cheng Xue Bao ; 40(3): 687-704, 2024 Mar 25.
Artigo em Chinês | MEDLINE | ID: mdl-38545971

RESUMO

Spider silk is a natural fiber known as "biosteel" with the strongest composite performance, such as high tensile strength and toughness. It is also equipped with excellent biocompatibility and shape memory ability, thus shows great potential in many fields such as biomedicine and tissue engineering. Spider silk is composed of macromolecular spidroin with rich structural diversity. The characteristics of the primary structure of natural spidroin, such as the high repeatability of amino acids in the core repetitive region, the high content of specific amino acids, the large molecular weight, and the high GC content of the spidroin gene, have brought great difficulties in heterologous expression. This review discusses focuses on the relationship between the featured motifs of the microcrystalline region in the repetitive unit of spidroin and its structure, as well as the spinning performance and the heterologous expression. The optimization design for the sequence of spidroin combined with heterologous expression strategy has greatly promoted the development of the biosynthesis of spider silk proteins. This review may facilitate the rational design and efficient synthesis of recombinant spidroin.


Assuntos
Fibroínas , Aranhas , Animais , Seda/genética , Seda/química , Fibroínas/genética , Fibroínas/química , Proteínas de Artrópodes , Materiais Biocompatíveis , Aminoácidos , Aranhas/genética
4.
Int J Biol Macromol ; 264(Pt 2): 130687, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38462112

RESUMO

Silk fibroin derived from silkworm cocoons exhibits excellent mechanical properties, good biocompatibility, and low immunogenicity. Previous studies showed that silk fibroin had an inhibitory effect on cells, suppressing proliferation and inducing apoptosis. However, the source of the toxicity and the mechanism of apoptosis induction are still unclear. In this study, we hypothesized that the toxicity of silk fibroin might originate from the crystalline region of the heavy chain of silk fibroin. We then verified the hypothesis and the specific induction mechanism. A target peptide segment was obtained from α-chymotrypsin. The potentially toxic mixture of silk fibroin peptides (SFPs) was separated by ion exchange, and the toxicity was tested by an MTT assay. The results showed that SFPs obtained after 4 h of enzymatic hydrolysis had significant cytotoxicity, and SFPs with isoelectric points of 4.0-6.8 (SFPα II) had a significant inhibitory effect on cell growth. LC-MS/MS analysis showed that SFPα II contained a large number of glycine-rich and alanine-rich repetitive sequence polypeptides from the heavy-chain crystallization region. A series of experiments showed that SFPα II mediated cell death through the apoptotic pathway by decreasing the expression of Bcl-2 protein and increasing the expression of Bax protein. SFPα II mainly affected the p53 pathway and the AMPK signaling pathway in HepG2 cells. SFPα II may indirectly increase the expression of Cers2 by inhibiting the phosphorylation of EGFR, which activated apoptotic signaling in the cellular mitochondrial pathway and inhibited the Akt/NF-κB pathway by increasing the expression of PPP2R2A.


Assuntos
Bombyx , Fibroínas , Animais , Fibroínas/farmacologia , Fibroínas/química , Cromatografia Líquida , Espectrometria de Massas em Tandem , Peptídeos/farmacologia , Peptídeos/química , Bombyx/química , Apoptose , Seda/química
5.
Biomed Phys Eng Express ; 10(3)2024 Mar 14.
Artigo em Inglês | MEDLINE | ID: mdl-38447212

RESUMO

The analysis of degradation in the presence of cell death and migration is a critical aspect of research in various biological fields, such as tissue engineering, regenerative medicine, and disease pathology. In present study, numerical study of degradation of scaffold were performed in present of cells, cell apoptosis and cell migration. A poly electrolyte complex (PEC) silk fibroin scaffold was used for degradation study. Degradation study in the presence of cells and migration were performed at fixed pH concentration 7.2. Similarly, degradation study of scaffold were performed at different pH cell apoptosis. A transient analysis of scaffold was evaluated in COMSOL 5.5 in presence of cryogenic temperature at different temperature gradient. The parameters; temperature, stress, strain tensor and deformation gradient associated with the degradation of polyelectrolyte complex scaffold were evaluated. Result shows that in both geometries minimum temperature had been achieved as 230.051 K at point P4 in series view and parallel view and at a point P3 for cell migration study for -5 k min-1and -1 k min-1, respectively. The maximum stress had been generated for 5.57 × 107N m-2for the temperature gradient of -2 K min-1at T cycle in the case of cell migration study. In contrast in series view the maximum stress 2.9 × 107 N m-2were observed at P4 which was higher as compare to P3. Similarly, for a parallel view, maximum stress (3.93 × 107 N m-2) was obtained for point P3. It had been observed that the maximum strain tensor 5.21 × 10-3, 5.15 × 10-3and 5.26 × 10-3was generated in series view at 230 k on a point P3 for - 1, -2 and -5 K min-1, respectively. Similarly, the maximum strain tensor 8.16 × 10-3, 8.09 × 10-3and 8.09 × 10-3was generated in parallel view at 230 k on a point P3 for -1, -2 and -5 K min-1, respectively. In the presence of cells, at a point P4 for temperature gradient of -1 and -2 K min-1, it had been closed to the scaffold wall, which had a different temperature profile than the point P3 and scaffold comes to the contact with the cells. The analysis of PEC scaffold degradation in the presence of cells, including cell apoptosis and migration, offers significant insights into the relationship between scaffold properties, cell behaviour, and tissue regeneration.


Assuntos
Fibroínas , Tecidos Suporte , Tecidos Suporte/química , Engenharia Tecidual/métodos , Fibroínas/química , Apoptose , Movimento Celular
6.
Carbohydr Polym ; 332: 121945, 2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38431423

RESUMO

Tissue-engineered is an effective method for repairing critical-size bone defects. The application of bioactive scaffold provides artificial matrix and suitable microenvironment for cell recruitment and extracellular matrix deposition, which can effectively accelerate the process of tissue regeneration. Among various scaffold properties, appropriate pore structure and distribution have been proven to play a crucial role in inducing cell infiltration differentiation and in-situ tissue regeneration. In this study, a chitosan (CS) /silk fibroin (SF) /bioactive glass (BG) composite scaffold with distinctive radially oriented pore structure was constructed. The composite scaffolds had stable physical and chemical properties, a unique pore structure of radial arrangement from the center to the periphery and excellent mechanical properties. In vitro biological studies indicated that the CS/SF/BG scaffold could promote osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and the expression of related genes due to the wide range of connected pore structures and released active elements. Furthermore, in vivo study showed CS/SF/BG scaffold with radial pores was more conducive to the repair of skull defects in rats with accelerated healing speed during the bone tissue remodeling process. These results demonstrated the developed CS/SF/BG scaffold would be a promising therapeutic strategy for the repair of bone defects regeneration.


Assuntos
Quitosana , Fibroínas , Ratos , Animais , Fibroínas/química , Tecidos Suporte/química , Osteogênese , Quitosana/química , Engenharia Tecidual/métodos , Regeneração Óssea
7.
Int J Biol Macromol ; 263(Pt 1): 129925, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38311129

RESUMO

Tissue engineering has emerged as a promising approach for addressing bone defects. Most of the traditional 3D printing materials predominantly relying on polymers and ceramics. Although these materials exhibit superior osteogenic effects, their gradual degradation poses a limitation. Digital light processing (DLP) 3D bioprinting that uses natural biomaterials as bioinks has become one of the promising strategies for bone regeneration. In this study, we introduce a hydrogel biomaterial derived from silk fibroin (SF). Notably, we present the novel integration of nano-hydroxyapatite (nHA) into the hydrogel, forming a composite hydrogel that rapidly cross-links upon initiation. Moreover, we demonstrate the loading of nHA through non-covalent bonds in SilMA. In vitro experiments reveal that composite hydrogel scaffolds with 10 % nHA exhibit enhanced osteogenic effects. Transcriptomic analysis indicates that the composite hydrogel promotes bone regeneration by inducing M2 macrophage polarization. Furthermore, rat femoral defect experiments validate the efficacy of SilMA/nHA10 in bone regeneration. This study synthesis of a simple and effective composite hydrogel bioink for bone regeneration, presenting a novel strategy for the future implementation of digital 3D printing technology in bone tissue engineering.


Assuntos
Fibroínas , Ratos , Animais , Fibroínas/farmacologia , Fibroínas/química , Durapatita/farmacologia , Durapatita/química , Tecidos Suporte/química , Hidrogéis/farmacologia , Hidrogéis/química , Materiais Biocompatíveis/farmacologia , Materiais Biocompatíveis/química , Engenharia Tecidual , Regeneração Óssea
8.
J Mater Chem B ; 12(9): 2384-2393, 2024 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-38349135

RESUMO

Oxidative stress is one of the obstacles preventing wound regeneration, especially for chronic wounds. Herein, designing a wound dressing with an anti-oxidant function holds great appeal for enhancing wound regeneration. In this study, a biocompatible and degradable nanofiber with a core-shell structure was fabricated via coaxial electrospinning, in which polycaprolactone (PCL) was applied as the core structure, while the shell was composed of a mixture of silk fibroin (SF) and tocopherol acetate (TA). The electrospun PST nanofibers were proven to have a network structure with significantly enhanced mechanical properties. The PSTs exhibited a diameter distribution with an average of 321 ± 134 nm, and the water contact angle of their surface is 124 ± 2°. The PSTs also exhibited good tissue compatibility, which can promote the adhesion and proliferation of L929 cells. Besides, the dissolution of silk fibroin encourages the release of TA, which could play a synergistic effect and regulate the oxidative stress effect in the damaged area, for it promotes the adhesion and proliferation of skin fibroblasts (L929), reduces the cytotoxicity of hydrogen peroxide to cells, and lowers the level of reactive oxygen species. The animal experiment indicated that the PSTs would promote the reconstruction of skin. These nanofibers are expected to repair skin ulcers related to diabetes.


Assuntos
Fibroínas , Nanofibras , Animais , Antioxidantes/farmacologia , Tecidos Suporte/química , Fibroínas/farmacologia , Fibroínas/química , Nanofibras/química , Bandagens
9.
Methods Mol Biol ; 2764: 249-263, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38393599

RESUMO

To imitate in vivo environment of cells, microfluidics offer controllable fashions at micro-scale and enable regulate flow-related parameters precisely, leveraging the current state of 3D systems to 4D level through the inclusion of flow and shear stress. In particular, integrating silk fibroin as an adhering layer with microfluidic chips enables to form more comprehensive and biocompatible network between cells since silk fibroin holds outstanding mechanical and biological properties such as easy processability, biocompatibility, controllable biodegradation, and versatile functionalization. In this chapter, we describe design and fabrication of a microfluidic chip, with silk fibroin-covered microchannels for the formation of 3D structures, such as MCF-7 (human breast cancer) cell spheroids as a model system. All the steps performed here are characterized by surface-sensitive tools and standard tissue culture methods. Overall, this strategy can be easily integrated into various high-tech application areas such as drug delivery systems, regenerative medicine, and tissue engineering in near future.


Assuntos
Neoplasias da Mama , Fibroínas , Humanos , Feminino , Fibroínas/química , Microfluídica , Engenharia Tecidual/métodos , Medicina Regenerativa , Materiais Biocompatíveis/química , Tecidos Suporte/química
10.
ACS Appl Mater Interfaces ; 16(7): 9210-9223, 2024 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-38330192

RESUMO

Biology resolves design requirements toward functional materials by creating nanostructured composites, where individual components are combined to maximize the macroscale material performance. A major challenge in utilizing such design principles is the trade-off between the preservation of individual component properties and emerging composite functionalities. Here, polysaccharide pectin and silk fibroin were investigated in their composite form with pectin as a thermal-responsive ion conductor and fibroin with exceptional mechanical strength. We show that segregative phase separation occurs upon mixing, and within a limited compositional range, domains ∼50 nm in size are formed and distributed homogeneously so that decent matrix collective properties are established. The composite is characterized by slight conformational changes in the silk domains, sequestering the hydrogen-bonded ß-sheets as well as the emergence of randomized pectin orientations. However, most dominant in the composite's properties is the introduction of dense domain interfaces, leading to increased hydration, surface hydrophilicity, and increased strain of the composite material. Using controlled surface charging in X-ray photoelectron spectroscopy, we further demonstrate Ca ions (Ca2+) diffusion in the pectin domains, with which the fingerprints of interactions at domain interfaces are revealed. Both the thermal response and the electrical conductance were found to be strongly dependent on the degree of composite hydration. Our results provide a fundamental understanding of the role of interfacial interactions and their potential applications in the design of material properties, polysaccharide-protein composites in particular.


Assuntos
Fibroínas , Nanoestruturas , Seda/química , Fibroínas/química , Polissacarídeos , Pectinas , Materiais Biocompatíveis/química
11.
Int J Biol Macromol ; 264(Pt 1): 130374, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38408575

RESUMO

Silk is a natural engineering material with a unique set of properties. The major constituent of silk is fibroin, a protein widely used in the biomedical field because of its mechanical strength, toughness and elasticity, as well as its biocompatibility and biodegradability. The domestication of silkworms allows large amounts of fibroin to be extracted inexpensively from silk cocoons. However, the industrial extraction process has drawbacks in terms of sustainability and the quality of the final medical product. The heterologous production of fibroin using recombinant DNA technology is a promising approach to address these issues, but the production of such recombinant proteins is challenging and further optimization is required due to the large size and repetitive structure of fibroin's DNA and amino acid sequence. In this review, we describe the structure-function relationship of fibroin, the current extraction process, and some insights into the sustainability of silk production for biomedical applications. We focus on recent advances in molecular biotechnology underpinning the production of recombinant fibroin, working toward a standardized, successful and sustainable process.


Assuntos
Bombyx , Fibroínas , Animais , Bombyx/química , Fibroínas/química , Materiais Biocompatíveis/química , Biotecnologia , Seda/química
12.
Int J Biol Macromol ; 263(Pt 2): 130380, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38395277

RESUMO

By mimicking in vivo bionic microenvironment and promoting osteogenic differentiation, the hybrid organic-inorganic nanofibrous membranes provide promising potential for guided bone regeneration (GBR) in the treatment of clinical bone defects. To develop a degradable and osteogenic membrane for GBR by combining the natural biomacromolecule silk fibroin (SF) and gelatine with the bioactive nano hydroxyapatite (nHA), the anhydride-modified gelatine-nano hydroxyapatite (GelMA-nHA) composites were synthesized in situ and introduced into silk fibroin to prepare nanofibrous membranes with different ratios using electrospinning and photocrosslinking. The nanofibrous membranes, particularly those with a mass ratio of 7:2:1, were found to exhibit satisfactory elongation at break up to 110 %, maintain the nanofibrous structure for up to 28 days, and rapidly form bone-like apatite within 3 days, thus offering advantages when it comes to guided bone regeneration. In vitro cell results showed that the SF/GelMA/nHA membranes had excellent biocompatibility and enhanced osteogenic differentiation of hBMSCs. In vivo studies revealed that the hybrid composite membranes can improve bone regeneration of critical-sized calvarial defects in rat model. Therefore, the novel hybrid nanofibrous membrane is proposed to be a alternative candidate for creating a bionic microenvironment that promotes bone regeneration, indicating their potential application to bone injury treatment.


Assuntos
Fibroínas , Nanofibras , Ratos , Animais , Fibroínas/farmacologia , Fibroínas/química , Osteogênese , Durapatita/farmacologia , Durapatita/química , Tecidos Suporte/química , Nanofibras/química , Biomimética , Regeneração Óssea , Seda/química
13.
Int J Biol Macromol ; 262(Pt 1): 129927, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38311130

RESUMO

To promote bone repair, it is desirable to develop three-dimensional multifunctional fiber scaffolds. The densely stacked and tightly arranged conventional two-dimensional electrospun fibers hinder cell penetration into the scaffold. Most of the existing three-dimensional structural materials are isotropic and monofunctional. In this research, a Janus nanofibrous scaffold based on silk fibroin/polycaprolactone (SF/PCL) was fabricated. SF-encapsulated SeNPs demonstrated stability and resistance to aggregation. The outside layer (SF/PCL/Se) of the Janus nanofiber scaffold displayed a structured arrangement of fibers, facilitating cell growth guidance and impeding cell invasion. The inside layer (SF/PCL/HA) featured a porous structure fostering cell adhesion. The Janus fiber scaffold containing SeNPs notably suppressed S. aureus and E. coli activities, correlating with SeNPs concentration. In vitro, findings indicated considerable enhancement in alkaline phosphatase (ALP) activity of MC3T3-E1 osteoblasts and upregulation of genes linked to osteogenic differentiation with exposure to the SF/PCL/HA/Se Janus nanofibrous scaffold. Moreover, in vivo, experiments demonstrated successful critical bone defect repair in mouse skulls using the SF/PCL/HA/Se Janus nanofiber scaffold. These findings highlight the potential of the SF/PCL-based Janus nanofibrous scaffold, integrating SeNPs and nHA, as a promising biomaterial in bone tissue engineering.


Assuntos
Fibroínas , Nanofibras , Camundongos , Animais , Fibroínas/farmacologia , Fibroínas/química , Tecidos Suporte/química , Osteogênese , Porosidade , Escherichia coli , Staphylococcus aureus , Engenharia Tecidual/métodos , Poliésteres/química , Regeneração Óssea , Nanofibras/química , Seda/química
14.
Colloids Surf B Biointerfaces ; 236: 113808, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38422669

RESUMO

In the cardiovascular field, coating containing copper used to catalyze NO (nitric oxide) production on non-degradable metal surfaces have shown unparalleled expected performance, but there are few studies on biodegradable metal surfaces. Magnesium-based biodegradable metals have been applied in cardiovascular field in large-scale because of their excellent properties. In this study, the coating of copper loaded in silk fibroin is fabricated on biodegradable ZE21B alloy. Importantly, the different content of copper is set to investigate the effects of on the degradation performance and cell behavior of magnesium alloy. Through electrochemical and immersion experiments, it is found that high content of copper will accelerate the corrosion of magnesium alloy. The reason is the spontaneous micro-batteries between copper and magnesium with the different standard electrode potentials, that is, the galvanic corrosion accelerates the corrosion of magnesium alloy. Moreover, the coating formed through silk fibroin by the right amount copper not only have a protective effect on the ZE21B alloy substrate, but also promotes the adhesion and proliferation of endothelial cells in blood vessel micro-environment. The production of NO catalyzed by copper ions makes this trend more significant, and inhibits the excessive proliferation of smooth muscle cells. These findings can provide guidance for the amount of copper in the coating on the surface of biodegradable magnesium alloy used for cardiovascular stent purpose.


Assuntos
Fibroínas , Fibroínas/farmacologia , Fibroínas/química , Cobre/farmacologia , Ligas/farmacologia , Ligas/química , Magnésio/farmacologia , Magnésio/química , Células Endoteliais , Materiais Revestidos Biocompatíveis/farmacologia , Materiais Revestidos Biocompatíveis/química , Corrosão
15.
Int J Biol Macromol ; 261(Pt 2): 129746, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38302025

RESUMO

In the development of small-diameter vascular grafts, it is crucial to achieve early-stage endothelialization to prevent thrombus formation and intimal hyperplasia. Silk fibroin (SF) from Bombyx mori is commonly used for such grafts. However, there is a need to expedite endothelialization post-implantation. In this study, we functionalized SF with Arg-Glu-Asp-Val (REDV) (SF + REDV) using cyanuric chloride to enhance endothelialization. The immobilization of REDV onto SF was confirmed and the amount of immobilized REDV could be calculated by 1H NMR. Furthermore, the conformational changes in Tyr, Ser, and Ala residues in [3-13C]Tyr- and [3-13C]Ser-SF due to REDV immobilization were monitored using 13C solid-state NMR. The REDV immobilized onto the SF film was found to be exposed on the film's surface, as confirmed by biotin-avidin system. Cell culture experiments, including adhesiveness, proliferation, and extensibility, were conducted using normal human umbilical vein endothelial cells (HUVEC) and normal human aortic smooth muscle cells (HAoSMC) on both SF and SF + REDV films to evaluate the impact of REDV on endothelialization. The results indicated a trend towards promoting HUVEC proliferation while inhibiting HAoSMC proliferation. Therefore, these findings suggest that SF + REDV may be more suitable than SF alone for coating small-diameter SF knitted tubes made of SF threads.


Assuntos
Bombyx , Fibroínas , Animais , Humanos , Fibroínas/química , Bombyx/química , Células Endoteliais , Peptídeos/química , Espectroscopia de Ressonância Magnética/métodos , Seda
16.
Sci Rep ; 14(1): 4428, 2024 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-38395958

RESUMO

Controlled release of proteins, such as growth factors, from biocompatible silk fibroin (SF) hydrogel is valuable for its use in tissue engineering, drug delivery, and other biological systems. To achieve this, we introduced silk fibroin-mimetic peptides (SFMPs) with the repeating unit (GAGAGS)n. Using green fluorescent protein (GFP) as a model protein, our results showed that SFMPs did not affect the GFP function when conjugated to it. The SFMP-GFP conjugates incorporated into SF hydrogel did not change the gelation time and allowed for controlled release of the GFP. By varying the length of SFMPs, we were able to modulate the release rate, with longer SFMPs resulting in a slower release, both in water at room temperature and PBS at 37 °C. Furthermore, the SF hydrogel with the SFMPs showed greater strength and stiffness. The increased ß-sheet fraction of the SF hydrogel, as revealed by FTIR analysis, explained the gel properties and protein release behavior. Our results suggest that the SFMPs effectively control protein release from SF hydrogel, with the potential to enhance its mechanical stability. The ability to modulate release rates by varying the SFMP length will benefit personalized and controlled protein delivery in various systems.


Assuntos
Fibroínas , Fibroínas/química , Hidrogéis/química , Preparações de Ação Retardada , Peptídeos , Sistemas de Liberação de Medicamentos , Seda/química
17.
ACS Appl Mater Interfaces ; 16(8): 9925-9943, 2024 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-38362893

RESUMO

Implantation of a phenotypically stable cartilage graft could represent a viable approach for repairing osteoarthritic (OA) cartilage lesions. In the present study, we investigated the effects of modulating the bone morphogenetic protein (BMP), transforming growth factor beta (TGFß), and interleukin-1 (IL-1) signaling cascades in human bone marrow stromal cell (hBMSC)-encapsulated silk fibroin gelatin (SF-G) bioink. The selected small molecules LDN193189, TGFß3, and IL1 receptor antagonist (IL1Ra) are covalently conjugated to SF-G biomaterial to ensure sustained release, increased bioavailability, and printability, confirmed by ATR-FTIR, release kinetics, and rheological analyses. The 3D bioprinted constructs with chondrogenically differentiated hBMSCs were incubated in an OA-inducing medium for 14 days and assessed through a detailed qPCR, immunofluorescence, and biochemical analyses. Despite substantial heterogeneity in the observations among the donors, the IL1Ra molecule illustrated the maximum efficiency in enhancing the expression of articular cartilage components, reducing the expression of hypertrophic markers (re-validated by the GeneMANIA tool), as well as reducing the production of inflammatory molecules by the hBMSCs. Therefore, this study demonstrated a novel strategy to develop a chemically decorated, printable and biomimetic SF-G bioink to produce hyaline cartilage grafts resistant to acquiring OA traits that can be used for the treatment of degenerated cartilage lesions.


Assuntos
Bioimpressão , Cartilagem Articular , Fibroínas , Humanos , Fibroínas/química , Cartilagem Articular/metabolismo , Materiais Biocompatíveis/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Diferenciação Celular , Gelatina/farmacologia , Gelatina/química , Tecidos Suporte/química , Engenharia Tecidual , Impressão Tridimensional
18.
Int J Biol Macromol ; 263(Pt 1): 130287, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38373567

RESUMO

Bioactive scaffolds accurately mimicking the structure and composition of the extracellular matrix have garnered significant interest in tissue engineering. In this study, we developed a platform utilizing natural silk nanofibrils, hyaluronic acid, and basic fibroblast growth factor for the purpose of promoting spinal cord regeneration by creating an optimal microenvironment. The bioactive scaffold exhibited notable characteristics such as high porosity and hydrophilicity, attributed to its unique nanostructure, high connectivity, and polysaccharide composition. Furthermore, the pore size of the scaffold can be adjusted within the range of 90 µm to 120 µm by varying the content of hyaluronic acid. In vitro, human umbilical vein endothelial cells were seeded into the scaffold, demonstrating enhanced cell viability. The scaffold facilitated cell proliferation and migration. In vivo experiments on rats indicated that the scaffold had a beneficial impact on spinal cord regeneration, creating a conducive environment for motor function recovery of the rats. This effect may be attributed to the scaffold's ability to stimulate axon growth and neuronal survival, as well as inhibit the formation of glial scars, as evidenced by the decreased expression of growth associated protein-43, microtubule-associated protein 2, and neurofilament-200. This study presents a promising method to develop a feasible bioscaffold for the treatment of spinal cord injury.


Assuntos
Fibroínas , Regeneração da Medula Espinal , Ratos , Animais , Humanos , Seda/química , Tecidos Suporte/química , Ácido Hialurônico/farmacologia , Fibroínas/farmacologia , Fibroínas/química , Engenharia Tecidual/métodos , Células Endoteliais da Veia Umbilical Humana
19.
Angew Chem Int Ed Engl ; 63(15): e202400595, 2024 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-38321642

RESUMO

Synthetic spidroin fibers have not yet attained the same level of toughness and stability as natural spider silks due to the complexity of composition and hierarchical structure. Particularly, understanding the intricate interactions between spidroin components in spider fiber is still elusive. Herein, we report modular design and preparation of spidroin-mimetic fibers composed of a conservative C-terminus spidroin module, two different natural ß-sheets modules, and a non-spidroin random-coil module. The resulting fibers exhibit a toughness of ~200 MJ/m3, reaching the highest value among the reported artificial spider silks. The interactions between two components of recombinant spidroins facilitate the intermolecular co-assembly of ß-sheets, thereby enhancing the mechanical strength and reducing batch-to-batch variability in the dual-component spidroin fibers. Additionally, the dual-component spidroin fibers offer potential applications in implantable or even edible devices. Therefore, our work presents a generic strategy to develop high-performance protein fibers for diverse translations in different scenarios.


Assuntos
Fibroínas , Aranhas , Animais , Fibroínas/química , Conformação Proteica em Folha beta , Seda/química
20.
Sci Rep ; 14(1): 3942, 2024 02 16.
Artigo em Inglês | MEDLINE | ID: mdl-38365964

RESUMO

Electrospun scaffolds play important roles in the fields of regenerative medicine and vascular tissue engineering. The aim of the research described here was to develop a vascular scaffold that mimics the structural and functional properties of natural vascular scaffolding. The mechanical properties of artificial vascular tissue represent a key issue for successful transplantation in small diameter engineering blood vessels. We blended silk fibroin (SF) and fibrin to fabricate a composite scaffold using electrospinning to overcome the shortcomings of fibrin with respect to its mechanical properties. Subsequently, we then carefully investigated the morphological, mechanical properties, hydrophilicity, hemocompatibility, degradation, cytocompatibility and biocompatibility of the SF/fibrin (0:100), SF/fibrin (15:85), SF/fibrin (25:75), and SF/fibrin (35:65) scaffolds. Based on these in vitro results, we implanted SF/fibrin (25:75) vascular scaffold subcutaneously and analyzed its in vivo degradation and histocompatibility. The fiber structure of the SF/fibrin hybrid scaffold was smooth and uniform, and its fiber diameters were relatively small. Compared with the fibrin scaffold, the SF/fibrin scaffold clearly displayed increased mechanical strength, but the hydrophilicity weakened correspondingly. All of the SF/fibrin scaffolds showed excellent blood compatibility and appropriate biodegradation rates. The SF/fibrin (25:75) scaffold increased the proliferation and adhesion of MSCs. The results of animal experiments confirmed that the degradation of the SF/fibrin (25:75) scaffold was faster than that of the SF scaffold and effectively promoted tissue regeneration and cell infiltration. All in all, the SF/fibrin (25:75) electrospun scaffold displayed balanced and controllable biomechanical properties, degradability, and good cell compatibility. Thus, this scaffold proved to be an ideal candidate material for artificial blood vessels.


Assuntos
Fibroínas , Engenharia Tecidual , Animais , Engenharia Tecidual/métodos , Fibroínas/farmacologia , Fibroínas/química , Tecidos Suporte/química , Fibrina , Seda/química
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