A hybrid structure based on silk fibroin/PVA nanofibers and alginate/gum tragacanth hydrogel embedded with cardamom extract.

Hybrid structures made of natural-synthetic polymers have been interested due to high biological features combining promising physical-mechanical properties. In this research, a hybrid dressing consisting of a silk fibroin (SF)/polyvinyl alcohol (PVA) nanofibers and sodium alginate (SA)/gum tragacanth (GT) hydrogel incorporating cardamom extract as an antibacterial agent was prepared. Accordingly, SF was extracted from cocoons followed by electrospinning in blend form with PVA (SF/PVA ratio: 1:1) under the voltage of 18 kV and the distances of 15 cm. The SEM images confirmed the formation of uniform, bead free fibers with the average diameter of 199 ± 28 nm. FTIR and XRD results revealed the successful extraction of SF and preparation of mixed fibrous mats. Next, cardamom oil extract-loaded SA/GT hydrogel was prepared and the nanofibrous structure was placed on the surface of hydrogel. SEM analysis depicted the uniform morphology of hybrid structure with desirable matching between two layers. TGA analysis showed desired thermal stability. The swelling ratio was found to be 1251% after 24 h for the hybrid structure and the drug was released without any initial burst. MTT assay and cell attachment results showed favorable biocompatibility and cell proliferation on samples containing extract, and antibacterial activity values of 85.35% against S. aureus and 75% against E. coli were obtained as well. The results showed that the engineered hybrid nanofibrous-hydrogel film structure incorporating cardamom oil extract could be a promising candidate for wound healing applications and skin tissue engineering.

Nanofiber-induced hierarchically-porous magnesium phosphate bone cements accelerate bone regeneration by inhibiting Notch signaling.

Magnesium phosphate bone cements (MPC) have been recognized as a viable alternative for bone defect repair due to their high mechanical strength and biodegradability. However, their poor porosity and permeability limit osteogenic cell ingrowth and vascularization, which is critical for bone regeneration. In the current study, we constructed a novel hierarchically-porous magnesium phosphate bone cement by incorporating extracellular matrix (ECM)-mimicking electrospun silk fibroin (SF) nanofibers. The SF-embedded MPC (SM) exhibited a heterogeneous and hierarchical structure, which effectively facilitated the rapid infiltration of oxygen and nutrients as well as cell ingrowth. Besides, the SF fibers improved the mechanical properties of MPC and neutralized the highly alkaline environment caused by excess magnesium oxide. Bone marrow stem cells (BMSCs) adhered excellently on SM, as illustrated by formation of more pseudopodia. CCK8 assay showed that SM promoted early proliferation of BMSCs. Our study also verified that SM increased the expression of OPN, RUNX2 and BMP2, suggesting enhanced osteogenic differentiation of BMSCs. We screened for osteogenesis-related pathways, including FAK signaing, Wnt signaling and Notch signaling, and found that SM aided in the process of bone regeneration by suppressing the Notch signaling pathway, proved by the downregulation of NICD1, Hes1 and Hey2. In addition, using a bone defect model of rat calvaria, the study revealed that SM exhibited enhanced osteogenesis, bone ingrowth and vascularization compared with MPC alone. No adverse effect was found after implantation of SM in vivo. Overall, our novel SM exhibited promising prospects for the treatment of critical-sized bone defects.

Facile In Situ Assembly of Nanofibers within Three-Dimensional Porous Matrices with Arbitrary Characteristics for Creating Biomimetic Architectures.

It is challenging to recapitulate the natural extracellular matrix's hierarchical nano/microfibrous three-dimensional (3D) structure with multilevel pores, good mechanical and hydrophilic properties, and excellent bioactivity for designing and developing advanced biomimetic materials. This work reports a new facile strategy for the scalable manufacturing of such a 3D architecture. Natural polymers in an aqueous solution are interpenetrated into a 3D microfibrous matrix with arbitrary shapes and property characteristics to self-assemble in situ into a nanofibrous network. The collagen fiber-like hierarchical structure and interconnected multilevel pores are achieved by self-assembly of the formed nanofibers within the 3D matrix, triggered by a simple cross-linking treatment. The as-prepared alginate/polypropylene biomimetic matrices are bioactive and have a tunable mechanical property (compressive modulus from ∼17 to ∼24 kPa) and a tunable hydrophilicity (water contact angle from ∼94° to 63°). This facile and versatile strategy allows eco-friendly and scalable manufacturing of diverse biomimetic matrices or modification of any existing porous matrices using different polymers.

The preparation and study on properties of calcium sulfate bone cement combined tuning silk fibroin nanofibers and vancomycin-loaded silk fibroin microspheres.

In this study, a bioactive composite material based on calcium sulfate hemihydrate (CSH) bone cement was studied, which use calcium sulfate dihydrate (CSD) as coagulant and silk fibroin nanofibers (SFF) solution as the curing liquid, further loaded vancomycin silk fibroin microspheres (SFM/VCM). The drug release effect of bone cements caused by tuning weight content of SFM/VCM (0.5, 1, 2%) and the concentration of silk fibroin solution (SFS) (20, 60, 100 mg/mL) used for preparation of SFM was studied in this article. Scanning electron microscope (SEM) demonstrated that the average diameter of microspheres gradually increased and the setting time was prolonged with the concentration of SFS increasing. X-ray diffraction (XRD) and Fourier transform infrared (FTIR) were used to analyze the composition of composite materials. The result of compressive strength revealed that the composites contained 0.5% SFM/VCM showed better mechanical performance independent on the concentration of microspheres and the cumulative drug release percentage of all composites were less than 55% after 4 weeks. The drug-loading bone cement possesses not only injectability but also sustained release capability which has a promising prospect in the field of bone substitute material.

Antibiotics drug release controlling and osteoblast adhesion from Titania nanotubes arrays using silk fibroin coating.

Bacterial infection, wide inflammation, and osteoporosis are the most common factors in the failure of orthopedic implants. The present study aims to design an orthopedic implant based on Titania nanotubes (TiO2-NTs) which not only have a high biocompatibility but also are characterized by anti-bacterial property. In order to improve the osseointegration of the TiO2-NTs structures (110-120 nm in diameter, 40 µm in length), they were used to coat the Titania implant by electrochemical anodizing. Vancomycin, which is soluble in water, was loaded as a main clinical drug to control intensive infections caused by positive gram bacteria. For the first time, Silk Fibroin (SF) Nanofibers coating was used to control drug release by the implementation of electrospinning on the TiO2-NTs surface. In order to investigate the anti-bacterial activities, S. aureus bacterium culture test was used. The cell culture of MG63 was conducted for both coated and non-coated samples of TiO2-NTs. The results showed that the SF Nanofibers coating not only controls the drug being freely released from TiO2-NTs but also effects adhesion and development of osteoblast cells. In this regard, this coating inhibits biofilm formation and development, as well as bacteria colonization due to anti-bacterial drug release. Therefore, this system can be considered as a promising alternative for orthopedic implants, preventing bone infection, osteomyelitis, bone cancer treatment, and other orthopedic diseases.

Creating Biomimetic Anisotropic Architectures with Co-Aligned Nanofibers and Macrochannels by Manipulating Ice Crystallization.

The continuous evolution of tissue engineering scaffolds has been driven by the desire to recapitulate structural features and functions of the natural extracellular matrix (ECM). However, it is still an extreme challenge to create a three-dimensional (3D) scaffold with both aligned nanofibers and aligned interconnected macrochannels to mimic the ECM of anisotropic tissues. Here, we develop a facile strategy to create such a scaffold composed of oriented nanofibers and interconnected macrochannels in the same direction, with various natural polymers typically used for tissue regeneration. The orientation of nanofibers and interconnected macrochannels can be easily tuned by manipulating ice crystallization. The scaffold demonstrates both structural and functional features similar to the natural ECM of anisotropic tissues. Taking silk fibroin as an example, the scaffold with radially oriented nanofibers and interconnected macrochannels is more efficient for capturing cells and promoting the growth of both nonadherent embryonic dorsal root ganglion neurons (DRGs) and adherent human umbilical vein endothelial cells (HUVECs) compared to the widely used scaffold types. Interestingly, DRGs and neurites on the SF scaffold demonstrate a 3D growth mode similar to that of natural nerve tissues. Furthermore, the coaligned nanofibers and macrochannels of the scaffold can direct HUVECs to assemble into blood vessel-like structures and their collagen deposition in their arrangement direction. The strategy could inspire the design and development of multifunctional 3D scaffolds with desirable structural features for engineering different tissues.

Impedimetric PSA aptasensor based on the use of a glassy carbon electrode modified with titanium oxide nanoparticles and silk fibroin nanofibers.

This article describes an impedimetric aptasensor for the prostate specific antigen (PSA), a widely accepted prostate cancer biomarker. A glassy carbon electrode (GCE) was modified with titanium oxide nanoparticles (TiO2) and silk fibroin nanofiber (SF) composite. The aptasensor was obtained by immobilizing a PSA-binding aptamer on the AuNP-modified with 6-mercapto-1-hexanol. The single fabrication steps were characterized by cyclic voltammetry and electrochemical impedance spectroscopy. The assay has two linear response ranges (from 2.5 fg.mL-1 to 25 pg.mL-1, and from 25 pg.mL-1 to 25 ng.mL-1) and a 0.8 fg.mL -1 detection limit. After optimization of experimental conditions, the sensor is highly selective for PSA over bovine serum albumin and lysozyme. It was successfully applied to the detection of PSA in spiked serum samples. Graphical abstract Schematic of the fabrication of an aptasensor for the prostate specific antigen (PSA). It is based on the use of a glassy carbon electrode modified with gold nanoparticles and titanium oxide-silk fibroin. The immobilization process of aptamer and interaction with PSA were followed by electrochemical impedance spectroscopy technique.