Silk Fibroin Particles Research Articles

Inhalable drug-loaded silk fibroin carriers for pulmonary drug delivery.

The design and development of engineered micro and nano-carriers offering superior therapeutic performance compared to traditional delivery forms, are crucial in pharmaceutical research. Aerosolization and inhalation of carriers with improved solubility/stability of insoluble drugs, has huge potential for targeted drug delivery (DD) in various pulmonary diseases. Indeed, dedicated carriers must meet specific dimensional rules for proper lung delivery. Particles between 2-10 μm in size are normally deposited in the tracheobronchial region, while particles of 0.5-2 μm may be properly deposited in the alveoli. In this work, we report the development of inhalable nanostructured carries made of a 'green' bio-inspired polymer from aqueous solutions, i.e. silk fibroin (SF), efficiently loaded with a hydrophobic drug, i.e. the thyroid hormone levothyroxine (L-T4), a drug for the treatment of idiopathic pulmonary fibrosis. The aim is to optimize a standard method for the synthesis of SF-based nanocarriers with controlled size and shape, where a fine control of their geometrical properties is aimed at efficiently controlling the pulmonary DD. L-T4 loaded SF particles were synthesized through a one-pot co-precipitation method. Optimized systems were determined by varying the chemo-physical parameters during the synthesis. Ethylenediaminetetraacetic acid (EDTA) was used to remove CaCO3 cores. The proposed synthesis routes have led to two SF structures, whose structural heterogeneity and nanostructured morphology have been demonstrated using fluorescence microscopy, DLS, SEM and EDX. Two systems with varying shape and size have been obtained: (i) a flat disk-like SF structure with an irregular surface and an in-plane length of about 1-2 μm; (ii) solid SF nanospheres, obtained using ethylene glycol as additive, showing two size populations (main diameters of 0.5 μm and 1.7 μm). Solid nanospherical systems, in particular, show a tendency to arrange into agglomerates that, when loosely bound into smaller particles, can facilitate the delivery at the alveoli. Both formulations exhibited similar drug loading efficiencies, evaluated by HPLC analysis. However, SF nanospheres showed greater in vitro drug release after 24 hours. The demonstrated control over the characteristics imparted to the proposed DD systems may be critical to select the most suitable size/shape to achieve high rates of delivery to the appropriate lung compartment.

Cellulose-Silk Fibroin Composite Particles for Hydrophilic Drug-Controlled Release System: Preparation and Characterization

The objective of this study was to investigate the dissolution of cellulose (C) powder as a potential material for particle formation when combined with a silk fibroin (SF) solution. Each polymer solution as well as a mixture of cellulose and silk fibroin in different ratios were prepared into particles using the solvent emulsion-diffusion method. The particle morphology was characterized by scanning electron microscopy (SEM) and by attenuated reflection Fourier-transformed infrared spectroscopy (ATR-FTIR) and thermogravimetric analyzer (TGA) for the C-SF molecular interaction and thermal properties, respectively. The results showed that the cellulose particles appeared as separate fibers and loosely bound together, while silk fibroin particles were spherical in shape with dense texture and smooth surface. The C/SF mixed particles with different ratios could be formed into spherical particles and a ratio of 1:3 has the most spherical and solid. The ATR-FTIR results indicated the absorption spectra of the main functional groups both cellulose and silk fibroin. Blue dextran helped to enhance the temperature of the maximum decomposition rate (Td,max) of the particles via hydrogen bond interaction between the blue dextran and polymers. A burst release of blue dextran was observed in the first 12 h due to the swelling of particle surfaces and the slowest release found in the silk fibroin particles. The C/SF composite showed different blue dextran- eleased profiles and the particle with a high content of silk fibroin-released blue dextran in slowly rate. The results of this work can be used as a basis for applying C/SF composite particles in a hydrophilic drug release-controlled system.

The recombinant BMP-2 loaded silk fibroin microspheres improved the bone phenotype of mild osteogenesis imperfecta mice.

Osteogenesis imperfecta (OI) is an inherited congenital disorder, characterized primarily by decreased bone mass and increased bone fragility. Bone morphogenetic protein-2 (BMP-2) is a potent cytokine capable of stimulating bone formation, however, its rapid degradation and unanticipated in vivo effects restrict its application. The sustained release characteristic of silk fibroin (SF) microspheres may potentially address the aforementioned challenges, nevertheless they have not previously been tested in OI treatment. In the current investigation, recombinant BMP-2 (rBMP-2) loaded SF (rBMP-2/SF) microspheres-based release carriers were prepared by physical adsorption. The SF microparticles were characterized by scanning electron microscopy (SEM) and were investigated for their cytotoxicity behavior as well as the release profile of rBMP-2. The rBMP-2/SF microspheres were administered via femoral intramedullary injection to two genotypes of OI-modeled mice daily for two weeks. The femoral microstructure and histological performance of OI mice were evaluated 2 weeks later. The findings suggested that rBMP-2/SF spheres with a rough surface and excellent cytocompatibility demonstrated an initial rapid release within the first three days (22.15 ± 2.88% of the loaded factor), followed by a transition to a slower and more consistent release rate, that persisted until the 15th day in an in vitro setting. The factor released from rBMP-2/SF particles exhibited favorable osteoinductive activity. Infusion of rBMP-2/SF microspheres, as opposed to blank SF spheres or rBMP-2 monotherapy, resulted in a noteworthy enhancement of femoral microstructure and promoted bone formation in OI-modeled mice. This research may offer a new therapeutic approach and insight into the management of OI. However, further investigation is required to determine the systematic safety and efficacy of rBMP-2/SF microspheres therapy for OI.

Silk Fibroin Particles as Carriers in the Development of Hemoglobin-Based Oxygen Carriers.

Oxygen therapeutics have a range of applications in transfusion medicine and disease treatment. Synthetic molecules and all-natural or semi-synthetic hemoglobin-based oxygen carriers (HBOCs) have seen success as potential circulating oxygen carriers. However, many early HBOC products stalled in development due to side effects from excess hemoglobin in the blood stream and hemoglobin entering the tissue. To overcome these issues, research has focused on increasing the molecular diameter of hemoglobin by polymerizing hemoglobin molecules or encapsulating hemoglobin in liposomal carriers. This work leverages the properties of silk fibroin, a cytocompatible and non-thrombogenic biopolymer, known to entrap protein-based cargo, to engineer a fully protein-based oxygen carrier. Herein, an all-aqueous solvent evaporation technique was used to form silk particles via phase separation from a bulk polyvinyl alcohol phase (PVA). Particles size was tuned, and particles were formed with and without hemoglobin. The encapsulation efficiency and ferrous state of hemoglobin were analyzed, resulting in 60% encapsulation efficiency and a maximum of 20% ferric hemoglobin, yielding 100 µg/mL active hemoglobin in certain sfHBOC formulations. The system did not elicit a strong inflammation response in vitro, demonstrating the potential for this particle system to serve as an injectable HBOC.

Gallium ions incorporated silk fibroin hydrogel with antibacterial efficacy for promoting healing of Pseudomonas aeruginosa-infected wound

The continual resistance to antibiotics and the generation of a series of bacterial infections has emerged as a global concern, which requires appropriate measures and therapeutics to address such a menace. Herein, we report on Silk fibroin (SF) hydrogel with good biocompatibility and biodegradability fabricated through the crosslinking of the SF of different concentrations with Gallium nitrate (Ga (NO3)3) against Pseudomonas aeruginosa. However, the SF: Ga = 500: 1 (w/w) (SF/Ga) demonstrated a good bactericidal and wound healing effect as a result of the moderate and prolonged release of the Ga3+ following the gradual degradation of the hydrogel. The Ga3+, known for its innovative nature acted as a crosslinked agent and a therapeutic agent employing the "Trojan horse" strategy to effectively deal with the bacteria. Also, the Ga3+, which is positively charged neutralizes the negative potential value of the SF particles to reduce the charge and further induce the β-sheet formation in the protein structure, a characteristic of gelation in SF. The morphology showed a fabricated homogenous structure with greater storage modulus- G' with low loss modulus- G'' modulus demonstrating the mechanical performance and the ability of the SF/Ga hydrogel to hold their shape, at the same time allowing for the gradual release of Ga3+. A demonstration of biocompatibility, biodegradability, bactericidal effect and wound healing in in vitro and in vivo present the SF/Ga hydrogel as an appropriate platform for therapeutic and for antibacterial wound dressing.

Synergized bioperformance bone scaffolds of poly(vinyl alcohol)/silk fibroin particles with TiO2 as the base bone-mimicking materials for oral and maxillofacial surgery

Synergized bioperformance bone scaffolds of poly(vinyl alcohol)/silk fibroin particles with TiO2 as the base bone-mimicking materials for oral and maxillofacial surgery

Self-Assembled Inhalable Immunomodulatory Silk Fibroin Nanocarriers for Enhanced Drug Loading and Intracellular Antibacterial Activity.

In this study, a pH-induced self-assembly-based method has been developed to form silk fibroin nanoparticles (SFN-2) with a higher drug loading capacity (21.0 ± 2.1%) and cellular uptake than that of silk fibroin particles produced by a conventional desolvation method (SFN-1). Using the self-assembly method, rifampicin-encapsulated silk fibroin nanoparticles (R-SFN-2) were prepared with a size of 165 ± 38 nm at an optimum pH of 3.8. In silico analysis reveals that at acidic pH, the amino acid side chain charge neutralization of acidic residues, especially GLU64, promotes the formation of additional favorable interactions between the silk fibroin and the drug. The SFN-2 also possess a good aerosol property with a mass median aerodynamic diameter of 3.82 ± 0.71 μm and fine particle fraction of 64.0 ± 1.4%. These SFN-2 particles were selectively endocytosed by macrophages through clathrin- and caveolae-mediated endocytosis with a higher uptake efficiency (66.2 ± 2.1%) and were found to exhibit a sustained drug release in the presence of macrophage intracellular lysates. The cytokine and biomarker expression analyses revealed that SFN-2 could exhibit an immunomodulatory effect by polarizing the macrophages to an initial M1 phase and later M2 phase. Further, R-SFN-2 also exhibited an enhanced and sustained intracellular antibacterial activity against Mycobacterium smegmatis-infected macrophages than free rifampicin. Thus, the self-assembled silk fibroin particles with immunomodulatory action combined with a good aerosol and intracellular drug release property can be an attractive choice as a carrier for developing pulmonary drug delivery systems.

Investigation of chip formation mechanism in ultra-precision diamond turning of silk fibroin film

The importance of silk fibroin particles (size from tens of nanometres to hundreds of microns) as effective drug carriers has been well identified due to their superior mechanical strength, biocompatibility and biodegradability. They are usually fabricated by methods that have difficulties in controlling the size and shape to meet the required functions. In addition, using chemical agents in the fabrication process may result in biological risks and degradation of the fibroin structures. In this study, ultra-precision diamond turning (UPDT) technology is researched as an alternative chemical-free manufacturing approach to generate silk particles in the form of cutting chips with controllable size and shape. The machinability in UPDT of silk fibroin film (hereinafter referred to as silk film) was discussed in terms of specific cutting force and chip morphology. The material removal in UPDT of silk film is taken place in the ductile regime. Experimental results reveal that a rise in the cutting speed could significantly reduce the specific cutting force, while decreasing the depth of cut (DOC) could increase the specific cutting force because of the size effect. Moreover, a sharp point tool using a less than 2.5 μm/rev feed rate is preferred to generate helical silk chips. The formation mechanism of shear bands and serrated chips in UPDT of silk film was investigated with the aid of a finite element and smoothed particle hydrodynamics (FE-SPH) hybrid numerical model with Cowper-Symonds (CS) material equation. The material parameters of silk film were determined for the first time at p = 7 and D = 1140 s−1. The simulated specific cutting force with this set of material parameters is 49.0% smaller than the measured one. The simulated chip morphology (e.g. shear band spacing) was also in good agreement with the measured values. The simulation study shows that the shear band formation is done via two joining parts: one part propagates from the cutting edge to the free surface, another part initiates on the free surface and evolves towards the cutting edge. The chip segment is formed by a microcrack propagating from the free surface to the cutting edge. The underlying mechanism of formation of the serrated chip is found to have its roots in the hierarchical structure of silk fibroin.

Osteo-conductive hydrogel scaffolds of poly(vinylalcohol) with silk fibroin particles for bone augmentation: Structural formation and in vitro testing

Bone augmentation is an effective approach to treat patients who have bone loss at the maxillofacial area. In this research, osteo-conductive hydrogel scaffolds of poly(vinylalcohol) (PVA) with silk fibroin particles (SFP) were fabricated. The SFP were formed by dropping a solution of silk fibroin into acetone at different volume ratios (v/v) of silk to acetone: 1:3 (SFP-3), 1:6 (SFP-6), 1:12 (SFP-12), and 1:24 (SFP-24). The various SFP solutions were mixed with a PVA solution before fabrication into hydrogels by freeze-thawing. Afterwards, the hydrogels were freeze-dried to fabricate the scaffolds. The particle size and charge, molecular organization, and morphology of the SFP were characterized and observed with dynamic light scattering, Fourier transform infrared spectroscopy, differential scanning calorimetry, and scanning electron microscopy (SEM). The morphologies of the hydrogel scaffolds were observed with SEM. Swelling percentage was used to assess the swelling behavior of the hydrogel scaffolds. The mechanical properties were also tested. The scaffolds were cultured with osteoblast cells to test the biological performance, cell viability and performance, alkaline phosphatase activity, calcium deposition, and total protein. The SFP-24 was the smallest in particle size. PVA hydrogel scaffolds with SFP-24 demonstrated low particle aggregation, good particle distribution within the scaffold, and a lower swelling percentage. PVA hydrogel scaffolds with SFP had higher mechanical stability than scaffolds without the SFP. Furthermore, the PVA hydrogel scaffold with SFP-24 had better biological performance. Finally, the results demonstrated that PVA hydrogel scaffolds with SFP-24 showed good osteo-conductive performance which is promising for bone augmentation.

Progress in Preparation of Silk Fibroin Microspheres for Biomedical Applications

As a natural biomaterial, silk fibroin (SF) holds great potential in biomedical applications with its broad availability, good biocompatibility, high mechanical strength, ease of fabrication, and controlled degradation. With emerging fabrication methods, nanoand microspheres made from SF have brought about unique opportunities in drug delivery, cell culture, and tissue engineering. For these applications, the size and distribution of silk fibroin particles (SFPs) are critical and require precise control during fabrication. Herein, we review common and emerging SFPs fabrication methods and their biomedical applications, and also the challenges and opportunities for SFPs in the near future. Lay Summary: The application of silk in textile has an extraordinarily long history and new biomedical applications emerged owing to the good biocompatibility and versatile fabrication options of its major protein component, silk fibroin. With the development of nanotechnology and microfabrication, silk fibroin has been fabricated into nano- or microspheres with precisely controlled shape and distribution. In this review, we summarize common and emerging silk fibroin particle fabrication methods and their biomedical applications, and also discuss their challenges and opportunities in the nearest future.

The Interactions of Quantum Dot-Labeled Silk Fibroin Micro/Nanoparticles with Cells.

When silk fibroin particles are used for controlled drug delivery, particle size plays a key role in the location of the carrier on the cells as well as the transport pathway, utilization efficiency, and therapeutic effect of the drugs. In this study, the interactions of different-sized silk fibroin particles and cell lines were investigated. Silk fibroin microparticles with dry size of 1.9 ± 0.4 μm (2.7 ± 0.3 μm in wet state) and silk fibroin nanoparticles with dry size of 51.5 ± 11.0 nm (174.8 ± 12.5 nm in wet state) were prepared by salting-out method and high-voltage electrospray method, respectively. CdSe/ZnS quantum dots were coupled to the surface of the micro/nanoparticles. Photostability observations indicated that the fluorescence stability of the quantum dots was much higher than that of fluorescein isothiocyanate. In vitro, microparticles and nanoparticles were co-cultured with human umbilical vein endothelial cells EA.hy 926 and cervical cancer cells HeLa, respectively. The fluorescence test and cell viability showed that the EA.hy926 cells tended to be adhered to the microparticle surfaces and the cell proliferation was significantly promoted, while the nanoparticles were more likely to be internalized in HeLa cells and the cell proliferation was notably inhibited. Our findings might provide useful information concerning effective drug delivery that microparticles may be preferred if the drugs need to be delivered to normal cell surface, while nanoparticles may be preferred if the drugs need to be transmitted in tumor cells.

Rheological and ion-conductive properties of injectable and self-healing hydrogels based on xanthan gum and silk fibroin

The hydrogels with injectable and self-healing properties were prepared from xanthan gum (XG) and silk fibroin (SF) by using sodium trimetaphosphate (STMP) as crosslinker. A three-stage model of oscillation-shear-oscillation experiment was designed to mimic injection process and to observe destruction and regeneration of the hydrogels after shear. The XG3-SF-STMP hydrogels immediately recovered to original storage modulus of 80.6%–93.8% on removing shear. The hydrogels were 3D printed into the self-supporting constructions of hydrogel fibers with connected porous structures, and the XG3-SF-STMP hydrogel fibers exhibited smaller width than XG3-STMP. Oscillation rheological behavior indicated that XG3-SF-STMP hydrogels formed rapidly and exhibited more solid-like gel behavior than XG3-STMP. The hydrogel structures were destroyed under a strain (100%) larger than critical strain, but were rebuilt under a small strain (1%) with recovery ratio of 91.36–93.96% within 120 s, suggesting a self-healing property. Introduction of SF particles into XG3-STMP crosslinked networks improved stiffness and retained recoverability. Carboxyl and phosphate groups in the hydrogel networks are beneficial for XG3-SF-STMP hydrogels to absorb enough liquid electrolytes, leading to effective ionic conductivity. The ion-conductive hydrogel with injectable, self-healing, controlled release and non-cytotoxic properties possesses a promising prospect for tissue engineering and drug release application.

Reinforcement of calcium phosphate cement using alkaline-treated silk fibroin.

BackgroundBone cement plays an important role in the treatment of osteoporotic vertebral compression fractures. Calcium phosphate cement (CPC) is a potential alternative to poly(methyl methacrylate), currently the gold standard of bone cements. However, the poor mechanical properties of CPCs limit their clinical applications. The objective of this study was to develop reinforced CPCs for minimally invasive orthopedic surgeries by compositing silk fibroin (SF) with α-tricalcium phosphate.MethodsSF solution was treated with calcium hydroxide and characterized by Zeta potential analyzer and Fourier transform infrared spectroscopy. The alkaline-treated SF (tSF) was com-posited with α-tricalcium phosphate to obtain tSF/CPC composite, which was characterized using mechanical tests, scanning electron microscopy, handling property and biocompatibility tests, and sheep vertebral augmentation tests.ResultsUpon treatment with calcium hydroxide, larger SF particles and more abundant negative charge appeared in tSF solution. The tSF/CPCs exhibited a compact structure, which consisted of numerous SF -CPC clusters and needle-like hydroxyapatite (HAp) crystals. In addition, high transition rate of HAp in tSF/CPCs was achieved. As a result, the mechanical property of tSF/ CPC composite cements was enhanced remarkably, with the compressive strength reaching as high as 56.3±1.1 MPa. Moreover, the tSF/CPC cements showed good injectability, anti-washout property, and decent biocompatibility. The tSF/CPCs could be used to augment defected sheep vertebrae to restore their mechanical strength.ConclusiontSF/CPC may be a promising composite bone cement for minimally invasive orthopedic surgeries.

Fabricated porous silk fibroin particles for pH-responsive drug delivery and targeting of tumor cells

To improve the roles of tumor drug, porous silk fibroin particles have been designed as drug carriers. Doxorubicin (DOX) is encapsulated during the fabrication process of the silk fibroin particles to form DOX loaded silk fibroin particles (SFPs-DOX). Folic acid (FA) is covalently grafted to the surface of the silk fibroin particles (FA-SFPs-DOX) as a target group to the folate receptor of tumor cells. The experimental results showed that under the conditions of low pH, high ionic strength and high enzyme concentration, the release of loaded drugs in particles has stimulus response under various conditions. The synthesized silk fibroin particles have good biocompatibility. The specific binding of fibroin particles to folate receptors provides the particles to with target functions to the tumor cells. The job develops potential application of silk fibroin particles as smart drug carriers for the targeted therapy of tumor.

Effect of solution viscosity on retardation of cell sedimentation in DLP 3D printing of gelatin methacrylate/silk fibroin bioink

To improve the cell suspension of the gelatin-methacrylate(GelMA) bioink, the viscosity of the ink solution was controlled by biocompatible silk fibroin(SF) particles. The viscosity of the GelMA solution increased about 1.4 × 102 folds by the addition of 1 w/v% SF particles. The increase of solution viscosity reduced the displacement of cells and retarded the cell sedimentation significantly. SF-GelMA bioink containing 1 w/v% SF was non-cytotoxic and used for the 3D bioprinting by directly projecting a feature onto the bioink. The cells encapsulated in the structures showed a high level of metabolic activity and demonstrated that SF-GelMA was a biocompatible material.

Effect of aqueous and particulate silk fibroin in a rat model of experimental colitis

Silk fibroin (SF) has anti-inflammatory properties and promotes wound healing. Moreover, SF particles act as carriers of active drugs against intestinal inflammation due to their capacity to deliver the compound to the damaged colonic tissue. The present work assesses the effect of SF in the trinitrobenzenesulfonic acid model of rat colitis that resembles human intestinal inflammation. SF (8mg/kg) was administered in aqueous solution orally and in two particulate formats by intrarectal route, following two technologies: spray drying to make microparticles and desolvation in organic solvent to produce nanoparticles. SF treatments ameliorated the colonic damage, reduced neutrophil infiltration and improved the compromised oxidative status of the colon. They also reduced the gene expression of pro-inflammatory cytokines like IL-1β and the anti-inflammatory cytokine IL-10. Moreover, they improved the intestinal wall integrity by increasing the gene expression of some of its markers (villin, trefoil factor-3 and mucins), thus accelerating the healing. The immunomodulatory properties of SF particles were also tested in vitro in macrophages: they activated the immune response in basal conditions without increasing it after a pro-inflammatory insult. In conclusion, SF particles could be useful as carriers to deliver active drugs to the damaged intestinal colon with additional anti-inflammatory and healing properties.

Three dimensional poly(ε-caprolactone) and silk fibroin nanocomposite fibrous matrix for artificial dermis

Ideal dermal substitutes should have comparable physicochemical and biological properties to the natural skin tissue. In this study, we report a novel strategy to “engineer” controlled 3D nanocomposite fibrous matrix of poly(ε-caprolactone) (PCL) and silk fibroin (SF) for an artificial dermis application. Using a custom-designed cold-plate electrospinning and automatic magnet agitation system, up to 6mm of the thickness was achieved resulting from the accumulation of ice crystal layers on the PCL nanofibers surface-modified with the SF particles. The sacrificed ice crystals induced interconnected macro-pores ranging from tens to hundreds μm. The agitation system introduced uniform distribution of the SF protein within/on the nanofibers, preventing the particles from precipitation and agglomeration. NIH 3T3 fibroblasts proliferated in vitro on the PCL and PCL/SF scaffolds for 7days, but there was no statistical difference between the groups. Conversely, In vivo rat model studies revealed that the wound healing rate and collagen deposition increased with the SF content within the nanocomposites. The unique 3D construct with the PCL/SF nanocomposite fibers provided desirable spatial cues, surface topography, and surface chemistry for the native cells to infiltrate into the scaffolds. The wound healing potential of the nanocomposites was comparable to the commercial Matriderm® artificial dermis.

Formulation of Biologically-Inspired Silk-Based Drug Carriers for Pulmonary Delivery Targeted for Lung Cancer.

The benefits of using silk fibroin, a major protein in silk, are widely established in many biomedical applications including tissue regeneration, bioactive coating and in vitro tissue models. The properties of silk such as biocompatibility and controlled degradation are utilized in this study to formulate for the first time as carriers for pulmonary drug delivery. Silk fibroin particles are spray dried or spray-freeze-dried to enable the delivery to the airways via dry powder inhalers. The addition of excipients such as mannitol is optimized for both the stabilization of protein during the spray-freezing process as well as for efficient dispersion using an in vitro aerosolisation impactor. Cisplatin is incorporated into the silk-based formulations with or without cross-linking, which show different release profiles. The particles show high aerosolisation performance through the measurement of in vitro lung deposition, which is at the level of commercially available dry powder inhalers. The silk-based particles are shown to be cytocompatible with A549 human lung epithelial cell line. The cytotoxicity of cisplatin is demonstrated to be enhanced when delivered using the cross-linked silk-based particles. These novel inhalable silk-based drug carriers have the potential to be used as anti-cancer drug delivery systems targeted for the lungs.

Effect of Silk Fibroin Reinforcement on the Properties of Potato Starch-Polyvinyl Alcohol Blend Films

Starch and polyvinyl alcohol composite films, reinforced with raw and methylmethacrylate-grafted silk fibroin particles, were prepared by the solution-casting method on leveled plates. Silk fibroin was used as reinforcement for starch and polyvinyl alcohol (St/PVA) blends in order to improve their mechanical and water-resistance properties. The composites were plasticized with citric acid and cross-linked with gluteraldehyde. The reinforced films showed an increase in tensile strength with decrease in elongation at break. The optimized samples were characterized by scanning electron microscopy and were studied for their antibacterial properties. The biodegradable behavior was studied by the soil burial method.

Silk Fibroin-Based Complex Particles with Bioactive Encrustation for Bone Morphogenetic Protein 2 Delivery

Application of bone morphogenetic protein 2 (BMP-2) currently faces its challenges, and its efficacy of delivery has to be improved. The proper dosage of the powerful bioactive molecule is still under discussion and needs to be investigated further. In this work, pure silk fibroin particles and particles with calcium carbonate encrustation (complex particles) are designed, developed, and functionalized by BMP-2. These are used to deliver the bioactive molecule to mesenchymal stem cells (MSCs) to induce osteogenic differentiation. Results are compared with those of control groups of BMP-2 carriers under the same condition. Silk fibroin-based particles with size and component variations are prepared by self-assembly, desolvation, and soft template formation to improve BMP-2 loading efficiency. Results show that the particles significantly enhance osteogenic differentiation of MSCs, which is evident in the high ALP enzyme activity as well as the increased level of expression of osteogenic genes. Specifically, the combination of calcium compound and BMP-2 in the silk fibroin-calcium carbonate complex particles synergistically enhances osteogenesis. Release tests and mathematical modeling are applied to describe BMP-2 dissolution profiles, and the release mechanism is based on diffusion and polymer chain relaxation. In summary, the particles show high efficacies of BMP-2 delivery, and introduction of the complex particle can progressively enhance osteogenesis.