Silk Nanoparticles Research Articles

Alpha mangostin loaded crosslinked silk fibroin-based nanoparticles for cancer chemotherapy

Silk fibroin has been utilized extensively for biomedical purposes, especially the drug delivery systems. This study introduced and characterized three novel α-mangostin loaded crosslinked fibroin nanoparticles (FNPs), using EDC or PEI as a crosslinker, for cancer treatment. All three formulas were spherical particles with a mean size of approximately 300 nm. By varying the type and/or amount of the crosslinkers, particle surface charge was controllable from −15 to +30 mV. Crosslinked FNPs exhibited higher drug entrapment efficiency (70%) and drug loading (7%) than non-crosslinked FNP. FT-IR, XRD, and DSC analytical methods confirmed that α-mangostin was entrapped in FNPs in molecular dispersion form. Compared to the free α-mangostin, the crosslinked FNPs increased the drug’s solubility up to threefold. They also showed sustained release characteristics of more than 3 days, and reduced free α-mangostin hematotoxicity by 90%. The α-mangostin loaded FNPs were physicochemically stable for up to 24 h when dispersed in intravenous diluent and for at least 6 months when preserved as lyophilized powder at 4 °C. In terms of anticancer efficacy, on both Caco-2 colorectal and MCF-7 breast adenocarcinoma cell lines, all formulas maintain α-mangostin’s apoptotic effect while exhibit greater cytotoxicity than the free drug. In conclusion, α-mangostin loaded crosslinked FNPs show high potential for cancer chemotherapy.

Coating of modified poly(ethylene terephthalate) fibers with sericin-capped silver nanoparticles for antimicrobial application

In this work, a kind of amine-type PET fibers was synthesized by reacting hexamethylenediamine (HMDA) with methacrylic acid-g-poly(ethylene terephthalate) (PET-g-MAA) fibers for the adsorption of silk sericin-capped silver nanoparticles (S-AgNPs) to produce antimicrobial fibers. Firstly, PET fibers were grafted MAA by using free radical polymerization technique and HMDA was covalently connected to the grafted PET fibers. Then, for S-AgNPs synthesis, 10 mL of AgNO3 solution (1 mM, 5 mM and 10 mM) was mixed with 10 mL of 1% sericin solution at pH 11. The obtained solution was stirred at room temperature for 24 h. The color change from transparent to yellow–brown indicated the formation of S-AgNPs. AgNPs formation was also determined by measuring the absorbance spectra of S-AgNPs between 300 and 600 nm using UV–Vis spectrophotometer. To determine the antimicrobial properties of S-AgNPs, agar-well diffusion tests were performed. 5 mM and 10 mM S-AgNPs groups showed antimicrobial activity on Escherichia coli and Staphylococcus aureus. After characterization of the synthesized S-AgNPs with UV–Vis spectrophotometer, Zetasizer, FTIR and TEM, the modified PET fibers were coated with S-AgNPs (5 mM and 10 mM). The S-AgNPs coated PET fibers were characterized by FTIR, SEM and X-ray fluorescence spectroscopy. The antimicrobial activities of the obtained PET fibers were investigated on S. aureus and E. coli bacteria by using disk diffusion test. It was found that the S-AgNPs coated modified PET fibers exhibited antimicrobial activities toward both gram-positive and gram-negative bacteria. The resulting polymeric PET fibers containing nano-silver can be used as an antimicrobial surface for many applications such as wound dressing.

Multi-bioresponsive silk fibroin-based nanoparticles with on-demand cytoplasmic drug release capacity for CD44-targeted alleviation of ulcerative colitis

The requirement for the favorable therapeutics against ulcerative colitis (UC) is that anti-inflammatory drugs can be specifically internalized by macrophages and subsequently be on-demand released to suppress inflammation. Herein, we developed a type of multi-bioresponsive anti-inflammatory drug (curcumin, CUR)-loaded nanoparticles (NPs) that were derived from natural silk fibroin and followed by surface functionalization with chondroitin sulfate (CS). The generated CS-CUR-NPs had a desired average particle size (175.4 nm), a uniform size distribution and negative surface charge (−35.5 mV). Strikingly, these NPs exhibited excellent bioresponsibility when triggered with the intrinsic stimuli (acidity, glutathione and reactive oxygen species) within activated macrophages, indicating that they could conceivably confer the on-demand intracellular drug release. Furthermore, we found that CS functionalization yielded notably targeted drug delivery to macrophages, and thereby enhanced the anti-inflammatory activities of NPs. Most importantly, animal experiments revealed that these nanotherapeutics could remarkably alleviate the symptoms of UC, maintain the homeostasis of intestinal microbiota and improve the survival rate of mice with UC through the route of oral administration or intravenous injection. Our results suggest that these facilely fabricated CS-CUR-NPs, which exhibit excellent biocompatibility, multi-bioresponsive drug release and macrophage-targeted capacity, could be exploited as a promising therapeutic platform for clinical UC treatment.

PEGylation-Dependent Metabolic Rewiring of Macrophages with Silk Fibroin Nanoparticles.

Silk fibroin nanoparticles are emerging as promising nanomedicines, but their full therapeutic potential is yet to be realized. These nanoparticles can be readily PEGylated to improve colloidal stability and to tune degradation and drug release profiles; however, the relationship between silk fibroin nanoparticle PEGylation and macrophage activation still requires elucidation. Here, we used in vitro assays and nuclear magnetic resonance based metabolomics to examine the inflammatory phenotype and metabolic profiles of macrophages following their exposure to unmodified or PEGylated silk fibroin nanoparticles. The macrophages internalized both types of nanoparticles, but they showed different phenotypic and metabolic responses to each nanoparticle type. Unmodified silk fibroin nanoparticles induced the upregulation of several processes, including production of proinflammatory mediators (e.g., cytokines), release of nitric oxide, and promotion of antioxidant activity. These responses were accompanied by changes in the macrophage metabolomic profiles that were consistent with a proinflammatory state and that indicated an increase in glycolysis and reprogramming of the tricarboxylic acid cycle and the creatine kinase/phosphocreatine pathway. By contrast, PEGylated silk fibroin nanoparticles induced milder changes to both inflammatory and metabolic profiles, suggesting that immunomodulation of macrophages with silk fibroin nanoparticles is PEGylation-dependent. Overall, PEGylation of silk fibroin nanoparticles reduced the inflammatory and metabolic responses initiated by macrophages, and this observation could be used to guide the therapeutic applications of these nanoparticles.

Investigating Recombinant Aciniform Silk Nanoparticles as Potential Drug Carriers and as Intermediates in Silk Fibrillogenesis

Spider silks form biomaterials including fibres, films, and nanospheres with excellent mechanical properties and biocompatibility. As such, silk materials have been investigated as a component of biomedical textiles for grafts and implants, as a scaffold for tissue regeneration, and as a drug delivery vehicle. Silk fibres generally form in vivo in a process where soluble proteins are forced through a narrowing spinning duct, giving rise to shear force. One theory of fibre formation is that silk proteins are stored in micellar form, with these micelles elongating while being forced together in the spinning duct, undergoing morphological and conformational transitions as shear forces are applied. Silk nanoparticles can be self-assembled, with concomitant or post-assembly incorporation of a variety of drug-like molecules into their interior. Here, we focus on the mechanism of formation of previously uncharacterized nanoparticles by recombinant aciniform spider silk proteins, their potential as drug carriers, and the role of these self-assembled species in the initial stages of fibre formation. The aciniform spidroin protein (AcSp1) from Argiope trifasciata consists of ≥14 repeat units of 200 amino acids (“W”, from wrapping silk) flanked by a non-repetitive N- and C-terminal domains. Recombinant proteins consisting of a single repeat (W1) spontaneously self-assemble into spherical micelle-like nanoparticles in potassium phosphate buffer. Insoluble W1 nanoparticles also form in highly concentrated phosphate buffer. Features of conformational transitions associated with nanoparticle formation from monomeric W1 are delineated here using tryptophan mutants of W1 through intrinsic tryptophan fluorescence and 19F NMR spectroscopy. Nanoparticle properties are correlated to these features using electron microscopy, dynamic light scattering, and Raman spectroscopy. Ultimately, reproducible and specific production of these nanoparticles will assist in the design and optimization of aciniform silk-based materials for biomedical and other materials applications.

Microfluidic-assisted silk nanoparticle tuning.

Silk is now making inroads into advanced pharmaceutical and biomedical applications. Both bottom-up and top-down approaches can be applied to silk and the resulting aqueous silk solution can be processed into a range of material formats, including nanoparticles. Here, we demonstrate the potential of microfluidics for the continuous production of silk nanoparticles with tuned particle characteristics. Our microfluidic-based design ensured efficient mixing of different solvent phases at the nanoliter scale, in addition to controlling the solvent ratio and flow rates. The total flow rate and aqueous : solvent ratios were important parameters affecting yield (1 mL min−1 > 12 mL min−1). The ratios also affected size and stability; a solvent : aqueous total flow ratio of 5 : 1 efficiently generated spherical nanoparticles 110 and 215 nm in size that were stable in water and had a high beta-sheet content. These 110 and 215 nm silk nanoparticles were not cytotoxic (IC50 > 100 μg mL−1) but showed size-dependent cellular trafficking. Overall, microfluidic-assisted silk nanoparticle manufacture is a promising platform that allows control of the silk nanoparticle properties by manipulation of the processing variables.

Evaluation of the Toxicity Effects of Silk Fibroin on Isolated Fibroblast and Huvec Cells.

Emerging line research showed that silk nanoparticles (NPs) have toxicity on the fibroblast and Huvec cells without any toxicity recognized mechanisms. Recently, it suggested peripheral arterial disease confounds almost eight million Americans. Also, due to the main effect of fibroblast in a production of extracellular matrix (ECM), adhesive molecules, glycoproteins and various cytokines, it decided to define the toxicity mechanistic of silk NPs in fibroblast and Huvec cells based on oxidative stress markers. Therefore, it investigated whether silk NPs is able to induce any abnormality in the fibroblast and Huvec cells based on reliable and documented oxidative stress methods. Our results indicated that silk NPs (0.5, 1 and 2 mg/mL) induces cellular and mitochondrial dysfunction including an increase in ROS production, lipid peroxidation, mitochondria membrane potential (MMP) collapse, and oxidation of thiol groups which caused to cytochrome c release. Besides, lysosomal integrity damage and decreased in ATP/ADP ratio proposed disruptive effect of silk NPs on the mitochondrial respiratory chain and cell death signaling induction.

Oral Delivery of Curcumin Using Silk Nano- and Microparticles.

The paper presents a study using silk nano- and microparticles as carriers to deliver curcumin (CUR), a multifunctional and efficient, yet water-insoluble, antioxidant, with the primary goal of enhancing bioavailability. CUR-loaded silk nanoparticles were prepared by emulsification, resulting in particle sizes ranging from 229 to 2286 nm in diameter, with loading efficiencies from 22 to 41%. The encapsulation mechanism involved hydrophobic interaction between the β-sheet structure fabricated from the silk and the phenol groups of curcumin. In vitro studies revealed that curcumin was released more slowly from the larger particles than the smaller particles. An HPLC-MS/MS method was developed to determine the content of curcumin in rat plasma. The particles were orally administered at a single dose in rats, and the pharmacokinetic parameters were evaluated and compared with those of plain curcumin crystals. After 24 h, the larger silk particles (CSN-800, CUR-silk nanoparticles with sizes ∼800 nm) showed longer plasma circulation time for CUR (Tmax was 1.76 ± 0.10 h) when compared with that of the smaller silk particles (CSN-200 with sizes ∼200 nm, Tmax was 1.17 ± 0.07 h). The Cmax of CSN-200 was 3-fold greater than that of CSN-800. By contrast to the groups of plain CUR (without silk particles), CSN-200 and CSN-800 increased CUR bioavailability 17- and 5- fold, respectively. When mixtures of CSN-200 and CSN-800 at different ratios were orally administered to rats, medium bioavailability (>13 times) and exposure time (T1/2 > 1.68 h) were obtained. These results elucidated the effect of CUR-silk particle sizes on bioavailability after oral delivery, as a basis for future applications.

Preparation of a Codelivery System Based on Vancomycin/Silk Scaffold Containing Silk Nanoparticle Loaded VEGF.

One of the main challenges of using biomaterials for inducing bone regeneration is the bacterial resistance before complete bone repair. Biomaterials with both antibacterial and bone regeneration properties are more promising for bone repair. In the present study, vascular endothelial growth factor (VEGF) was loaded on silk fibroin nanoparticles (SFNPs) and then embedded in silk scaffold containing vancomycin to form a dual drug release system. The chemical and physical properties of the fabricated structure were confirmed by Fourier transform infrared, scanning electron microscopy, and ζ-potential analysis. The size of spherical SFNPs was ∼92 nm. The release kinetics of vancomycin and VEGF showed that ∼99.56% of vancomycin and ∼14% of VEGF were released during 21 and 28 days, respectively. The bioactivity of VEGF was ∼75%. Disk diffusion test confirmed the ability of this drug delivery system against methicillin-resistant Staphylococcus aureus (MRSA). Moreover, expression of the endothelial markers (FLK-1, vWF, and VE-cadherin), alkaline phosphatase, and matrix mineral production were higher in VEGF loaded groups. Taken together, the results indicated that the fabricated codelivery system was able to simultaneously deliver antibiotic and angiogenic factor, which can be considered as a potential candidate for the treatment of contaminated bone injuries.

Protein Polymer-Based Nanoparticles: Fabrication and Medical Applications.

Nanoparticles are particles that range in size from about 1–1000 nanometers in diameter, about one thousand times smaller than the average cell in a human body. Their small size, flexible fabrication, and high surface-area-to-volume ratio make them ideal systems for drug delivery. Nanoparticles can be made from a variety of materials including metals, polysaccharides, and proteins. Biological protein-based nanoparticles such as silk, keratin, collagen, elastin, corn zein, and soy protein-based nanoparticles are advantageous in having biodegradability, bioavailability, and relatively low cost. Many protein nanoparticles are easy to process and can be modified to achieve desired specifications such as size, morphology, and weight. Protein nanoparticles are used in a variety of settings and are replacing many materials that are not biocompatible and have a negative impact on the environment. Here we attempt to review the literature pertaining to protein-based nanoparticles with a focus on their application in drug delivery and biomedical fields. Additional detail on governing nanoparticle parameters, specific protein nanoparticle applications, and fabrication methods are also provided.

Self-stabilized silk sericin-based nanoparticles: In vivo biocompatibility and reduced doxorubicin-induced toxicity

A variety of colloid stabilizers and cryoprotectants confer improved nanoparticle (NP) colloidal stability and redisperability. However, discounted tumor targetability, delivery efficacy and possible side effects limit the application in vascular delivery of NPs. Here we present water-soluble silk sericin (SS) not only as a material for the preparation of NPs, but also both a dispersion stabilizer and a cryoprotectant. In the absence of any stabilizers, SS-based NPs (SSC@NPs) can resist the adsorption of serum proteins, preventing the formation of particle agglomerates. Following freeze-drying without addition of cryoprotectants, SSC@NPs powder can be easily resuspended into NP dispersion with a nearly monodispersed distribution. Additionally, SSC@NPs do not result in acute toxicity in mice at a dose of 400 mg/kg with a slow injection. Moreover, doxorubicin (DOX)-loaded SSC@NPs (DOX-SSC@NPs) diminish the biodistribution of DOX in the heart, mitigating DOX-induced cardiotoxicity of mice without compromising therapeutic efficacy. Our results suggest that the self-stabilized SSC@NPs could be a secure and effective drug carrier for intravenous administration when deprived of protective agents. Statement of SignificanceDuring manufacturing process such as freeze-drying, or interaction with complex fluids like blood, NPs for systemic drug delivery need to be highly dispersible and structurally intact. In this work, we have demonstrated the self-stability of SSC@NPs subjected to biological media and freeze-drying. This study represents the first work showing water-soluble SS could both act as a dispersion stabilizer and a cryoprotectant due to its hydrophilicity. Plus, good in vivo biocompatibility of SSC@NPs has been confirmed. Therefore, it may be promising that water-soluble SS can be generally used as a safe biomaterial against serum adsorption.

Degradation Behavior of Silk Nanoparticles-Enzyme Responsiveness.

Silk nanoparticles are viewed as promising vectors for intracellular drug delivery as they can be taken up into cells by endocytosis and trafficked to lysosomes, where lysosomal enzymes and the low pH trigger payload release. However, the subsequent degradation of the silk nanoparticles themselves still requires study. Here, we report the responsiveness of native and PEGylated silk nanoparticles to degradation following exposure to proteolytic enzymes (protease XIV and α-chymotrypsin) and papain, a cysteine protease. Both native and PEGylated silk nanoparticles showed similar degradation behavior over a 20 day exposure period (degradation rate: protease XIV > papain ≫ α-chymotrypsin). Within 1 day, the silk nanoparticles were rapidly degraded by protease XIV, resulting in a ∼50% mass loss, an increase in particle size, and a reduction in the amorphous content of the silk secondary structure. By contrast, 10 days of papain treatment was necessary to observe any significant change in nanoparticle properties, and α-chymotrypsin treatment had no effect on silk nanoparticle characteristics over the 20-day study period. Silk nanoparticles were also exposed ex vivo to mammalian lysosomal enzyme preparations to mimic the complex lysosomal microenvironment. Preliminary results indicated a 45% reduction in the silk nanoparticle size over a 5-day exposure. Overall, the results demonstrate that silk nanoparticles undergo enzymatic degradation, but the extent and kinetics are enzyme-specific.

Silk nanoparticles: from inert supports to bioactive natural carriers for drug delivery.

Silk proteins have been studied and employed for the production of drug delivery (nano)systems. They show excellent biocompatibility, controllable biodegradability and non-immunogenicity and, if needed, their properties can be modulated by blending with other polymers. Silk fibroin (SF), which forms the inner core of silk, is a (bio)material officially recognized by the Food and Drug Administration for human applications. Conversely, the potential of silk sericin (SS), which forms the external shell of silk, could still be considered under evaluation. At the best of our knowledge, nanoparticles based on silk sericin "alone" cannot be produced, due to its physicochemical instability influenced by extreme pH, high water solubility and temperature; for these reasons, it almost always needs to be combined with other polymers for the development of drug delivery systems. In this review, we focused on silk proteins as bioactive natural carriers, since they show not only optimal features as inert excipients, but also remarkable intrinsic biological activities. SF has anti-inflammatory properties, while SS presents antioxidant, anti-tyrosine, anti-aging, anti-elastase and anti-bacterial features. Here, we give an overview on SF or SS silk-based nanosystems, with particular attention on the production techniques.

Silk nanoparticles: proof of lysosomotropic anticancer drug delivery at single-cell resolution

Silk nanoparticles are expected to improve chemotherapeutic drug targeting to solid tumours by exploiting tumour pathophysiology, modifying the cellular pharmacokinetics of the payload and ultimately resulting in trafficking to lysosomes and triggering drug release. However, experimental proof for lysosomotropic drug delivery by silk nanoparticles in live cells is lacking and the importance of lysosomal pH and enzymes controlling drug release is currently unknown. Here, we demonstrate, in live single human breast cancer cells, the role of the lysosomal environment in determining silk nanoparticle-mediated drug release. MCF-7 human breast cancer cells endocytosed and trafficked drug-loaded native and PEGylated silk nanoparticles (∼100 nm in diameter) to lysosomes, with subsequent drug release from the respective carriers and nuclear translocation within 5 h of dosing. A combination of low pH and enzymatic degradation facilitated drug release from the silk nanoparticles; perturbation of the acidic lysosomal pH and inhibition of serine, cysteine and threonine proteases resulted in a 42% ± 2.2% and 33% ± 3% reduction in nuclear-associated drug accumulation for native and PEGylated silk nanoparticles, respectively. Overall, this study demonstrates the importance of lysosomal activity for anticancer drug release from silk nanoparticles, thereby providing direct evidence for lysosomotropic drug delivery in live cells.

Biocompatibility assessment of silk nanoparticles: hemocompatibility and internalization by human blood cells

Many nanoparticles are designed for use as potential nanomedicines for parenteral administration. However, emerging evidence suggests that hemocompatibility is important, but is highly particle- and test-bed dependent. Thus, knowledge of bulk material properties does not predict the hemocompatibility of uncharacterized nanoparticles, including silk nanoparticles. This study compares the hemocompatibility of silk versus silica nanoparticles, using whole human blood under quasi-static and flow conditions. Substantial hemocompatibility differences are noted for some nanoparticles in quasi-static versus dynamic studies; i.e., the inflammatory response to silk nanoparticles is significantly lower under flow versus quasi-static conditions. Silk nanoparticles also have very low coagulant properties - an observation that scales from the macro- to the nano-level. These nanoparticle hemocompatibility studies are complemented by preliminary live cell measurements to evaluate the endocytosis and trafficking of nanoparticles in human blood cells. Overall, this study demonstrates that nanoparticle hemocompatibility is affected by several factors, including the test bed design.

Metabolic Reprogramming of Macrophages Exposed to Silk, Poly(lactic-co-glycolic acid), and Silica Nanoparticles.

Monitoring macrophage metabolism in response to nanoparticle exposure provides new insights into biological outcomes, such as inflammation or toxicity, and supports the design of tailored nanomedicines. This paper describes the metabolic signature of macrophages exposed to nanoparticles ranging in diameter from 100 to 125 nm and made from silk, poly(lactic-co-glycolic acid) or silica. Nanoparticles of this size and type are currently at various stages of preclinical and clinical development for drug delivery applications. 1 H NMR analysis of cell extracts and culture media is used to quantify the changes in the intracellular and extracellular metabolomes of macrophages in response to nanoparticle exposure. Increased glycolytic activity, an altered tricarboxylic acid cycle, and reduced ATP generation are consistent with a proinflammatory phenotype. Furthermore, amino acids possibly arising from autophagy, the creatine kinase/phosphocreatine system, and a few osmolytes and antioxidants emerge as important players in the metabolic reprogramming of macrophages exposed to nanoparticles. This metabolic signature is a common response to all nanoparticles tested; however, the direction and magnitude of some variations are clearly nanoparticle specific, indicating material-induced biological specificity. Overall, metabolic reprogramming of macrophages can be achieved with nanoparticle treatments, modulated through the choice of the material, and monitored using 1 H NMR metabolomics.

Stem cell-extracellular vesicles as drug delivery systems: New frontiers for silk/curcumin nanoparticles

The aim of this work was to develop a novel carrier-in-carrier system based on stem cell-extracellular vesicles loaded of silk/curcumin nanoparticles by endogenous technique. Silk nanoparticles were produced by desolvation method and curcumin has been selected as drug model because of its limited water solubility and poor bioavailability. Nanoparticles were stable, with spherical geometry, 100nm in average diameter and the drug content reached about 30%. Cellular uptake studies, performed on mesenchymal stem cells (MSCs), showed the accumulation of nanoparticles in the cytosol around the nuclear membrane, without cytotoxic effects. Finally, MSCs were able to release extracellular vesicles entrapping silk/curcumin nanoparticles. This combined biological-technological approach represents a novel class of nanosystems, combining beneficial effects of both regenerative cell therapies and pharmaceutical nanomedicine, avoiding the use of viable replicating stem cells.

Silk nanoparticles—an emerging anticancer nanomedicine

Silk is a sustainable and ecologically friendly biopolymer with a robust clinical track record in humans for load bearing applications, in part due to its excellent mechanical properties and biocompatibility. Our ability to take bottom-up and top-down approaches for the generation of silk (inspired) biopolymers has been critical in supporting the evolution of silk materials and formats, including silk nanoparticles for drug delivery. Silk nanoparticles are emerging as interesting contenders for drug delivery and are well placed to advance the nanomedicine field. This review covers the use of Bombyx mori and recombinant silks as an anticancer nanomedicine, highlighting the emerging trends and developments as well as critically assessing the current opportunities and challenges by providing a context specific assessment of this multidisciplinary field.

Study on Antheraea perny Silk Fibroin Nanoparticles Carried Insulin

Chinese oak tasar Antheraea pernyi silk fibroin nanoparticles are a promising biomaterial for drug delivery because of its good properties, such as biodegradability and biocompatibility. In recent decades, self-assembling nanoparticles derived from Antheraea perny silk fibroin have risen too much interest in application of drug delivery. In this paper, the Ca2+ in calcium gluconate induced self-assembling nanoparticles. It was fabricated by blending aqueous Antheraea perny silk fibroin solution, calcium ion and insulin in a proper mixing ratio. The structure of the silk nanoparticles carried insulin was characterized by X-ray diffraction, FTIR and DTA. The size and the morphology of silk nanoparticles were tested using scanning electron microscope and particulate size description analysers. The drug content, covering rate and in vitro cumulative release of silk nanoparticles were also determinate. The results demonstrated that the Ca2+ induced selfassembling nanoparticles hold the particle size of 200~500 nm. The phenomenon of the sustained release is comparatively obvious in the silk nanoparticles. To summarize, this research will open up a new experimental basis on the usage of macromolecular drug, insulin which can properly meet the satisfaction of the patients desired for sustained-release insulin.

Monodisperse phase transfer and surface bioengineering of metal nanoparticles via a silk fibroin protein corona.

Uniform hydrophobic nanoparticles synthesized in nonpolar solvents possess excellent physio-chemical properties, showing great potential in biomedical applications. However, the presence of hydrophobic ligands on their surfaces limits their use under physiological conditions. Inspired by protein coronas present at the nano-bio interface, here we report a facile and universal method for phase transfer and surface bioengineering of hydrophobic nanoparticles using β-sheet-rich silk fibroin, a FDA-approved natural protein. Due to its amphiphilicity and high mechanical stiffness, the β-sheet-rich silk fibroin not only readily drags nanoparticles from an organic phase into aqueous media but also endows them with excellent monodispersity and long-term stability. The silk fibroin-coated nanoparticles can retain the magnetic and optical properties of the original nanoparticles, acting effectively as probes for biomedical imaging and biosensing. Furthermore, hydrophobic drugs can be easily adsorbed onto the protein coating via hydrophobic interaction, allowing the construction of promising theranostic nanoagents. Given these unique features, the strategy developed here possesses great promise in facilitating biomedical applications of hydrophobic nanomaterials.