Silk Fibroin Nanoparticles Research Articles

The Effect of Sterilization on the Characteristics of Silk Fibroin Nanoparticles.

In recent years, silk fibroin nanoparticles (SFNs) have been consolidated as drug delivery systems (DDSs) with multiple applications in personalized medicine. The design of a simple, inexpensive, and scalable preparation method is an objective pursued by many research groups. When the objective is to produce nanoparticles suitable for biomedical uses, their sterility is essential. To achieve sufficient control of all the crucial stages in the process and knowledge of their implications for the final characteristics of the nanoparticles, the present work focused on the final stage of sterilization. In this work, the sterilization of SFNs was studied by comparing the effect of different available treatments on the characteristics of the nanoparticles. Two different sterilization methods, gamma irradiation and autoclaving, were tested, and optimal conditions were identified to achieve the sterilization of SFNs by gamma irradiation. The minimum irradiation dose to achieve sterilization of the nanoparticle suspension without changes in the nanoparticle size, polydispersity, or Z-potential was determined to be 5 kiloGrays (kGy). These simple and safe methods were successfully implemented for the sterilization of SFNs in aqueous suspension and facilitate the application of these nanoparticles in medicine.

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.

BioMIPs: molecularly imprinted silk fibroin nanoparticles to recognize the iron regulating hormone hepcidin

The possibility to prepare molecularly imprinted nanoparticles from silk fibroin was recently demonstrated starting from methacrylated silk fibroin and choosing a protein as template. Here, we attempted the imprinting of fibroin-based molecularly imprinted polymers (MIPs), called bioMIPs, using as a template hepcidin that is a iron-metabolism regulator-peptide, possessing a hairpin structure. A homogeneous population (PDI < 0.2) of bioMIPs with size ~50 nm was produced. The bioMIPs were selective for the template; the estimated dissociation constant for hepcidin was KD = 3.6 ± 0.5 10−7 M and the average number of binding sites per bioMIP was equal to 2. The bioMIPs used in a competitive assay for hepcidin in serum showed a detection range of 1.01 10−7– 6.82 10−7 M and a limit of detection of 3.29 10−8 M.Graphical abstract

3D-printable chitosan/silk fibroin/cellulose nanoparticle scaffolds for bone regeneration via M2 macrophage polarization

Biopolymers-induced immune microenvironment exhibited prominent effects on bone regeneration. Osteo-immunomodulatory responses of cellulose nanoparticles incorporated chitosan hydrogel scaffolds have not yet been reported. The objective of this study was to monitor the synergistic effects of silk fibroin and cellulose nanoparticles on the immune-modulatory behavior of chitosan biopolymer scaffolds. 3D-printed biodegradable cellulose nanoparticles-reinforced chitosan/silk fibroin (CS/SF/CNPs) scaffolds were fabricated and characterized by different spectroscopic techniques. The improved rheological and recovery strength was observed in CS/SF/CNPs hydrogels than pure polymer hydrogels. A significant shift from M1 → M2 macrophages polarization occurred in the CS/SF/CNPs scaffolds treated groups than the control after 3 days of incubation, showing its immune-modulatory potential. Osteo-immunomodulatory effects of the fabricated scaffolds were analyzed on human bone marrow-derived mesenchymal stem cells (hBMSCs), with macrophages-derived conditioned media (M-CM). Enhanced bone regeneration was observed in the calvaria defect rat model, indicating that the fabricated scaffolds are promising materials for bone-healing applications.

Silk fibroin-anastrozole loaded prolonged-release biodegradable nanomedicine: A promising drug delivery system for breast cancer therapy

Although several methods are being used for the inhibition, cure, and regimen of breast cancer, many drawbacks are underlying. The idea of this study was to evolve biodegradable, anastrozole-loaded silk fibroin nanoparticles for breast cancer therapy. The nanoparticles were formulated via solvent change method using various concentrations of silk fibroin, and mannitol (stabilizer). Nanoparticles were developed and characterized in respect of particle size, zeta potential, polydispersity, and encapsulation efficiency followed by optimization using 25 fractional factorial design in Design Expert software-12 (Stat-Ease, USA). Thereafter, using variance analysis (ANOVA), the optimized formulation was selected (Sol 2) and subjected to compatibility studies, thermal analysis, surface morphology, x-ray diffraction, transmission electron spectroscopy, drug release profile, and accelerated stability study. The optimized SF-ANS-NP formulation reported a particle size of 181.70 ± 1.3 nm. At 168th h, a sustained release of 94.15% was recorded. The accelerated stability analysis values have proven to be stable for 90 days. The findings showed that anastrozole nanoparticles combined with silk fibroin might be a potentially viable delivery strategy for long-term breast cancer therapy.

Fabrication of Silk Scaffold Containing Simvastatin-Loaded Silk Fibroin Nanoparticles for Regenerating Bone Defects

Background:In the present study, a tissue engineered SF scaffold containing simvastatin-loaded SFNPs were used to stimulate the regeneration of the defected bone.Methods:At first, the porous SF scaffold was prepared using freeze-drying. Then simvastatin-loaded SFNPs were made by dissolvation method and embedded in the SF scaffold. Afterwards, the scaffold and the NPs were characterized in terms of physicochemical properties and the ability to release the simvastatin small molecule.Results:The results exhibited that the SF scaffold had a porous structure suitable for releasing the small molecule and inducing the proliferation and attachment of osteoblast cells. SFNPs containing simvastatin had spherical morphology and were 174 ± 4 nm in size with -24.5 zeta potential. Simvastatin was also successfully encapsulated within the SFNPs with 68% encapsulation efficiency. Moreover, the small molecule revealed a sustained release profile from the NPs during 35 days. The results obtained from the in vitro cell-based studies indicated that simvastatin-loaded SFNPs embedded in the scaffold had acceptable capacity to promote the proliferation and ALP production of osteoblast cells while inducing osteogenic matrix precipitation. Conclusion:The SF scaffold containing simvastatin-loaded SFNPs could have a good potential to be used as a bone tissue-engineered construct.

Poly (vinyl alcohol)/Silk Fibroin/Ag NPs composite nanofibers for bone tissue engineering.

In this work, electrospinning was used for the preparation of composite nanofibrous scaffold, of polyvinyl alcohol (PVA), silk fibroin (SF) extract of Bombyx mori cocoons and silver nanoparticles (Ag NPs), as a substrate for bone tissue engineering. The PVA pristine was prepared at a concentration of 10% wt. The composite nanofibers scaffolds of PVA was prepared with silk fibroin and silver nanoparticles, in relation of PVA: SF (90:10) (v/v) respectively. The formation and presence of AgNPs was confirmed by ultraviolet-visible spectroscopy (Uv-vis). The diameter distribution of the nanofibers was narrow by SEM using Image J software. The chemical composition was determined by FTIR spectra. The wettability was determined using water contact angle. The results showed the average nanofiber diameter of PVA10 pristine was 108.18 nm and to PVA10/SF/Ag NPs was 106.62 nm, no significant changes were noted in the mean diameter, but there were changes in its morphology. The average nanofiber diameter increase with the concentration of PVA at PVA15/SF/Ag NPs was 189.12 nm to PVA18/SF/Ag NPs was 224,23 nm. FTIR spectra indicated characteristic absorption peaks related to the chemical structure of PVA, fibroin and Ag NPs, it demonstrated good interactions between them, caused by strong intermolecular hydrogen bonds. The contact angle of the scaffolds PVA 10%wt decrease with the incorporation of fibroin and show hydrophilic characteristics. The achievements indicate the potential of the nanofibers of PVA15/SF/Ag NPs as a possible substitute for bone tissue engineering.Clinical Relevance-This establishes a possible substrate of PVA/SF/Ag NPs that exhibit desired properties such as porosity and high surface area to volume ratio for bone tissue engineering.

Extraction and Encapsulation of Phenolic Compounds of Tunisian Rosemary (Rosmarinus officinalis L.) Extracts in Silk Fibroin Nanoparticles.

Rosemary (Rosmarinus officinalis L.) is known to be an effective potential source of natural antioxidants which confer benefits to human health. Their bioactive properties are mainly due to phenolic compounds but these molecules are highly vulnerable to oxidants, light, heat, pH, water and enzymatic activities. Therefore, the stability and shelf life of phenolic compounds should be increased by being protected from chemical and physical damage by means of encapsulation prior to application. Encapsulation is becoming increasingly important in the pharmaceutical, food, cosmetics, textile, personal care, chemical, biotechnology, and medicinal industries due to its potential for stabilization and delivery of delicate and precious bioactive compounds. The aim of the present work was to describe the polyphenolic profile of Tunisian Rosemary, collected from two different bioclimatic areas, and further loading in silk fibroin nanoparticles. The loaded nanoparticles were characterized in terms of morphology, size, polydispersity, Z-potential, secondary structure of the protein, encapsulation efficiency, loading content, and antioxidant activity. On one hand, HPLC analysis revealed the presence of 18 polyphenolic compounds of whichcarnosic acid and carnosol were found to be the most abundant compounds (46.3 to 76.4 and 22.4 to 43.5 mg of compound per gram of dry plant weight (mg/g DPW) respectively), Total phenolic content (TPC) ranged from 85.8 to 137.3 mg of gallic acid equivalent (GAE)/g DPW in post-distilled rosemary extracts andantioxidant activity reached the values of 5.9 to 8.3 µmol of ascorbic acid equivalent (AAE)/g DPW). On the other hand loaded nanoparticles were almost spherical and presented nanometric size and negative Z-potential. Although the encapsulation efficiency in silk fibroin nanoparticles and the drug loading content were low in the conditions of the assay, the encapsulated polyphenols retained near 85% of the radical scavenging activity against DPPH· after 24 h. of incubation at 37 °C. The results showed that post-distilled rosemary residues had an effective potential as natural antioxidants due to their significant antioxidant activity and seemed to be useful in both pharmaceutical and food industries with beneficial properties that might confer benefits to human health and these silk fibroin nanoparticles loaded with rosemary extracts are thus a promising combination for several applications in food technology or nanomedicine.

Silk Fibroin Nanoparticles Functionalized with Fibronectin for Release of Vascular Endothelial Growth Factor to Enhance Angiogenesis

ABSTRACT This work aimed at enhancing the angiogenesis in the presence of silk fibroin nanoparticles functionalized with fibronectin (FN-SF NPs) for controlled release of vascular endothelial growth factor (VEGF). The FN-SF NP samples were synthesized via the desolvation method and then VEGF was encapsulated into them. The physicochemical properties of the VEGF-loaded FN-SF NPs were evaluated by field-emission scanning electron microscopy (FE-SEM), dynamic light scattering (DLS), and Fourier transform infrared spectroscopy (FTIR). Results showed the spherical morphology, size of ~90 nm with a polydispersity index of 0.21, and the presence of a characteristic peak in 1063 cm−1 owing to functionalizing between SF NPs and FN. Afterward, the VEGF release from the SF NPs with and without FN functionalization was carried out utilizing enzyme-linked immunosorbent assay (ELISA) and the outcomes revealed that the FN-SF NPs have slower VEGF release than that of unfunctionalized sample with difference of approximately 10%. Eventually, the bioactivity of VEGF released from SF NPs, and FN-SF NPs was assessed using human umbilical vein endothelial cells (HUVECs) culture and the results exhibited a remarkable increase of their proliferation on FN-SF NPs owing to the presence of VEGF even after 10 days.

Biomimetic Nucleation of Metal-Organic Frameworks on Silk Fibroin Nanoparticles for Designing Core-Shell-Structured pH-Responsive Anticancer Drug Carriers.

Silk fibroin (SF) is a biomacromolecule that can be assembled into nanostructures and induce biomimetic nucleation of inorganic materials. Zeolitic imidazolate framework-8 (ZIF-8), a metal-organic framework (MOF), can be dissolved selectively under acidic pH. Here, we integrated SF and ZIF-8 to develop novel drug carriers that selectively release drug in the acidic intracellular environment of cancer cells. Specifically, SF was assembled into nanoparticles (SF-NPs), which were then loaded with an antitumor drug, doxorubicin (DOX), to form DSF-NPs. Due to the SF-mediated organization of ZIF-8 precursors such as zinc ions, the DSF-NPs further templated the nucleation of ZIF-8 onto their surface to generate core-shell-structured NPs (termed DSF@Z-NPs) with ZIF-8 as a shell and DSF-NP as a core. We found that the DSF@Z-NPs, highly stable under neutral conditions, could be uptaken by breast cancer cells, release DOX selectively owing to dissolution of ZIF-8 shells in the acidic intracellular environment in a controlled manner, and induce cell apoptosis. We also confirmed that the DSF@Z-NPs could inhibit tumor growth more efficiently to reach a higher survival rate than their controls by inducing cell apoptosis in vivo. Our study suggests that SF and MOF could be combined to design a new type of cancer therapeutics.

Pectin-cellulose hydrogel, silk fibroin and magnesium hydroxide nanoparticles hybrid nanocomposites for biomedical applications

Natural polymers are at the center of materials development for biomedical and biotechnological applications based on their biocompatibility, low-toxicity and biodegradability. In this study, a novel nanobiocomposite based on cross-linked pectin-cellulose hydrogel, silk fibroin, and Mg(OH)2 nanoparticles was designed and synthesized. After extensive physical-chemical characterization, the biological response of pectin-cellulose/silk fibroin/Mg(OH)2 nanobiocomposite scaffolds was evaluated by cell viability, red blood cells hemolytic and anti-biofilm assays. After 3 days and 7 days, the cell viability of this nanobiocomposite scaffold was 65.5% and 60.5% respectively. The hemolytic effect was below 20%. Furthermore, the presence of silk fibroin and Mg(OH)2 nanoparticles allowed to enhance the anti-biofilm activity, inhibiting the P. aeruginosa biofilm formation.

Antitumor Activity of Rosmarinic Acid-Loaded Silk Fibroin Nanoparticles on HeLa and MCF-7 Cells.

Rosmarinic acid (RA), one of the most important polyphenol-based antioxidants, has drawn increasing attention because of its remarkable bioactive properties, including anti-inflammatory, anticancer and antibacterial activities. The aim of this study was to synthesize and characterize RA-loaded silk fibroin nanoparticles (RA-SFNs) in terms of their physical–chemical features and composition, and to investigate their antitumor activity against human cervical carcinoma and breast cancer cell lines (HeLa and MCF-7). Compared with the free form, RA bioavailability was enhanced when the drug was adsorbed onto the surface of the silk fibroin nanoparticles (SFNs). The resulting particle diameter was 255 nm, with a polydispersity index of 0.187, and the Z-potential was −17 mV. The drug loading content of the RA-SFNs was 9.4 wt.%. Evaluation of the in vitro drug release of RA from RA-SFNs pointed to a rapid release in physiological conditions (50% of the total drug content was released in 0.5 h). Unloaded SFNs exhibited good biocompatibility, with no significant cytotoxicity observed during the first 48 h against HeLa and MCF-7 cancer cells. In contrast, cell death increased in a concentration-dependent manner after treatment with RA-SFNs, reaching an IC50 value of 1.568 and 1.377 mg/mL on HeLa and MCF-7, respectively. For both cell lines, the IC50 of free RA was higher. The cellular uptake of the nanoparticles studied was increased when RA was loaded on them. The cell cycle and apoptosis studies revealed that RA-SFNs inhibit cell proliferation and induce apoptosis on HeLa and MCF-7 cell lines. It is concluded, therefore, that the RA delivery platform based on SFNs improves the antitumor potential of RA in the case of the above cancers.

Development of a Biomimetic Peptide-Based Nanoformulation Against the Breast Cancer

Nanotechnology has enabled the preparation of various materials for overcoming the rapid clearance of drugs, nonspecific uptake or actions, and poor tumor penetration. Based on the significance of using biomimetic substances, silk fibroin nanoparticles (SF-NPs) have been increasingly prepared for the delivery of therapeutics. Meanwhile, aggregation and low stability in the biological medium may negatively affect their efficiency. This prompted us to coat SF-NPs with polydopamine (PDA), and for efficient accumulation and increasing therapeutic efficiency against breast cancer, paclitaxel (PTX)-loaded PDA-coated SF-NPs were conjugated with targeting peptide, iRGD (iRGD-PDA-PTX-SF-NPs). The peptide impacts on the cellular uptake, cytotoxicity, tumor penetrability of NPs, and their antitumor effects were evaluated. iRGD-PDA-PTX-SF-NPs with suitable physicochemical characteristics and drug loading released PTX in a controlled manner, and efficient cellular uptake was observed. Improved pharmacological profile of PTX was revealed by increased anticancer effects in vitro and in tumor-bearing Balb/c mice, including the delayed growth of the tumor and enhanced rate of survival. The prepared NPs showed no toxic effects against the healthy tissues indicating the histocompatibility and safety of these biomimetic and long-circulating nanoplatforms. The peptide-based SF-NPs could be considered as promising biomimetic nanoformulation against breast cancer.

Silk fibroin nanoparticles enhance quercetin immunomodulatory properties in DSS-induced mouse colitis

Inflammatory bowel disease (IBD) is a chronic and idiopathic inflammatory disorder affecting the gastrointestinal tract. The pharmacological treatments used currently for its treatment lack efficacy, so new therapeutic strategies should be developed. In this context, flavonoids loaded in biopolymeric nanoparticles can be considered as novel promising candidates. The aim of the present study was to evaluate the intestinal anti-inflammatory effects of quercetin when is administered loaded in silk fibroin nanoparticles (QSFN) in the dextran sulphate sodium experimental model of mouse colitis, which displays some similarities to human IBD. Previously characterized quercetin-loaded silk fibroin nanoparticles (QSFN). QSFN showed a reversible aggregation profile induced by the acidification of the solution but did not affect the loaded quercetin. Daily administration of QSFN significantly reduced disease activity index values compared to the control colitic group. This beneficial effect was not only corroborated by the histological examination of the colonic specimens but also the improvement of the colonic expression of the different proinflammatory cytokines (Tnf-α, Il-1β, Il-6, Mcp-1, Icam-1, Nlrp3 and iNOS). Therefore, these data suggest that QSFN could be a promising alternative to current treatments as a drug delivery system for IBD treatment.

Bioinspired silk fibroin nano-delivery systems protect against 5-FU induced gastrointestinal mucositis in a mouse model and display antitumor effects on HT-29 colorectal cancer cells in vitro

Colorectal cancer (CRC), is the second cause of cancer-related deaths worldwide is one of the most prevalent types of cancers. Conventional treatment continues to rely on surgery, chemotherapy, and radiotherapy, but for advanced cases, adjuvant chemotherapy remains the main approach for improving surgical outcomes and lower the disease recurrence probability. Chemotherapy-induced gastrointestinal (GI) toxicity is the main dose-limiting factor for many chemotherapeutic regimens, including 5-FU, and one of the biggest oncological challenges. Up to 40% of the patients receiving 5-FU get mucositis, 10–15% of which develop severe symptoms. In this context, our study aimed to develop a bioinspired nanosized drug delivery system as a strategy to reduce 5-FU associated side effects, such as GI mucositis. To this end, SF-based nanoparticles were prepared and characterized in terms of size and morphology, as well as in terms of in vitro antitumoral activity on a biomimetic colorectal cancer model by investigation of apoptosis, DNA fragmentation, and release of reactive oxygen species. Additionally, the capacity of the SF-based nanocarriers to offer intestinal protection against 5-FU-induced GI mucositis was evaluated in vivo using a mouse model that mimics the chemotherapy-associated gut mucositis occurring in colorectal cancer. Our studies show that silk fibroin nanoparticles efficiently deliver 5-FU to tumor cells in vitro while protecting against drug-induced GI mucositis in a mouse model.

Modification of Polyacrylonitrile Fibers with Silk Fibroin Nanoparticles. Dyeing of Pan Fiber with Fibroin Nanoparticles by Acid Dyes

Modification of Polyacrylonitrile Fibers with Silk Fibroin Nanoparticles. Dyeing of Pan Fiber with Fibroin Nanoparticles by Acid Dyes

Molecularly Imprinted Silk Fibroin Nanoparticles.

Nanosized biomimetics prepared by the strategy of molecular imprinting, that is, the stamping of recognition sites by means of a template-assisted synthesis, are demonstrating potential as plastic antibodies in medicine, proving effective for cell imaging and targeted therapies. Most molecularly imprinted nanoparticles (MIP-NPs) are currently made of soft matter, such as polyacrylamide and derivatives. Yet, MIP-NPs biocompatibility is crucial for their effective translation into clinical uses. Here, we propose the original idea to synthesize fully biocompatible molecularly imprinted nanoparticles starting from the natural polymer silk fibroin (MIP SF-NPs), which is nontoxic and highly biocompatible. The conditions to produce MIP SF-NPs of different sizes (dmean ∼ 50 nm; dmean ∼ 100 nm) were set using the response surface method. The stamping of a single, high affinity (KD = 57 × 10–9 M), and selective recognition site per silk fibroin nanoparticle was demonstrated, together with the confirmation of nontoxicity. Additionally, MIP SF-NPs were used to decorate silk microfibers and silk nanofibers, providing a general means to add entailed biofunctionalities to materials.

Effect of Yerba Mate and Silk Fibroin Nanoparticles on the Migration Properties in Ethanolic Food Simulants and Composting Disintegrability of Recycled PLA Nanocomposites.

The main objective of the present research is to study the effect of the incorporation of low amounts of silk fibroin nanoparticles (SFNs) and yerba mate nanoparticles (YMNs) on the migration phenomenon into ethanolic food simulants as well as on the disintegrability under composting conditions of mechanically recycled polylactic acid (PLA). Recycled PLA was obtained under simulated recycling conditions by melt processing virgin PLA into films and further subjecting them to an accelerated aging process, which involved photochemical, thermal, and hydrothermal aging steps followed by an intense washing step. SFNs were extracted from Bombyx mori cocoons and YMNs from yerba mate waste. Then, recycled PLA was melted, reprocessed, and reinforced with either 1%wt. of SFNs or YMNs, by melt extrusion, and further processed into films by compression molding. The obtained nanocomposites were exposed to ethanolic food simulants (ethanol 10% v/v, simulant A and ethanol 50% v/v, simulant D1) and the structural, thermal, and mechanical properties were studied before and after the exposure to the food simulants. The migration levels in both food simulants were below the overall migration limits required for food contact materials. The materials were disintegrated under simulated composting conditions at the laboratory scale level and it was observed that the nanoparticles delayed the disintegration rate of the recycled PLA matrix, but nanocomposites were fully disintegrated in less than one month.

Genetic hybridization of highly active exogenous functional proteins into silk-based materials using “light-clothing” strategy

Genetic hybridization of highly active exogenous functional proteins into silk-based materials using “light-clothing” strategy

Enhanced Cytotoxic Activity of Docetaxel-Loaded Silk Fibroin Nanoparticles against Breast Cancer Cells.

Despite decades of research, breast cancer therapy remains a great challenge. Docetaxel is an antimicrotubule agent that is effectively used for the treatment of breast cancer. However, its clinical use is significantly hampered by its low water solubility and systemic toxicity. The current study was designed to prepare docetaxel (DXL)-loaded silk-fibroin-based nanoparticles (SF-NPs) and to screen their potential antitumor activity against breast cancer cell lines. DXL-loaded SF-NPs were prepared using a nanoprecipitation technique and were evaluated for particle size, zeta potential, entrapment efficiency, and in vitro release profile. In addition, DXL-loaded SF-NPs were screened for in vitro cytotoxicity, cellular uptake, and apoptotic potential against MCF-7 and MDA-MB-231 breast cancer cell lines. The prepared DXL-loaded SF-NPs were 178 to 198 nm in diameter with a net negative surface charge and entrapment efficiency ranging from 56% to 72%. In vitro release studies exhibited a biphasic release profile of DXL from SF-NPs with sustained drug release for 72 h. In vitro cell studies revealed that entrapment of DXL within SF-NPs significantly improved cytotoxic potential against breast cancer cell lines, compared to the free drug, and enhanced cellular uptake of DXL by breast cancer cells. Furthermore, the accumulation in the G2/M phase was significantly higher in cells treated with DXL-loaded SF-NPs than in cells treated with free DXL. Collectively, the superior antitumor activities of DXL-loaded SF-NPs against breast cancer cells, compared to free DXL, could be ascribed to improved apoptosis and cell cycle arrest. Our results highlighted the feasibility of using silk fibroin nanoparticles as a nontoxic biocompatible delivery vehicle for enhanced therapeutic outcomes in breast cancer.