Silk Fibroin Nanoparticles: Synthesis and Applications as Drug Nanocarriers

The use of nanoparticles in biomedical fields is a very promising scientific area and has aroused the interest of researchers in the search for new biodegradable, biocompatible and non-toxic materials. This chapter is based on the features of the biopolymer silk fibroin and its applications in nanomedicine. Silk fibroin, obtained from the Bombyx mori silkworm, is a natural polymeric biomaterial whose main features are its amphiphilic chemistry, biocompatibility, biodegradability, excellent mechanical properties in various material formats, and processing flexibility. All of these properties make silk fibroin a useful candidate to act as nanocarrier. In this chapter, the structure of silk fibroin, its biocompatibility and degradability are reviewed. In addition, an intensive review on the silk fibroin nanoparticle synthesis methods is also presented. Finally, the application of the silk fibroin nanoparticles for drug delivery acting as nanocarriers is detailed.

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.

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.

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.