Crosslinked Fibroin Nanoparticles: Investigations on Biostability, Cytotoxicity, and Cellular Internalization

Abstract

Recently, crosslinked fibroin nanoparticles (FNP) using the crosslinker 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) or the polymer poly(ethylenimine) (PEI) have been developed and showed potentials as novel drug delivery systems. Thus, this study further investigated the biological properties of these crosslinked FNP by labeling them with fluorescein isothiocyanate (FITC) for in vitro studies. All formulations possessed a mean particle size of approximately 300 nm and a tunable zeta potential (−20 to + 30 mV) dependent on the amount/type of crosslinkers. The FITC-bound FNP showed no significant difference in physical properties compared to the blank FNP. They possessed a binding efficacy of 3.3% w/w, and no FITC was released in sink condition up to 8 h. All formulations were colloidal stable in the sheep whole blood. The degradation rate of these FNP in blood could be controlled depending on their crosslink degree. Moreover, no potential toxicity in erythrocytes, Caco-2, HepG2, and 9L cells was noted for all formulations at particle concentrations of < 1 mg/mL. Finally, all FNP were internalized into the Caco-2 cells after 3 h incubation. The uptake rate of the positively charged particles was significantly higher than the negatively charged ones. In summary, the crosslinked FNP were safe and showed high potentials as versatile systems for biomedical applications.

Silk fibroin hydrogel containing Sesbania sesban L. extract for rheumatoid arthritis treatment

“…Fibroin was extracted and purified from the raw Bombyx mori silkworm cocoons, following our previous studies (Pham et al., 2018 , 2019 , 2020 ). For this, the cocoons were degummed (i.e.…”

“…The cell viability was determined using the MTS assay, with slight modification from a previous study (Pham et al. 2020 ). To this end, RAW 264.7 cells were seeded into a 96-well plate at a concentration of 5 × 10 3 cells/well.…”

“… 2011 ; Oliveira et al., 2020 , 2020 ). Among numerous hydrogel biomaterials, silk fibroin, commonly extracted from Bombyx mori silkworm, has gained increasingly attention due to its biocompatibility, biodegradability, versatility in chemical modification, controlled-release property, among others (Pham et al., 2019 , 2020 , 2021 ). Fibroin hydrogels have been proposed and developed for various biomedical applications (Oliveira et al., 2020 ; Zheng & Zuo 2021 ; Ziadlou et al.…”

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

“…The uptake studies were performed using A549 and RAW264.7 macrophage cell lines. 3,16 Briefly, 1 × 10 5 cells were seeded onto a sterile tissue culture-treated 48-well plate and cultured overnight to reach 80% confluency. Then, the cells were treated with 100 μg/mL of C6-SFN and incubated for 2 h at 37 °C and 5% CO 2 condition.…”

Engineered protein nanodrug as an emerging therapeutic tool

“…Fibroin is the main protein that exists in silk fibers. Due to its low immunogenicity, biodegradability, mechanical strength, significant productivity, and low cost, fibroin is an attractive material for nanoparticle generation [11,[56][57][58], especially for vehicles of sensitive biologicals such as insulin and growth factors. Kundu et al reported the fabrication of silk fibroin spherical nanoparticles as depots for VEGF [59].…”