Facile and Controllable Fabrication of Protein-Only Nanoparticles through Photo-Induced Crosslinking of Albumin and Their Application as DOX Carriers.

Xiangyu Long,Fangling Ji,Lingyun Jia,Chao Zhang,Jun Ren

doi:10.3390/nano9050797

Xiangyu Long, Fangling Ji + Show 3 more

Open Access

https://doi.org/10.3390/nano9050797

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Abstract

Protein-based nanoparticles, as an alternative to conventional polymer-based nanoparticles, offer great advantages in biomedical applications owing to their functional and biocompatible characteristics. However, the route of fabrication towards protein-based nanoparticles faces substantial challenges, including limitations in size control and unavoidable usage of toxic crosslinkers or organic solvents, which may raise safety concerns related to products and their degradation components. In the present study, a photo-induced crosslinking approach was developed to prepare stable, size-controlled protein-only nanoparticles. The facile one-step reaction irradiated by visible light enables the formation of monodispersed bovine serum albumin nanoparticles (BSA NPs) within several minutes through a tyrosine photo-redox reaction, requiring no cross-linking agents. The size of the BSA NPs could be precisely manipulated (from 20 to 100 nm) by controlling the duration time of illumination. The resultant BSA NPs exhibited spherical morphology, and the α-helix structure in BSA was preserved. Further study demonstrated that the 35 nm doxorubicin (DOX)-loaded BSA NPs achieved a drug loading content of 6.3%, encapsulation efficiency of 70.7%, and a controlled release profile with responsivity to both pH and reducing conditions. Importantly, the in vitro drug delivery experiment demonstrated efficient cellular internalizations of the DOX-loaded BSA NPs and inhibitory activities on MCF-7 and HeLa cells. This method shows the promise of being a platform for the green synthesis of protein-only nanoparticles for biomedical applications.

Highlights

Therapeutic nanoparticles (NPs) are attracting significant attention for their great promise in numerous medical applications, including drug delivery, imaging, vaccine formulations, and biodetection [1,2]
DOX/Fluorescein isothiocyanate (FITC)-bovine serum albumin nanoparticles (BSA NPs) were obtained for further use
Bovine serum albumin (BSA) NPs were prepared at physiological conditions by illuminating the BSA solution that contained Ru(II) and ammonium persulphate (APS) with a 200 W incandescent lamp

Summary

Introduction

Therapeutic nanoparticles (NPs) are attracting significant attention for their great promise in numerous medical applications, including drug delivery, imaging, vaccine formulations, and biodetection [1,2]. Harsh conditions would lead the denaturation and unpredictable biocompatibility of albumin; in the solution without organic solvents, random crosslinking of albumin molecules may lead to aggregation and unsatisfactory size control, especially for sub-100 nm particles Other ways such as heating aggregation may produce hydrophobicity-driven protein assembly with weak stability [17]. The photoredox system has been applied to analyze the aggregation of the amyloid β-protein (Aβ), and it has been widely used to crosslink tyrosine-rich proteins such as gelatin and fibrinogen into hydrogels for tissue engineering materials [23,24,25,26] For these applications, the photochemical method is featured by high efficiency and mild reaction conditions. Scheme 1.SIclhleumster1a.tIilolunstoraftitohneofathberifcaabrtiicoantioonfodf rdurugg--lloaaddeeddbobvoinveinseerusmerualmbumailnbunamnoinpanrtaicnleosp(BaSrAticles (BSA NPs) and tNhPesi)raanpdpthliecirataipopnlicaastioandaos xaodrouxobriucbinici(nD(DOOXX))vveehhiicclel.e

Materials and Reagents

Methods

Characterizations

Cellular Internalization Assay

Cytotoxicity Assay

Preparation and Characterization of BSA NPs