Regular ArticleGold nanoparticle-guarded large-pore mesoporous silica nanocomposites for delivery and controlled release of cytochrome c
Graphical abstract
Introduction
Proteins play critical roles in disease prevention, cancer therapies and health promotion. Compared with small-molecular drugs and genetic drugs, protein-based pharmaceuticals could directly reverse the disordered functions of cells with their advantages of definite biological function, high pharmacological activity, specificity and low side effects. Therefore, proteins have been widely used in medical applications, such as cancer treatment [1], vaccination [2] and regenerative medicine [3]. However, there are still several challenges to be addressed, such as in vivo instability caused by fast enzymatic degradation and blood clearance, and poor internalization owing to the permselectivity of plasma membrane.
In the past decades, a variety of nanocarriers have been developed to maintain the biological activity and to achieve targeted delivery of protein pharmaceuticals, such as nanogels [3], inorganic nanoparticles [6], polymers [7] and lipids [4], [5], [6], [7], [8]. Among them, mesoporous silica nanoparticle (MSN) is one of the superior inorganic nanocarriers. MSNs have been widely used for drug delivery, due to its adjustable pore size, flexible surface modification, inert chemical properties, as well as favorable biocompatibility and low biotoxicity [9], [10], [11]. However, the pore size of contemporary MSNs is up to 3 nm, which is insufficient to encapsulate protein pharmaceuticals. Recently, in order to load proteins into the mesopores, MSNs with expanded pore size larger than the protein dimensions has been developed to meet the requirement of protein delivery [12], [13], [14]. Kros et al. [15] prepared MSNs with 10 ± 1 nm of disk-shaped cavities, which provided a scaffold with high encapsulation efficiency of proteins. Kim et al. [16] reported the synthesis of extra-large pore mesoporous silica nanoparticles (XL-MSNs) to transport ovalbumin and CpG oligonucleotide into dendritic cells and evaluated their potential applications as preventive cancer vaccine.
Although the expanded pores enable MSNs to load and deliver various cargos, cargos are easy to escape from the loose and open mesopores of MSNs, resulting in undesired spontaneous drug leakage before reaching targeting cells. Recently, there have been several strategies to inhibit cargo leakage [17], [18], [19], [20], [21]. Ma et al. presented a multifunctional nanoplatform integrating PbS/CdS quantum dots and Fe3O4 nanoparticles (NPs) within large-pore MSNs (LPMSNs) enabled by simple thiol modification [22]. Salis et al. showed that MSNs with a hexagonal structure could be conjugated to two relevant proteins (e.g. bovine serum albumin and lysozyme) through Schiff-base bond [19]. Nevertheless, MSNs with large pores that are able to efficiently load and controllably release protein pharmaceuticals in the microenvironment of targeting cells are highly desired. To address these demands, we designed a gold nanoparticle (AuNP)-guarded LPMSNs (LPMSN@AuNP) with pH-triggered release of cytochrome c (CC, a crucial death regulatory protein). Ethyl acetate was used as a pore expander to obtain LPMSNs. A pH-sensitive silane coupling agent (DMMA-APTES) was grafted on the surface of LPMSNs to endow them with pH-triggered charge reversal in acidic lysosome microenvironment. Amino-terminated polyethylene glycol (PEG)-modified AuNPs (AuNP-PEG5k-NH2) were subsequently adsorbed on the surface of LPMSNs through electrostatic attraction as gate-guards to avoid the leakage of CC that can trigger apoptosis in cancer cells (Scheme 1) [23], [24]. The capability of nanocomposites to deliver proteins into cancer cells and charge reversal-induced protein release were investigated by flow cytometry and confocal laser scanning microscopy (CLSM). Furthermore, HeLa xenograft model mice were established to evaluate the antitumor effects of the nanocomposites.
Section snippets
Materials
Tetraethoxysilane (TEOS, AP), triethylamine, succinic anhydride (SA, 99%) and dimethyl sulphoxide (DMSO, AP) were purchased from Sinopharm Chemical Reagent Co., Ltd, (Shanghai, China). Hydrogen tetrachloroaurate(III) trihydrate (HAuCl4·3H2O, 99.9%), 3-aminopropyl triethoxysilane (APTES, 95%), cetyltrimethyl ammonium bromide (CTAB), fluorescein isothiocyanate (FITC, 90%) and 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) were purchased from Sigma-Aldrich (St. Louis, MO, U.S.A.).
Synthesis and Characterization of nanocomposites
LPMSNs were prepared through hydrolysis of TEOS with a large amount of ethyl acetate to enlarge the mesopores and CuS NPs as seed. LPMSNs with different morphologies and pore sizes were obtained by varying the amount of ethyl acetate (Fig. S1 in Supporting Information (SI)). When a large amount of ethyl acetate (10 mL) was added, ethyl acetate formed large emulsions in the CTAB micelles, which contributed to the formation of large mesopores (Fig. 1a and b) [16], [25]. In addition, the absence
Conclusions
In summary, we have developed a novel method to prepare gold nanoparticle-guarded large-pore mesoporous silica nanocomposites for protein delivery. Compared with previously reported approaches [12], [13], [14], [15], [16], [17], [18], [19], [20], [21], [22], LPMSN@AuNPs nanocomposites not only provide the capability of protein encapsulation within the expanded pores, but also endow themselves intelligent response to tumor microenvironment for efficient protein delivery. Specifically, AuNPs with
CRediT authorship contribution statement
Chen Guo: Investigation, Formal analysis, Writing - original draft. Yamin Zhang: Investigation, Formal analysis, Writing - original draft. Yuce Li: Investigation. Lianbin Zhang: Investigation, Validation. Hao Jiang: Conceptualization, Supervision, Funding acquisition, Writing - review & editing. Juan Tao: Supervision, Writing - review & editing. Jintao Zhu: Conceptualization, Validation, Funding acquisition, Writing - review & editing.
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgments
This work was supported by the National Natural Science Foundation of China (81772913 and 51525302) and the Fundamental Research Funds for the Central Universities (2172019kfyXJJS070). We thank the support from Prof. Guanxin Shen in the Immunological Laboratory of Tongji Medical College, HUST. We also thank the HUST Analytical and Testing Center for allowing us to use its facilities.
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These authors contributed equally to this paper.