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

Marta G Fuster,Mercedes G Montalbán,Gloria Víllora,Guzmán Carissimi

doi:10.3390/polym13183169

Abstract

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.

Highlights

During recent years, natural compounds and their derivatives have increasingly been investigated for their curative properties and for their potential in the development of new drugs [1,2,3]
As can be seen from the distribution (Figure 1A), both systems present a narrow monomodal distribution with a polydispersity index (PdI) of 0.110 and 0.187 for SFFigNursea1n. d(AR) SAiz-eSdFiNstrsi,buretisopnebcatsievdeolyn.thTehientZen-saitvyeorfaSgFeNasnadnddRifAfu-SsFiNosnwciothefafinciinesnertt of the values values ofof r25S5FnNmScsoFrNwarensledaartnei1do.n19R6c3Ao4μ-eSfn2Ffi/mNcises,.nartnefsdopr 2eth.c9et9itvwμeol2ys/yfssoterrmetshspeaenscdatm(iBv)eeZlpy-p,aorwatemhniteilaetledtrhisse.trAiRbsuArtei-oSgnaFbrNdassseZdp-orpneostteheennittneiatdelnsity of and electrophoretic mobility, silk fibroin nanoparticles (SFNs) showed a value of −30 mV and −2.408 μmcm/Vs, respectively, while the Rosmarinic acid (RA)-SFNs presented values of −17 mV and −1.333 μmcm/Vs
In light of the results of the apoptosis analysis, we can conclude that Rosmarinic Acid-Loaded Silk Fibroin Nanoparticles (RA-SFNs) induce apoptosis on both cell lines leading to a more significant antiproliferative effect on HeLa cells because the increase of the cells on apoptotic phase is higher than in MCF-7 cells. These results are in Polymers 2021, 13, x FOR PEER REVIEWgood agreement with the in vitro cytotoxicity assays which revealed that RA-SFNs in HeL1a8 of 21 cells had doble cytotoxic effects than in MCF-7 cells confirming that HeLa cells are more sensitive to RA than MCF-7 cells

Summary

Introduction

Natural compounds and their derivatives have increasingly been investigated for their curative properties and for their potential in the development of new drugs [1,2,3]. The encapsulation of RA in drug nanocarriers is a promising option for increasing the therapeutical performance of RA because the longer circulation times achieved in blood allow prolonged biological activity and a greater probability of accumulating in inflamed tissues, in which vascular permeability is increased [16]. With these purposes in mind, RA has been loaded or encapsulated in solid lipid nanoparticles [14,17,18], natural [15,19,20,21] or synthetic [16,22] polymer nanoparticles and inorganic nanoparticles [23]. Among the different approaches explored to date, nanoparticles based on biopolymers are of particular interest as drug nanocarriers because they are biodegradable, natural and environmentally-friendly [24,25]

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