Nanoencapsulation of pomegranate bioactive compounds for breast cancer chemoprevention

Amit B Shirode,1,2,* Dhruba J Bharali,3,* Sameera Nallanthighal,1,2 Justin K Coon,1,2 Shaker A Mousa,3 Ramune Reliene1,2 1Department of Environmental Health Sciences, University at Albany, State University of New York, Albany, NY, USA; 2Cancer Research Center, University at Albany, Rensselaer, NY, USA; 3Pharmaceutical Research Institute, Albany College of Pharmacy and Health Sciences, Albany, NY, USA *These authors contributed equally to this work Abstract: Pomegranate polyphenols are potent antioxidants and chemopreventive agents but have low bioavailability and a short half-life. For example, punicalagin (PU), the major polyphenol in pomegranates, is not absorbed in its intact form but is hydrolyzed to ellagic acid (EA) moieties and rapidly metabolized into short-lived metabolites of EA. We hypothesized that encapsulation of pomegranate polyphenols into biodegradable sustained release nanoparticles (NPs) may circumvent these limitations. We describe here the development, characterization, and bioactivity assessment of novel formulations of poly(D,L-lactic-co-glycolic acid)–poly(ethylene glycol) (PLGA–PEG) NPs loaded with pomegranate extract (PE) or individual polyphenols such as PU or EA. Monodispersed, spherical 150–200 nm average diameter NPs were prepared by the double emulsion–solvent evaporation method. Uptake of Alexa Fluor-488-labeled NPs was evaluated in MCF-7 breast cancer cells over a 24-hour time course. Confocal fluorescent microscopy revealed that PLGA–PEG NPs were efficiently taken up, and the uptake reached the maximum at 24 hours. In addition, we examined the antiproliferative effects of PE-, PU-, and/or EA-loaded NPs in MCF-7 and Hs578T breast cancer cells. We found that PE, PU, and EA nanoprototypes had a 2- to 12-fold enhanced effect on cell growth inhibition compared to their free counterparts, while void NPs did not affect cell growth. PU-NPs were the most potent nanoprototype of pomegranates. Thus, PU may be the polyphenol of choice for further chemoprevention studies with pomegranate nanoprototypes. These data demonstrate that nanotechnology-enabled delivery of pomegranate polyphenols enhances their anticancer effects in breast cancer cells. Thus, pomegranate polyphenols are promising agents for nanochemoprevention of breast cancer. Keywords: PLGA–PEG nanoparticles, pomegranate extract, punicalagin, ellagic acid, MCF-7 cells, Hs578T cell

Hydroalcoholic extracts from the bark of Quercus suber L. (Cork): optimization of extraction conditions, chemical composition and antioxidant potential

Cork is the bark of the tree Quercus suber L. which ï¬ nds use in diverse applications. However, a signiï¬ cant percentage is still rejected and burned for energy production, despite containing valuable molecules for materials processing and with important biological activities. Herein, the optimization of the extraction process to obtain these molecules, using mild solvents and conditions, is described within a bioreï¬ nery perspective. The extracts were obtained by direct contact solvent extractions with water, ethanol and its mixtures for different time and temperatures, and evaluated for chemical composition, total phenolic content (TPC) and antioxidant properties [by DPPH radical scavenging, ferric reducing antioxidant power (FRAP), Trolox equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) assays]. The results showed that the extraction process is accelerated and higher yields are achieved with the increase in temperature without chemical degradation or compromising the antioxidant capacity. For all solvents, at reï¬ ux temperature, more than 90% of the extractable material is obtained within 6 h (80% within 1 h). The highest TPC and antioxidant capacity are observed for the extracts obtained with mixtures of water and ethanol of similar volumes. The antioxidant capacity measured by DPPH, FRAP and TEAC assays was found to be proportional to the extract TPC, while ORAC is favored for higher percentages of ethanol on the extracting solvent. The main constituents of these extracts are the ellagitannins, vescalagin, castalagin and b-O-ethylvescalagin, along with other phenolic acids (mainly ellagic and gallic acids) and various ï¬ avonols. The extracts stability was monitored up to 1 year of storage with neither reduction in the antioxidant capacity nor chemical degradation. These results show that extracts with strong antioxidant potential and high content of bioactive molecules can be obtained from the processing of waste streams. Cork is a sustainable forest product and the development of new ï¬ elds of application contributes toward a zero waste cycle for a complete material bioreï¬ nery.The authors are grateful to Amorim Cork Composites for providing the cork powder and for the financing provided by the COMPETE/QREN/EU funding program through project BioActiveCork (QREN FCOMP-01-0202-FEDER-005455). Ivo M. Aroso and João P. Fernandes Fig. 6 Comparison between fresh and 1 year stored extracts for a TPC and b DPPH scavenging capacity Wood Sci Technol123 acknowledge the financial support from FCT through grants SFRH/BD/42273/2007 and SFRH/BD/73162/2010, respectively. Funding was also granted from the European Union’s Seventh Framework Programme (FP7/2007–2013) under Grant Agreement No. REGPOT-CT2012-316331-POLARIS and from Project ‘‘Novel smart and biomimetic materials for innovative regenerative medicine approaches (Ref.: RL1 - ABMR - NORTE-01-0124-FEDER-000016)’’ co-financed by North Portugal Regional Operational Programme (ON.2 – O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF). Conflict of Interest: The authors declare that they have no conflict of interest.info:eu-repo/semantics/publishedVersio