Formulation Design, Statistical Optimization, and In Vitro Evaluation of a Naringenin Nanoemulsion to Enhance Apoptotic Activity in A549 Lung Cancer Cells

Overview

Journal Pharmaceuticals (Basel)

Publisher MDPI

Specialty Chemistry

Date 2020 Jul 19

PMID 32679917

Citations 37

Authors

Shadab Md

Nabil A Alhakamy

Hibah M Aldawsari

Mohammad Husain

Sabna Kotta

Samaa T Abdullah

Usama A Fahmy

Mohamed A Alfaleh

Hani Z Asfour

Affiliations

Soon will be listed here.

Abstract

Naringenin (NAR), a flavonoid mainly found in citrus and grapefruits, has proven anti-cancer activities. However, the poor water solubility and low bioavailability of NAR limits its use as a therapeutic agent. The aim of this study was to develop and optimize stable naringenin nanoemulsions (NAR-NE) using a Box-Behnken experimental design to obtain a formulation with a higher efficiency. Anticancer activity of optimized NAR-NE was evaluated in A549 lung cancer cells using cell viability, flow-cytometric assays, and enzyme-linked immunosorbent assay. The stabilized nanoemulsion, which showed a spherical surface morphology, had a globule size of 85.6 ± 2.1 nm, a polydispersity index of 0.263 ± 0.02, a zeta potential of -9.6 ± 1.2 mV, and a drug content of 97.34 ± 1.3%. The NAR release from the nanoemulsion showed an initial burst release followed by a stable and controlled release for a longer period of 24 h. The nanoemulsion exhibited excellent thermodynamic and physical stability against phase separation and storage. The NAR-NE showed concentration-dependent cytotoxicity in A549 lung cancer cells, which was greater than that of free NAR. The percentage of apoptotic cells and cell cycle arrest at the G2/M and pre-G1 phases induced by NAR-NE were significantly higher than those produced by free NAR ( < 0.05). NAR-NEs were more effective than the NAR solution in reducing Bcl2 expression, while increasing pro-apoptotic Bax and caspase-3 activity. Therefore, stabilized NAR-NE could be a suitable drug delivery system to enhance the effects of NAR in the treatment of lung cancer.

Citing Articles

Emerging Nanoparticle-Based Diagnostics and Therapeutics for Cancer: Innovations and Challenges.

Puttasiddaiah R, Basavegowda N, Lakshmanagowda N, Raghavendra V, Sagar N, Sridhar K Pharmaceutics. 2025; 17(1).

PMID: 39861718 PMC: 11768644. DOI: 10.3390/pharmaceutics17010070.

Synthesis of silver nanoparticles embedded into melamine polyaminal networks as antibacterial and anticancer active agents.

Alotaibi M, Almalki B, Tashkandi N, Basingab F, Abdullah S, Alkayal N Sci Rep. 2024; 14(1):20008.

PMID: 39198544 PMC: 11358378. DOI: 10.1038/s41598-024-70606-0.

Oral co-polymeric raft-forming nano gels for targeted empagliflozin delivery against stomach cancer (SGC7901).

Alhakamy N, Abdullah S, Md S, Ansari A, Bhattamisra S, Ibrahim I Heliyon. 2024; 10(13):e34074.

PMID: 39071709 PMC: 11279758. DOI: 10.1016/j.heliyon.2024.e34074.

Preparation, optimization, and characterization of genistein-ginseng long-acting polymeric gel as a breast cancer treatment alternative.

Abdullah S, Md S, Altamimi A, Alahdal H, Ali R, Alkreathy H Discov Oncol. 2024; 15(1):257.

PMID: 38960937 PMC: 11222347. DOI: 10.1007/s12672-024-01132-8.

Therapeutic Implications of Dietary Polyphenols-Loaded Nanoemulsions in Cancer Therapy.

Tomar R, Das S, Balaga V, Tambe S, Sahoo J, Rath S ACS Appl Bio Mater. 2024; 7(4):2036-2053.

PMID: 38525971 PMC: 11530091. DOI: 10.1021/acsabm.3c01205.

References

Akhter M, Kumar S, Nomani S . Sonication tailored enhance cytotoxicity of naringenin nanoparticle in pancreatic cancer: design, optimization, and studies. Drug Dev Ind Pharm. 2020; 46(4):659-672. DOI: 10.1080/03639045.2020.1747485. View

Ostertag F, Weiss J, McClements D . Low-energy formation of edible nanoemulsions: factors influencing droplet size produced by emulsion phase inversion. J Colloid Interface Sci. 2012; 388(1):95-102. DOI: 10.1016/j.jcis.2012.07.089. View

Maity S, Mukhopadhyay P, Kundu P, Chakraborti A . Alginate coated chitosan core-shell nanoparticles for efficient oral delivery of naringenin in diabetic animals-An in vitro and in vivo approach. Carbohydr Polym. 2017; 170:124-132. DOI: 10.1016/j.carbpol.2017.04.066. View

Arul D, Subramanian P . Naringenin (citrus flavonone) induces growth inhibition, cell cycle arrest and apoptosis in human hepatocellular carcinoma cells. Pathol Oncol Res. 2013; 19(4):763-70. DOI: 10.1007/s12253-013-9641-1. View

Sandhu P, Kumar R, Beg S, Jain S, Kushwah V, Katare O . Natural lipids enriched self-nano-emulsifying systems for effective co-delivery of tamoxifen and naringenin: Systematic approach for improved breast cancer therapeutics. Nanomedicine. 2017; 13(5):1703-1713. DOI: 10.1016/j.nano.2017.03.003. View

Tsai M, Huang Y, Fang J, Fu Y, Wu P . Preparation and evaluation of submicron-carriers for naringenin topical application. Int J Pharm. 2015; 481(1-2):84-90. DOI: 10.1016/j.ijpharm.2015.01.034. View

Zhang Y, Shang Z, Gao C, Du M, Xu S, Song H . Nanoemulsion for solubilization, stabilization, and in vitro release of pterostilbene for oral delivery. AAPS PharmSciTech. 2014; 15(4):1000-8. PMC: 4113613. DOI: 10.1208/s12249-014-0129-4. View

Tiwari R, Dubey V, Kesavan K . Ocular Self-Microemulsifying Drug Delivery System of Prednisolone Improves Therapeutic Effectiveness in the Treatment of Experimental Uveitis. Ocul Immunol Inflamm. 2017; 27(2):303-311. DOI: 10.1080/09273948.2017.1333622. View

Mayer S, Weiss J, McClements D . Vitamin E-enriched nanoemulsions formed by emulsion phase inversion: factors influencing droplet size and stability. J Colloid Interface Sci. 2013; 402:122-30. DOI: 10.1016/j.jcis.2013.04.016. View

10.

Kumar S, Birundha K, Kaveri K, Devi K . Antioxidant studies of chitosan nanoparticles containing naringenin and their cytotoxicity effects in lung cancer cells. Int J Biol Macromol. 2015; 78:87-95. DOI: 10.1016/j.ijbiomac.2015.03.045. View

11.

Chang H, Chang Y, Lai S, Chen K, Wang K, Chiu T . Naringenin inhibits migration of lung cancer cells via the inhibition of matrix metalloproteinases-2 and -9. Exp Ther Med. 2017; 13(2):739-744. PMC: 5348669. DOI: 10.3892/etm.2016.3994. View

12.

Khan I, Bhardwaj M, Shukla S, Lee H, Oh M, Bajpai V . Carvacrol encapsulated nanocarrier/ nanoemulsion abrogates angiogenesis by downregulating COX-2, VEGF and CD31 in vitro and in vivo in a lung adenocarcinoma model. Colloids Surf B Biointerfaces. 2019; 181:612-622. DOI: 10.1016/j.colsurfb.2019.06.016. View

13.

Parashar P, Rathor M, Dwivedi M, Saraf S . Hyaluronic Acid Decorated Naringenin Nanoparticles: Appraisal of Chemopreventive and Curative Potential for Lung Cancer. Pharmaceutics. 2018; 10(1). PMC: 5874846. DOI: 10.3390/pharmaceutics10010033. View

14.

Faramarzi L, Dadashpour M, Sadeghzadeh H, Mahdavi M, Zarghami N . Enhanced anti-proliferative and pro-apoptotic effects of metformin encapsulated PLGA-PEG nanoparticles on SKOV3 human ovarian carcinoma cells. Artif Cells Nanomed Biotechnol. 2019; 47(1):737-746. DOI: 10.1080/21691401.2019.1573737. View

15.

Fuster M, Carissimi G, Montalban M, Villora G . Improving Anticancer Therapy with Naringenin-Loaded Silk Fibroin Nanoparticles. Nanomaterials (Basel). 2020; 10(4). PMC: 7221656. DOI: 10.3390/nano10040718. View

16.

Rajamani S, Radhakrishnan A, Sengodan T, Thangavelu S . Augmented anticancer activity of naringenin-loaded TPGS polymeric nanosuspension for drug resistive MCF-7 human breast cancer cells. Drug Dev Ind Pharm. 2018; 44(11):1752-1761. DOI: 10.1080/03639045.2018.1496445. View

17.

Muralidharan R, Babu A, Amreddy N, Basalingappa K, Mehta M, Chen A . Folate receptor-targeted nanoparticle delivery of HuR-RNAi suppresses lung cancer cell proliferation and migration. J Nanobiotechnology. 2016; 14(1):47. PMC: 4915183. DOI: 10.1186/s12951-016-0201-1. View

18.

Aldawsari H, Gorain B, Alhakamy N, Md S . Role of therapeutic agents on repolarisation of tumour-associated macrophage to halt lung cancer progression. J Drug Target. 2019; 28(2):166-175. DOI: 10.1080/1061186X.2019.1648478. View

19.

Ahmad N, Ahmad R, Ahmad F, Ahmad W, Alam M, Amir M . Poloxamer-chitosan-based Naringenin nanoformulation used in brain targeting for the treatment of cerebral ischemia. Saudi J Biol Sci. 2020; 27(1):500-517. PMC: 6933235. DOI: 10.1016/j.sjbs.2019.11.008. View

20.

Md S, Gan S, Haw Y, Ho C, Wong S, Choudhury H . In vitro neuroprotective effects of naringenin nanoemulsion against β-amyloid toxicity through the regulation of amyloidogenesis and tau phosphorylation. Int J Biol Macromol. 2018; 118(Pt A):1211-1219. DOI: 10.1016/j.ijbiomac.2018.06.190. View