Design and Synthesis of Novel Thymoquinone-Zein Nanoparticles; Evaluation of the Inhibitory Effect on Candida albicans and Biofilm Formation in Vitro

Document Type : Original Research

Authors
S. Rahsepar 1
S. Roudbar mohammadi‎ 1
H. Delavari 2
M. Roudbari 3
Z. Mohammad Hassan 4
1 Department of Medical Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2 Department of Materials Engineering, Faculty of Engineering, Tarbiat Modares University, Tehran, Iran
3 Department of Parasitology and Mycology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
4 Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University
10.52547/iem.8.2.169
Abstract
Backgrounds: Candida albicans (C. albicans) as a fungal pathogen is part of the normal flora of the human body, which could cause various infections in patients with defective immune systems. Nowadays, there is a need to design and synthesis new drug formulations to overcome drug resistance in this genus. Thymoquinone (TQ) is the main ingredient in Nigella sativa, which has considerable antifungal properties. The aim of this study was to investigate the inhibitory effects of thymoquinone-zein nanoparticles (TQ-ZNPs) on C. albicans.

Materials & Methods: In the current study, TQ was encapsulated in zein (as a biodegradable carrier) and polyethylene glycol (PEG). The antifungal activity of TQ-ZNPs against C. albicans (ATCC 10231; standard strain) and their inhibitory effects on biofilm formation were examined using standardized broth microdilution and MTT assays, respectively. The total oxidant status (TOS) of C. albicans was assessed using colorimetric method, and the toxic effect of nanoparticles on peripheral blood mononuclear cells (PBMCs) was evaluated by MTT assay.

Findings: The minimum inhibitory concentration (MIC) of TQ-ZNPs was significantly reduced compared to that of free TQ. MIC values of TQ-ZNPs and free TQ were determined to be 7.4 and 50 µg/mL, respectively. Biofilm formation was inhibited, and oxidant production by fungal cells was increased. The findings of this study showed that TQ-ZNPs had no toxic effect on PBMCs.

Conclusion: This study results revealed that the synthesized nanoparticles had a good antifungal activity without any toxicity. The results demonstrated the superior efficiency of TQ-ZNPs over free TQ. Hence, this structure could be used to load hydrophobic drugs. However, more studies are needed to evaluate the beneficial properties of TQ-ZNPs.

Keywords

Subjects

1. Nobile CJ, Johnson ADJArom. Candida albicans biofilms and human disease. 2015;69:71-92.
2. Waller JCJS. The Kingdom of Plants. 2008;322(5903):860-.
3. Singh A, Ahmad I, Akhter S, Jain GK, Iqbal Z, Talegaonkar S, et al. Nanocarrier based formulation of Thymoquinone improves oral delivery: stability assessment, in vitro and in vivo studies. 2013;102:822-32.
4. Abdel-Fattah A-FM, Matsumoto K, Watanabe HJEjop. Antinociceptive effects of Nigella sativa oil and its major component, thymoquinone, in mice. 2000;400(1):89-97.
5. Nemmar A, Al‐Salam S, Zia S, Marzouqi F, Al‐Dhaheri A, Subramaniyan D, et al. Contrasting actions of diesel exhaust particles on the pulmonary and cardiovascular systems and the effects of thymoquinone. 2011;164(7):1871-82.
6. Raghunandhakumar S, Paramasivam A, Senthilraja S, Naveenkumar C, Asokkumar S, Binuclara J, et al. Thymoquinone inhibits cell proliferation through regulation of G1/S phase cell cycle transition in N-nitrosodiethylamine-induced experimental rat hepatocellular carcinoma. 2013;223(1):60-72.
7. Erboga M, Kanter M, Aktas C, Sener U, Erboga ZF, Donmez YB, et al. Thymoquinone ameliorates cadmium-induced nephrotoxicity, apoptosis, and oxidative stress in rats is based on its anti-apoptotic and anti-oxidant properties. 2016;170(1):165-72.
8. Awad AS, Abd Al Haleem EN, El-Bakly WM, Sherief MAJN-Ssaop. Thymoquinone alleviates nonalcoholic fatty liver disease in rats via suppression of oxidative stress, inflammation, apoptosis. 2016;389(4):381-91.
9. Liu H, Liu HY, Jiang YN, Li NJMmr. Protective effect of thymoquinone improves cardiovascular function, and attenuates oxidative stress, inflammation and apoptosis by mediating the PI3K/Akt pathway in diabetic rats. 2016;13(3):2836-42.
10. Khader M, Eckl P, Bresgen NJJoe. Effects of aqueous extracts of medicinal plants on MNNG-treated rat hepatocytes in primary cultures. 2007;112(1):199-202.
11. Simon H-U, Haj-Yehia A, Levi-Schaffer FJA. Role of reactive oxygen species (ROS) in apoptosis induction. 2000;5(5):415-8.
12. Shukla R, Cheryan MJIc, products. Zein: the industrial protein from corn. 2001;13(3):171-92.
13. Zhang Y, Cui L, Chen Y, Zhang H, Zhong J, Sun Y, et al. Zein-based nanofibres for drug delivery: classes and current applications. 2015;21(22):3199-207.
14. Lai L, Guo HJIjop. Preparation of new 5-fluorouracil-loaded zein nanoparticles for liver targeting. 2011;404(1-2):317-23.
15. Wayne PJCdM-A. Reference method for broth dilution antifungal susceptibility testing of yeasts, approved standard. 2002.
16. Chandra J, Kuhn DM, Mukherjee PK, Hoyer LL, McCormick T, Ghannoum MAJJob. Biofilm formation by the fungal pathogen Candida albicans: development, architecture, and drug resistance. 2001;183(18):5385-94.
17. Motavalli M, Khodadadi I, Fallah M, Maghsood AHJPr. Effect of oxidative stress on vital indicators of Acanthamoeba castellanii (T4 genotype). 2018;117(9):2957-62.
18. Nikoomanesh F, Roudbarmohammadi S, Khoobi M, Haghighi F, Roudbary MJAc, nanomedicine,, biotechnology. Design and synthesis of mucoadhesive nanogel containing farnesol: investigation of the effect on HWP1, SAP6 and Rim101 genes expression of Candida albicans in vitro. 2019;47(1):64-72.
19. De Rosa FG, Corcione S, Filippini C, Raviolo S, Fossati L, Montrucchio C, et al. The effect on mortality of fluconazole or echinocandins treatment in candidemia in internal medicine wards. 2015;10(5):e0125149.
20. Canela HMS, Cardoso B, Vitali LH, Coelho HC, Martinez R, Ferreira MEdSJM. Prevalence, virulence factors and antifungal susceptibility of Candida spp. isolated from bloodstream infections in a tertiary care hospital in Brazil. 2018;61(1):11-21.
21. Tumbarello M, Posteraro B, Trecarichi EM, Fiori B, Rossi M, Porta R, et al. Biofilm production by Candida species and inadequate antifungal therapy as predictors of mortality for patients with candidemia. 2007;45(6):1843-50.
22. Ramage G, Martínez JP, López-Ribot JLJFyr. Candida biofilms on implanted biomaterials: a clinically significant problem. 2006;6(7):979-86.
23. Ramage G, Bachmann S, Patterson TF, Wickes BL, López-Ribot JLJJoAC. Investigation of multidrug efflux pumps in relation to fluconazole resistance in Candida albicans biofilms. 2002;49(6):973-80.
24. Moghim H, Taghipoor S, Shahinfard N, Kheiri S, Panahi RJJoHP. Antifungal effects of Zataria multiflora and Nigella sativa extracts against Candida albicans. 2015;4.
25. Najee H, Kamerzan C, Marutescu L, Gheorghe I, Popa M, Gradisteanu G, et al. Antifungal activity of some medicinal plant extracts against Candida albicans nosocomial isolates. 2018;23(6):14073-6.
26. Almshawit H, Macreadie IJMr. Fungicidal effect of thymoquinone involves generation of oxidative stress in Candida glabrata. 2017;195:81-8.
27. Randhawa MA, Gondal MA, Al-Zahrani A-HJ, Rashid SG, Ali AJJoES, Health PA. Synthesis, morphology and antifungal activity of nano-particulated amphotericin-B, ketoconazole and thymoquinone against Candida albicans yeasts and Candida biofilm. 2015;50(2):119-24.
28. Bhattacharjee M, Upadhyay P, Sarker S, Basu A, Das S, Ghosh A, et al. Combinatorial therapy of Thymoquinone and Emodin synergistically enhances apoptosis, attenuates cell migration and reduces stemness efficiently in breast cancer. 2020;1864(11):129695.
29. Bhattacharya S, Ahir M, Patra P, Mukherjee S, Ghosh S, Mazumdar M, et al. PEGylated-thymoquinone-nanoparticle mediated retardation of breast cancer cell migration by deregulation of cytoskeletal actin polymerization through miR-34a. 2015;51:91-107.
30. İşcan G, İşcan A, Demirci FJNpc. Anticandidal effects of thymoquinone: Mode of action determined by transmission electron microscopy (TEM). 2016;11(7):1934578X1601100726.
31. El-Najjar N, Chatila M, Moukadem H, Vuorela H, Ocker M, Gandesiri M, et al. Reactive oxygen species mediate thymoquinone-induced apoptosis and activate ERK and JNK signaling. 2010;15(2):183-95.
Infection Epidemiology and Microbiology
Volume 8, Issue 2
June 2022
Pages 169-176