Characterization, cell toxicity, and antimicrobial activity of a carvacrol-encapsulating nanoliposomal system against Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli

Document Type : Research Paper

Authors

1 Department of Medical Biotechnology, School of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran

2 Department of Medical Biotechnology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

3 Department of Agricultural Biotechnology, College of Agriculture, Isfahan University of Technology (IUT), Isfahan, Iran

4 Biotechnology Research Center, International Campus, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

5 Natural Products and Medicinal Plants Research Center, North Khorasan University of Medical Sciences, Bojnurd, Iran

6 Department of Advanced Technologies, School of Medicine, North Khorasan University of Medical Sciences, Bojnurd, Iran

Abstract

Objective(s): Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli are the main pathogenic bacteria involved in severe, polymicrobial, and multidrug-resistant infections. For these infections to be overcome, lipid-based nanoparticles and nanoformulations such as liposomes have demonstrated marked potential in fighting bacterial infections by delivering antibacterial drugs, fusing with bacterial membranes, and promoting the direct delivery of antibacterial agents to bacteria. This study assesses the antibacterial effects of various formulations of carvacrol (CV)-encapsulating nanoliposomes on S. aureus, P. aeruginosa, and E. coli. The study further evaluates the cytotoxicity of the fabricated nanoliposomal system against human foreskin fibroblast (HFF) cells.
Materials and Methods: Various formulations of the liposomal nanosystem were first prepared through the thin film hydration method utilizing different concentrations of soy phosphatidylcholine (SPC), cholesterol (Chol), and Tween 60. The formulations were then evaluated for drug entrapment efficiency and release profiles, and the optimum formulation was determined for the experiments. The optimum formulation was then structurally analyzed, and the cytotoxicity of free CV and encapsulated CV on both bacteria and HFF cells was evaluated. 
Results: Microbial tests revealed that CV-LPs outperform free CV regarding their antibacterial effects on the studied bacterial strains, with the maximum inhibitory effect exerted on S. aureus, followed by E. coli and P. aeruginosa. 
Conclusion: Furthermore, the MTT assay indicated that the cytotoxicity of CV against normal HFF cells was remarkably declined when it was encapsulated in the liposomal nanosystem.

Keywords

References

1.     Beygi M, Oroojalian F, Hosseini SS, Mokhtarzadeh A, Kesharwani P, Sahebkar A. Recent progress in functionalized and targeted polymersomes and chimeric polymeric nanotheranostic platforms for cancer therapy. Prog Mater Sci. 2023:101209.
2.    Oroojalian F, Beygi M, Baradaran B, Mokhtarzadeh A, Shahbazi MA. Immune cell membrane‐coated biomimetic nanoparticles for targeted cancer therapy. Small. 2021;17(12):2006484.
3.    Zolghadri S, Beygi M, Mohammad TF, Alijanianzadeh M, Pillaiyar T, Garcia-Molina P, et al. Targeting tyrosinase in hyperpigmentation: current status, Limitations and future promises. Biochem Pharmacol. 2023:115574.
4.    Oroojalian F, Karimzadeh S, Javanbakht S, Hejazi M, Baradaran B, Webster TJ, et al. Current trends in stimuli-responsive nanotheranostics based on metal–organic frameworks for cancer therapy. Mater Today. 2022;57:192-224.
5.    Karimi MA, Dadmehr M, Hosseini M, Korouzhdehi B, Oroojalian F. Sensitive detection of methylated DNA and methyltransferase activity based on the lighting up of FAM-labeled DNA quenched fluorescence by gold nanoparticles. RSC Adv. 2019;9(21):12063-12069.
6.    Hosseini Motlagh NS, Sharifi E, Hghiralsadat BF, Sardari Zarchi M, Yaghoubi F, Oroojalian F. Investigation and comparison of laser and ultrasound effects on the temperature increasing of the solutions containing graphene oxide nanoparticles for thermal treatment of osteosarcoma cancer cells. Nanomed J. 2023;10(4):313-322.
7.    Mir M, Permana AD, Ahmed N, Khan GM, ur Rehman A, Donnelly RF. Enhancement in site-specific delivery of carvacrol for potential treatment of infected wounds using infection responsive nanoparticles loaded into dissolving microneedles: A proof of concept study. Eur J Pharm Biopharm. 2020;147:57-68.
8.    Costa MF, Durço AO, Rabelo TK, Barreto RdSS, Guimarães AG. Effects of Carvacrol, Thymol and essential oils containing such monoterpenes on wound healing: A systematic review. J Pharm Pharmacol. 2019;71(2):141-155.
9.    Trevisan DAC, Campanerut-Sa PAZ, da Silva AF, Batista AFP, Seixas FAV, Peralta RM, et al. Action of carvacrol in Salmonella Typhimurium biofilm: A proteomic study. J Appl Biomed. 2020;18(4):106-114.
10.    Trevisan DAC, Silva AFd, Negri M, Abreu Filho BAd, Machinski Junior M, Patussi EV, et al. Antibacterial and antibiofilm activity of carvacrol against Salmonella enterica serotype Typhimurium. Braz J Pharm Sci. 2018;54.
11.    Omidi M, Malakoutian M, Choolaei M, Oroojalian F, Haghiralsadat F, Yazdian F. A Label-Free detection of biomolecules using micromechanical biosensors. Chin Phys Lett. 2013;30(6):068701.
12.    Azizi M, Tavana M, Farsi M, Oroojalian F. Yield performance of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W. Curt.: Fr.) P. Karst.(higher Basidiomycetes), using different waste materials as substrates. Int J Med Mushrooms. 2012;14(5).
13.    Azizi M, Chizzola R, Ghani A, Oroojalian F. Composition at different development stages of the essential oil of four Achillea species grown in Iran. Nat Prod Commun. 2010;5(2):1934578X1000500224.
14.    Chizzola R, Saeidnejad AH, Azizi M, Oroojalian F, Mardani H. Bunium persicum: variability in essential oil and antioxidants activity of fruits from different Iranian wild populations. Genet Resour Crop Evol. 2014;61:1621-1631.
15.    Ebrahimpour M, Akhlaghi M, Hemati M, Ghazanfary S, Shahriary S, Ghalekohneh SJ, et al. In vitro evaluation and comparison of anticancer, antimicrobial, and antifungal properties of thyme niosomes containing essential oil. Nanomed J. 2022;9(4).
16.    Rashidi A, Omidi M, Choolaei M, Nazarzadeh M, Yadegari A, Haghierosadat F, et al. Electromechanical properties of vertically aligned carbon nanotube. Adv Mat Res. 2013;705:332-336.
17.    Mir M, Ahmed N, Permana AD, Rodgers AM, Donnelly RF, Rehman AU. Enhancement in site-specific delivery of carvacrol against methicillin resistant Staphylococcus aureus induced skin infections using enzyme responsive nanoparticles: a proof of concept study. Pharmaceutics. 2019;11(11):606.
18.    Nia AH, Behnam B, Taghavi S, Oroojalian F, Eshghi H, Shier WT, et al. Evaluation of chemical modification effects on DNA plasmid transfection efficiency of single-walled carbon nanotube–succinate–polyethylenimine conjugates as non-viral gene carriers. Med Chem Comm. 2017;8(2):364-375.
19.    Pishavar E, Oroojalian F, Ramezani M, Hashemi M. Cholesterol‐conjugated PEGylated PAMAM as an efficient nanocarrier for plasmid encoding interleukin‐12 immunogene delivery toward colon cancer cells. Biotechnol Prog.. 2020;36(3):e2952.
20.    Oroojalian F, Rezayan AH, Mehrnejad F, Nia AH, Shier WT, Abnous K, et al. Efficient megalin targeted delivery to renal proximal tubular cells mediated by modified-polymyxin B-polyethylenimine based nano-gene-carriers. Mater Sci Eng C. 2017;79:770-782.
21.    Rahimizadeh M, Eshghi H, Shiri A, Ghadamyari Z, Matin MM, Oroojalian F, et al. Fe (HSO 4) 3 as an efficient catalyst for diazotization and diazo coupling reactions. J Korean Chem Soc. 2012;56(6):716-9.
22.    Oroojalian F, Rezayan AH, Shier WT, Abnous K, Ramezani M. Megalin-targeted enhanced transfection efficiency in cultured human HK-2 renal tubular proximal cells using aminoglycoside-carboxyalkyl-polyethylenimine-containing nanoplexes. Int J Pharm. 2017;523(1):102-20.
23.    Gholami Z, Dadmehr M, Jelodar NB, Hosseini M, Parizi AP. One-pot biosynthesis of CdS quantum dots through in vitro regeneration of hairy roots of Rhaphanus sativus L. and their apoptosis effect on MCF-7 and AGS cancerous human cell lines. Mater Res Express. 2020;7(1):015056.
24.    Pourpirali R, Mahmoudnezhad A, Oroojalian F, Zarghami N, Pilehvar Y. Prolonged proliferation and delayed senescence of the adipose-derived stem cells grown on the electrospun composite nanofiber co-encapsulated with TiO2 nanoparticles and metformin-loaded mesoporous silica nanoparticles. Int J Pharm. 2021;604:120733.
25.    Alves PM, Al-Badi E, Withycombe C, Jones PM, Purdy KJ, Maddocks SE. Interaction between Staphylococcus aureus and Pseudomonas aeruginosa is beneficial for colonisation and pathogenicity in a mixed biofilm. Pathog Dis. 2018;76(1).
26.    Giacometti A, Cirioni O, Schimizzi A, Del Prete M, Barchiesi F, D’Errico M, et al. Epidemiology and microbiology of surgical wound infections. J Clin Microbiol. 2000;38(2):918-922.
27.    Puca V, Marulli RZ, Grande R, Vitale I, Niro A, Molinaro G, et al. Microbial species isolated from infected wounds and antimicrobial resistance analysis: Data emerging from a three-years retrospective study. Antibiotics. 2021;10(10):1162.
28.    Moghaddam FA, Ebrahimian M, Oroojalian F, Yazdian-Robati R, Kalalinia F, Tayebi L, et al. Effect of thymoquinone-loaded lipid–polymer nanoparticles as an oral delivery system on anticancer efficiency of doxorubicin. J Nanostructure Chem. 2021:1-12.
29.    Negut I, Grumezescu V, Grumezescu AM. Treatment Strategies for Infected Wounds. Molecules. 2018;23(9).
30.    Di Sotto A, Paolicelli P, Nardoni M, Abete L, Garzoli S, Di Giacomo S, et al. SPC liposomes as possible delivery systems for improving bioavailability of the natural sesquiterpene β-caryophyllene: lamellarity and drug-loading as key features for a rational drug delivery design. Pharmaceutics. 2018;10(4):274.
31.    El-Gizawy SA, Nouh A, Saber S, Kira AY. Deferoxamine-loaded transfersomes accelerates healing of pressure ulcers in streptozotocin-induced diabetic rats. J Drug Deliv Sci Technol. 2020;58:101732.
32.    Ruwizhi N, Aderibigbe BA. The efficacy of cholesterol-based carriers in drug delivery. Molecules. 2020;25(18):4330.
33.    Khan I, Needham R, Yousaf S, Houacine C, Islam Y, Bnyan R, et al. Impact of phospholipids, surfactants and cholesterol selection on the performance of transfersomes vesicles using medical nebulizers for pulmonary drug delivery. J Drug Deliv Sci Technol. 2021;66:102822.
34.    Kamboj S, Saini V, Bala S. Formulation and characterization of drug loaded nonionic surfactant vesicles (niosomes) for oral bioavailability enhancement. Sci World J. 2014;2014.
35.    Panwar P, Pandey B, Lakhera P, Singh K. Preparation, characterization, and in vitro release study of albendazole-encapsulated nanosize liposomes. Int J Nanomedicine. 2010:101-108.
36.    Maja L, Željko K, Mateja P. Sustainable technologies for liposome preparation. J Supercrit Fluids. 2020;165:104984.
37.    Danaei M, Dehghankhold M, Ataei S, Hasanzadeh Davarani F, Javanmard R, Dokhani A, et al. Impact of particle size and polydispersity index on the clinical applications of lipidic nanocarrier systems. Pharmaceutics. 2018;10(2):57.
38.    Neves LF, Duan J, Voelker A, Khanal A, McNally L, Steinbach-Rankins J, et al. Preparation and optimisation of anionic liposomes for delivery of small peptides and cDNA to human corneal epithelial cells. J Microencapsul. 2016;33(4):391-399.
39.    Liolios C, Gortzi O, Lalas S, Tsaknis J, Chinou I. Liposomal incorporation of carvacrol and thymol isolated from the essential oil of Origanum dictamnus L. and in vitro antimicrobial activity. Food Chem. 2009;112(1):77-83.
40.    Cacciatore FA, Dalmás M, Maders C, Isaía HA, Brandelli A, da Silva Malheiros P. Carvacrol encapsulation into nanostructures: Characterization and antimicrobial activity against foodborne pathogens adhered to stainless steel. Food Res Int. 2020;133:109143.
41.    Gobin M, Proust R, Lack S, Duciel L, Des Courtils C, Pauthe E, et al. A combination of the natural molecules gallic acid and carvacrol eradicates P. aeruginosa and S. aureus mature biofilms. Int J Mol Sci. 2022;23(13):7118.
42.    Lambert R, Skandamis PN, Coote PJ, Nychas GJ. A study of the minimum inhibitory concentration and mode of action of oregano essential oil, thymol and carvacrol. J Appl Microbiol. 2001;91(3):453-462.
43.    Yao N, Xu Q, He J-K, Pan M, Hou Z-F, Liu D-D, et al. Evaluation of Origanum vulgare essential oil and its active ingredients as potential drugs for the treatment of toxoplasmosis. Front Cell Infect Microbiol. 2021;11:793089.
Nanomedicine Journal
Volume 11, Issue 1
January 2024
Pages 93-106

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