AAPS PharmSciTech

, Volume 19, Issue 8, pp 3490–3500 | Cite as

Optimization and Characterization of Thymoquinone-Loaded Liposomes with Enhanced Topical Anti-inflammatory Activity

  • Mahmoud Mostafa
  • Eman AlaaeldinEmail author
  • Usama Farghaly Aly
  • Hatem A. Sarhan
Research Article Theme: Advances in Topical Delivery of Drugs
Part of the following topical collections:
  1. Theme: Advances in Topical Delivery of Drugs


Thymoquinone, the major constituent of Nigella sativa oil has been found to have a promising topical anti-inflammatory activity; however, exaggerated heat and photo-sensitivity and lipophilicity prevent the best use of this promising product. The present work aimed to formulate an ideal thymoquinone liposomal system for topical delivery. Different liposomal systems were developed using thin film hydration method by applying different cholesterol molar concentrations, different total lipid molar concentrations, and different drug-to-lipid ratios. Morphological characterization of the prepared formulae was performed using polarized light, scanning electron microscope, and transmission electron microscope. The optimized formula (F12) was selected on the basis of enhanced permeation through the skin and was incorporated into chitosan gel for topical application. The gel formulation was clear with suitable skin permeation and exhibited acceptable rheological properties. Using carrageenan-induced paw edema in rats, the developed chitosan gel (F12) showed significant superior in vivo anti-inflammatory activity over the chitosan gel of the TQ (p < 0.05) and comparable effect to the marketed indomethacin gel. As a conclusion, results revealed the potential of formulating thymoquinone as liposomal formulation in enhancing the anti-inflammatory effect compared to the TQ solution.


liposomes thymoquinone skin enhanced drug deposition, enhanced stability 



The authors wish to thank the Lipoid GmbH company (Germany) for the generous gift of Phospholipon 90H. Also, we are indebted to Dr. Rehab Refaee (Department of Histology, Faculty of Medicine, Minia University) for her contribution concerning the histology of skin observation.

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.


  1. 1.
    Ali B, Blunden G. Pharmacological and toxicological properties of Nigella sativa. Phytother Res. 2003;17(4):299–305.PubMedGoogle Scholar
  2. 2.
    Aljabre S. In vitro antifungal activity of thymoqyuinone against Scopulariopsis brevicaulis. Arab J Pharm Sci. 2005;3:27–33.Google Scholar
  3. 3.
    Kundu JK, Liu L, Shin J-W, Surh Y-J. Thymoquinone inhibits phorbol ester-induced activation of NF-κB and expression of COX-2, and induces expression of cytoprotective enzymes in mouse skin in vivo. Biochem Biophys Res Commun. 2013;438(4):721–7.PubMedGoogle Scholar
  4. 4.
    Yousefi M, Barikbin B, Kamalinejad M, Abolhasani E, Ebadi A, Younespour S, et al. Comparison of therapeutic effect of topical Nigella with betamethasone and Eucerin in hand eczema. J Eur Acad Dermatol Venereol. 2013;27(12):1498–504.PubMedGoogle Scholar
  5. 5.
    Ivankovic S, Stojkovic R, Jukic M, Milos M, Milos M, Jurin M. The antitumor activity of thymoquinone and thymohydroquinone in vitro and in vivo. Exp Oncol. 2006;28(3):220–4.PubMedGoogle Scholar
  6. 6.
    Houghton PJ, Zarka R, de las Heras B, Hoult J. Fixed oil of Nigella sativa and derived thymoquinone inhibit eicosanoid generation in leukocytes and membrane lipid peroxidation. Planta Med. 1995;61(01):33–6.PubMedGoogle Scholar
  7. 7.
    El Gazzar M, El Mezayen R, Marecki JC, Nicolls MR, Canastar A, Dreskin SC. Anti-inflammatory effect of thymoquinone in a mouse model of allergic lung inflammation. Int Immunopharmacol. 2006;6(7):1135–42.PubMedGoogle Scholar
  8. 8.
    Salmani JMM, Asghar S, Lv H, Zhou J. Aqueous solubility and degradation kinetics of the phytochemical anticancer thymoquinone; probing the effects of solvents, pH and light. Molecules. 2014;19(5):5925–39.PubMedPubMedCentralGoogle Scholar
  9. 9.
    Mezei M, Gulasekharam V. Liposomes-a selective drug delivery system for the topical route of administration I. Lotion dosage form. Life Sci. 1980;26(18):1473–7.PubMedGoogle Scholar
  10. 10.
    Doppalapudi S, Jain A, Chopra DK, Khan W. Psoralen loaded liposomal nanocarriers for improved skin penetration and efficacy of topical PUVA in psoriasis. Eur J Pharm Sci. 2017;96:515–29.PubMedGoogle Scholar
  11. 11.
    Dragicevic-Curic N, Gräfe S, Gitter B, Winter S, Fahr A. Surface charged temoporfin-loaded flexible vesicles: in vitro skin penetration studies and stability. Int J Pharm. 2010;384(1–2):100–8.PubMedGoogle Scholar
  12. 12.
    Agarwal R, Katare O, Vyas S. Preparation and in vitro evaluation of liposomal/niosomal delivery systems for antipsoriatic drug dithranol. Int J Pharm. 2001;228(1):43–52.PubMedGoogle Scholar
  13. 13.
    Bangham A, Standish MM, Watkins J. Diffusion of univalent ions across the lamellae of swollen phospholipids. J Mol Biol. 1965;13(1):238–IN27.PubMedGoogle Scholar
  14. 14.
    Bangham A, Standish M, Weissmann G. The action of steroids and streptolysin S on the permeability of phospholipid structures to cations. J Mol Biol. 1965;13(1):253–IN28.PubMedGoogle Scholar
  15. 15.
    Odeh F, Ismail SI, Abu-Dahab R, Mahmoud IS, Al Bawab A. Thymoquinone in liposomes: a study of loading efficiency and biological activity towards breast cancer. Drug Deliv. 2012;19(8):371–7.PubMedGoogle Scholar
  16. 16.
    Lopez-Pinto J, Gonzalez-Rodriguez M, Rabasco A. Effect of cholesterol and ethanol on dermal delivery from DPPC liposomes. Int J Pharm. 2005;298(1):1–12.PubMedGoogle Scholar
  17. 17.
    New RC. Preparation of liposomes. In: New RRC, editors. Liposomes, a practical approach. Oxford etc: IRZ press; 1990. p. 33–104.Google Scholar
  18. 18.
    Ruozi B, Tosi G, Forni F, Fresta M, Vandelli MA. Atomic force microscopy and photon correlation spectroscopy: two techniques for rapid characterization of liposomes. Eur J Pharm Sci. 2005;25(1):81–9.PubMedGoogle Scholar
  19. 19.
    Değim Z, Çelebi N, Alemdaroğlu C, Deveci M, Öztürk S, Özoğul C. Evaluation of chitosan gel containing liposome-loaded epidermal growth factor on burn wound healing. Int Wound J. 2011;8(4):343–54.PubMedGoogle Scholar
  20. 20.
    Qiu Y, Gao Y, Hu K, Li F. Enhancement of skin permeation of docetaxel: a novel approach combining microneedle and elastic liposomes. J Control Release. 2008;129(2):144–50.PubMedGoogle Scholar
  21. 21.
    Erdurmus M, Yagci R, Yilmaz B, Hepsen IF, Turkmen C, Aydin B, et al. Inhibitory effects of topical thymoquinone on corneal neovascularization. Cornea. 2007;26(6):715–9.PubMedGoogle Scholar
  22. 22.
    Winter CA, Risley EA, Nuss GW. Carrageenin-induced edema in hind paw of the rat as an assay for antiinflammatory drugs. Exp Biol Med. 1962;111(3):544–7.Google Scholar
  23. 23.
    Adeyemi O, Okpo S, Ogunti O. Analgesic and anti-inflammatory effects of the aqueous extract of leaves of Persea americana Mill (Lauraceae). Fitoterapia. 2002;73(5):375–80.PubMedGoogle Scholar
  24. 24.
    Morris CJ. Carrageenan-induced paw edema in the rat and mouse. Methods Mol Biol. 2003;225:115–21.Google Scholar
  25. 25.
    Moncada S, Ferreira SH, Vane J. Prostaglandins, aspirin-like drugs and the oedema of inflammation. Nature 246(5430):217–219.PubMedGoogle Scholar
  26. 26.
    Bowd AD. Animal care courses: helping fulfill the mandate of animal care committees in Canada. J Appl Anim Welf Sci. 1998;1(4):353–60.PubMedGoogle Scholar
  27. 27.
    Bamgbose S, Noamesi B. Studies on cryptolepine. Planta Med. 1981;41(04):392–6.PubMedGoogle Scholar
  28. 28.
    Duffy JC, Dearden JC, Rostron C. Design, synthesis and biological testing of a novel series of anti-inflammatory drugs. J Pharm Pharmacol. 2001;53(11):1505–14.PubMedGoogle Scholar
  29. 29.
    Nežić L, Škrbić R, Dobrić S, Stojiljković MP, Jaćević V, Šatara SS, et al. Simvastatin and indomethacin have similar anti-inflammatory activity in a rat model of acute local inflammation. Basic Clin Pharmacol Toxicol. 2009;104(3):185–91.PubMedGoogle Scholar
  30. 30.
    Sinko PJ, Singh Y. Martin’s physical pharmacy and pharmaceutical sciences: physical chemical and biopharmaceutical principles in the pharmaceutical sciences. Sixth edition; 2011.Google Scholar
  31. 31.
    Mayer LD, Tai LC, Ko DS, Masin D, Ginsberg RS, Cullis PR, et al. Influence of vesicle size, lipid composition, and drug-to-lipid ratio on the biological activity of liposomal doxorubicin in mice. Cancer Res. 1989;49(21):5922–30.PubMedGoogle Scholar
  32. 32.
    Smola M, Vandamme T, Sokolowski A. Nanocarriers as pulmonary drug delivery systems to treat and to diagnose respiratory and non respiratory diseases. Int J Nanomedicine. 2008;3(1):1–19.PubMedPubMedCentralGoogle Scholar
  33. 33.
    Fang J-Y, Hong C-T, Chiu W-T, Wang Y-Y. Effect of liposomes and niosomes on skin permeation of enoxacin. Int J Pharm. 2001;219(1):61–72.PubMedGoogle Scholar
  34. 34.
    Zucker D, Marcus D, Barenholz Y, Goldblum A. Liposome drugs' loading efficiency: a working model based on loading conditions and drug's physicochemical properties. J Control Release. 2009;139(1):73–80.PubMedGoogle Scholar
  35. 35.
    Hu L, Tang X, Cui F. Solid lipid nanoparticles (SLNs) to improve oral bioavailability of poorly soluble drugs. J Pharm Pharmacol. 2004;56(12):1527–35.PubMedGoogle Scholar
  36. 36.
    McIntosh TJ. The effect of cholesterol on the structure of phosphatidylcholine bilayers. Biochim Biophys Acta Biomembr. 1978;513(1):43–58.Google Scholar
  37. 37.
    New R. Characterization of liposomes. In: New R, (editor) Liposomes—a practical approach. Oxford: IRL Press; 1990. p. 105–62.Google Scholar
  38. 38.
    Kirby C, Clarke J, Gregoriadis G. Effect of the cholesterol content of small unilamellar liposomes on their stability in vivo and in vitro. Biochem J. 1980;186(2):591–8.PubMedPubMedCentralGoogle Scholar
  39. 39.
    du Plessis J, Ramachandran C, Weiner N, Müller D. The influence of particle size of liposomes on the deposition of drug into skin. Int J Pharm. 1994;103(3):277–82.Google Scholar
  40. 40.
    Vargha-Butler E, Hurst E. Study of liposomal drug delivery systems 1. Surface characterization of steroid loaded MLV liposomes. Colloids Surf B: Biointerfaces. 1995;3(5):287–95.Google Scholar
  41. 41.
    Jones MN. The surface properties of phospholipid liposome systems and their characterisation. Adv Colloid Interf Sci. 1995;54:93–128.Google Scholar
  42. 42.
    Jones MN. Surface properties and interactions of vesicles. Curr Opin Colloid Interface Sci. 1996;1(1):91–100.Google Scholar
  43. 43.
    Chen D, Xia D, Li X, Zhu Q, Yu H, Zhu C, et al. Comparative study of Pluronic® F127-modified liposomes and chitosan-modified liposomes for mucus penetration and oral absorption of cyclosporine A in rats. Int J Pharm. 2013;449:1):1–9.PubMedGoogle Scholar
  44. 44.
    Li X, Chen D, Le C, Zhu C, Gan Y, Hovgaard L, et al. Novel mucus-penetrating liposomes as a potential oral drug delivery system: preparation, in vitro characterization, and enhanced cellular uptake. Int J Nanomed. 2011;6(3):151–62.Google Scholar
  45. 45.
    Miyazaki S, Tobiyama T, Takada M, Attwood D. Percutaneous absorption of indomethacin from pluronic F127 gels in rats. J Pharm Pharmacol. 1995;47(6):455–7.PubMedGoogle Scholar
  46. 46.
    Kyrikou I, Daliani I, Mavromoustakos T, Maswadeh H, Demetzos C, Hatziantoniou S, et al. The modulation of thermal properties of vinblastine by cholesterol in membrane bilayers. Biochim Biophys Acta Biomembr. 2004;1661(1):1–8.Google Scholar
  47. 47.
    Demetzos C. Differential scanning calorimetry (DSC): a tool to study the thermal behavior of lipid bilayers and liposomal stability. J Liposome Res. 2008;18(3):159–73.PubMedGoogle Scholar
  48. 48.
    Ghaffar KA, Marasini N, Giddam AK, Batzloff MR, Good MF, Skwarczynski M, et al. The role of size in development of mucosal liposome-lipopeptide vaccine candidates against group A Streptococcus. Med Chem. 2017;13(1):22–7.Google Scholar
  49. 49.
    Orienti I, Luppi B, Zecchi V. Chitosan and its N-carboxyethyl and N-aminoethyl derivatives as vehicles for topical formulations. J Cosmet Sci. 1999;50(5):307–14.Google Scholar
  50. 50.
    Illum L, Farraj NF, Davis SS. Chitosan as a novel nasal delivery system for peptide drugs. Pharm Res. 1994;11(8):1186–9.PubMedGoogle Scholar
  51. 51.
    Varshosaz J, Jaffari F, Karimzadeh S. Development of bioadhesive chitosan gels for topical delivery of lidocaine. Sci Pharm. 2006;74(4):209–32.Google Scholar
  52. 52.
    Paranjothy K. Gels as topical applications. Indian Drugs. 1994;31:224.Google Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2018

Authors and Affiliations

  • Mahmoud Mostafa
    • 1
  • Eman Alaaeldin
    • 1
    Email author
  • Usama Farghaly Aly
    • 1
  • Hatem A. Sarhan
    • 1
  1. 1.Department of PharmaceuticsThe University of MiniaMiniaEgypt

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