Development of Nanoemulsion Formulation Containing Ylang Ylang Essential Oil for Topical Applications, Evaluation of In Vitro Cytotoxicity and ADMET Profile

Year 2024, Volume: 11 Issue: 3, 1181 - 1196, 30.08.2024

Nesrin Karabatak , Bahar Gök , Yasemin Budama-kılınc

https://doi.org/10.18596/jotcsa.1418645

Abstract

Ultraviolet (UV) rays damage DNA, causing adverse effects such as photoaging and cancer on the skin. For the well-being of individuals, there is a need to develop innovative skin products with high effectiveness using protective and therapeutic agents. In this context, in our study, A nanoemulsion (NE) formulation containing Ylang-ylang essential oil (YO), which has many biological active properties such as antimicrobial, antioxidant, anti-inflammatory, and anticancer, was produced by the ultrasonic emulsification method and characterized. The thermodynamic stability was evaluated, and its in vitro release profile determined the dialysis membrane technique. The cytotoxic effect of YO-NE was examined with the in vitro method on the HacaT cell line using the MTT method and in silico method using the ADMET profile. Dynamic light scattering (DLS) results showed that the average droplet size of the YO-NE formulation was 184.1±2.307 nm, the polydispersity index (PdI) was 0.151±0.006, and the Zeta potential (ζ) -10.8 ± 0.400 mV. As a result of in vitro release studies, it was observed that 99.98± 1.00% of YO release from NE occurred within 5 hours. Based on the thermodynamic stability test results, it was determined that the developed formulation did not show sedimentation or phase separation. Also, it remained stable under three different storage conditions in the three-month stability test. Cytotoxicity results revealed that the YO-NE formulation was safe. All the results indicated that the YO-NE formulation might be considered a non-toxic product candidate with physicochemical properties suitable for topical use.

Keywords

Nanoemulsion, Ylang Ylang Essential Oil, Cytotoxicity, Topical Application

Supporting Institution

Yildiz Technical University Scientific Research Foundation [Project ID: FCD-2021-4509]

Project Number

FCD-2021-4509

Thanks

This work was supported by Yildiz Technical University Scientific Research Foundation [FCD-2021-4509]. In this study, the infrastructure of the Applied Nanotechnology and Antibody Production Laboratory established with TUBITAK support (project numbers: 115S132, 117S097 and 5200110) was used. The authors would also like to thank TUBITAK for their support.

References

• 1. Kammeyer A, Luiten RM. Oxidation events and skin aging. Ageing Res Rev [Internet]. 2015 May;21:16–29. Available from: <URL>.
• 2. Nanni V, Canuti L, Gismondi A, Canini A. Hydroalcoholic extract of Spartium junceum L. flowers inhibits growth and melanogenesis in B16-F10 cells by inducing senescence. Phytomedicine [Internet]. 2018 Jul;46:1–10. Available from: <URL>.
• 3. Rabe JH, Mamelak AJ, McElgunn PJS, Morison WL, Sauder DN. Photoaging: Mechanisms and repair. J Am Acad Dermatol [Internet]. 2006 Jul;55(1):1–19. Available from: <URL>.
• 4. Zhu S, Zhao Z, Qin W, Liu T, Yang Y, Wang Z, et al. The Nanostructured lipid carrier gel of Oroxylin A reduced UV-induced skin oxidative stress damage. Colloids Surfaces B Biointerfaces [Internet]. 2022 Aug;216:112578. Available from: <URL>.
• 5. González S, Fernández-Lorente M, Gilaberte-Calzada Y. The latest on skin photoprotection. Clin Dermatol [Internet]. 2008 Nov;26(6):614–26. Available from: <URL>.
• 6. Mukherjee S, Date A, Patravale V, Korting HC, Roeder A, Weindl G. Retinoids in the treatment of skin aging: an overview of clinical efficacy and safety. Clin Interv Aging [Internet]. 2006 Dec;1(4):327–48. Available from: <URL>.
• 7. Pacheco MT, Silva ACG, Nascimento TL, Diniz DGA, Valadares MC, Lima EM. Protective effect of sucupira oil nanoemulsion against oxidative stress in UVA-irradiated HaCaT cells. J Pharm Pharmacol [Internet]. 2019 Sep 3;71(10):1532–43. Available from: <URL>.
• 8. Guzmán E, Lucia A. Essential Oils and Their Individual Components in Cosmetic Products. Cosmetics [Internet]. 2021 Dec 3;8(4):114. Available from: <URL>.
• 9. Sedky NK, Abdel-Kader NM, Issa MY, Abdelhady MMM, Shamma SN, Bakowsky U, et al. Co-Delivery of Ylang Ylang Oil of Cananga odorata and Oxaliplatin Using Intelligent pH-Sensitive Lipid-Based Nanovesicles for the Effective Treatment of Triple-Negative Breast Cancer. Int J Mol Sci [Internet]. 2023 May 7;24(9):8392. Available from: <URL>.
• 10. Goodrich KR. Floral scent in Annonaceae. Bot J Linn Soc [Internet]. 2012 May;169(1):262–79. Available from: <URL>.
• 11. Kalagatur NK, Mudili V, Kamasani JR, Siddaiah C. Discrete and combined effects of Ylang-Ylang (Cananga odorata) essential oil and gamma irradiation on growth and mycotoxins production by Fusarium graminearum in maize. Food Control [Internet]. 2018 Dec;94:276–83. Available from: <URL>.
• 12. Upadhyay N, Singh VK, Dwivedy AK, Chaudhari AK, Dubey NK. Assessment of nanoencapsulated Cananga odorata essential oil in chitosan nanopolymer as a green approach to boost the antifungal, antioxidant and in situ efficacy. Int J Biol Macromol [Internet]. 2021 Feb;171:480–90. Available from: <URL>.
• 13. Youness RA, Al-Mahallawi AM, Mahmoud FH, Atta H, Braoudaki M, Fahmy SA. Oral Delivery of Psoralidin by Mucoadhesive Surface-Modified Bilosomes Showed Boosted Apoptotic and Necrotic Effects Against Breast and Lung Cancer Cells. Polymers (Basel) [Internet]. 2023 Mar 15;15(6):1464. Available from: <URL>.
• 14. Fahmy SA, Ponte F, Fawzy IM, Sicilia E, Azzazy HMES. Betaine host–guest complexation with a calixarene receptor: enhanced in vitro anticancer effect. RSC Adv [Internet]. 2021;11(40):24673–80. Available from: <URL>.
• 15. Azzazy HMES, Sawy AM, Abdelnaser A, Meselhy MR, Shoeib T, Fahmy SA. Peganum harmala Alkaloids and Tannic Acid Encapsulated in PAMAM Dendrimers: Improved Anticancer Activities as Compared to Doxorubicin. ACS Appl Polym Mater [Internet]. 2022 Oct 14;4(10):7228–39. Available from: <URL>.
• 16. Fahmy SA, Mahdy NK, Al Mulla H, ElMeshad AN, Issa MY, Azzazy HMES. PLGA/PEG Nanoparticles Loaded with Cyclodextrin-Peganum harmala Alkaloid Complex and Ascorbic Acid with Promising Antimicrobial Activities. Pharmaceutics [Internet]. 2022 Jan 7;14(1):142. Available from: <URL>.
• 17. Kong M, Park HJ. Stability investigation of hyaluronic acid based nanoemulsion and its potential as transdermal carrier. Carbohydr Polym [Internet]. 2011 Jan 30;83(3):1303–10. Available from: <URL>.
• 18. Soriano-Ruiz JL, Calpena-Capmany AC, Cañadas-Enrich C, Febrer NB de, Suñer-Carbó J, Souto EB, et al. Biopharmaceutical profile of a clotrimazole nanoemulsion: Evaluation on skin and mucosae as anticandidal agent. Int J Pharm [Internet]. 2019 Jan;554:105–15. Available from: <URL>.
• 19. Thomas L, Zakir F, Mirza MA, Anwer MK, Ahmad FJ, Iqbal Z. Development of Curcumin loaded chitosan polymer based nanoemulsion gel: In vitro, ex vivo evaluation and in vivo wound healing studies. Int J Biol Macromol [Internet]. 2017 Aug;101:569–79. Available from: <URL>.
• 20. Echeverría J, Albuquerque R. Nanoemulsions of Essential Oils: New Tool for Control of Vector-Borne Diseases and In Vitro Effects on Some Parasitic Agents. Medicines [Internet]. 2019 Mar 27;6(2):42. Available from: <URL>.
• 21. Jaiswal M, Dudhe R, Sharma PK. Nanoemulsion: an advanced mode of drug delivery system. 3 Biotech [Internet]. 2015 Apr 8;5(2):123–7. Available from: <URL>.
• 22. Sharadha M, Gowda D V, Vishal Gupta N, Akhila AR. An overview on topical drug delivery system–updated review. Int J Res Pharm Sci [Internet]. 2020;11(1):368–85. Available from: <URL>.
• 23. Ghosh V, Saranya S, Mukherjee A, Chandrasekaran N. Cinnamon Oil Nanoemulsion Formulation by Ultrasonic Emulsification: Investigation of Its Bactericidal Activity. J Nanosci Nanotechnol [Internet]. 2013 Jan 1;13(1):114–22. Available from: <URL>.
• 24. Gul U, Khan MI, Madni A, Sohail MF, Rehman M, Rasul A, et al. Olive oil and clove oil-based nanoemulsion for topical delivery of terbinafine hydrochloride: in vitro and ex vivo evaluation. Drug Deliv [Internet]. 2022 Dec 31;29(1):600–12. Available from: <URL>.
• 25. Ozkan B, Altuntas E, Cakir Koc R, Budama-Kilinc Y. Development of piperine nanoemulsions: an alternative topical application for hypopigmentation. Drug Dev Ind Pharm [Internet]. 2022 Mar 4;48(3):117–27. Available from: <URL>.
• 26. Budama-Kilinc Y, Gok B, Kecel-Gunduz S, Altuntas E. Development of nanoformulation for hyperpigmentation disorders: Experimental evaluations, in vitro efficacy and in silico molecular docking studies. Arab J Chem [Internet]. 2022 Dec;15(12):104362. Available from: <URL>.
• 27. Gaba B, Khan T, Haider MF, Alam T, Baboota S, Parvez S, et al. Vitamin E Loaded Naringenin Nanoemulsion via Intranasal Delivery for the Management of Oxidative Stress in a 6-OHDA Parkinson’s Disease Model. Biomed Res Int [Internet]. 2019 Apr 14;2019:2382563. Available from: <URL>.
• 28. Ercin E, Kecel-Gunduz S, Gok B, Aydin T, Budama-Kilinc Y, Kartal M. Laurus nobilis L. Essential Oil-Loaded PLGA as a Nanoformulation Candidate for Cancer Treatment. Molecules [Internet]. 2022 Mar 15;27(6):1899. Available from: <URL>.
• 29. Rashid SA, Bashir S, Naseem F, Farid A, Rather IA, Hakeem KR. Olive Oil Based Methotrexate Loaded Topical Nanoemulsion Gel for the Treatment of Imiquimod Induced Psoriasis-like Skin Inflammation in an Animal Model. Biology (Basel) [Internet]. 2021 Oct 31;10(11):1121. Available from: <URL>.
• 30. Budama‐Kilinc Y, Kecel‐Gunduz S, Ozdemir B, Bicak B, Akman G, Arvas B, et al. New nanodrug design for cancer therapy: Its synthesis, formulation, in vitro and in silico evaluations. Arch Pharm (Weinheim) [Internet]. 2020 Nov 5;353(11):2000137. Available from: <URL>.
• 31. Gok B, Budama-Kilinc Y, Kecel-Gunduz S. Anti-aging activity of Syn-Ake peptide by in silico approaches and in vitro tests. J Biomol Struct Dyn [Internet]. 2024 Jul 2;42(10):5015–29. Available from: <URL>.
• 32. Gunaseelan S, Balupillai A, Govindasamy K, Ramasamy K, Muthusamy G, Shanmugam M, et al. Linalool prevents oxidative stress activated protein kinases in single UVB-exposed human skin cells. Slominski AT, editor. PLoS One [Internet]. 2017 May 3;12(5):e0176699. Available from: <URL>.
• 33. Haque MI, Khalipha ABR, Sakib MR, Prottay AAS, Islam MA, Zannat R, et al. In-Silico Molecular Docking Study of Linalool Against 3ELJ for Cell Cycle Arrest and Apoptosis in HEPG2 Cells. Int J Evergr Sci Res [Internet]. 2021;3(2):105–18. Available from: <URL>.
• 34. Steliopoulos P, Wüst M, Adam KP, Mosandl A. Biosynthesis of the sesquiterpene germacrene D in Solidago canadensis: 13C and 2H labeling studies. Phytochemistry [Internet]. 2002 May;60(1):13–20. Available from: <URL>.
• 35. Prosser I, Altug IG, Phillips AL, König WA, Bouwmeester HJ, Beale MH. Enantiospecific (+)- and (−)-germacrene D synthases, cloned from goldenrod, reveal a functionally active variant of the universal isoprenoid-biosynthesis aspartate-rich motif. Arch Biochem Biophys [Internet]. 2004 Dec;432(2):136–44. Available from: <URL>.
• 36. Daina A, Michielin O, Zoete V. SwissADME: a free web tool to evaluate pharmacokinetics, drug-likeness and medicinal chemistry friendliness of small molecules. Sci Rep [Internet]. 2017 Mar 3;7(1):42717. Available from: <URL>.
• 37. Pires DE V, Blundell TL, Ascher DB. pkCSM: Predicting Small-Molecule Pharmacokinetic and Toxicity Properties Using Graph-Based Signatures. J Med Chem [Internet]. 2015 May 14;58(9):4066–72. Available from: <URL>.
• 38. Lipinski CA, Lombardo F, Dominy BW, Feeney PJ. Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv Drug Deliv Rev [Internet]. 2001 Mar;46(1–3):3–26. Available from: <URL>.
• 39. Rakhecha B, Agnihotri P, Dakal TC, Saquib M, Monu, Biswas S. Anti-inflammatory activity of nicotine isolated from Brassica oleracea in rheumatoid arthritis. Biosci Rep [Internet]. 2022 Apr 29;42(4):BSR20211392. Available from: <URL>.
• 40. Mojumdar M, Kabir MSH, Hasan MS, Ahmed T, Rahman MR, Akter Y, et al. Molecular docking and pass prediction for analgesic activity of some isolated compounds from Acalypha indica L. and ADME/T property analysis of the compounds. World J Pharm Res [Internet]. 2016;5(7):1761–70. Available from: <URL>.
• 41. Zhao D, Ge Y, Xiang X, Dong H, Qin W, Zhang Q. Structure and stability characterization of pea protein isolate-xylan conjugate-stabilized nanoemulsions prepared using ultrasound homogenization. Ultrason Sonochem [Internet]. 2022 Nov;90:106195. Available from: <URL>.
• 42. Manzoor M, Sharma P, Murtaza M, Jaiswal AK, Jaglan S. Fabrication, characterization, and interventions of protein, polysaccharide and lipid-based nanoemulsions in food and nutraceutical delivery applications: A review. Int J Biol Macromol [Internet]. 2023 Jun;241:124485. Available from: <URL>.
• 43. Kapoor A, Preet S. Evaluation of Acaricidal Activity of Cinnamomum camphora (F. Lauraceae) Essential Oil Nanoemulsion Against Cattle Tick Rhipicephalus microplus. Adv Zool Bot [Internet]. 2023 Apr;11(2):121–8. Available from: <URL>.
• 44. Rupa E, Li J, Arif M, Yaxi H, Puja A, Chan A, et al. Cordyceps militaris Fungus Extracts-Mediated Nanoemulsion for Improvement Antioxidant, Antimicrobial, and Anti-Inflammatory Activities. Molecules [Internet]. 2020 Dec 4;25(23):5733. Available from: <URL>.
• 45. Ayllón-Gutiérrez R, López-Maldonado EA, Macías-Alonso M, González Marrero J, Díaz-Rubio L, Córdova-Guerrero I. Evaluation of the Stability of a 1,8-Cineole Nanoemulsion and Its Fumigant Toxicity Effect against the Pests Tetranychus urticae, Rhopalosiphum maidis and Bemisia tabaci. Insects [Internet]. 2023 Jul 24;14(7):663. Available from: <URL>.
• 46. Liu Q, Huang H, Chen H, Lin J, Wang Q. Food-Grade Nanoemulsions: Preparation, Stability and Application in Encapsulation of Bioactive Compounds. Molecules [Internet]. 2019 Nov 21;24(23):4242. Available from: <URL>.
• 47. Gorjian H, Mihankhah P, Khaligh NG. Influence of tween nature and type on physicochemical properties and stability of spearmint essential oil (Mentha spicata L.) stabilized with basil seed mucilage nanoemulsion. J Mol Liq [Internet]. 2022 Aug;359:119379. Available from: <URL>.
• 48. Mohammed NK, Muhialdin BJ, Meor Hussin AS. Characterization of nanoemulsion of Nigella sativa oil and its application in ice cream. Food Sci Nutr [Internet]. 2020 Jun 21;8(6):2608–18. Available from: <URL>.
• 49. Kampa J, Frazier R, Rodriguez-Garcia J. Physical and Chemical Characterisation of Conventional and Nano/Emulsions: Influence of Vegetable Oils from Different Origins. Foods [Internet]. 2022 Feb 25;11(5):681. Available from: <URL>.
• 50. Lima TS, Silva MFS, Nunes XP, Colombo A V, Oliveira HP, Goto PL, et al. Cineole-containing nanoemulsion: Development, stability, and antibacterial activity. Chem Phys Lipids [Internet]. 2021 Sep;239:105113. Available from: <URL>.
• 51. Özdemir S, Üner B, Karaküçük A, Çelik B, Sümer E, Taş Ç. Nanoemulsions as a Promising Carrier for Topical Delivery of Etodolac: Formulation Development and Characterization. Pharmaceutics [Internet]. 2023 Oct 23;15(10):2510. Available from: <URL>.
• 52. Gündel S da S, de Godoi SN, Santos RCV, da Silva JT, Leite LB de M, Amaral AC, et al. In vivo antifungal activity of nanoemulsions containing eucalyptus or lemongrass essential oils in murine model of vulvovaginal candidiasis. J Drug Deliv Sci Technol [Internet]. 2020 Jun;57:101762. Available from: <URL>.
• 53. Lunardi CN, Gomes AJ, Rocha FS, De Tommaso J, Patience GS. Experimental methods in chemical engineering: Zeta potential. Can J Chem Eng [Internet]. 2021 Mar 12;99(3):627–39. Available from: <URL>.
• 54. Dheyab MA, Aziz AA, Jameel MS, Noqta OA, Khaniabadi PM, Mehrdel B. Simple rapid stabilization method through citric acid modification for magnetite nanoparticles. Sci Rep [Internet]. 2020 Jul 1;10(1):10793. Available from: <URL>.
• 55. Kildaci I, Budama-Kilinc Y, Kecel-Gunduz S, Altuntas E. Linseed Oil Nanoemulsions for treatment of Atopic Dermatitis disease: Formulation, characterization, in vitro and in silico evaluations. J Drug Deliv Sci Technol [Internet]. 2021 Aug;64:102652. Available from: <URL>.
• 56. Gok B, Budama Kilinc Y. Chlorogenic acid nanoemulsion for staphylococcus aureus causing skin infection: Synthesis, characterization and evaluation of antibacterial efficac. Sigma J Eng Nat Sci [Internet]. 2022 Aug;41(2):322–30. Available from: <URL>.
• 57. Marhamati M, Ranjbar G, Rezaie M. Effects of emulsifiers on the physicochemical stability of Oil-in-water Nanoemulsions: A critical review. J Mol Liq [Internet]. 2021 Oct;340:117218. Available from: <URL>.
• 58. Rashid SA, Bashir S, Ullah H, Khan DH, Shah PA, Danish MZ, et al. Development, characterization and optimization of methotrexate-olive oil nano-emulsion for topical application. Pak J Pharm Sci [Internet]. 2021;34. Available from: <URL>.
• 59. Hammodi ID, N. Abd Alhammid S. Preparation and Characterization of Topical Letrozole Nanoemulsion for Breast Cancer. Iraqi J Pharm Sci [Internet]. 2020 Jun 25;29(1):195–206. Available from: <URL>.
• 60. Roselan MA, Ashari SE, Faujan NH, Mohd Faudzi SM, Mohamad R. An Improved Nanoemulsion Formulation Containing Kojic Monooleate: Optimization, Characterization and In Vitro Studies. Molecules [Internet]. 2020 Jun 4;25(11):2616. Available from: <URL>.
• 61. Wang L, Guan X, Zheng C, Wang N, Lu H, Huang Z. New Low-Energy Method for Nanoemulsion Formation: pH Regulation Based on Fatty Acid/Amine Complexes. Langmuir [Internet]. 2020 Sep 1;36(34):10082–90. Available from: <URL>.
• 62. Altuntaş E, Yener G. Formulation and Evaluation of Thermoreversible In Situ Nasal Gels Containing Mometasone Furoate for Allergic Rhinitis. AAPS PharmSciTech [Internet]. 2017 Oct 9;18(7):2673–82. Available from: <URL>.
• 63. Mat Hadzir N, Basri M, Abdul Rahman MB, Salleh AB, Raja Abdul Rahman RNZ, Basri H. Phase Behaviour and Formation of Fatty Acid Esters Nanoemulsions Containing Piroxicam. AAPS PharmSciTech [Internet]. 2013 Mar 6;14(1):456–63. Available from: <URL>.
• 64. Guttoff M, Saberi AH, McClements DJ. Formation of vitamin D nanoemulsion-based delivery systems by spontaneous emulsification: Factors affecting particle size and stability. Food Chem [Internet]. 2015 Mar;171:117–22. Available from: <URL>.
• 65. Bernardi DS, Pereira TA, Maciel NR, Bortoloto J, Viera GS, Oliveira GC, et al. Formation and stability of oil-in-water nanoemulsions containing rice bran oil: in vitro and in vivo assessments. J Nanobiotechnology [Internet]. 2011;9(1):44. Available from: <URL>.
• 66. Altuntaş E, Yener G. Anti-aging potential of a cream containing herbal oils and honey: Formulation and in vivo evaluation of effectiveness using non-invasive biophysical techniques. IOSR J Pharm Biol Sci [Internet]. 2015;10(6):51–60. Available from: <URL>.
• 67. Shen Y, Ni ZJ, Thakur K, Zhang JG, Hu F, Wei ZJ. Preparation and characterization of clove essential oil loaded nanoemulsion and pickering emulsion activated pullulan-gelatin based edible film. Int J Biol Macromol [Internet]. 2021 Jun;181:528–39. Available from: <URL>.
• 68. Jerobin J, Sureshkumar RS, Anjali CH, Mukherjee A, Chandrasekaran N. Biodegradable polymer based encapsulation of neem oil nanoemulsion for controlled release of Aza-A. Carbohydr Polym [Internet]. 2012 Nov;90(4):1750–6. Available from: <URL>.
• 69. Fopase R, Pathode SR, Sharma S, Datta P, Pandey LM. Lipopeptide and essential oil based nanoemulsion for controlled drug delivery. Polym Technol Mater [Internet]. 2020 Dec 11;59(18):2076–86. Available from: <URL>.
• 70. Rajitha P, Shammika P, Aiswarya S, Gopikrishnan A, Jayakumar R, Sabitha M. Chaulmoogra oil based methotrexate loaded topical nanoemulsion for the treatment of psoriasis. J Drug Deliv Sci Technol [Internet]. 2019 Feb;49:463–76. Available from: <URL>.
• 71. Mohd Narawi M, Chiu HI, Yong YK, Mohamad Zain NN, Ramachandran MR, Tham CL, et al. Biocompatible Nutmeg Oil-Loaded Nanoemulsion as Phyto-Repellent. Front Pharmacol [Internet]. 2020 Mar 17;11:214. Available from: <URL>.
• 72. Shah J, Nair AB, Jacob S, Patel RK, Shah H, Shehata TM, et al. Nanoemulsion Based Vehicle for Effective Ocular Delivery of Moxifloxacin Using Experimental Design and Pharmacokinetic Study in Rabbits. Pharmaceutics [Internet]. 2019 May 11;11(5):230. Available from: <URL>.
• 73. Botros SR, Hussein AK, Mansour HF. A Novel Nanoemulsion Intermediate Gel as a Promising Approach for Delivery of Itraconazole: Design, In Vitro and Ex Vivo Appraisal. AAPS PharmSciTech [Internet]. 2020 Oct 6;21(7):272. Available from: <URL>.
• 74. Roggia I, Gomes P, Dalcin AJF, Ourique AF, Mânica da Cruz IB, Ribeiro EE, et al. Profiling and Evaluation of the Effect of Guarana-Loaded Liposomes on Different Skin Cell Lines: An In Vitro Study. Cosmetics [Internet]. 2023 May 12;10(3):79. Available from: <URL>.
• 75. Serrano I, Alhinho B, Cunha E, Tavares L, Trindade A, Oliveira M. Bacteriostatic and Antibiofilm Efficacy of a Nisin Z Solution against Co-Cultures of Staphylococcus aureus and Pseudomonas aeruginosa from Diabetic Foot Infections. Life [Internet]. 2023 Feb 11;13(2):504. Available from: <URL>.
• 76. Standard I. Biological evaluation of medical devices—Part 5: Tests for in vitro cytotoxicity. Geneve, Switz Int Organ Stand. 2009;
• 77. Caldeira LR, Fernandes FR, Costa DF, Frézard F, Afonso LCC, Ferreira LAM. Nanoemulsions loaded with amphotericin B: A new approach for the treatment of leishmaniasis. Eur J Pharm Sci [Internet]. 2015 Apr;70:125–31. Available from: <URL>.
• 78. Musa SH, Razali FN, Shamsudin N, Salim N, Basri M. Novel topical nano-colloidal carrier loaded with cyclosporine: Biological evaluation potentially for psoriasis treatment. J Drug Deliv Sci Technol [Internet]. 2021 Jun;63:102440. Available from: <URL>.
• 79. Meghani N, Patel P, Kansara K, Ranjan S, Dasgupta N, Ramalingam C, et al. Formulation of vitamin D encapsulated cinnamon oil nanoemulsion: Its potential anti-cancerous activity in human alveolar carcinoma cells. Colloids Surfaces B Biointerfaces [Internet]. 2018 Jun;166:349–57. Available from: <URL>.
• 80. Liao S, Yang G, Wang Z, Ou Y, Huang S, Li B, et al. Ultrasonic preparation of Tween-essential oil (Zanthoxylum schinifolium Sieb. et Zucc) oil/water nanoemulsion: Improved stability and alleviation of Staphylococcus epidermidis biofilm. Ind Crops Prod [Internet]. 2022 Nov;188:115654. Available from: <URL>.
• 81. Singh I, Morris A. Performance of transdermal therapeutic systems: Effects of biological factors. Int J Pharm Investig [Internet]. 2011;1(1):4. Available from: <URL>.
• 82. James JP, Ail PD, Crasta L, Kamath RS, Shura MH, T.J S. In Silico ADMET and Molecular Interaction Profiles of Phytochemicals from Medicinal Plants in Dakshina Kannada. J Heal Allied Sci NU [Internet]. 2024 Apr 19;14(2):190–201. Available from: <URL>.
• 83. Drioiche A, Ailli A, Remok F, Saidi S, Gourich AA, Asbabou A, et al. Analysis of the Chemical Composition and Evaluation of the Antioxidant, Antimicrobial, Anticoagulant, and Antidiabetic Properties of Pistacia lentiscus from Boulemane as a Natural Nutraceutical Preservative. Biomedicines [Internet]. 2023 Aug 24;11(9):2372. Available from: <URL>.
• 84. Jamuna S, Rathinavel A, Mohammed Sadullah S, Devaraj S. In silico approach to study the metabolism and biological activities of oligomeric proanthocyanidin complexes. Indian J Pharmacol [Internet]. 2018;50(5):242–50. Available from: <URL>.
• 85. Li XJ, Yang YJ, Li YS, Zhang WK, Tang HB. α-Pinene, linalool, and 1-octanol contribute to the topical anti-inflammatory and analgesic activities of frankincense by inhibiting COX-2. J Ethnopharmacol [Internet]. 2016 Feb;179:22–6. Available from: <URL>.
• 86. Amanpour A, Çelebi F, Kahraman IG, Çelik F. Diyet İnflamatuar İndeksi, İnflamasyon ve Beslenme. Türkiye Sağlık Bilim ve Araştırmaları Derg [Internet]. 2022 Dec 29;5(3):59–80. Available from: <URL>.
• 87. Ulbrich H, Eriksson EE, Lindbom L. Leukocyte and endothelial cell adhesion molecules as targets for therapeutic interventions in inflammatory disease. Trends Pharmacol Sci [Internet]. 2003 Dec;24(12):640–7. Available from: <URL>.