Download PDF
Planta Med 2021; 87(03): 253-266
DOI: 10.1055/a-1330-8765
Formulation and Delivery Systems of Natural Products
Original Papers

Formulation of a Semisolid Emulsion Containing Leptospermum scoparium Essential Oil and Evaluation of In Vitro Antimicrobial and Antibiofilm Efficacy

Gemma C. Porter‡
1   Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, Dunedin, New Zealand
,
Syarida H. Safii‡
2   Department of Restorative Dentistry, University of Malaya, Kuala Lumpur
,
Natalie J. Medlicott
3   School of Pharmacy, University of Otago, Dunedin, New Zealand
,
Warwick J. Duncan
1   Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, Dunedin, New Zealand
,
Geoffrey R. Tompkins
1   Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, Dunedin, New Zealand
,
Dawn E. Coates
1   Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, Dunedin, New Zealand
› Author AffiliationsSupported by: Universiti Malaya
Supported by: Lottery Health (New Zealand)
Supported by: New Zealand Dental Research Foundation
› Further Information
    Permissions and Reprints

Abstract

Manuka oil, an essential oil derived from the Leptospermum scoparium, has been traditionally used for wound care and as a topical antibacterial, antifungal, and anti-inflammatory. However, the essential oil is not well retained at mucosal sites, such as the oral cavity, where the benefits of the aforementioned properties could be utilized toward the treatment of persistent biofilms. Within this study, L. scoparium essential oil was incorporated into a semisolid emulsion for improved delivery. The safety profile of L. scoparium essential oil on human gingival fibroblasts was determined via cell viability, cytotoxicity, and caspase activation. The minimal bactericidal concentration of L. scoparium essential oil was determined, and the emulsionʼs antibiofilm effects visualized using confocal laser scanning microscopy. L. scoparium essential oil demonstrated a lower IC50 (0.02% at 48 h) when compared to the clinical control chlorhexidine (0.002% at 48 h) and displayed lower cumulative cytotoxicity. Higher concentrations of L. scoparium essential oil (≥ 0.1%) at 6 h resulted in higher caspase 3/7 activation, suggesting an apoptotic pathway of cell death. A minimal bactericidal concentration of 0.1% w/w was observed for 6 oral bacteria and 0.01% w/v for Porphyromonas gingivalis. Textural and rheometric analysis indicated increased stability of emulsion with a 1 : 3 ratio of L. scoparium essential oil: Oryza sativa carrier oil. The optimized 5% w/w L. scoparium essential oil emulsion showed increased bactericidal penetrative effects on Streptococci gordonii biofilms compared to oil alone and to chlorhexidine controls. This study has demonstrated the safety, formulation, and antimicrobial activity of L. scoparium essential oil emulsion for potential antibacterial applications at mucosal sites.

Key words

Myrtaceae - Leptospermum Scoparium - essential oil - antimicrobial - emulsion - periodontal disease

Supporting Information

    The rheological investigations of semisolid emulsions, including the storage modulus (G’), loss modulus (G”), and loss tangent (tan δ) as a function of frequency, for carrier emulsions and L. scoparium essential oil emulsions at equivalent final oil concentrations, are available in Table S1 as supporting information.

  • Supporting Information


Publication History

Received: 30 January 2020

Accepted after revision: 27 November 2020

Article published online:
12 January 2021

© 2021. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

  • References

  • 1 Porter NG, Wilkins AL. Chemical, physical and antimicrobial properties of essential oils of Leptospermum scoparium and Kunzea ericoides. Phytochemistry 1998; 50: 407-415
    CrossrefPubMedGoogle Scholar
  • 2 Lis-Balchin M, Hart SL, Deans SG. Pharmacological and antimicrobial studies on different tea tree oils (Melaleuca alternifolia, Leptospermum scoparium or Manuka and Kunzea ericoides or Kanuka), originating in Australia and New Zealand. Phyther Res 2000; 14: 623-629
    CrossrefPubMedGoogle Scholar
  • 3 Derraik GB. New Zealand Mānuka (Leptospermum scoparium; Myrtaceae): a brief account of its natural history and human perceptions. New Zeal Gard J 2008; 11: 25-27
    PubMedGoogle Scholar
  • 4 Douglas MH, Van Klink JW, Smallfield BM, Perry NB, Anderson RE, Johnstone P, Weavers RT. Essential oils from New Zealand manuka: Triketone and other chemotypes of Leptospermum scoparium . Phytochemistry 2004; 65: 1255-1264
    CrossrefPubMedGoogle Scholar
  • 5 Lis-Balchin M, Hart SL. An investigation of the actions of the essential oils of Manuka (Leptospermum scoparium) and Kanuka (Kunzea ericoides), Myrtaceae on guinea-pig smooth muscle. J Pharm Pharmacol 1998; 50: 809-811
    CrossrefPubMedGoogle Scholar
  • 6 Johnston M, McBride M, Dahiya D, Owusu-Apenten R, Nigam PS. Antibacterial activity of Manuka honey and its components: an overview. AIMS Microbiol 2018; 4: 655-664
    CrossrefPubMedGoogle Scholar
  • 7 Chen CC, Yan SH, Yen MY, Wu PF, Liao WT, Huang TS, Wen ZH, David Wang HM. Investigations of kanuka and manuka essential oils for in vitro treatment of disease and cellular inflammation caused by infectious microorganisms. J Microbiol Immunol Infect 2016; 49: 104-111
    CrossrefPubMedGoogle Scholar
  • 8 Nazir MA. Prevalence of periodontal disease, its association with systemic diseases and prevention. Int J Health Sci (Qassim) 2017; 11: 72-80
    PubMedGoogle Scholar
  • 9 Ferreira MC, Dias-Pereira AC, Branco-de-Almeida LS, Martins CC, Paiva SM. Impact of periodontal disease on quality of life: a systematic review. J Periodontal Res 2017; 52: 651-665
    CrossrefPubMedGoogle Scholar
  • 10 Cullinan MP, Seymour GJ. Periodontal disease and systemic illness: will the evidence ever be enough?. Periodontol 2000 2013; 62: 271-286
    CrossrefPubMedGoogle Scholar
  • 11 Caffesse RG, Echeverría JJ. Treatment trends in periodontics. Periodontol 2000 2019; 79: 7-14
    CrossrefPubMedGoogle Scholar
  • 12 Miastkowska M, Michalczyk A, Figacz K, Sikora E. Nanoformulations as a modern form of biofungicide. J Environ Heal Sci Eng 2020; 18: 119-128
    CrossrefPubMedGoogle Scholar
  • 13 Lu WC, Huang DW, Wang CCR, Yeh CH, Tsai JC, Huang YT, Li PH. Preparation, characterization, and antimicrobial activity of nanoemulsions incorporating citral essential oil. J Food Drug Anal 2018; 26: 82-89
    CrossrefPubMedGoogle Scholar
  • 14 Vervelle A, Mouhyi J, Del Corso M, Hippolyte MP, Sammartino G, Dohan Ehrenfest DM. [Mouthwash solutions with microencapsuled natural extracts: efficiency for dental plaque and gingivitis]. Rev Stomatol Chir Maxillofac 2010; 111: 148-151
    CrossrefPubMedGoogle Scholar
  • 15 Perry NB, Brennan NJ, Van Klink JW, Harris W, Douglas MH, McGimpsey JA, Smallfield BM, Anderson RE. Essential oils from New Zealand manuka and kanuka: chemotaxonomy of Leptospermum . Phytochemistry 1997; 44: 1485-1494
    CrossrefPubMedGoogle Scholar
  • 16 Schnitzler P, Wiesenhofer K, Reichling J. Comparative study on the cytotoxicity of different Myrtaceae essential oils on cultured Vero and RC-37 cells. Pharmazie 2008; 63: 830-835
    PubMedGoogle Scholar
  • 17 Hayes AJ, Leach DN, Markham JL, Markovic B. In vitro cytotoxicity of Australian tea tree oil using human cell lines. J Essent Oil Res 1997; 9: 575-582 Accessed March 3, 2020 at: https://www.tandfonline.com/action/journalInformation?journalCode=tjeo20
    CrossrefPubMedGoogle Scholar
  • 18 Russo R, Corasaniti MT, Bagetta G, Morrone LA. Exploitation of cytotoxicity of some essential oils for translation in cancer therapy. eCAM 2015; DOI: 10.1155/2015/397821.
    CrossrefPubMedGoogle Scholar
  • 19 Mikus J, Harkenthal M, Steverding D, Reichling J. In vitro effect of essential oils and isolated mono- and sesquiterpenes on Leishmania major and Trypanosoma brucei. Planta Med 2000; 66: 366-368 doi:10.1055/s-2000-8548
    Article in Thieme ConnectPubMedGoogle Scholar
  • 20 Pavithra PS, Mehta A, Verma RS. Induction of apoptosis by essential oil from P. missionis in skin epidermoid cancer cells. Phytomedicine 2018; 50: 184-195
    CrossrefPubMedGoogle Scholar
  • 21 Benford HL, Frith JC, Auriola S, Mönkkönen J, Rogers MJ. Farnesol and geranylgeraniol prevent activation of caspases by aminobisphosphonates: biochemical evidence for two distinct pharmacological classes of bisphosphonate drugs. Mol Pharmacol 1999; 56: 131-140
    CrossrefPubMedGoogle Scholar
  • 22 Takarada K, Kimizuka R, Takahashi N, Honma K, Okuda K, Kato T. A comparison of the antibacterial efficacies of essential oils against oral pathogens. Oral Microbiol Immunol 2004; 19: 61-64 doi:10.1046/j.0902-0055.2003.00111.x
    CrossrefPubMedGoogle Scholar
  • 23 Gibbons S. Phytochemicals for bacterial resistance–strengths, weaknesses and opportunities. Planta Med 2008; 74: 594-602
    Article in Thieme ConnectPubMedGoogle Scholar
  • 24 Budzyńska A, Wieckowska-Szakiel M, Sadowska B, Kalemba D, Rózalska B. Antibiofilm activity of selected plant essential oils and their major components. Polish J Microbiol 2011; 60: 35-41
    CrossrefPubMedGoogle Scholar
  • 25 Chantrapornchai W, Clydesdale FM, McClements DJ. Understanding Colors in Emulsions. In: Culver CA, Wrolstad RE. eds. Color Quality of fresh and processed Foods. Washington D. C.: ACS Publications; 2008: 364-387
    Google Scholar
  • 26 Cheaburu-Yilmaz CN, Karasulu HY, Yilmaz O. Nanoscaled dispersed Systems used in Drug-Delivery Applications. In: Vasile C. ed. Polymeric Nanomaterials in Nanotherapeutics. Amsterdam, Netherlands: Elsevier; 2019: 437-468
    CrossrefGoogle Scholar
  • 27 Guvendiren M, Lu HD, Burdick JA. Shear-thinning hydrogels for biomedical applications. Soft Matter 2012; 8: 260-272
    CrossrefPubMedGoogle Scholar
  • 28 Hanning SM, Medlicott NJ. Oil-based compositions as saliva substitutes: a pilot study to investigate in-mouth retention. Int J Pharm 2016; 501: 265-270
    CrossrefPubMedGoogle Scholar
  • 29 Bruschi ML, Jones DS, Panzeri H, Gremião MPD, De Freitas O, Lara EHG. Semisolid systems containing propolis for the treatment of periodontal disease: in vitro release kinetics, syringeability, rheological, textural, and mucoadhesive properties. J Pharm Sci 2007; 96: 2074-2089
    CrossrefPubMedGoogle Scholar
  • 30 Jones DS, Irwin CR, Woolfson AD, Djokic J, Adams V. Physicochemical characterization and preliminary in vivo efficacy of bioadhesive, semisolid formulations containing flurbiprofen for the treatment of gingivitis. J Pharm Sci 1999; 88: 592-598
    CrossrefPubMedGoogle Scholar
  • 31 Kelly HM, Deasy PB, Ziaka E, Claffey N. Formulation and preliminary in vivo dog studies of a novel drug delivery system for the treatment of periodontitis. Int J Pharm 2004; 274: 167-183
    CrossrefPubMedGoogle Scholar
  • 32 Song CY, Nam EH, Park SH, Hwang CY. In vitro efficacy of the essential oil from Leptospermum scoparium (manuka) on antimicrobial susceptibility and biofilm formation in Staphylococcus pseudintermedius isolates from dogs. Vet Dermatol 2013; 24: 404-408 e87 doi:10.1111/vde.12045
    CrossrefPubMedGoogle Scholar
  • 33 Brady A, Loughlin R, Gilpin D, Kearney P, Tunney M. In vitro activity of tea-tree oil against clinical skin isolates of meticillin-resistant and -sensitive Staphylococcus aureus and coagulase-negative staphylococci growing planktonically and as biofilms. J Med Microbiol 2006; 55: 1375-1380
    CrossrefPubMedGoogle Scholar
  • 34 Hope CK, Clements D, Wilson M. Determining the spatial distribution of viable and nonviable bacteria in hydrated microcosm dental plaques by viability profiling. J Appl Microbiol 2002; 93: 448-455
    CrossrefPubMedGoogle Scholar
  • 35 Takenaka S, Trivedi HM, Corbin A, Pitts B, Stewart PS. Direct visualization of spatial and temporal patterns of antimicrobial action within model oral biofilms. Appl Environ Microbiol 2008; 74: 1869-1875
    CrossrefPubMedGoogle Scholar
  • 36 Sparkman OD. Identification of essential oil components by gas chromatography/mass spectroscopy Robert P. Adams. J Am Soc Mass Spectrom 1997; 8: 671-672
    CrossrefPubMedGoogle Scholar
  • 37 NIST/ERA/NIH Mass Spectral Library. Software/Data Version (NIST08); NIST Standard Reference Database. Gaithersburg: National Institute of Standards and Technology; 2008
    Google Scholar
  • 38 Zafar S, Coates DE, Cullinan MP, Drummond BK, Milne T, Seymour GJ. Zoledronic acid and geranylgeraniol regulate cellular behaviour and angiogenic gene expression in human gingival fibroblasts. J Oral Pathol Med 2014; 43: 711-721
    CrossrefPubMedGoogle Scholar
  • 39 Carson CF, Riley TV. Antimicrobial activity of the major components of the essential oil of Melaleuca alternifolia . J Appl Bacteriol 1995; 78: 264-269
    CrossrefPubMedGoogle Scholar
  • 40 CLSI. Performance Standards for antimicrobial Disk Susceptibility Tests. 13th ed. CLSI standard MO2. Watne, PA: Clinical and Laboratory Standards Institute; 2018
    Google Scholar
  • 41 Hanning SM, Yu T, Jones DS, Andrews GP, Kieser JA, Medlicott NJ. Lecithin-based emulsions for potential use as saliva substitutes in patients with xerostomia–viscoelastic properties. Int J Pharm 2013; 456: 560-568
    CrossrefPubMedGoogle Scholar
  • 42 Anderl JN, Franklin MJ, Stewart PS. Role of antibiotic penetration limitation in Klebsiella pneumoniae biofilm resistance to ampicillin and ciprofloxacin. Antimicrob Agents Chemother 2000; 44: 1818-1824
    CrossrefPubMedGoogle Scholar
  • 43 Singh R, Ray P, Das A, Sharma M. Penetration of antibiotics through Staphylococcus aureus and Staphylococcus epidermidis biofilms. J Antimicrob Chemother 2010; 65: 1955-1958
    CrossrefPubMedGoogle Scholar
  • 44 Heydorn A, Nielsen AT, Hentzer M, Sternberg C, Givskov M, Ersboll BK, Molin S. Quantification of biofilm structures by the novel computer program COMSTAT. Microbiology 2000; 146: 2395-2407
    CrossrefPubMedGoogle Scholar
  • 45 Vorregaard M. Comstat2–a Modern 3D image Analysis Environment for Biofilms. Denmark: Tech Univ Denmark Kongens Lyngby; 2008: 5-30
    Google Scholar