Development and efficacy of tryptophol-containing emulgel for reducing subcutaneous fungal nodules from Scedosporium apiospermum eumycetoma

Thitinan Kitisin , Watcharamat Muangkaew, Sumate Ampawong, Nichapa Sansurin, Natthawut Thitipramote, Passanesh Sukphopetch

Abstract


Background and purpose: Subcutaneous infections caused by Scedosporium apiospermum present as chronic eumycetomatous manifestations in both immunocompromised and immunocompetent individuals. Serious adverse effects/toxicities from the long-term use of antifungal drugs and antifungal resistance have been reported in patients with S. apiospermum infections. The present study aimed to determine the anti-S. apiospermum activities of fungal quorum sensing molecule known as tryptophol (TOH) and to develop a TOH-containing emulgel for treating S. apiospermum eumycetoma.

Experimental approach: Anti-S. apiospermum activities of TOH were determined and compared with voriconazole. Effects of TOH on S. apiospermum biofilm formation and human foreskin fibroblast (HFF)-1 cell cytotoxicity were determined. Moreover, TOH-containing emulgel was developed and physical properties, in vitro, and in vivo antifungal activities against S. apiospermum eumycetoma were evaluated.

Findings/Results: The minimal concentration of TOH at 100 µM exhibited anti-S. apiospermum activities by reducing growth rate, germination rate, and biofilm formation with less cytotoxicity to HFF-1 cells than voriconazole. Further study on the development of an emulgel revealed that TOH-containing emulgel exhibited excellent physical properties including homogeneity, consistency, and stability. Treatment by TOH-containing emulgel significantly reduced subcutaneous mass in a mouse model of S. apiospermum eumycetoma. The histopathological assessment showed marked improvement after 14 days of TOH-containing emulgel treatment.

Conclusion and implications: TOH could be used as an anti-fungal agent against S. apiospermum infections. A novel and stable TOH-containing emulgel was developed with excellent anti-S. apiospermum activities suggesting the utilization of TOH-containing emulgel as an innovative therapeutic approach in the treatment of S. apiospermum eumycetoma.

 

 


Keywords


Emulgel; Eumycetoma; Quorum sensing molecule, Scedosporium apiospermum; Tryptophol.

Full Text:

PDF

References


Chen SC, Halliday CL, Hoenigl M, Cornely OA, Meyer W. Scedosporium and Lomentospora infections: contemporary microbiological tools for the diagnosis of invasive disease. J Fungi (Basel). 2021;7(1):23. DOI: 10.3390/jof7010023.

Guarro J, Kantarcioglu AS, Horré R, Rodriguez-Tudela JL, Cuenca Estrella M, Berenguer J, et al. Scedosporium apiospermum: changing clinical spectrum of a therapy-refractory opportunist. Med Mycol. 2006;44(4):295-327. DOI: 10.1080/13693780600752507.

Cortez KJ, Roilides E, Quiroz-Telles F, Meletiadis J, Antachopoulos C, Knudsen T, et al. Infections caused by Scedosporium spp. Clin Microbiol Rev. 2008;21(1):157-197.DOI: 10.1128/CMR.00039-07.

Luplertlop N. Pseudallescheria/Scedosporium complex species: from saprobic to pathogenic fungus. J Mycol Med. 2018;28(2):249-256. DOI: 10.1016/j.mycmed.2018.02.015.

Mello TP, Aor AC, Oliveira SS, Branquinha MH, Santos AL. Conidial germination in Scedosporium apiospermum, S. aurantiacum, S. minutisporum and Lomentospora prolificans: influence of growth conditions and antifungal susceptibility profiles. Mem Inst Oswaldo Cruz. 2016;111(7):484-494.DOI: 10.1590/0074-02760160200.

Gulati V, Bakare S, Tibrewal S, Ismail N, Sayani J, Baghla DP. A rare presentation of concurrent Scedosporium apiospermum and Madurella griseaeumycetoma in an immunocompetent host. Case Rep Pathol. 2012;2012:154201.DOI: 10.1155/2012/154201.

Kitisin T, Muangkaew W, Ampawong S, Sukphopetch P. Tryptophol coating reduces catheter-related cerebral and pulmonary infections by Scedosporium apiospermum. Infect Drug Resist. 2020;13:2495-2508.DOI: 10.2147/IDR.S255489.

Kitisin T, Muangkaew W, Ampawong S, Chutoam P, Thanomsridetchai N, Tangwattanachuleeporn M, et al. Isolation of fungal communities and identification of Scedosporium species complex with pathogenic potentials from a pigsty in Phra Nakhon Si Ayutthaya, Thailand. New Microbiol. 2021;44(1):33-41.PMID: 33582822.

Tammer I, Tintelnot K, Braun-Dullaeus RC, Mawrin C, Scherlach C, Schlüter D, et al. Infections due to Pseudallescheria/Scedosporium species in patients with advanced HIV disease--a diagnostic and therapeutic challenge. Int J Infect Dis. 2011;15(6):e422-e429.DOI: 10.1016/j.ijid.2011.03.004.

Wiederhold NP, Lewis RE. Antifungal activity against Scedosporium species and novel assays to assess antifungal pharmacodynamics against filamentous fungi. Med Mycol. 2009;47(4):422-432.DOI: 10.1080/13693780802510224.

Roemer T, Krysan DJ. Antifungal drug development: challenges, unmet clinical needs, and new approaches. Cold Spring Harb Perspect Med. 2014;4(5):a019703.DOI: 10.1101/cshperspect.a019703.

Scolding P, Fahal A, Yotsu RR. Drug therapy for mycetoma. Cochrane Database Syst Rev. 2018;2018(7):CD013082.DOI: 10.1002/14651858.CD013082.

Welsh O, Al-Abdely HM, Salinas-Carmona MC, Fahal AH. Mycetoma medical therapy. PLoS Negl Trop Dis. 2014;8(10):e3218.DOI: 10.1371/journal.pntd.0003218.

Wongsuk T, Pumeesat P, Luplertlop N. Fungal quorum sensing molecules: role in fungal morphogenesis and pathogenicity. J Basic Microbiol. 2016;56(5):440-447.DOI: 10.1002/jobm.201500759.

Albuquerque P, Casadevall A. Quorum sensing in fungi--a review. Med Mycol. 2012;50(4):337-345.

DOI: 10.3109/13693786.2011.652201.

Kruppa M. Quorum sensing and Candida albicans. Mycoses. 2009;52(1):1-10.DOI: 10.1111/j.1439-0507.2008.01626.x.

Palmieriand A, Petrini M. Tryptophol and derivatives: natural occurrence and applications to the synthesis of bioactive compounds. Nat Prod Rep. 2019:36(3),490-530.DOI: 10.1039/c8np00032h.

Rao RP, Hunter A, Kashpur O, Normanly J. Aberrant synthesis of indole-3-acetic acid in Saccharomyces cerevisiae triggers morphogenic transition, a virulence trait of pathogenic fungi. Genetics. 2010;185(1):211-220.DOI: 10.1534/genetics.109.112854.

Singkum P, Muangkaew W, Suwanmanee S, Pumeesat P, Wongsuk T, Luplertlop N. Suppression of the pathogenicity of Candida albicans by the quorum-sensing molecules farnesol and tryptophol. J Gen Appl Microbiol. 2020;65(6):277-283.DOI: 10.2323/jgam.2018.12.002.

Lachenmeier DW. Safety evaluation of topical applications of ethanol on the skin and inside the oral cavity. J Occup Med Toxicol. 2008;3(1):26-41. DOI: 10.1186/1745-6673-3-26.

Ali Khan B, Ullah S, Khan MK, Alshahrani SM, Braga VA. Formulation and evaluation of Ocimum basilicum-based emulgel for wound healing using animal model. Saudi Pharm J. 2020;28(12):1842-1850.DOI: 10.1016/j.jsps.2020.11.011.

Jagdale S, Pawar S. Gellified emulsion of ofloxacin for transdermal drug delivery system. Adv Pharm Bull. 2017;7(2):229-239.DOI: 10.15171/apb.2017.028.

Luplertlop N, Muangkaew W, Pumeesat P, Suwanmanee S, Singkum P. Distribution of Scedosporium species in soil from areas with high human population density and tourist popularity in six geographic regions in Thailand. PLoS One. 2019;14(1):e0210942.DOI: 10.1371/journal.pone.0210942.

Rollin-Pinheiro R, de Meirelles JV, Vila TVM, Fonseca BB, Alves V, Frases S, et al. Biofilm formation by Pseudallescheria/Scedosporium species: a comparative study. Front Microbiol. 2017;8:1568.DOI: 10.3389/fmicb.2017.01568.

Mello TP, Aor AC, Gonçalves DS, Seabra SH, Branquinha MH, Santos AL. Assessment of biofilm formation by Scedosporium apiospermum, S. aurantiacum, S. minutisporum and Lomentospora prolificans. Biofouling. 2016;32(7):737-749.DOI: 10.1080/08927014.2016.1192610.

Pierce CG, Uppuluri P, Tristan AR, Wormley FL Jr, Mowat E, Ramage G, et al. A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing. Nat Protoc. 2008;3(9):1494-1500.DOI: 10.1038/nport.2008.141.

Riss TL, Moravec RA, Niles AL, Duellman S, Benink HA, Worzella TJ, et al. Cell Viability Assays. In: Markossian S, Grossman A, Brimacombe K, et al. editors. Assay Guidance Manual. USA: Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2004. Available from: https://www.ncbi.nlm.nih.gov/books/NBK144065/.

Rayamajhi M, Zhang Y, Miao EA. Detection of pyroptosis by measuring released lactate dehydrogenase activity. Methods Mol Biol. 2013;1040:85-90.DOI: 10.1007/978-1-62703-523-1_7.

Chen H, Chang X, Du D, Li J, Xu H, Yang X. Microemulsion-based hydrogel formulation of ibuprofen for topical delivery. Int J Pharm. 2006;315(1-2):52-58.DOI: 10.1016/j.ijpharm.2006.02.015.

Shahin M, Hady SA, Hammad M, Mortada N. Novel jojoba oil-based emulsion gel formulations for clotrimazole delivery. AAPS PharmSciTech. 2011;12(1):239-247.DOI: 10.1208/s12249-011-9583-4.

Hussain A, Samad A, Singh SK, Ahsan MN, Haque MW, Faruk A, et al. Nanoemulsion gel-based topical delivery of an antifungal drug, in vitro activity and in vivo activity evaluation. Drug Deliv. 2016;23(2):642-647.DOI: 10.3109/10717544.2014.933284.

Ogidi OC, Oyetayo VO, Akinyele BJ. In vitro evaluation of antimicrobial efficacy of extracts obtained from raw and fermented wild macrofungus, Lenzites quercina. Int J Microbiol. 2015;2015:106308.DOI: 10.1155/2015/106308.

Ampawong S, Luplertlop N. Experimental scedosporiosis induces cerebral oedema associated with abscess regarding aquaporin-4 and Nrf-2 depletions. Biomed Res Int. 2019;2019:6076571.DOI: 10.1155/2019/6076571.

Luplertlop N, Pumeesat P, Muangkaew W, Wongsuk T, Alastruey-Izquierdo A. Environmental screening for the Scedosporium apiospermumspecies complex in public parks in Bangkok, Thailand. PLoS One. 2016;11(7):e0159869.DOI: 10.1371/journal.pone.0159869.

Ramirez-Garcia A, Pellon A, Rementeria A, Buldain I, Barreto-Bergter E, Rollin-Pinheiro R, et al. Scedosporium and Lomentospora: an updated overview of underrated opportunists. Med Mycol. 2018;56(suppl_1):102-125.DOI: 10.1093/mmy/myx113.

Tóth EJ, Nagy GR, Homa M, Ábrók M, Kiss IÉ, Nagy G, et al. Recurrent Scedosporium apiospermum mycetoma successfully treated by surgical excision and terbinafine treatment: a case report and review of the literature. Ann Clin Microbiol Antimicrob. 2017;16(1):31.DOI: 10.1186/s12941-017-0195-z.

Goldman C, Akiyama MJ, Torres J, Louie E, Meehan SA. Scedosporium apiospermum infections and the role of combination antifungal therapy and GM-CSF: a case report and review of the literature. Med Mycol Case Rep. 2016;11:40-43.DOI: 10.1016/j.mmcr.2016.04.005.

Lackner M, de Hoog GS, Verweij PE, Najafzadeh MJ, Curfs-Breuker I, Klaassen CH, et al. Species-specific antifungal susceptibility patterns of Scedosporium and Pseudallescheria species. Antimicrob Agents Chemother. 2012;56(5):2635-2642.DOI: 10.1128/AAC.05910-11.

Girmenia C, Luzi G, Monaco M, Martino P. Use of voriconazole in treatment of Scedosporium apiospermum infection: case report. J Clin Microbiol. 1998;36(5):1436-1438.DOI: 10.1128/JCM.36.5.1436-1438.1998.

Anchisi C, Maccioni AM, Sinico C, Valenti D. Stability studies of new cosmetic formulations with vegetable extracts as functional agents. Farmaco. 2001;56(5-7):427-431.DOI: 10.1016/s0014-827x(01)01055-2.

Rao M, Sukre G, Aghav S, Kumar M. Optimization of metronidazole emulgel. J Pharm (Cairo). 2013;2013:501082.DOI: 10.1155/2013/501082.

Oliveira Fde M, Unis G, Hochhegger B, Severo LC. Scedosporium apiospermum eumycetoma successfully treated with oral voriconazole: report of a case and review of the Brazilian reports on scedosporiosis. Rev Inst Med Trop Sao Paulo. 2013;55(2):121-123.DOI: 10.1590/s0036-46652013000200010.

Zeng J, Kamei K, Zheng Y, Nishimura K. Susceptibility of Pseudallescheria boydii and Scedosporium apiospermum to new antifungal agents. Nihon Ishinkin Gakkai Zasshi. 2004;45(2):101-104.DOI: 10.3314/jjmm.45.101.

Walsh TJ, Lutsar I, Driscoll T, Dupont B, Roden M, Ghahramani P, et al. Voriconazole in the treatment of aspergillosis, scedosporiosis and other invasive fungal infections in children. Pediatr Infect Dis J.

;21(3):240-248.DOI: 10.1097/00006454-200203000-00015.

Perfect JR, Marr KA, Walsh TJ, Greenberg RN, DuPont B, de la Torre-Cisneros J, et al. Voriconazole treatment for less-common, emerging, or refractory fungal infections. Clin Infect Dis. 2003;36(9):1122-1131.DOI: 10.1086/374557.

Johnson LB, Kauffman CA. Voriconazole: a new triazole antifungal agent. Clin Infect Dis. 2003;36(5):630-637.DOI: 10.1086/367933.

Bertin J, Guo Y, Wang L, Srinivasula SM, Jacobson MD, Poyet JL, et al. CARD9 is a novel caspase recruitment domain-containing protein that interacts with BCL10/CLAP and activates NF-kappa B. J Biol Chem. 2000;275(52): 41082-41086.DOI: 10.1074/jbc.C000726200.

Eno CO, Zhao G, Olberding KE, Li C. The Bcl-2 proteins Noxa and Bcl-xL co-ordinately regulate oxidative stress-induced apoptosis. Biochem J. 2012;444(1):69-78.DOI: 10.1042/BJ20112023.


Refbacks

  • There are currently no refbacks.


Creative Commons LicenseThis work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.