DOI QR코드

DOI QR Code

Embryonic Zebrafish Model - A Well-Established Method for Rapidly Assessing the Toxicity of Homeopathic Drugs - Toxicity Evaluation of Homeopathic Drugs Using Zebrafish Embryo Model -

  • Gupta, Himanshu R (Department of Medical Genetics, MGMIUDBS, Mahatma Gandhi Mission Institute of Health Sciences) ;
  • Patil, Yogesh (MGMCET & Central Research Laboratory, Mahatma Gandhi Mission Institute for Health Sciences) ;
  • Singh, Dipty (National Centre for Preclinical Reproductive and Genetic Toxicology (NIRRH), National Institute of Research in Reproductive Health (ICMR)) ;
  • Thakur, Mansee (Mahatma Gandhi Mission College of Engineering and Technology & Department of Medical Biotechnology, Central Research Laboratory, MGM Medical College, MGMIHS)
  • Received : 2016.09.07
  • Accepted : 2016.12.01
  • Published : 2016.12.31

Abstract

Objectives: Advancements in nanotechnology have led to nanoparticle (NP) use in various fields of medicine. Although the potential of NPs is promising, the lack of documented evidence on the toxicological effects of NPs is concerning. A few studies have documented that homeopathy uses NPs. Unfortunately, very few sound scientific studies have explored the toxic effects of homeopathic drugs. Citing this lack of high-quality scientific evidence, regulatory agencies have been reluctant to endorse homeopathic treatment as an alternative or adjunct treatment. This study aimed to enhance our insight into the impact of commercially-available homeopathic drugs, to study the presence of NPs in those drugs and any deleterious effects they might have, and to determine the distribution pattern of NPs in zebrafish embryos (Danio rerio). Methods: Homeopathic dilutions were studied using high-resolution transmission electron microscopy with selected area electron diffraction (SAED). For the toxicity assessment on Zebrafish, embryos were exposed to a test solution from 4 - 6 hours post-fertilization, and embryos/larvae were assessed up to 5 days post-fertilization (dpf ) for viability and morphology. Toxicity was recorded in terms of mortality, hatching delay, phenotypic defects and metal accumulation. Around 5 dpf was found to be the optimum developmental stage for evaluation. Results: The present study aimed to conclusively prove the presence of NPs in all high dilutions of homeopathic drugs. Embryonic zebrafish were exposed to three homeopathic drugs with two potencies (30CH, 200CH) during early embryogenesis. The resulting morphological and cellular responses were observed. Exposure to these potencies produced no visibly significant malformations, pericardial edema, and mortality and no necrotic and apoptotic cellular death. Conclusion: Our findings clearly demonstrate that no toxic effects were observed for these three homeopathic drugs at the potencies and exposure times used in this study. The embryonic zebrafish model is recommended as a well-established method for rapidly assessing the toxicity of homeopathic drugs.

References

  1. Bellavite P, Andrioli G, Lussignoli S, Signorini A, Orto-lani R, Conforti A. A scientific reappraisal of the 'principle of similarity'. Med Hypotheses. 1997;49(3):203-12. https://doi.org/10.1016/S0306-9877(97)90204-8
  2. Bellavite P, Signorini A. The emerging science of homeopathy. Berkeley (CA): North Atlantic; 2002. 424 p.
  3. Teixeira MZ. 'Paradoxical strategy for treating chronic diseases': a therapeutic model used in homeopathy for more than two centuries, Homeopathy. 2005;94(4):265-6.
  4. Roduner E. Size matters: why nanomaterials are different. Chem Soc Rev. 2006;35(7):583-92. https://doi.org/10.1039/b502142c
  5. Chikramane PS, Suresh AK, Bellare JR, Kane SG. Extreme homeopathic dilutions retain starting mate-rials: a nanoparticulate perspective. Homeopathy. 2010;99(4):231-42. https://doi.org/10.1016/j.homp.2010.05.006
  6. Sharma A, Purkait B. Identification of medicinally active ingredient in ultradiluted Digitalis purpurea: fluorescence spectroscopic and cyclic-voltammet-ric study. J Anal Methods Chem. 2012;2012:DOI: 10.1155/2012/109058. https://doi.org/10.1155/2012/109058
  7. Bellavite P, Signorini A. The emerging science of homeopathy: complexity, biodynamics and nanopharma-cology. California: North Atlantic Books; 1995. 424 p.
  8. Dantas F, Rampes H. Do homeopathic medicines provoke adverse effects? a systematic review. Br Homeopath J. 2000;89(S1):S35-8. https://doi.org/10.1054/homp.1999.0378
  9. Posadzki P, Alotaibi A, Ernst E. Adverse effects of homeopathy: a systematic review of published case reports and case series. Int J Clin Pract. 2012;66(12):1178-88. https://doi.org/10.1111/ijcp.12026
  10. Vickers AJ. Independent replication of pre-clinical re-search in homoeopathy: a systematic review. Forsch Komplementarmed. 2004;6(6):311-20. https://doi.org/10.1159/000021286
  11. Barve R, Chaughule R. Size-dependent in vivo/in vitro results of homoeopathic herbal extracts. J Nanostruct Chem. 2013;3(1):18. https://doi.org/10.1186/2193-8865-3-18
  12. Goldsmith P. Zebrafish as a pharmacological tool: the how, why and when. Curr Opin Pharmacol. 2004;4(5):504-12. https://doi.org/10.1016/j.coph.2004.04.005
  13. Rubinstein AL. Zebrafish assays for drug toxicity screening. Expert Opin Drug Metab Toxicol. 2006;2(2):231-40. https://doi.org/10.1517/17425255.2.2.231
  14. Winter MJ, Redfern WS, Hayfield AJ, Owen SF, Valentin JP, Hutchinson TH. Validation of a larval zebrafish locomotor assay for assessing the seizure liability of early-stage development drugs. J Pharmacol Toxicol Methods. 2008;57(3):176-87. https://doi.org/10.1016/j.vascn.2008.01.004
  15. Postlethwait JH, Woods IG, Ngo-Hazelett P, Yan YL, Kelly PD, Chu F, et al. Zebrafish comparative genomics and the origins of vertebrate chromosomes. Genome Res. 2000;10(12):1890-902. https://doi.org/10.1101/gr.164800
  16. Redfern WS, Waldron G, Winter MJ, Butler P, Holbrook N, Wallis R, et al. Zebrafish assays as early safety pharmacology screens: paradigm shift or red herring?. J Pharmacol Toxicol Methods. 2008;58(2):110-7. https://doi.org/10.1016/j.vascn.2008.05.006
  17. Parng C, Seng WL, Semino C, McGrath P. Zebrafish: a preclinical model for drug screening. Assay Drug Dev Technol. 2002;1(1):41-8. https://doi.org/10.1089/154065802761001293
  18. Ali S, van Mil HGJ, Richardson MK. Large-scale assessment of the zebrafish embryo as a possible predictive model in toxicity testing. Plos one. 2011;6(6):DOI: 10.1371/journal.pone.0021076. https://doi.org/10.1371/journal.pone.0021076
  19. McGrath P, Li CQ. Zebrafish: a predictive model for assessing drug-induced toxicity. Drug Discov Today. 2008;13(9-10):394-401. https://doi.org/10.1016/j.drudis.2008.03.002
  20. Bell IR, Muralidharan S, Schwartz GE. Nanoparti-cle characterization of traditional homeopathical-ly-manufactured silver (Argentum metallicum) medicines and placebo controls. J Nanomed Nanotechnol. 2015;6(4):DOI: 10.4172/2157-7439.1000311. https://doi.org/10.4172/2157-7439.1000311
  21. Westerfield M. A guide for the laboratory use of zebraf-ish danio (brachydanio) rerio. Eugene:University of Oregon Press; 2000.
  22. Test no. 236: fish embryo acute toxicity (FET) test [internet]. OECD; 2013 [Nov, 2016]. Available from: http://www.oecd-ilibrary.org/docserver/down-load/9713161e.pdf?expires=1481098481&id=id&ac-cname=guest&checksum=0EFE0447C6B022B5CA-F2AAEB18B9F3CC.
  23. Hu YL, Qi W, Han F, Shao JZ, Gao JQ. Toxicity evaluation of biodegradable chitosan nanoparticles using a zebrafish embryo model. Int J Nanomedicine. 2011;6:3351-9.
  24. Chikramane PS, Kalita D, Suresh AK, Kane SG, Bellare JR. Why extreme dilutions reach non-zero asymptotes: a nanoparticulate hypothesis based on froth flotation. Langmuir. 2012;28(45):15864-75. https://doi.org/10.1021/la303477s
  25. Upadhyay RP, Nayak C. Homeopathy emerging as na-nomedicine. International Journal of High Dilution Re-search. 2011;10(37):299-310.
  26. Demangeat JL. NMR relaxation evidence for sol-ute-induced nanosized superstructures in ultramo-lecular aqueous dilutions of silica-lactose. J Mol Liq. 2010;155(2-3):71-9. https://doi.org/10.1016/j.molliq.2010.05.010
  27. Rajendran ES. Field emission scanning electron microscopic (FESEM) and energy dispersive spectroscopic (EDS) studies of centesimal scale potencies of the homeopathic drug Lycopodium clavatum. AJHM. 2015;108(1):9-18.
  28. Konovalov A, Ryzhkina I. Highly diluted aqueous solutions: formation of nano-sized molecular assemblies (nanoassociates). Geochem Int. 2014;52(13):1207-26. https://doi.org/10.1134/S0016702914130072
  29. Linde K, Jonas WB, Melchart D, Worku F, Wagner H, Ei-tel F. Critical review and meta-analysis of serial agitated dilutions in experimental toxicology. Hum Exp Toxicol. 1994;13(7):481-92. https://doi.org/10.1177/096032719401300706
  30. Makri A, Goveia M, Balbus J, Parkin R. Children's susceptibility to chemicals: a review by developmental stage. J Toxicol Environ Health, Part B. 2004;7(6):417-35. https://doi.org/10.1080/10937400490512465
  31. Fun with homeopaths and meta-analyses of homeopathy trials [internet]. Science Based Medicine; 2008. Available from: https://www.sciencebasedmedicine.org/contact-us/.