Open Access Open Access  Restricted Access Subscription Access
Open Access Open Access Open Access  Restricted Access Restricted Access Subscription Access

Evaluation of Homeopathic Drugs on Glucocorticoid Induced Osteoporosis (GIOP) Zebrafish Model


Affiliations
1 Department of Medical Genetics, MGM School of Biomedical Sciences, MGM Medical College Building, MGM Institute of Health Sciences, Kamothe, Navi Mumbai (M.S.), India
2 Department of Medical Biotechnology and Central Research Laboratory, MGMCET, MGM School of Biomedical Sciences, MGM Medical College Building, MGM Institute of Health Sciences, Kamothe, Navi Mumbai (M.S.), India
     

   Subscribe/Renew Journal


Background: Homeopathic remedies are proposed as source for traditional preventive medicines. In this context our study focuses to evaluate efficacy of homoeopathic drug with reference to GIOP in Zebrafish vertebrate model. The overall approach behind this study was to determine whether homeopathic drugs can be used along with of allopathic medicines as an alternate therapy. Methods: In the present study, an attempt is made to find out the efficacy of homoeopathic medicines-Argentum Metallicum, Calcaria Carbonica and Sepia in two dilutions 6CH and 30 CH on GIOP model of zebra fish in two dilutions. Exposure was studied at 3 different points - 4, 14 and 28 days exposure (treatment with test drugs) and staining intensities were measured using Image J software. Control groups used were untreated control larvae, GIOP model untreated post 11 dpf and GIOP model treated with Alendronate. The medium was changed every day at the same time. Staining intensities measured for Alizarin Red and Calcein dye stained images. Results: Staining intensities of Alizarin red and Calcein staining of the treated group on statistical analysis showed that the means of GIOP and other treated groups were not statistically significant at p<0.05 except in the case of 28 day study for Calcarea Carbonica 30C and Sepia 30C. Though screening was processed using Alizarin red and Calcein staining, our quantification screening with calcein labeling indeed facilitated the process and diminished labor of handling. Conclusion: Our findings show that, despite some physiological differences between mammals and teleosts, the zebra fish represents an effective model for screening of bone defects. This model has been used for preliminary studies on homeopathy drugs prescribed for treatment of Osteoporosis. The proposed calcein staining protocol can represents a powerful tool for in vivo monitoring of mineralized structures.

Keywords

Homeopathy, Osteoporosis, Zebra Fish, Prednisolone, Alizarin Red, Calcein.
Subscription Login to verify subscription
User
Notifications
Font Size


  • Apschner, A., Schulte-Merker, S. and Witten, P.E. (2011). Chapter 10 - Not All Bones are Created Equal – Using Zebrafish and Other Teleost Species in Osteogenesis Research. In: Detrich HW, Westerfield M, Zon LI (eds).Methods in Cell Biology Massachusetts: Academic Press, pp. 239–255
  • Barrett, R., Chappell, C., Quick, M. and Fleming, A. (2006). A rapid, high content, in vivo model of glucocortico-idinduced osteoporosis. Biotechnol. J., 1 (6) : 651–655.
  • Bellavite, P., Ortolani, R. and Conforti, A. (2006). Immunology and homeopathy. 3. Experimental studies on animal models. Evid Based Complement Alternat Med., 3 : 171-186.
  • Bird, N.C. and Mabee, P.M. (2003).Developmental morphology of the axial skeleton of the zebrafish, Daniorerio (Ostariophysi: Cyprinidae). Developmental Dynamics, 228 (3) : 337-57
  • Du, S.J., Frenkel, V., Kindschi, G. and Zohar, Y. (2001). Visualizing normal and defective bone development in Zebrafish embryos using the fluorescent chromophore calcein. Dev. Biol., 238 : 239–246.
  • Fleming, A. and Alderton, W.K. (2013). Drug discovery today. Disease Models, 10 : e43-e50.
  • George, S., Xia, T., Rallo, R., Zhao, Y., Ji, Z., Lin, S., Wang, X., Zhang, H., France, B., Schoenfeld, D., Damoiseaux, R., Liu, R., Lin, S., Bradley, K.A., Cohen, Y. and Nel, A.E. (2011). Use of a high-throughput screening approach coupled with in vivo zebrafish embryo screening to develop hazard ranking for engineered nanomaterials. ACS Nano, 5 : 1805–1817.
  • Ge, J., Wang, X. and Cui, F. (2006). Microstructural characteristics and nanomechanical properties across the thickness of the wild-type zebrafish skeletal bone. Materials Sci. and Engg.: C., 26(4):710-715.
  • Gorman, K.F. and Breden, F. (2007). Teleosts as models for human vertebral stability and deformity. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacol., 145(1):28-38.
  • Gupta, H.R., Patil, Y., Singh, D. and Thakur, M. (2016). Embryonic zebrafish model-A well-established method for rapidly assessing the toxicity of homeopathic drugs : Toxicity evaluation of homeopathic drugs using zebrafish embryo model. J. Pharmacopuncture, 19 (4) : 319.
  • Kimmel, C.B., Miller, C.T., Kruze, G., Ullmann, B., BreMiller, R.A., Larison, K.D. and Snyder, H.C. (1998). The shaping of pharyngeal cartilages during early development of the zebrafish. Developmental Biol., 203 (2) : 245-263.
  • Nicenboim, J., Malkinson, G., Lupo, T., Asaf, L., Sela, Y., Mayseless, O., Gibbs-Bar, L., Senderovich, N., Hashimshony, T., Shin, M., Jerafi-Vider, A., Avraham-Davidi, I., Krupalnik, V., Hofi, R., Almog, G., Astin, J.W., Golani, O., Ben-Dor, S., Crosier, P. S., Herzog, W. , Lawson, N. D., Hanna, J.H., Yanai, I. and Yaniv, K. (2015). Lymphatic vessels arise from specialized angioblasts within a venous niche. Nature, 522 : 56–61.
  • Pasqualetti, S., Congiu, T., Banfi, G. and Mariotti, M. (2015). Alendronate rescued osteoporotic phenotype in a model of glucocorticoid-induced osteoporosis in adult zebrafish scale. Internat. J. Exp. Pathol., 96(1):11-20
  • Recidoro, A.M., Roof, A.C., Schmitt, M., Worton, L.E., Petrie, T., Strand, N., Ausk, B.J., Srinivasan, S., Moon, R.T., Gardiner, E.M., Kaminsky, W., Bain, S.D., Allan, C.H., Gross, T.S. and Kwon, R.Y. (2014). Botulinum toxin induces muscle paralysis and inhibits bone regeneration in zebrafish. J. Bone Miner. Res., 29 : 2346–2356.
  • Schilling, T.F. and Kimmel, C.B. (1997). Musculoskeletal patterning in the pharyngeal segments of the zebrafish embryo.Development, 124(15) : 2945-2960.
  • Springer, V.G. and Johnson, G.D. (2000).Use and advantages of ethanol solution of alizarin red S dye for staining bone in fishes. Copeia, 2000 (1) : 300-301.
  • Vandamme, T.F. (2014). Use of rodents as models of human diseases. J. Pharm. Bioallied Sci., 6 (2) : 9-7.
  • Van Wijk, R. and Albrecht, H. (2007). Proving and therapeutic experiments in the HomBRex basic homeopathy research database. Homeopathy, 96:252
  • Vilmann, H. (1968). The in vivo staining of bone with alizarin red S. J Anat., 105(3) : 533–545.
  • Westerfield M. (2000). The zebrafish book: a guide for the laboratory use of zebrafish. http://zfin. Org/zf_info/zfbook/zfbk.html. 2000.
  • World Health Organization (WHO). The WHO Strategy on Research for Health. Available from: [Last accessed on 2016 May 23]. Back to cited text no. 1
  • Central Council for Research in Homoeopathy. Dossier (2013). Available from: http://www.ccrhindia.org/dossier/content/page26.html. [Last accessed on 2016 May 24].

Abstract Views: 261

PDF Views: 9




  • Evaluation of Homeopathic Drugs on Glucocorticoid Induced Osteoporosis (GIOP) Zebrafish Model

Abstract Views: 261  |  PDF Views: 9

Authors

Himanshu Gupta
Department of Medical Genetics, MGM School of Biomedical Sciences, MGM Medical College Building, MGM Institute of Health Sciences, Kamothe, Navi Mumbai (M.S.), India
Mansee Thakur
Department of Medical Biotechnology and Central Research Laboratory, MGMCET, MGM School of Biomedical Sciences, MGM Medical College Building, MGM Institute of Health Sciences, Kamothe, Navi Mumbai (M.S.), India

Abstract


Background: Homeopathic remedies are proposed as source for traditional preventive medicines. In this context our study focuses to evaluate efficacy of homoeopathic drug with reference to GIOP in Zebrafish vertebrate model. The overall approach behind this study was to determine whether homeopathic drugs can be used along with of allopathic medicines as an alternate therapy. Methods: In the present study, an attempt is made to find out the efficacy of homoeopathic medicines-Argentum Metallicum, Calcaria Carbonica and Sepia in two dilutions 6CH and 30 CH on GIOP model of zebra fish in two dilutions. Exposure was studied at 3 different points - 4, 14 and 28 days exposure (treatment with test drugs) and staining intensities were measured using Image J software. Control groups used were untreated control larvae, GIOP model untreated post 11 dpf and GIOP model treated with Alendronate. The medium was changed every day at the same time. Staining intensities measured for Alizarin Red and Calcein dye stained images. Results: Staining intensities of Alizarin red and Calcein staining of the treated group on statistical analysis showed that the means of GIOP and other treated groups were not statistically significant at p<0.05 except in the case of 28 day study for Calcarea Carbonica 30C and Sepia 30C. Though screening was processed using Alizarin red and Calcein staining, our quantification screening with calcein labeling indeed facilitated the process and diminished labor of handling. Conclusion: Our findings show that, despite some physiological differences between mammals and teleosts, the zebra fish represents an effective model for screening of bone defects. This model has been used for preliminary studies on homeopathy drugs prescribed for treatment of Osteoporosis. The proposed calcein staining protocol can represents a powerful tool for in vivo monitoring of mineralized structures.

Keywords


Homeopathy, Osteoporosis, Zebra Fish, Prednisolone, Alizarin Red, Calcein.

References