Homeopathy Efficacy on Neuropathic Pain

2877 words (12 pages) Essay in Medical

23/09/19 Medical Reference this

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 “Explain whyMagar et al. (2018) has attracted so much controversy since publication”

 

Neuropathic pain is caused by lesion or disease of the somatosensory system, but there are still some limitations relating to its safe and effective medication (Carrasco et al., 2018). One piece of research trying to prove the homeopathy efficacy on neuropathic pain is “Ultra-diluted Toxicodendron pubescens attenuated pro inflammatory cytokines and ROS meditated neuropathic pain in rats” (Magar et al., 2018). However, it has attracted so much criticism after being published in Scientific Reports. At least 5 news articles and 4 technical blogs have recently questioned it (from the Nature website), although the co-author Patil claimed the work was done “with utmost integrity” (Guglielmi, 2018, pp. 174). Before the root of the controversy is further discussed, the basic terminology and concept will be defined in this essay. From Australian Government Review (2015), ultra-dilution is one common technique in homeopathy – a “like cures like” treatment when a substance causing the symptoms is identified and the more the substance is diluted, the more its impact helps to cure the symptoms. Toxicodendron pubescensor RhusTox (RT), known as Atlantic poison oak, which has similar effects to poison ivy, can cause severe dermatitis in sensitive individuals (PLANTS Database, 2015). In 2018, Magar and his colleagues indicated that RT extract was comparable with an analgesic medicine – gabapentin in reducing inflammation and nerve pain after being diluted into different concentrations (from 10-2 to 10-36). For in-vitro experiments, the human cancer cells U-87 were pre-treated by Lipopolysaccharide (LPS) to induce Reactive Oxygen Species (ROS) before determining the production of ROS, cytokines (TNF-α, IL-1β, and IL-6), superoxide dismutase (SOD) and catalase activity (with the positive control H2O2). The anti-inflammation and immunomodulation of RT were stated from a decrease of aggressive oxidative and pro-inflammatory elements in cells. For in-vivo experiments, the researchers carried out the tests of allodynia and histopathology study. They stimulated the sciatic nerve by squeezing the rats’ paws and provided once daily ultra-diluted RT extract for a group of eight rats and the prescribed drug for another group while supplying one saline solution for a control group. The results showed a lower quantity of the inflammatory and pain messengers in the sciatic nerve, which was similar to the capability of gabapentin.

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The three features leading to the article’s controversy, which will be analysed in the subsequent paragraphs, are the defective experimental design, the inaccurate data, and the unusually brief review process before publication. Firstly, there were concerns about the reliability of the experimental design with regard to the chemicals, the protocols and the measurements. For example, the cytokine sets using in the current study (Magar et al., 2018) had the exact category numbers (even the same batches) as those in the previous study (Chanchal et al., 2016). Although the central subjects of both articles were rats – not mice, it was observable that the Immunoassay Product Guide (from eBiosciences Incorporation) only had category 887324 (not 837324) for mouse TNF-α sets and category 887064 (not 837064) for mouse IL-6 sets (Table 1). This error may have resulted in the inaccurate cytokine assessments. When comparing to the ARRIVE guidelines (Kilkenny et al., 2012), the experiment protocols lacked some essential aspects such as the total number of animals with specific developmental stages, the number of independent replications of each experiment and the definite steps to diminish the subjective prejudice. The authors did not define the blinded groups, and there was no evidence that randomized experiments were obtained because the researchers (who evaluated the rats) were aware which groups received the medicinal treatment.

Cytokine

Ready-SET-Go®

The current study (2018)

The previous study (2016)

Immunoassay Product Guide

(from eBiosciences Incorporation)

Not available (N/A) rat or mouse

Mouse

Mouse

Rat

TNF-α

83 7324-22

83 7324-22

88-7324

88-7340

IL-6

83 7064-22

83 7064-22

88-7064

N/A

IL-1β

88 7013-22

88 7013-22

88-7013

88-6010

Note: In 2017, eBiosciences Incorporation became a part of Thermo Fisher Scientific Company so eBioscience Ready-SET-Go! Kits altered to Invitrogen uncoated ELISA and still remained the same strengths despite the changed names and packages (according to manufacturer guides)

 

Table 1. Comparison of category numbers of Cytokine Ready-SET-Go® between the current study (2018), the previous study (2016), and Immunoassay Product Guide (from eBiosciences Incorporation) revealed the inaccurate cytokine assessments.

Moreover, the behavioural investigation was not explained adequately because the group representation or sample selection, the standard preparation, and the measurement method were vague. For instance, it was not clear why the authors choose to compare just one concentration of RT (10-12) with one concentration of gabapentin (60 mg/ kg). The rationale of suspending gabapentin 60 mg/kg/day in 0.5% carboxymethyl cellulose (CMC) was also omitted, whereas the reported anti-neuropathic dose of gabapentin was 50 mg/kg/day in the previous study (Chanchal et al., 2016). Also, the volume of tissue supernatant and Griess reagent were adjusted from the equal amount in (Kumar et al., 2014) to the ratio of (50:500) with no explanation.

Secondly, the growing suspicion about the paper’s integrity basically evolved from discrepancies between the text and the figures, the imprecise data, and the insufficient interpretation. The text did not match the results displayed in the RT dilution, the measurement time and the magnification scale figures. It was detected by Bucci that Figures 1B, 1C, and 1D exhibited the slower RT concentrations of 10-2, 10-4, 10-6, and 10-8 (instead of 10−8, 10−12, 10−24, and 10-30 in the Results section), which confused the oxidative stress consequences (Guglielmi, 2018). In Figure 3, there were the wrong proportions of the time axis and the recorded days 1, 3, 6, 9, 11 and 14 (having 5 intervals of 2, 3, 3, 2, and 3 days) was incompatible with the planning days 1, 3, 7, 11 and 14. Histological variations in sciatic nerve were noted at the scale 1000 μm in the Results section, which was conflicted with “100 µM” in Figure 6. There were at least 6 findings of inconsistent data as well (Table 2).

Data from article figure/ table

Data from article text (Results section)

LPS (Figure 1F)

Control + LPS (legend of Figure 1F legend)

RT 1 x 10-6, 1 x 10-12, 1 x 10-24, 1 x 10-30 (Figure 2A-D)

RT 1 x 10-8, 1 x 10-12, 1 x 10-24, 1 x 10-30

gm (Figure 3C)

sec

m/s (Figure 4)

mm/sec

pg/ml (Figure 5A-C)

pg/mg

µM (Figure 6)

µm

 

Table 2. Comparison of data from article figure/ table and text showed inconsistent data.

Furthermore, there was a range of incorrect data such as wavelength numbers or copied graphics. No biochemical parameters were measured at the wavelength 405 nm which was replicated in Figures 1 and 2 (Table 3). Bucci also proved that Figure 1H was identical (for the same graph and label) of Figure 1G, while those figures were different in the legend, and the exact duplicate between Figure 1J and Figure 1G when they were supposed to be dissimilar (Guglielmi, 2018). Data interpretation was inadequate when the IL-10 exhibition in Figure 2 or the absence of the H2O2 columns in Figure 1B, 1C and Figure 2A-D was not fully explained, the one-way ANOVA was mentioned as an analysis method in the figure legends (not in detail), and the p-value was abused to overestimate the statistical significance in all experiments for therapeutic conclusion.

Biochemical parameters

Measurement wavelength (nm)

This article

Earlier references

SOD

405 (Figure 1)

560*

Catalase

405 (Figure 1)

240*

Cytokines (TNF-α, IL-1β, IL-6)

405 (Figure 2)

450**

NO

540 (in text)

540***

MDA

N/A

532*

GSH

N/A

412*

Note: *Chanchal et al., 2016; **Manufacturer guides; ***Kumar et al., 2014

Table 3. Comparison of measurement wavelengths for each biological parameter in this article and earlier references revealed unreliable data.

Thirdly, the unusual review process, which possibly resulted from careless proof-reading conducted over a short period of time, raised some arguments among the community. It was received on 14th July, accepted on 28th August, and published on 10th September, and an editor’s warning for its conclusion was announced on 1st October. The article with homeopathy theme publishing in a scientific journal was also suspected because “The available evidence is not compelling and fails to demonstrate that homeopathy is an effective treatment for any of the reported clinical conditions” (Australian Government Review, 2015, p.72). All the inaccuracy in the article (whether due to typographical mistakes or not) should have been noticed by the peer-reviewers to make the necessary adjustments so it exposed the negligent checking and editing from both authors and reviewers.

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Finally, this article, with its flawed design, incorrect data, and poor review, is not a unique case being disputed as an unreliable homeopathy publication. For example, a study of the effects of a highly-diluted antiserum published in 1988 in Nature was argued by many scientists (Davenas et al., 1988). More investigation should be carried out on the homeopathy study and other research which lacked certain scientific standards. The authors, the reviewers, and the readers should always be vigilant over the existing articles, and need to cooperate on verifying the true value of each paper before, during, and after it is published.

REFERENCES

Articles

  • Carrasco, C., Naziroǧlu, M., Rodríguez, A.B., & Pariente, J.A. (2018). Neuropathic Pain: Delving into the Oxidative Origin and the Possible Implication of Transient Receptor Potential Channels. Frontiers in physiology9, 95.
  • Chanchal, S.K., Mahajan, U.B., Siddharth, S., Reddy, N., Goyal, S.N., Patil, P.H., Bommanahalli, B.P., Kundu, C.N., Patil, C.R. & Ojha, S. (2016). In vivo and in vitro protective effects of omeprazole against neuropathic pain. Scientific reports6, 30007.
  • Davenas, E., Beauvais, F., Amara, J., Oberbaum, M., Robinzon, B., Miadonna, A., Tedeschi, A., Pomeranz, B., Fortner, P., Belon, P., Sainte-Laudy, J., Poitevin, B., & Benveniste, J. (1988). Human basophil degranulation triggered by very dilute antiserum against IgE. Nature, 333(6176), 816-818.
  • Guglielmi, G. (2018). Homeopathy advocates have championed the paper, but scientists doubt its claims. Nature, 562(7726), 173-174.
  • Karwa, P. (2010). Effects of Thalidomide on Polymorphonuclear cell functions in vitro. International Journal of PharmTech Research, 2(4), 2445-2449.
  • Kilkenny C., Browne W.J., Cuthill I.C., Emerson M., & Altman D. G. (2012). Improving bioscience research reporting: the ARRIVE guidelines for reporting animal researchOsteoarthritis Cartilage, 20(4), 256-260.
  • Kumar, A., Meena, S. & Pottabathini, R. (2014). Effect of Ashwagandha (Withania somnifera) against chronic constriction injury induced behavioral and biochemical alterations: possible involvement of nitric oxide mechanism. International Journal of Nutrition, Pharmacology, Neurological Diseases, 4, 131.
  • Magar, S., Nayak, D., Mahajan, U.B., Patil, K.R., Shinde, S.D., Goyal, S.N., Swaminarayan, S., Patil, C.R., Ojha, S. & Kundu, C.N. (2018). Ultra-diluted Toxicodendron pubescens attenuates pro-inflammatory cytokines and ROS-mediated neuropathic pain in rats. Scientific reports8(1), 13562.

Web Documents

Other resources

Australian Government Department of Health. (2015). Homeopathy overview report. Review of the Australian Government Rebate on Natural Therapies for Private Health Insurance, 69-75. Australia: Department of Health.

APPENDIX: 6 figures with highly suspicious points (Sources: Magar et al., 2018)

 

Figure 1. Effect of RT on LPS mediated ROS, SOD and catalase activity in U-87 cells.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Figure 2. Effect of RT on LPS stimulated pro-inflammatory cytokines in U-87 cells.
Figure 3. RT improved the cold, warm and mechanical stimuli induced allodynia in CCI induced neuropathic pain in rats.

 

Figure 4. RT improved the MNCV in CCI-induced neuropathic pain in rats.

 

 

 

 

 

 

 

 

 

Figure 5. RT reduced the release of pro-inflammatory cytokines in CCI-induced neuropathic pain.

 

 

 

Figure 6. RT inhibited the CCI-induced histological alterations in the longitudinal sections of sciatic nerve in rats.

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