This research work was contributed by all authors mentioned in this presentation. However the research paper published by journal of Homeopathy (publisher: thieme) might not have been informed about the correct sequence of authorship. (https://www.thieme-connect.com/products/ejournals/abstract/10.1055/s-0043-1776039) Author had already presented this work in LMHI 2021 conference when affiliated with Dr. D.Y.Patil HMC, Pune. Dr. Aditya D Patil is not affiliated with Dr. D Y Patil HMC from 11/02/2022. The work carried out was from 02/03/2021 to 11/02/2022). Hereby expressing the thoughts of dishonesty for the all authors by ignoring the informed consent for publishing this work. Background: The ongoing COVID-19 pandemic caused by the outbreak of SARS-CoV-2 is a global emergency wherein implementation of social measures along with boosting prophylactic immunity is considered to be effective. In India, Arsenicum Album 30C (AA30) is one of the key homeopathic medicine intervened in order to enhance the prophylactic immunity against SARS-CoV-2 during the pandemic. Although, AA30 has been found effective in improving the CD4/CD8 cell ratio which aids in defence against SARS-CoV-2 infections, its detailed mechanism of action remains elusive. Material and methods: Immunomodulatory potential of AA30 was investigated in the stem cell model isolated from dental pulp (DPMSCs). Cytotoxicity of the AA30 was determined by the MTT and BrdU incorporation assay at the 1, 3, 5, 10, 20, 30 and 50%. The effect of AA30 on the cytokine release (CCL2, TNF –alpha, IL -1beta, CXCL 10, IL-2, IL-4, TGF- beta, CXCL-8, IL-12, IFN-gamma, IL-10, IL-6 and IL-17) of DPMSCs were assessed by the fluorescence activated cell sorter (FACS). Results: AA30 exhibited significant increase in the proliferation of DPMSCs at the concentration range 10-50% (p<0.05 whereas none of the diluted dose were cytotoxic to the cells as evidenced by the MTT and BrdU incorporation assay. Interestingly, treatment if AA30 resulted into increased levels of anti inflammatory cytokines (IL4 and IL10, p<0.05) whereas levels of pro-inflammatory cytokines were down regulated (IL 1 beta, TNF alfa, p<0.05). Conclusion: Our data unequivocally demonstrates for the first time that AA30 is non cytotoxic in nature and exhibits immunomodulatory potential. Our results support current intervention policy to use AA30 as a prophylactic candidate to boost immunity against SARS-CoV-2. Further studies are required to elucidate its role in the management of SARS-CoV-2 infection. Keywords: Arsenicum Album 30C (AA30), Immunomodulator, Inflammatory cytokines, Prophylactic, SARS-CoV-2 infection

1. 1
Re-Accredited by NAAC with a CGPA of 3.62 on a 4-point scale at " A " Grade
An ISO 9001:2015 & 14001:2015 CERTIFIED UNIVERSITY

2. Arsenicum Album 30C exhibits immunomodulatory
potential in stem cell model
Dr. Parth Aphale1, Dr. Aditya Patil2, , Dr. Dharmendra B. Sharma3, Dr. Ramesh Bhonde4,
Dr. Avinash Sanap5, Dr. Supriya Kheur6
Re-Accredited by NAAC with a CGPA of 3.62 on a 4-point scale at " A " Grade
An ISO 9001:2015 & 14001:2015 CERTIFIED UNIVERSITY

3. Fig.1-Yang, L., Xie, X., Tu, Z. et al. The signal pathways and treatment of cytokine storm in COVID-
19. Sig Transduct Target Ther 6, 255 (2021). https://doi.org/10.1038/s41392-021-00679-0
Fig.2-https://www.sinobiological.com/resource/cytokines/all-proinflammatory-cytokines
FIG.3 Nguyen, Julie & Austin, Evan & Huang, Alisen & Mamalis, Andrew & Jagdeo, Jared. (2020). The IL-
4/IL-13 axis in skin fibrosis and scarring: mechanistic concepts and therapeutic targets. Archives of
Dermatological Research. 312. 10.1007/s00403-019-01972-3.

4. AIM
•To study the immunomodulatory
activity of homoeopathic medicine
Arsenicum Album by employing stem
cell model.

5. 1. To observe the action of Arsenicum Album on the cytokine release
(CCL2, TNF –alpha, IL -1beta, CXCL 10, IL-2, IL-4, TGF- beta, CXCL-8, IL-
12, IFN-gamma, IL-10, IL-6 and IL-17) through DPMSCs
2. To study the cytotoxicity of Arsenicum Album 30C
employing stem cells.
3.To provide evidence-based science and confirm the role of
Arsenicum Album as immunomodulator

6. • Does Arsenicum Album 30 shows any
immunomodulatory activity as claimed
by the advisory released by Ministry of
AYUSH, Govt. of India?
RESEARCH
QUESTION.1
• Can Stem cell model be used efficiently
to prove this immunomodulatory
activity of Arsenicum Album 30C?
Research
Question.2

7. • Arsenicum Album 30C does not show any
immunomodulatory activity evident
through stem cell model via inflammatory
markers.
Null
Hypothesis
• Arsenicum Album 30C shows promising
immunomodulatory activity evident through
stem cell model via inflammatory markers.
Alternative
Hypothesis

8. • Regenerative Medicine Laboratory at Dr.
D.Y. Patil Dental College, Pimpri, Pune
PLACE OF
STUDY
• 1 year
DURATION OF
STUDY
• Arsenicum Album 30C (AA30) on Mesenchymal
stem cells
INTERVENTION

10. 0
20
40
60
80
100
120
140
160
180
Control 1 3 5 10 20 30 50
% Viability
AA30 exhibited significant increase in the
proliferation of DPMSCs at the
concentration range 10-50% (p<0.05
whereas none of the diluted dose were
cytotoxic to the cells as evidenced by the
MTT and BrdU incorporation assay.

11. TNF-alpha
IL-1Beta
CXCL-10 IL-17
IL-6
Levels of pro-inflammatory cytokines were down regulated CCL2, IL-6 p<0.05).
CCL2

12. TGF-beta
Interestingly, treatment if AA30 resulted into
increased levels of anti inflammatory cytokine (IL10 at
20% conc., p<0.05) TGF- Beta and IL-4 secretion was
also regulated and maintained.
47.4
47.6
47.8
48
48.2
48.4
48.6
48.8
49
49.2
Medium
Control
Control AA 10 AA 20 AA 50
IL-4
IL-4
51
51.5
52
52.5
53
53.5
54
54.5
Medium
Control
Control AA 10 AA 20 AA 50
IL-10
IL-10

13. ANTI-
INFLAMMATORY
CYTOKINES
PROINFLAMMATORY
CYTOKINES
FIG.5-Castelli V, Cimini A and Ferri C (2020) Cytokine Storm in COVID-19: “When You Come Out of the Storm, You
Won’t Be the Same Person Who Walked in”. Front. Immunol. 11:2132. doi: 10.3389/fimmu.2020.02132
FIG.6-Zou, Sheng. (2017). Five Balances in the Management of Rheumatoid Arthritis. Journal of
Biosciences and Medicines. 05. 10-21. 10.4236/jbm.2017.59002.

14. MANAGEMENT
• Hon’ble Chancellor, P.D. Patil Sir, Pro- Chancellor, Dr. Bhagyashree Patil, Trustee, Dr. Smita Jadhav, Vice
Chancellor, Prof. Dr. N.J. Pawar
Regeneretive
Medicine
Laboratory
• Dr. Ramesh Bhonde, Director, Research, Dr. D.Y. Patil Vidyapeeth, Pune
• Dr. Avinash Sanap, Research Associate, Regenerative Medicine Laboratory, Dr. D.Y. Patil Vidyapeeth
Ethics Committee
• All esteemed members of the Institutional Ethics Committee- Stem cell Research
Principal
• Dr. Dharmendra B. Sharma, Principal, Dr. D.Y. Patil Homoeopathic Medical College and Research Centre