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Homeopathy
DOI: 10.1055/s-0043-1776039
Original Research Article

Stem Cell Secretome Modulated by Arsenicum album 30C Ameliorates Lipopolysaccharide-induced Cytokine Storm in Blood Mononuclear Cells in vitro

Parth Aphale   *
1   Dr D. Y. Patil Homeopathic Medical College and Research Centre, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune, Maharashtra, India
,
Avinash Sanap   *
2   Regenerative Medicine Laboratory, Dr. D. Y. Patil Dental College and Hospital, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune, Maharashtra, India
,
Dharmendra Sharma
1   Dr D. Y. Patil Homeopathic Medical College and Research Centre, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune, Maharashtra, India
,
Avinash Kharat
2   Regenerative Medicine Laboratory, Dr. D. Y. Patil Dental College and Hospital, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune, Maharashtra, India
,
Supriya Kheur
2   Regenerative Medicine Laboratory, Dr. D. Y. Patil Dental College and Hospital, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune, Maharashtra, India
,
Chinmay Gawade
1   Dr D. Y. Patil Homeopathic Medical College and Research Centre, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune, Maharashtra, India
,
Indumati Somasundaram
3   Department of Biotechnology Engineering, Kolhapur Institute of Technology's College of Engineering, Kolhapur, India
,
Ramesh Bhonde
2   Regenerative Medicine Laboratory, Dr. D. Y. Patil Dental College and Hospital, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune, Maharashtra, India
› Author Affiliations Funding In-house funding was given by Dr. D.Y. Patil Vidyapeeth, Pimpri, Pune (India), to carry out the study (DPU/554(9)/2020).
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Abstract

Background The therapeutic effectiveness of mesenchymal stem cells (MSCs) and their secretome can be enhanced by means of physical, chemical and biological preconditioning. Arsenicum album 30C (AA30) has been one of the leading homeopathic medicines used in prophylaxis against SARS-CoV-2 infection.

Aims This study aimed to investigate whether AA30 preconditioning could influence the growth factors and cytokine profile of the human dental pulp-derived MSC (DPD-MSC) secretome. Also, to test the efficacy of the AA30-preconditioned DPD-MSC secretome in ameliorating the lipopolysaccharide (LPS)-induced cytokine storm in human peripheral blood mononuclear cells (PBMCs) as an in-vitro cellular model.

Methods The cytotoxicity of AA30 was assessed in DPD-MSCs by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Growth factors and cytokine levels in the AA30-preconditioned DPD-MSC secretome were analysed by fluorescence-activated cell sorting (FACS) analysis. The angiogenic potential of the AA30-preconditioned DPD-MSC secretome was assessed by chick yolk-sac membrane (YSM) assay. Culture medium with 0.001% ethanol was used as vehicle control. The efficacy of the AA30-preconditioned DPD-MSC secretome in ameliorating the cytokine storm was assessed in LPS pre-treated PBMCs. The mRNA and protein expression of inflammatory markers such as IL-1β, IL-6 and IL-10 were analysed by using RT-PCR and FACS analysis respectively.

Results AA30 did not exhibit cytotoxicity in the concentration range of 1% to 50%. Furthermore, the AA30-preconditioned DPD-MSC secretome exhibited a significant increase in the levels of angiogenic factors, such as human angiopoietin-2, EPO and PDGF-AA, and decreased levels of cytokines, such as TNF-α, CXCL-8 and IL-6. The AA30-preconditioned DPD-MSC secretome showed augmented angiogenesis compared to vehicle controls. The DPD-MSC secretome ameliorated LPS-induced mRNA and protein expression of IL-1β, IL-6 and IL-10 in PBMCs.

Conclusion The AA30-preconditioned DPD-MSC secretome augmented angiogenesis and ameliorated the LPS-induced cytokine storm in human PBMCs in vitro. Our data demonstrate that AA30 preconditioning enhances the therapeutic potency of MSCs and their secretome.

Keywords

Arsenicum album 30C - COVID-19 - Cytokine storm - Inflammation - Mesenchymal Stem Cells

Data Availability

The data of the current study will be available from the corresponding author upon reasonable request.


Author Contributions

Study conception and design: A.S., P.A., R.B. Data collection: AS. Analysis and interpretation of results: A.S., R.B., P.A. Draft manuscript preparation: A.S., C.G., R.B. Infrastructure access and analysis: D.S., A.K., S.K, I.S.


* Equal first authors.


Supplementary Material



Publication History

Received: 21 March 2023

Accepted: 14 August 2023

Article published online:
07 December 2023

© 2023. Faculty of Homeopathy. This article is published by Thieme.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

  • References

  • 1 World Health Organization. WHO Director-General's opening remarks at the media briefing on COVID-19—March 2020. Accessed August 29, 2023, at: https://www.who.int/director-general/speeches/detail/who-director-general-s-opening-remarks-at-the-media-briefing-on-covid-19---11-march-2020
    PubMedGoogle Scholar
  • 2 Mangalmurti N, Hunter CA. Cytokine storms: understanding COVID-19. Immunity 2020; 53: 19-25
    CrossrefPubMedGoogle Scholar
  • 3 de la Rica R, Borges M, Gonzalez-Freire M. COVID-19: in the eye of the cytokine storm. Front Immunol 2020; 11: 558898
    CrossrefPubMedGoogle Scholar
  • 4 Ragab D, Salah Eldin H, Taeimah M, Khattab R, Salem R. The COVID-19 cytokine storm; what we know so far. Front Immunol 2020; 11: 1446
    CrossrefPubMedGoogle Scholar
  • 5 Ali MJ, Hanif M, Haider MA. et al. Treatment options for COVID-19: a review. Front Med (Lausanne) 2020; 7: 480
    CrossrefPubMedGoogle Scholar
  • 6 Nugraha RV, Ridwansyah H, Ghozali M, Khairani AF, Atik N. Traditional herbal medicine candidates as complementary treatments for COVID-19: a review of their mechanisms, pros and cons. Evid Based Complement Alternat Med 2020; 2020: 2560645
    CrossrefPubMedGoogle Scholar
  • 7 Clapers ML, Velat M, Clapers JM, Vidal-Jové J, Mora J. Analysis of homeopathic genus epidemicus for COVID-19 patients in Spain. Homeopathy 2021; 110: 236-243
    Article in Thieme ConnectPubMedGoogle Scholar
  • 8 Chaudhary A, Khurana A. A review on the role of homeopathy in epidemics with some reflections on COVID-19 (SARS-CoV-2). Indian J Res Homeopathy 2020; 14: 100-109
    CrossrefPubMedGoogle Scholar
  • 9 Ministry of AYUSH, Government of India. Guidelines for Homeopathy Practitioners for prophylaxis and symptomatic management of COVID-19 patients in home isolation. Available at https://ayushportal.nic.in/pdf/31.Ayush.pdf
    PubMed
  • 10 Wei X, Yang X, Han ZP, Qu FF, Shao L, Shi YF. Mesenchymal stem cells: a new trend for cell therapy. Acta Pharmacol Sin 2013; 34: 747-754
    CrossrefPubMedGoogle Scholar
  • 11 Eleuteri S, Fierabracci A. Insights into the secretome of mesenchymal stem cells and its potential applications. Int J Mol Sci 2019; 20: 4597
    CrossrefPubMedGoogle Scholar
  • 12 Attia N, Mashal M. Mesenchymal stem cells: the past present and future. Adv Exp Med Biol 2021; 1312: 107-129
    CrossrefPubMedGoogle Scholar
  • 13 Zhou C, Zhang B, Yang Y. et al. Stem cell-derived exosomes: emerging therapeutic opportunities for wound healing. Stem Cell Res Ther 2023; 14: 107
    CrossrefPubMedGoogle Scholar
  • 14 Li M, Jiang Y, Hou Q, Zhao Y, Zhong L, Fu X. Potential pre-activation strategies for improving therapeutic efficacy of mesenchymal stem cells: current status and future prospects. Stem Cell Res Ther 2022; 13: 146
    CrossrefPubMedGoogle Scholar
  • 15 Hu C, Li L. Preconditioning influences mesenchymal stem cell properties in vitro and in vivo. J Cell Mol Med 2018; 22: 1428-1442
    CrossrefPubMedGoogle Scholar
  • 16 Xu AL, Rodriguez II LA, Walker III KP. et al. Mesenchymal stem cells reconditioned in their own serum exhibit augmented therapeutic properties in the setting of acute respiratory distress syndrome. Stem Cells Transl Med 2019; 8: 1092-1106
    CrossrefPubMedGoogle Scholar
  • 17 Udalamaththa VL, Jayasinghe CD, Udagama PV. Potential role of herbal remedies in stem cell therapy: proliferation and differentiation of human mesenchymal stromal cells. Stem Cell Res Ther 2016; 7: 110
    CrossrefPubMedGoogle Scholar
  • 18 Antebi B, Rodriguez II LA, Walker III KP. et al. Short-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cells. Stem Cell Res Ther 2018; 9: 265
    CrossrefPubMedGoogle Scholar
  • 19 Desai S, Jagtap J, Sainani S, Bhonde R. Guiding stem cells for cutaneous repair. Curr Res Pharmacol Drug Discov 2022; 4: 100145
    CrossrefPubMedGoogle Scholar
  • 20 Sanap A, Kheur S, Kharat A, Bhonde R. Ascorbic acid and IFNγ preconditioning enhance the potency of human mesenchymal stem cells to ameliorate LPS induced cytokine storm. Int Immunopharmacol 2023; 122: 110643
    CrossrefPubMedGoogle Scholar
  • 21 Xu R, Feng Z, Wang FS. Mesenchymal stem cell treatment for COVID-19. EBioMedicine 2022; 77: 103920
    CrossrefPubMedGoogle Scholar
  • 22 Wang L, Li Y, Xu M. et al. Regulation of inflammatory cytokine storms by mesenchymal stem cells. Front Immunol 2021; 12: 726909
    CrossrefPubMedGoogle Scholar
  • 23 Mizukami A, Swiech K. Mesenchymal stromal cells: from discovery to manufacturing and commercialization. Stem Cells Int 2018; 2018: 4083921
    CrossrefPubMedGoogle Scholar
  • 24 Li B, Ouchi T, Cao Y, Zhao Z, Men Y. Dental-derived mesenchymal stem cells: state of the art. Front Cell Dev Biol 2021; 9: 654559
    CrossrefPubMedGoogle Scholar
  • 25 Tumpara S, Gründing AR, Sivaraman K. et al. Boosted pro-inflammatory activity in human PBMCs by lipopolysaccharide and SARS-CoV-2 spike protein is regulated by α-1 antitrypsin. Int J Mol Sci 2021; 22: 7941
    CrossrefPubMedGoogle Scholar
  • 26 Patil VR, Kharat AH, Kulkarni DG, Kheur SM, Bhonde RR. Long term explant culture for harvesting homogeneous population of human dental pulp stem cells. Cell Biol Int 2018; 42: 1602-1610
    CrossrefPubMedGoogle Scholar
  • 27 Sanap A, Joshi K, Shah T, Tillu G, Bhonde R. Pre-conditioning of mesenchymal stem cells with piper longum L. augments osteogenic differentiation. J Ethnopharmacol 2021; 273: 113999
    CrossrefPubMedGoogle Scholar
  • 28 Sanap A, Chandravanshi B, Shah T. et al. Herbal pre-conditioning induces proliferation and delays senescence in Wharton's Jelly Mesenchymal Stem Cells. Biomed Pharmacother 2017; 93: 772-778
    CrossrefPubMedGoogle Scholar
  • 29 Shekatkar MR, Kheur SM, Kharat AH. et al. Assessment of angiogenic potential of mesenchymal stem cells derived conditioned medium from various oral sources. J Clin Transl Res 2022; 8: 323-338
    PubMedGoogle Scholar
  • 30 Bhonde R, Sanap A, Joshi K. Mesenchymal stem cells as a platform for research on traditional medicine. J Ayurveda Integr Med 2021; 12: 722-728
    CrossrefPubMedGoogle Scholar
  • 31 Paudyal V, Sun S, Hussain R, Abutaleb MH, Hedima EW. Complementary and alternative medicines use in COVID-19: a global perspective on practice, policy and research. Res Social Adm Pharm 2022; 18: 2524-2528
    CrossrefPubMedGoogle Scholar
  • 32 Clinical Trial Registry of India. Effectiveness of Arsenicum album 30c in prevention of COVID-19 in individuals residing in hot spots of red zones—a multicentric, randomized, cluster level, controlled trial. CTRI/2020/05/025205; 2020
    PubMedGoogle Scholar
  • 33 Nayak D, Devarajan K, Pal PP. et al. Efficacy of Arsenicum album 30C in the prevention of COVID-19 in individuals residing in containment areas—a prospective, multicentre, cluster-randomized, parallel arm, community based, open-label study. Complement Med Res 2022 ; (e-pub ahead of print)
    PubMedGoogle Scholar
  • 34 Croci S, Bonacini M, Dolci G. et al. Human dental pulp stem cells modulate cytokine production in vitro by peripheral blood mononuclear cells from coronavirus disease 2019 patients. Front Cell Dev Biol 2021; 8: 609204
    CrossrefPubMedGoogle Scholar
  • 35 Beghini DG, Horita SI, Henriques-Pons A. Mesenchymal stem cells in the treatment of COVID-19, a promising future. Cells 2021; 10: 2588
    CrossrefPubMedGoogle Scholar
  • 36 Banerjee P, Bhattacharyya SS, Pathak S, Naoual B, Belon P, Khuda-Bukhsh AR. Comparative efficacy of two microdoses of a potentized homeopathic drug, arsenicum album, to ameliorate toxicity induced by repeated sublethal injections of arsenic trioxide in mice. Pathobiology 2008; 75: 156-170
    CrossrefPubMedGoogle Scholar
  • 37 Khuda-Bukhsh AR, Roy-Karmakar S, Banerjee A. et al. A follow-up study on the efficacy of the homeopathic remedy Arsenicum album in volunteers living in high risk arsenic contaminated areas. Evid Based Complement Alternat Med 2011; 2011: 129214
    PubMedGoogle Scholar
  • 38 Das D, De A, Dutta S, Biswas R, Boujedaini N, Khuda-Bukhsh AR. Potentized homeopathic drug Arsenicum album 30c positively modulates protein biomarkers and gene expressions in Saccharomyces cerevisae exposed to arsenate. J Chin Integr Med 2011; 9: 752-760
    CrossrefPubMedGoogle Scholar
  • 39 Theruvath AH, Raveendran R, Philips CA. Dangerous placebo during the COVID-19 pandemic: a series of homeopathic Arsenicum album-induced liver injury. Cureus 2022; 14: e26062
    PubMedGoogle Scholar
  • 40 Vabret N, Britton GJ, Gruber C. et al. Immunology of COVID-19: current state of the science. Immunity 2020; 52: 910-941
    CrossrefPubMedGoogle Scholar
  • 41 Sabaka P, Koščálová A, Straka I. et al. Role of interleukin 6 as a predictive factor for a severe course of COVID-19: retrospective data analysis of patients from a long-term care facility during Covid-19 outbreak. BMC Infect Dis 2021; 21: 308
    CrossrefPubMedGoogle Scholar
  • 42 Shen WX, Luo RC, Wang JQ, Chen ZS. Features of cytokine storm identified by distinguishing clinical manifestations in COVID-19. Front Public Health 2021; 9: 671788
    CrossrefPubMedGoogle Scholar
  • 43 Coperchini F, Chiovato L, Croce L, Magri F, Rotondi M. The cytokine storm in COVID-19: an overview of the involvement of the chemokine/chemokine-receptor system. Cytokine Growth Factor Rev 2020; 53: 25-32
    CrossrefPubMedGoogle Scholar
  • 44 Diao B, Wang C, Tan Y. et al. Reduction and functional exhaustion of T cells in patients with coronavirus disease 2019. Front Immunol 2020; 11: 827
    CrossrefPubMedGoogle Scholar
  • 45 Soin AS, Kumar K, Choudhary NS. et al. Tocilizumab plus standard care versus standard care in patients in India with moderate to severe COVID-19-associated cytokine release syndrome (COVINTOC): an open-label, multicentre, randomised, controlled, phase 3 trial. Lancet Respir Med 2021; 9: 511-521
    CrossrefPubMedGoogle Scholar
  • 46 Lescure FX, Honda H, Fowler RA. et al. Sarilumab in patients admitted to hospital with severe or critical COVID-19: a randomised, double-blind, placebo-controlled, phase 3 trial. Lancet Respir Med 2021; 9: 522-532
    CrossrefPubMedGoogle Scholar
  • 47 Islam H, Chamberlain TC, Mui AL, Little JP. Elevated interleukin-10 levels in COVID-19: potentiation of pro-inflammatory responses or impaired anti-inflammatory action?. Front Immunol 2021; 12: 677008
    CrossrefPubMedGoogle Scholar
  • 48 Bellamri N, Morzadec C, Fardel O, Vernhet L. Arsenic and the immune system. Curr Opin Toxicol 2018; 10: 60-68
    CrossrefPubMedGoogle Scholar
  • 49 Biswas R, Ghosh P, Banerjee N. et al. Analysis of T-cell proliferation and cytokine secretion in the individuals exposed to arsenic. Hum Exp Toxicol 2008; 27: 381-386
    CrossrefPubMedGoogle Scholar
  • 50 Charan N, Lavanya N, Praveen B, Praveen A, Sridevi A, Narasimha G. Antiviral activity of antimony and arsenic oxides. Der Pharma Chem. 2012; 4: 687-689
    PubMedGoogle Scholar