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Animals : an open access journal from MDPI2024; 14(18); doi: 10.3390/ani14182681

Revealing the Therapeutic Potential of Muscle-Derived Mesenchymal Stem/Stromal Cells: An In Vitro Model for Equine Laminitis Based on Activated Neutrophils, Anoxia-Reoxygenation, and Myeloperoxidase.

Abstract: Laminitis in horses is a crippling condition marked by the deterioration of the dermal-epidermal interface, leading to intense lameness and discomfort, often necessitating euthanasia. This study aimed to establish an in vitro model of laminitis using a continuous keratinocyte cell line exposed to anoxia-reoxygenation and an activated neutrophil supernatant. A significant decrease in the keratinocytes' metabolism was noted during the reoxygenation period, indicative of cellular stress. Adding muscle-derived mesenchymal stem/stromal cells during the reoxygenation demonstrated a protective effect, restoring the keratinocytes' metabolic activity. Moreover, the incubation of the keratinocytes with either an activated neutrophil supernatant or myeloperoxidase alone induced increased keratinocyte myeloperoxidase activity, which was modulated by stem cells. These findings underscore the potential of muscle-derived mesenchymal stem/stromal cells in mitigating inflammation and restoring keratinocyte metabolism, offering insights for future cell therapy research in laminitis treatment.
Publication Date: 2024-09-14 PubMed ID: 39335269PubMed Central: PMC11428732DOI: 10.3390/ani14182681Google Scholar: Lookup
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  • Journal Article

Summary

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The research article describes an experiment to investigate the use of muscle-derived mesenchymal stem/stromal cells (MDMSCs) as a potential treatment for laminitis – a painful disease affecting horses. The scientists discovered that these cells can reduce inflammation and restore cell metabolism, hinting at their potential application in future therapies for this condition.

Objective and Methods of the Study

  • The study was conducted to examine the therapeutic potential of MDMSCs in treating laminitis in horses, a crippling condition often leading to euthanasia.
  • The researchers developed an in-vitro model. In this model, keratinocyte cells (which make up a substantial part of the dermal-epidermal junction) were exposed to anoxia-reoxygenation and activated neutrophil supernatant – conditions that mimic the environment during a laminitis episode.

Findings on Keratinocyte Metabolism

  • The scientists observed a significant decrease in the metabolism of the keratinocyte cells during the reoxygenation period, reflecting a state of cellular stress analogous to what occurs during a laminitis attack.
  • When MDMSCs were added during reoxygenation, they had a protective effect, restoring the metabolic activity of the keratinocytes.

Impact of Activated Neutrophil Supernatant and Myeloperoxidase

  • The researchers also noticed an increase in keratinocyte’s myeloperoxidase (MPO) activity when they were incubated with either activated neutrophil supernatant or MPO alone. MPO is an enzyme found in neutrophils and it’s been associated with inflammatory conditions. This suggests that these elements contribute to the inflammation seen in laminitis.
  • However, this increased MPO activity was reduced when MDMSCs were present, showing the stem cells’ ability to mitigate inflammation.

Implications for Laminitis Treatment

  • These findings highlight the potential for MDMSCs in reducing inflammation and restoring cell metabolism under stress conditions – which are crucial features needed for a laminitis treatment.
  • The study provides a valuable foundation for future research into cell therapy options for managing and treating laminitis in horses.

Cite This Article

APA
Serteyn D, Storms N, Mouithys-Mickalad A, Sandersen C, Niesten A, Duysens J, Graide H, Ceusters J, Franck T. (2024). Revealing the Therapeutic Potential of Muscle-Derived Mesenchymal Stem/Stromal Cells: An In Vitro Model for Equine Laminitis Based on Activated Neutrophils, Anoxia-Reoxygenation, and Myeloperoxidase. Animals (Basel), 14(18). https://doi.org/10.3390/ani14182681

Publication

ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 14
Issue: 18

Researcher Affiliations

Serteyn, Didier
  • Department of Equine Clinical Sciences, University of Liège, 4000 Liège, Belgium.
  • Center for Oxygen Research and Development, B6, University of Liège, FARAH, Quartier Vallée 2 Avenue de Cureghem 5D, 4000 Liège, Belgium.
Storms, Nazaré
  • Department of Equine Clinical Sciences, University of Liège, 4000 Liège, Belgium.
Mouithys-Mickalad, Ange
  • Center for Oxygen Research and Development, B6, University of Liège, FARAH, Quartier Vallée 2 Avenue de Cureghem 5D, 4000 Liège, Belgium.
Sandersen, Charlotte
  • Department of Equine Clinical Sciences, University of Liège, 4000 Liège, Belgium.
  • Center for Oxygen Research and Development, B6, University of Liège, FARAH, Quartier Vallée 2 Avenue de Cureghem 5D, 4000 Liège, Belgium.
Niesten, Ariane
  • Center for Oxygen Research and Development, B6, University of Liège, FARAH, Quartier Vallée 2 Avenue de Cureghem 5D, 4000 Liège, Belgium.
Duysens, Julien
  • Center for Oxygen Research and Development, B6, University of Liège, FARAH, Quartier Vallée 2 Avenue de Cureghem 5D, 4000 Liège, Belgium.
Graide, Hélène
  • Center for Oxygen Research and Development, B6, University of Liège, FARAH, Quartier Vallée 2 Avenue de Cureghem 5D, 4000 Liège, Belgium.
Ceusters, Justine
  • Center for Oxygen Research and Development, B6, University of Liège, FARAH, Quartier Vallée 2 Avenue de Cureghem 5D, 4000 Liège, Belgium.
Franck, Thierry
  • Center for Oxygen Research and Development, B6, University of Liège, FARAH, Quartier Vallée 2 Avenue de Cureghem 5D, 4000 Liège, Belgium.

Grant Funding

  • Mitotrans / French Community of Belgium

Conflict of Interest Statement

D. Serteyn and J. Ceusters are the co-inventors of a patent related to muscle-derived stem cells. This patent is licensed by the University of Liège to Revatis, a spin-off company in which D. Serteyn and J. Ceusters serve as scientific advisors. The other co-authors declare no conflicts of interest.

References

This article includes 36 references
  1. Menzies-Gow N.J., Stevens K., Barr A., Camm I., Pfeiffer D., Marr C.M.. Severity and outcome of equine pasture-associated laminitis managed in first opinion practice in the UK.. Vet. Rec. 2010;167:364–369.
    doi: 10.1136/vr.c3206pubmed: 20817896google scholar: lookup
  2. Katz L.M., Bailey S.R.. A review of recent advances and current hypotheses on the pathogenesis of acute laminitis.. Equine Vet. J. 2012;44:752–761.
  3. van Eps A.W.. Acute laminitis: Medical and supportive therapy.. Vet. Clin. North Am. Equine Pract. 2010;26:103–114.
    doi: 10.1016/j.cveq.2009.12.011pubmed: 20381739google scholar: lookup
  4. Peroni J.F., Harrison W.E., Moore J.N., Graves J.E., Lewis S.J., Krunkosky T.M., Robertson T.P.. Black walnut extract-induced laminitis in horses is associated with heterogeneous dysfunction of the laminar microvasculature.. Equine Vet. J. 2005;37:546–551.
    doi: 10.2746/042516405775314781pubmed: 16295933google scholar: lookup
  5. Hurley D.J., Parks R.J., Reber A.J., Donovan D.C., Okinaga T., Vandenplas M.L., Peroni J., Moore J.. Dynamic changes in circulating leukocytes during the induction of equine laminitis with black walnut extract.. Vet. Immunol. Immunopathol. 2006;110:195–206.
    doi: 10.1016/j.vetimm.2005.09.015pubmed: 16290066google scholar: lookup
  6. Loftus J.P., Belknap J.K., Stankiewicz K.M., Black S.J.. Laminar xanthine oxidase, superoxide dismutase and catalase activities in the prodromal stage of black-walnut induced equine laminitis.. Equine Vet. J. 2007;39:48–53.
    doi: 10.2746/042516406X151320pubmed: 17228595google scholar: lookup
  7. Leise B.. The role of neutrophils in equine laminitis.. Cell Tissue Res. 2018;371:541–550.
    doi: 10.1007/s00441-018-2788-zpubmed: 29397426google scholar: lookup
  8. Galantino-Homer H., Brooks S.A.. Genetics and Signaling Pathways of Laminitis.. Vet. Clin. N. Am. Equine Pract. 2020;36:379–394.
    doi: 10.1016/j.cveq.2020.04.001pubmed: 32654786google scholar: lookup
  9. Riggs L.M., Franck T., Moore J.N., Krunkosky T.M., Hurley D.J., Peroni J.F., de la Rebière G., Serteyn D.A.. Neutrophil myeloperoxidase measurements in plasma, laminar tissue, and skin of horses given black walnut extract.. Am. J. Vet. Res. 2007;68:81–86.
    doi: 10.2460/ajvr.68.1.81pubmed: 17199423google scholar: lookup
  10. Storms N., de la Rebière G., Franck T., Mouithys Mickalad A., Sandersen C., Ceusters J., Serteyn D.. Neutrophil extracellular traps and active myeloperoxidase concentrate in lamellar tissue of equids with naturally occurring laminitis.. Vet. Immunol. Immunopathol. 2024;270:110738.
    doi: 10.1016/j.vetimm.2024.110738pubmed: 38452577google scholar: lookup
  11. Aratani Y.. Myeloperoxidase: Its role for host defense, inflammation, and neutrophil function.. Arch. Biochem. Biophys. 2018;640:47–52.
    doi: 10.1016/j.abb.2018.01.004pubmed: 29336940google scholar: lookup
  12. French K.R., Pollitt C.C.. Equine laminitis: Loss of hemidesmosomes in hoof secondary epidermal lamellae correlates to dose in an oligofructose induction model: An ultrastructural study.. Equine Vet. J. 2004;36:230–235.
    doi: 10.2746/0425164044877125pubmed: 15147130google scholar: lookup
  13. Shepherd H.M., Gauthier J.M., Terada Y., Li W., Krupnick A.S., Gelman A.E., Kreisel D.. Updated Views on Neutrophil Responses in Ischemia-Reperfusion Injury.. Transplantation. 2022;106:2314–2324.
    doi: 10.1097/TP.0000000000004221pmc: PMC9712152pubmed: 35749228google scholar: lookup
  14. Serteyn D., de la Rebière de Pouyade G., Sandersen C., Salciccia A., Grulke S.. Muscle Mitochondrial Dysfunction in Horses Affected by Acute Laminitis.. Bioenergetics. 2014;3:1000120.
    doi: 10.4172/2167-7662.1000120google scholar: lookup
  15. He L., Liu R., Yue H., Zhang X., Pan X., Sun Y., Shi J., Zhu G., Qin C., Guo Y.. Interaction between neutrophil extracellular traps and cardiomyocytes contributes to atrial fibrillation progression.. Signal Transduct. Target. Ther. 2023;8:279.
    doi: 10.1038/s41392-023-01497-2pmc: PMC10368710pubmed: 37491321google scholar: lookup
  16. Sørensen O., Borregaard N.. Neutrophil extracellular trap —The dark side of neutrophils.. J. Clin. Investig. 2016;126:1612–1620.
    doi: 10.1172/JCI84538pmc: PMC4855925pubmed: 27135878google scholar: lookup
  17. Delgado-Rizo V., Martínez-Guzmán M., Iñiguez-Gutierrez L., García-Orozco A., Alvarado-Navarro A., Fafutis-Morris M.. Neutrophil Extracellular Traps and Its Implications in Inflammation: An Overview.. Front. Immunol. 2017;8:81.
    doi: 10.3389/fimmu.2017.00081pmc: PMC5292617pubmed: 28220120google scholar: lookup
  18. Tablin A., Li R.. A comparative review of neutrophil extracellular traps in sepsis. Front. Vet. Sci. 2018;5:291.
    pmc: PMC6280561pubmed: 30547040
  19. Zhang F., Li Y., Wu J., Zhang J., Cao P., Sun Z., Wang W.. The role of extracellular traps in ischemia reperfusion injury.. Front. Immunol. 2022;13:1022380.
    doi: 10.3389/fimmu.2022.1022380pmc: PMC9533173pubmed: 36211432google scholar: lookup
  20. Jennaro T.S., Puskarich M.A., Evans C.R., Karnovsky A., Flott T.L., McLellan L.A., Jones A.E., Stringer K.A.. Sustained Perturbation of Metabolism and Metabolic Subphenotypes Are Associated With Mortality and Protein Markers of the Host Response.. Crit. Care Explor. 2023;5:e0881.
  21. Miceli V., Bulati M., Gallo A., Iannolo G., Busà R., Conaldi P.G., Zito G.. Role of Mesenchymal Stem/Stromal Cells in Modulating Ischemia/Reperfusion Injury: Current State of the Art and Future Perspectives.. Biomedicines. 2023;11:689.
  22. Rowart P., Erpicum P., Detry O., Weekers L., Grégoire C., Lechanteur C., Briquet A., Beguin Y., Krzesinski J.M., Jouret F.. Mesenchymal Stromal Cell Therapy in Ischemia/Reperfusion Injury.. J. Immunol. Res. 2015;2015:602597.
    doi: 10.1155/2015/602597pmc: PMC4518154pubmed: 26258151google scholar: lookup
  23. Yao J., Zheng J., Cai J., Zeng K., Zhou C., Zhang J., Li S., Li H., Chen L., He L.. Extracellular vesicles derived from human umbilical cord mesenchymal stem cells alleviate rat hepatic ischemia-reperfusion injury by suppressing oxidative stress and neutrophil inflammatory response.. FASEB J. 2019;33:1695–1710.
    doi: 10.1096/fj.201800131RRpubmed: 30226809google scholar: lookup
  24. Magaña-Guerrero F.S., Domínguez-López A., Martínez-Aboytes P., Buentello-Volante B., Garfias Y.. Human Amniotic Membrane Mesenchymal Stem Cells inhibit Neutrophil Extracellular Traps through TSG-6.. Sci. Rep. 2017;7:12426.
    doi: 10.1038/s41598-017-10962-2pmc: PMC5622031pubmed: 28963485google scholar: lookup
  25. Ceusters J., Lejeune J.P., Sandersen C., Niesten A., Lagneaux L., Serteyn D.. From skeletal muscle to stem cells: An innovative and minimally-invasive process for multiple species.. Sci. Rep. 2017;7:696.
    doi: 10.1038/s41598-017-00803-7pmc: PMC5429713pubmed: 28386120google scholar: lookup
  26. Barrett J., MacDonald S.. Use of Biologics and Stem Cells in the Treatment of Other Inflammatory Diseases in the Horse.. Vet. Clin. N. Am. Equine Pract. 2023;39:553–563.
    doi: 10.1016/j.cveq.2023.07.004pubmed: 37607855google scholar: lookup
  27. Pycock J.F., Allen W.E., Morris T.H.. Rapid, single-step isolation of equine neutrophils on a discontinuous Percoll density gradient.. Res. Vet. Sci. 1987;42:411–412.
    doi: 10.1016/S0034-5288(18)30727-6pubmed: 3616152google scholar: lookup
  28. Braissant O., Astasov-Frauenhoffer M., Waltimo T., Bonkat G.. A review of methods to determine viability, vitality, and metabolic rates in microbiology.. Front. Microbiol. 2020;11:547458.
    doi: 10.3389/fmicb.2020.547458pmc: PMC7705206pubmed: 33281753google scholar: lookup
  29. Ghasemi M., Turnbull T., Sebastian S., Kempson I.. The MTT Assay: Utility, Limitations, Pitfalls, and Interpretation in Bulk and Single-Cell Analysis.. Int. J. Mol. Sci. 2021;22:12827.
    doi: 10.3390/ijms222312827pmc: PMC8657538pubmed: 34884632google scholar: lookup
  30. Stockert J.C., Horobin R.W., Colombo L.L., Blázquez-Castro A.. Tetrazolium salts and formazan products in cell biology: Viability assessment, fluorescence imaging, and labeling perspectives.. Acta Histochem. 2018;120:159–167.
    doi: 10.1016/j.acthis.2018.02.005pubmed: 29496266google scholar: lookup
  31. Marin W., Marin D., Ao X., Liu Y.. Mitochondria as a therapeutic target for cardiac ischemia-reperfusion injury (Review). Int. J. Mol. Med. 2021;47:485–499.
    doi: 10.3892/ijmm.2020.4823pmc: PMC7797474pubmed: 33416090google scholar: lookup
  32. Huang R., Zhang C., Xiang Z., Lin T., Ling J., Hu H.. Role of mitochondria in renal ischemia-reperfusion injury.. FEBS J. 2024.
    doi: 10.1111/febs.17130pubmed: 38567754google scholar: lookup
  33. Burn G.L., Foti A., Marsman G., Patel D.F., Zychlinsky A.. The Neutrophil.. Immunity. 2021;54:1377–1391.
    doi: 10.1016/j.immuni.2021.06.006pubmed: 34260886google scholar: lookup
  34. Rodriguez A.M., Nakhle J., Griessinger E., Vignais M.L.. Intercellular mitochondria trafficking highlighting the dual role of mesenchymal stem cells as both sensors and rescuers of tissue injury.. Cell Cycle. 2018;17:712–721.
  35. Liu D., Gao Y., Liu J., Huang Y., Yin J., Feng Y., Shi L., Meloni B.P., Zhang C., Zheng M.. Intercellular mitochondrial transfer as a means of tissue revitalization.. Signal Transduct. Target. Ther. 2021;6:65.
    doi: 10.1038/s41392-020-00440-zpmc: PMC7884415pubmed: 33589598google scholar: lookup
  36. Franck T., Storms N., Ceusters J., Sandersen C., de la Rebière G., Mouithys-Mickalad A., Serteyn D.. Targeting Mitochondria 2023 Abstract Book.. J. Mitochondria Plast. Endosymbiosis. 2023;1((Suppl. S1)):2270281.