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Home / Equine Research Bank / J Vis Exp / Nerve Stimulator-guided Injection of Autologous Stem Cells N...
Journal of visualized experiments : JoVE2018; (139); 58023; doi: 10.3791/58023

Nerve Stimulator-guided Injection of Autologous Stem Cells Near the Equine Left Recurrent Laryngeal Nerve.

Abstract: Recurrent laryngeal neuropathy (RLN) commonly affects horses and is characterized by abnormal respiratory sounds and exercise intolerance. The recurrent laryngeal nerve shows lesions of demyelination. The benefit of applying stem cells to demyelinated nerves has been demonstrated in various animal models. The aim of the study was to test the feasibility and safety of a peri-neuronal injection of autologous muscle-derived mesenchymal stem cells to the left recurrent laryngeal nerve in healthy horses by using an electrical nerve stimulator. Muscle-derived stems cell are obtained from five healthy Standardbred horses by sampling 20 mg of muscle tissue with a semi-automatic 14 G biopsy needle from the triceps muscle. Movements of the larynx are monitored via upper-airway video endoscopy. The left recurrent laryngeal nerve is approached with an insulated nerve block needle. Nerve stimulation is applied, starting at 2 mA, and the successful abduction of the left arytenoid is monitored. The stimulation intensity is reduced progressively. When a loss of the motor response is observed at 0.5 mA, 10 autologous muscle-derived stem cells are injected. Two examiners, who are blinded to the time point, score the laryngeal function of the horses prior to the treatment and at day 1, day 7, and day 28 after the injection of the cells. In a sixth horse, 1 mL of 2% lidocaine is injected to further confirm the correct positioning of the needle. This leads to a temporary paralysis of the left arytenoid cartilage. This study proves that the recurrent laryngeal nerve can be approached with the help of an electrical nerve stimulator and that the electrical stimulation of the nerve is well tolerated by the horses. No modification of the laryngeal function was observed in any of the horses after the injection of the stem cells. Further studies should be conducted to describe the effects of a peri-neuronal injection of autologous muscle-derived mesenchymal stem cells to horses suffering from RLN.
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Publication Date: 2018-09-26 PubMed ID: 30320737PubMed Central: PMC6235316DOI: 10.3791/58023Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research investigated the feasibility and safety of using a nerve stimulator-guided injection of stem cells directly to a nerve in horses, which is a proposed treatment for a common equine neurological disorder.

Research Objectives

  • The central aim of this study was to explore the feasibility and safety of injecting autologous muscle-derived mesenchymal stem cells into the left recurrent laryngeal nerve in healthy horses. The researchers delivered these cells using an electrical nerve stimulator to guide the nerve’s approach.
  • The intent behind the use of stem cells was to leverage their potential in aiding the recovery of demyelinated nerves, as demonstrated in other animal models.

Methods

  • Stem cells were sourced from the muscle tissues of five healthy Standardbred horses. These tissues were sampled using a semi-automatic biopsy needle.
  • The researchers then monitored the movements of the horses’ larynx using upper-airway video endoscopy.
  • They approached the left recurrent laryngeal nerve with an insulated nerve block needle, applying nerve stimulation that began at 2 mA.
  • Upon observing a successful abduction of the left arytenoid (part of the larynx), they gradually reduced the stimulation intensity until a loss of motor response was noticed at 0.5 mA, at which point they injected the stem cells.

Results

  • After the procedure, two independent examiners, who lacked knowledge about the timepoint of the treatment, evaluated the laryngeal function of the horses. Assessments occurred before the treatment, as well as 1, 7, and 28 days after the stem cell injection.
  • The study found that using an electrical nerve stimulator to approach the recurrent laryngeal nerve is doable and that the electrical stimulation of the nerve is well tolerated by the horses.
  • No changes in laryngeal function were observed in any of the horses after the stem cell injection.

Further Investigation

  • The researchers suggest that more studies should be performed to understand the impact of injecting autologous muscle-derived mesenchymal stem cells in horses affected by Recurrent laryngeal neuropathy (RLN).
  • As a control, one additional horse received an injection of lidocaine which temporarily paralyzed the left arytenoid cartilage, further affirming the proper positioning of the injection needle.

Cite This Article

APA
Sandersen C, Ceusters J, Fourez A, Tosi I, Graide H, Lejeune JP, Serteyn D. (2018). Nerve Stimulator-guided Injection of Autologous Stem Cells Near the Equine Left Recurrent Laryngeal Nerve. J Vis Exp(139), 58023. https://doi.org/10.3791/58023

Publication

Journal of visualized experiments : JoVE
ISSN: 1940-087X
NlmUniqueID: 101313252
Country: United States
Language: English
Issue: 139
PII: 58023

Researcher Affiliations

Sandersen, Charlotte
  • Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Liege; charlotte.sandersen@uliege.be.
Ceusters, Justine
  • Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Liege.
Fourez, Alexia
  • Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Liege.
Tosi, Irene
  • Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Liege.
Graide, Helene
  • Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Liege.
Lejeune, Jean-Philippe
  • Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Liege; Mont-le-Soie Equine Research Centre.
Serteyn, Didier
  • Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Liege; Mont-le-Soie Equine Research Centre.

MeSH Terms

  • Animals
  • Electric Stimulation / instrumentation
  • Electric Stimulation / methods
  • Horse Diseases / pathology
  • Horse Diseases / physiopathology
  • Horses
  • Injections / methods
  • Injections / veterinary
  • Larynx / physiopathology
  • Neurosurgical Procedures / methods
  • Neurosurgical Procedures / veterinary
  • Recurrent Laryngeal Nerve / pathology
  • Recurrent Laryngeal Nerve / physiopathology
  • Stem Cell Transplantation / methods
  • Stem Cell Transplantation / veterinary
  • Stem Cells

References

This article includes 26 references
  1. Duncan ID, Griffiths IR, Madrid RE. A light and electron microscopic study of the neuropathy of equine idiopathic laryngeal hemiplegia.. Neuropathol Appl Neurobiol 1978 Nov-Dec;4(6):483-501.
    pubmed: 745676doi: 10.1111/j.1365-2990.1978.tb01358.xgoogle scholar: lookup
  2. Cahill JI, Goulden BE. Equine laryngeal hemiplegia. Part IV. Muscle pathology.. N Z Vet J 1986 Nov;34(11):186-90.
    pubmed: 16031233doi: 10.1080/00480169.1986.35343google scholar: lookup
  3. Dixon PM, McGorum BC, Railton DI, Hawe C, Tremaine WH, Pickles K, McCann J. Laryngeal paralysis: a study of 375 cases in a mixed-breed population of horses.. Equine Vet J 2001 Sep;33(5):452-8.
    pubmed: 11558739doi: 10.2746/042516401776254790google scholar: lookup
  4. Hahn C, Dixon PM, Robinson E, Wade JF. Review of the pathological changes in equine recurrent laryngeal neuropathy. Havemeyer Monograph Series No. 11 2003; pp. 9–11.
  5. Biasutti S, Dart AJ, Jeffcott LB. A review of recent developments in the clinical application of prosthetic laryngoplasty for recurrent laryngeal neuropathy: Indications, complications and outcome. Equine Veterinary Education 2016;29(6):337–345.
  6. Barnett TP, O'Leary JM, Parkin TD, Dixon PM, Barakzai SZ. Long-term maintenance of arytenoid cartilage abduction and stability during exercise after laryngoplasty in 33 horses.. Vet Surg 2013 Apr;42(3):291-5.
    pubmed: 23452305doi: 10.1111/j.1532-950x.2013.01109.xgoogle scholar: lookup
  7. Ceusters J, Lejeune JP, 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 Apr 6;7(1):696.
    pmc: PMC5429713pubmed: 28386120doi: 10.1038/s41598-017-00803-7google scholar: lookup
  8. Ding DC, Shyu WC, Lin SZ. Mesenchymal stem cells.. Cell Transplant 2011;20(1):5-14.
    pubmed: 21396235doi: 10.3727/096368910xgoogle scholar: lookup
  9. Jiang L, Jones S, Jia X. Stem Cell Transplantation for Peripheral Nerve Regeneration: Current Options and Opportunities.. Int J Mol Sci 2017 Jan 5;18(1).
    pmc: PMC5297728pubmed: 28067783doi: 10.3390/ijms18010094google scholar: lookup
  10. Tohill M, Mantovani C, Wiberg M, Terenghi G. Rat bone marrow mesenchymal stem cells express glial markers and stimulate nerve regeneration.. Neurosci Lett 2004 May 27;362(3):200-3.
    pubmed: 15158014doi: 10.1016/j.neulet.2004.03.077google scholar: lookup
  11. Ji JF, He BP, Dheen ST, Tay SS. Interactions of chemokines and chemokine receptors mediate the migration of mesenchymal stem cells to the impaired site in the brain after hypoglossal nerve injury.. Stem Cells 2004;22(3):415-27.
    pubmed: 15153618doi: 10.1634/stemcells.22-3-415google scholar: lookup
  12. Urmey WF. Using the nerve stimulator for peripheral or plexus nerve blocks.. Minerva Anestesiol 2006 Jun;72(6):467-71.
    pubmed: 16682917
  13. De Andrés J, Sala-Blanch X. Peripheral nerve stimulation in the practice of brachial plexus anesthesia: a review.. Reg Anesth Pain Med 2001 Sep-Oct;26(5):478-83.
    pubmed: 11561271doi: 10.1053/rapm.2001.26485google scholar: lookup
  14. Robinson NE. Consensus statements on equine recurrent laryngeal neuropathy: conclusions of the Havemeyer Workshop. Equine Veterinary Education 2004;16(6):333–336.
  15. . Rank Score Tests. The BMJ 2018.
  16. Hahn CN, Matiasek K, Dixon PM, Molony V, Rodenacker K, Mayhew IG. Histological and ultrastructural evidence that recurrent laryngeal neuropathy is a bilateral mononeuropathy limited to recurrent laryngeal nerves.. Equine Vet J 2008 Nov;40(7):666-72.
    pubmed: 19165936doi: 10.2746/042516408x335711google scholar: lookup
  17. Orlic D, Kajstura J, Chimenti S, Jakoniuk I, Anderson SM, Li B, Pickel J, McKay R, Nadal-Ginard B, Bodine DM, Leri A, Anversa P. Bone marrow cells regenerate infarcted myocardium.. Nature 2001 Apr 5;410(6829):701-5.
    pubmed: 11287958doi: 10.1038/35070587google scholar: lookup
  18. Kopen GC, Prockop DJ, Phinney DG. Marrow stromal cells migrate throughout forebrain and cerebellum, and they differentiate into astrocytes after injection into neonatal mouse brains.. Proc Natl Acad Sci U S A 1999 Sep 14;96(19):10711-6.
    pmc: PMC17948pubmed: 10485891doi: 10.1073/pnas.96.19.10711google scholar: lookup
  19. Dezawa M, Takahashi I, Esaki M, Takano M, Sawada H. Sciatic nerve regeneration in rats induced by transplantation of in vitro differentiated bone-marrow stromal cells.. Eur J Neurosci 2001 Dec;14(11):1771-6.
    pubmed: 11860471doi: 10.1046/j.0953-816x.2001.01814.xgoogle scholar: lookup
  20. Akiyama Y, Radtke C, Honmou O, Kocsis JD. Remyelination of the spinal cord following intravenous delivery of bone marrow cells.. Glia 2002 Sep;39(3):229-36.
    pmc: PMC2605380pubmed: 12203389doi: 10.1002/glia.10102google scholar: lookup
  21. Mahanthappa NK, Anton ES, Matthew WD. Glial growth factor 2, a soluble neuregulin, directly increases Schwann cell motility and indirectly promotes neurite outgrowth.. J Neurosci 1996 Aug 1;16(15):4673-83.
    pmc: PMC6579021pubmed: 8764655doi: 10.1523/jneurosci.16-15-04673.1996google scholar: lookup
  22. Sanchez-Ramos J, Song S, Cardozo-Pelaez F, Hazzi C, Stedeford T, Willing A, Freeman TB, Saporta S, Janssen W, Patel N, Cooper DR, Sanberg PR. Adult bone marrow stromal cells differentiate into neural cells in vitro.. Exp Neurol 2000 Aug;164(2):247-56.
    pubmed: 10915564doi: 10.1006/exnr.2000.7389google scholar: lookup
  23. Farzamfar S, Naseri-Nosar M, Ghanavatinejad A, Vaez A, Zarnani AH, Salehi M. Sciatic nerve regeneration by transplantation of menstrual blood-derived stem cells.. Mol Biol Rep 2017 Oct;44(5):407-412.
    pubmed: 28980161doi: 10.1007/s11033-017-4124-1google scholar: lookup
  24. Morrison SJ, Shah NM, Anderson DJ. Regulatory mechanisms in stem cell biology.. Cell 1997 Feb 7;88(3):287-98.
    pubmed: 9039255doi: 10.1016/s0092-8674(00)81867-xgoogle scholar: lookup
  25. Brini AT, Amodeo G, Ferreira LM, Milani A, Niada S, Moschetti G, Franchi S, Borsani E, Rodella LF, Panerai AE, Sacerdote P. Therapeutic effect of human adipose-derived stem cells and their secretome in experimental diabetic pain.. Sci Rep 2017 Aug 29;7(1):9904.
    pmc: PMC5575274pubmed: 28851944doi: 10.1038/s41598-017-09487-5google scholar: lookup
  26. Liu W, Yu F, Zhou Z, Li YC, Fan D, Zhu K. Autologous Bone Marrow-Derived Stem Cells for Treating Diabetic Neuropathy in Metabolic Syndrome.. Biomed Res Int 2017;2017:8945310.
    pmc: PMC5643093pubmed: 29098161doi: 10.1155/2017/8945310google scholar: lookup

Citations

This article has been cited 1 times.
  1. Cequier A, Sanz C, Rodellar C, Barrachina L. The Usefulness of Mesenchymal Stem Cells beyond the Musculoskeletal System in Horses. Animals (Basel) 2021 Mar 25;11(4).
    doi: 10.3390/ani11040931pubmed: 33805967google scholar: lookup
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