FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Frontiers in veterinary science2023; 10; 1117829; doi: 10.3389/fvets.2023.1117829

Cytokines in equine platelet lysate and related blood products.

Abstract: In equine medicine, the use of regenerative therapeutics has gained growing attention, but is still a new and complex field with room for improvement. Platelet lysate (PL) can be used as therapeutic agent but is also a promising supplement for the culture of multipotent mesenchymal stromal cells. To enable a targeted use of PL both in clinic and laboratory, it is crucial to learn more details on its effective ingredients. While so far, mainly growth factor components have been analyzed in platelet-based products such as PL, the current study focuses on the content of cytokines in serum, plasma, platelet concentrate and PL. Blood was harvested from 20 clinically healthy horses and subjected to blood count and chemistry analysis, as well as to further processing to PL. Plasma and platelet concentrate were produced by a buffy-coat-based method and PL was produced from the concentrate by freeze-thawing. Samples from each horse were analyzed regarding interleukin (IL)-1β, -4, -6 and -10, interferon-γ and tumor necrosis factor-α concentrations using sandwich ELISAs. Cytokine concentrations in serum, plasma, concentrate and PL were similar and correlated significantly. However, there was a large inter-individual variability in cytokine concentrations between the different donor horses. The samples from some donor animals had overall very high cytokine concentrations, while samples from other donors had no measurable cytokine ingredient. This pattern was observed for all cytokines. There was a noticeable link between high cytokine concentrations in the blood products and abnormal findings in blood chemistry. Cytokine concentrations in samples from horses with abnormal findings were significantly higher than in samples from the remaining horses. The interindividual differences in cytokine concentrations could be highly relevant when using PL for therapy and cell culture, as the mode of action of the PL is likely changed depending on the presence of pro- and anti-inflammatory cytokines. Blood chemistry might be useful to predict cytokine concentrations in blood products.
Copyright © 2023 Moellerberndt, Hagen, Niebert, Büttner and Burk.
Get Full Text
Publication Date: 2023-03-09 PubMed ID: 36968472PubMed Central: PMC10033973DOI: 10.3389/fvets.2023.1117829Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

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 article studies the content of specific proteins called cytokines in horse blood products such as platelet lysate, serum, plasma, and platelet concentrate. The article explores the relationship between cytokine concentrations in these products, their relationship with abnormal blood chemistry findings, and the potential implications for therapeutic use and cell culture.

Objective and Methodology of the Research

  • The research aims to investigate cytokine contents in various equine blood products. Cytokines are proteins involved in cell signaling and can trigger both inflammatory and anti-inflammatory responses. The products focused on are platelet lysate (PL), serum, plasma, and platelet concentrate.
  • The study was carried out using blood from 20 healthy horses. The blood underwent various tests including blood count, chemistry analysis, and further processing to PL. The plasma and platelet concentrate were generated by using a method known as the buffy-coat-based method, and the PL was produced from the concentrate through freeze-thawing.
  • The samples from each horse were analyzed for concentrations of different cytokines, including interleukin (IL)-1β, -4, -6, and -10, interferon-γ, and tumor necrosis factor-α, using method called sandwich ELISAs.

Findings from the Research

  • The study found that the cytokine concentrations in serum, plasma, concentrate, and PL were similar and correlated significantly. However, there was substantial variability in cytokine concentrations between different donor horses.
  • Some samples had very high concentrations of cytokines, while others had no measurable cytokine elements. This heterogeneity was observed across all types of cytokines assessed.
  • Fascinatingly, a significant connection was discovered between high cytokine concentrations in blood products and abnormal blood chemistry findings. In other words, horses with abnormal blood chemistry tended to have blood products with higher cytokine concentrations compared to horses with normal chemistry results.

Implications of the Findings

  • Given the role of cytokines in causing inflammation and in regulating immune responses, the different concentrations in individual horses could have important implications when using PL for therapy and cell culture.
  • The therapeutic effectiveness and how PL works could alter depending on the levels of pro-inflammatory and anti-inflammatory cytokines. This necessitates examining and understanding the cytokine content of these blood products before they are used therapeutically.
  • Furthermore, blood chemistry may be a useful indicator to predict cytokine concentrations in blood products. This could offer an additional method for clinicians to tailor their treatment plans more effectively.

Cite This Article

APA
Moellerberndt J, Hagen A, Niebert S, Büttner K, Burk J. (2023). Cytokines in equine platelet lysate and related blood products. Front Vet Sci, 10, 1117829. https://doi.org/10.3389/fvets.2023.1117829

Publication

Frontiers in veterinary science
ISSN: 2297-1769
NlmUniqueID: 101666658
Country: Switzerland
Language: English
Volume: 10
Pages: 1117829
PII: 1117829

Researcher Affiliations

Moellerberndt, Julia
  • Equine Clinic (Surgery, Orthopedics), Justus-Liebig-University Giessen, Giessen, Germany.
Hagen, Alina
  • Equine Clinic (Surgery, Orthopedics), Justus-Liebig-University Giessen, Giessen, Germany.
Niebert, Sabine
  • Equine Clinic (Surgery, Orthopedics), Justus-Liebig-University Giessen, Giessen, Germany.
Büttner, Kathrin
  • Unit for Biomathematics and Data Processing, Faculty of Veterinary Medicine, Justus-Liebig-University Giessen, Giessen, Germany.
Burk, Janina
  • Equine Clinic (Surgery, Orthopedics), Justus-Liebig-University Giessen, Giessen, Germany.

Conflict of Interest Statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

References

This article includes 44 references
  1. Golebiewska EM, Poole AW. Platelet secretion: From haemostasis to wound healing and beyond.. Blood Rev 2015 May;29(3):153-62.
    doi: 10.1016/j.blre.2014.10.003pmc: PMC4452143pubmed: 25468720google scholar: lookup
  2. Scully D, Naseem KM, Matsakas A. Platelet biology in regenerative medicine of skeletal muscle.. Acta Physiol (Oxf) 2018 Jul;223(3):e13071.
    doi: 10.1111/apha.13071pubmed: 29633517google scholar: lookup
  3. Jiang P, Zhang Y, Ru B, Yang Y, Vu T, Paul R, Mirza A, Altan-Bonnet G, Liu L, Ruppin E, Wakefield L, Wucherpfennig KW. Systematic investigation of cytokine signaling activity at the tissue and single-cell levels.. Nat Methods 2021 Oct;18(10):1181-1191.
    doi: 10.1038/s41592-021-01274-5pmc: PMC8493809pubmed: 34594031google scholar: lookup
  4. Burnouf T, Goubran HA, Chen TM, Ou KL, El-Ekiaby M, Radosevic M. Blood-derived biomaterials and platelet growth factors in regenerative medicine.. Blood Rev 2013 Mar;27(2):77-89.
    doi: 10.1016/j.blre.2013.02.001pubmed: 23434399google scholar: lookup
  5. Santos SCNDS, Sigurjonsson ÓE, Custódio CA, Mano JFCDL. Blood Plasma Derivatives for Tissue Engineering and Regenerative Medicine Therapies.. Tissue Eng Part B Rev 2018 Dec;24(6):454-462.
    doi: 10.1089/ten.teb.2018.0008pmc: PMC6443031pubmed: 29737237google scholar: lookup
  6. Hagen A, Lehmann H, Aurich S, Bauer N, Melzer M, Moellerberndt J, Patané V, Schnabel CL, Burk J. Scalable Production of Equine Platelet Lysate for Multipotent Mesenchymal Stromal Cell Culture.. Front Bioeng Biotechnol 2020;8:613621.
    doi: 10.3389/fbioe.2020.613621pmc: PMC7859354pubmed: 33553119google scholar: lookup
  7. Strandberg G, Sellberg F, Sommar P, Ronaghi M, Lubenow N, Knutson F, Berglund D. Standardizing the freeze-thaw preparation of growth factors from platelet lysate.. Transfusion 2017 Apr;57(4):1058-1065.
    doi: 10.1111/trf.13998pubmed: 28182293google scholar: lookup
  8. Altaie A, Owston H, Jones E. Use of platelet lysate for bone regeneration - are we ready for clinical translation?. World J Stem Cells 2016 Feb 26;8(2):47-55.
    doi: 10.4252/wjsc.v8.i2.47pmc: PMC4766250pubmed: 26981170google scholar: lookup
  9. Burnouf T, Strunk D, Koh MB, Schallmoser K. Human platelet lysate: Replacing fetal bovine serum as a gold standard for human cell propagation?. Biomaterials 2016 Jan;76:371-87.
    doi: 10.1016/j.biomaterials.2015.10.065pubmed: 26561934google scholar: lookup
  10. Bieback K. Platelet lysate as replacement for fetal bovine serum in mesenchymal stromal cell cultures.. Transfus Med Hemother 2013 Oct;40(5):326-35.
    doi: 10.1159/000354061pmc: PMC3822281pubmed: 24273486google scholar: lookup
  11. Fekete N, Gadelorge M, Fürst D, Maurer C, Dausend J, Fleury-Cappellesso S, Mailänder V, Lotfi R, Ignatius A, Sensebé L, Bourin P, Schrezenmeier H, Rojewski MT. Platelet lysate from whole blood-derived pooled platelet concentrates and apheresis-derived platelet concentrates for the isolation and expansion of human bone marrow mesenchymal stromal cells: production process, content and identification of active components.. Cytotherapy 2012 May;14(5):540-54.
    doi: 10.3109/14653249.2012.655420pmc: PMC3400099pubmed: 22296115google scholar: lookup
  12. Zamani M, Yaghoubi Y, Movassaghpour A, Shakouri K, Mehdizadeh A, Pishgahi A, Yousefi M. Novel therapeutic approaches in utilizing platelet lysate in regenerative medicine: Are we ready for clinical use?. J Cell Physiol 2019 Aug;234(10):17172-17186.
    doi: 10.1002/jcp.28496pubmed: 30912141google scholar: lookup
  13. Abu-Ameerh MA, Jafar HD, Hasan MH, Al Bdour MD, Msallam M, Ababneh OH, Alhattab DM, Al-Kurdi B, Awidi AA, Awidi AS. Platelet lysate promotes re-epithelialization of persistent epithelial defects: a pilot study.. Int Ophthalmol 2019 Jul;39(7):1483-1490.
    doi: 10.1007/s10792-018-0968-1pubmed: 29978342google scholar: lookup
  14. Textor JA, Clark KC, Walker NJ, Aristizobal FA, Kol A, LeJeune SS, Bledsoe A, Davidyan A, Gray SN, Bohannon-Worsley LK, Woolard KD, Borjesson DL. Allogeneic Stem Cells Alter Gene Expression and Improve Healing of Distal Limb Wounds in Horses.. Stem Cells Transl Med 2018 Jan;7(1):98-108.
    doi: 10.1002/sctm.17-0071pmc: PMC5746157pubmed: 29063737google scholar: lookup
  15. Sovkova V, Vocetkova K, Rampichova M, Mickova A, Buzgo M, Lukasova V, Dankova J, Filova E, Necas A, Amler E. Platelet lysate as a serum replacement for skin cell culture on biomimetic PCL nanofibers.. Platelets 2018 Jun;29(4):395-405.
    doi: 10.1080/09537104.2017.1316838pubmed: 28649896google scholar: lookup
  16. Jochems CE, van der Valk JB, Stafleu FR, Baumans V. The use of fetal bovine serum: ethical or scientific problem?. Altern Lab Anim 2002 Mar-Apr;30(2):219-27.
    doi: 10.1177/026119290203000208pubmed: 11971757google scholar: lookup
  17. van der Valk J, Mellor D, Brands R, Fischer R, Gruber F, Gstraunthaler G, Hellebrekers L, Hyllner J, Jonker FH, Prieto P, Thalen M, Baumans V. The humane collection of fetal bovine serum and possibilities for serum-free cell and tissue culture.. Toxicol In Vitro 2004 Feb;18(1):1-12.
    doi: 10.1016/j.tiv.2003.08.009pubmed: 14630056google scholar: lookup
  18. Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop Dj, Horwitz E. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement.. Cytotherapy 2006;8(4):315-7.
    doi: 10.1080/14653240600855905pubmed: 16923606google scholar: lookup
  19. Hagen A, Niebert S, Brandt VP, Holland H, Melzer M, Wehrend A, Burk J. Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate.. Front Vet Sci 2022;9:890302.
    doi: 10.3389/fvets.2022.890302pmc: PMC9395693pubmed: 36016806google scholar: lookup
  20. Gilbertie JM, Long JM, Schubert AG, Berglund AK, Schaer TP, Schnabel LV. Pooled Platelet-Rich Plasma Lysate Therapy Increases Synoviocyte Proliferation and Hyaluronic Acid Production While Protecting Chondrocytes From Synoviocyte-Derived Inflammatory Mediators.. Front Vet Sci 2018;5:150.
    doi: 10.3389/fvets.2018.00150pmc: PMC6039577pubmed: 30023361google scholar: lookup
  21. Mussano F, Genova T, Munaron L, Petrillo S, Erovigni F, Carossa S. Cytokine, chemokine, and growth factor profile of platelet-rich plasma.. Platelets 2016 Jul;27(5):467-71.
    doi: 10.3109/09537104.2016.1143922pubmed: 26950533google scholar: lookup
  22. Hillmann A, Paebst F, Brehm W, Piehler D, Schubert S, Tárnok A, Burk J. A novel direct co-culture assay analyzed by multicolor flow cytometry reveals context- and cell type-specific immunomodulatory effects of equine mesenchymal stromal cells.. PLoS One 2019;14(6):e0218949.
    doi: 10.1371/journal.pone.0218949pmc: PMC6597077pubmed: 31247035google scholar: lookup
  23. Bauer N, Keresztes M. Blutuntersuchung. Handbuch Pferdepraxis 4th edition, Stuttgart: Enke (2016). p. 27–32.
  24. Heinemann D. Ex Vivo In Vitro Studies of the Impact of Zylexis® on the Immune Response Following Vaccination with Resequin® NN Plus with Special Consideration of the EHV-1/-4 Specific Antibody Response, the Immune Status the Cytokine Gene Expression Patterns in Horse Blood. Inaugural-Dissertation University of Berlin, Veterinary Medicine (2008).
  25. Yoshihara R, Shiozawa S, Imai Y, Fujita T. Tumor necrosis factor alpha and interferon gamma inhibit proliferation and alkaline phosphatase activity of human osteoblastic SaOS-2 cell line.. Lymphokine Res 1990 Spring;9(1):59-66.
    pubmed: 2109156
  26. Le-Thi-Phuong T, Thirion G, Coutelier JP. Distinct gamma interferon-production pathways in mice infected with lactate dehydrogenase-elevating virus.. J Gen Virol 2007 Nov;88(Pt 11):3063-3066.
    doi: 10.1099/vir.0.83242-0pubmed: 17947531google scholar: lookup
  27. Peng M, Yin N, Chhangawala S, Xu K, Leslie CS, Li MO. Aerobic glycolysis promotes T helper 1 cell differentiation through an epigenetic mechanism.. Science 2016 Oct 28;354(6311):481-484.
    doi: 10.1126/science.aaf6284pmc: PMC5539971pubmed: 27708054google scholar: lookup
  28. Farrar MA, Schreiber RD. The molecular cell biology of interferon-gamma and its receptor.. Annu Rev Immunol 1993;11:571-611.
    doi: 10.1146/annurev.iy.11.040193.003035pubmed: 8476573google scholar: lookup
  29. Jansen JE, Aschenbrenner D, Uhlig HH, Coles MC, Gaffney EA. A method for the inference of cytokine interaction networks.. PLoS Comput Biol 2022 Jun;18(6):e1010112.
    doi: 10.1371/journal.pcbi.1010112pmc: PMC9216621pubmed: 35731827google scholar: lookup
  30. Liu C, Chu D, Kalantar-Zadeh K, George J, Young HA, Liu G. Cytokines: From Clinical Significance to Quantification.. Adv Sci (Weinh) 2021 Aug;8(15):e2004433.
    doi: 10.1002/advs.202004433pmc: PMC8336501pubmed: 34114369google scholar: lookup
  31. Samara O, Alajlouni J, Al Najar M, Saleh M, Ryalat N, Gharaibeh A. Intra-articular autologous platelet lysates produce positive mri structural changes in early and intermediate knee osteoarthrosis. Pakistan J Radiol (2017) 27:14–8.
  32. Al-Ajlouni J, Awidi A, Samara O, Al-Najar M, Tarwanah E, Saleh M, Awidi M, Hassan FA, Samih M, Bener A, Dweik M. Safety and Efficacy of Autologous Intra-articular Platelet Lysates in Early and Intermediate Knee Osteoarthrosis in Humans: A Prospective Open-Label Study.. Clin J Sport Med 2015 Nov;25(6):524-8.
    doi: 10.1097/JSM.0000000000000166pubmed: 25387167google scholar: lookup
  33. Tyrnenopoulou P, Diakakis N, Karayannopoulou M, Savvas I, Koliakos G. Evaluation of intra-articular injection of autologous platelet lysate (PL) in horses with osteoarthritis of the distal interphalangeal joint.. Vet Q 2016 Jun;36(2):56-62.
    doi: 10.1080/01652176.2016.1141257pubmed: 26828234google scholar: lookup
  34. Markazi R, Soltani-Zangbar MS, Zamani M, Eghbal-Fard S, Motavalli R, Kamrani A, Dolati S, Ahmadi M, Aghebati-Maleki L, Mehdizadeh A, Eslamian F, Pishgahi A, Yousefi M. Platelet lysate and tendon healing: comparative analysis of autologous frozen-thawed PRP and ketorolac tromethamine in the treatment of patients with rotator cuff tendinopathy.. Growth Factors 2022 Nov;40(5-6):163-174.
    doi: 10.1080/08977194.2022.2093198pubmed: 36026559google scholar: lookup
  35. Chen LW, Huang CJ, Tu WH, Lu CJ, Sun YC, Lin SY, Chen WL. The corneal epitheliotrophic abilities of lyophilized powder form human platelet lysates.. PLoS One 2018;13(3):e0194345.
    doi: 10.1371/journal.pone.0194345pmc: PMC5856377pubmed: 29547658google scholar: lookup
  36. Huang CJ, Sun YC, Christopher K, Pai AS, Lu CJ, Hu FR, Lin SY, Chen WL. Comparison of corneal epitheliotrophic capacities among human platelet lysates and other blood derivatives.. PLoS One 2017;12(2):e0171008.
    doi: 10.1371/journal.pone.0171008pmc: PMC5289502pubmed: 28152010google scholar: lookup
  37. Geremicca W, Fonte C, Vecchio S. Blood components for topical use in tissue regeneration: evaluation of corneal lesions treated with platelet lysate and considerations on repair mechanisms.. Blood Transfus 2010 Apr;8(2):107-12.
    doi: 10.2450/2009.0091-09pmc: PMC2851214pubmed: 20383304google scholar: lookup
  38. Etulain J. Platelets in wound healing and regenerative medicine.. Platelets 2018 Sep;29(6):556-568.
    doi: 10.1080/09537104.2018.1430357pubmed: 29442539google scholar: lookup
  39. Nurden AT, Nurden P, Sanchez M, Andia I, Anitua E. Platelets and wound healing.. Front Biosci 2008 May 1;13:3532-48.
    doi: 10.2741/2947pubmed: 18508453google scholar: lookup
  40. Anitua E, Andia I, Ardanza B, Nurden P, Nurden AT. Autologous platelets as a source of proteins for healing and tissue regeneration.. Thromb Haemost 2004 Jan;91(1):4-15.
    doi: 10.1160/TH03-07-0440pubmed: 14691563google scholar: lookup
  41. Naskou MC, Sumner SM, Chocallo A, Kemelmakher H, Thoresen M, Copland I, Galipeau J, Peroni JF. Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells.. Stem Cell Res Ther 2018 Mar 22;9(1):75.
    doi: 10.1186/s13287-018-0823-3pmc: PMC5863827pubmed: 29566772google scholar: lookup
  42. Yaneselli K, Barrachina L, Remacha AR, Algorta A, Vitoria A, Cequier A, Romero A, Vázquez FJ, Maisonnave J, Rodellar C. Effect of allogeneic platelet lysate on equine bone marrow derived mesenchymal stem cell characteristics, including immunogenic and immunomodulatory gene expression profile.. Vet Immunol Immunopathol 2019 Nov;217:109944.
    doi: 10.1016/j.vetimm.2019.109944pubmed: 31563725google scholar: lookup
  43. Barrachina L, Remacha AR, Romero A, Vázquez FJ, Albareda J, Prades M, Gosálvez J, Roy R, Zaragoza P, Martín-Burriel I, Rodellar C. Priming Equine Bone Marrow-Derived Mesenchymal Stem Cells with Proinflammatory Cytokines: Implications in Immunomodulation-Immunogenicity Balance, Cell Viability, and Differentiation Potential.. Stem Cells Dev 2017 Jan 1;26(1):15-24.
    doi: 10.1089/scd.2016.0209pubmed: 27712399google scholar: lookup
  44. Brandt L, Schubert S, Scheibe P, Brehm W, Franzen J, Gross C, Burk J. Tenogenic Properties of Mesenchymal Progenitor Cells Are Compromised in an Inflammatory Environment.. Int J Mol Sci 2018 Aug 28;19(9).
    doi: 10.3390/ijms19092549pmc: PMC6163784pubmed: 30154348google scholar: lookup

Citations

This article has been cited 7 times.
  1. Seidel SRT, Fülber J, Barbosa ÂP, Penatti NMA, Demasi M, Baccarin RYA. Medium-term storage of platelet-derived orthobiologics: a feasible alternative for equine practice. Front Vet Sci 2025;12:1720164.
    doi: 10.3389/fvets.2025.1720164pubmed: 41602624google scholar: lookup
  2. Roth SP, Liso G, Brehm W, Wagner B, Schnabel CL, Troillet A. Selected cytokine and chemokine concentrations in equine autologous conditioned serum are similar under defined and practically relevant storage conditions. Front Vet Sci 2025;12:1588240.
    doi: 10.3389/fvets.2025.1588240pubmed: 40496923google scholar: lookup
  3. Barshick MR, Ely KM, Mogge KC, Chance LM, Johnson SE. Methylsulfonylmethane (MSM) Supplementation in Adult Horses Supports Improved Skeletal Muscle Inflammatory Gene Expression Following Exercise. Animals (Basel) 2025 Jan 14;15(2).
    doi: 10.3390/ani15020215pubmed: 39858215google scholar: lookup
  4. Egli P, Boone L, Huber L, Higgins C, Gaonkar PP, Arrington J, Naskou MC, Peroni J, Gordon J, Lascola KM. Pilot study characterizing a single pooled preparation of equine platelet lysate for nebulization in the horse. Front Vet Sci 2024;11:1488942.
    doi: 10.3389/fvets.2024.1488942pubmed: 39726585google scholar: lookup
  5. Yaneselli K, Ávila G, Rossi A, Rial A, Castro S, Estradé MJ, Suárez G, Algorta A. Impact of different formulations of platelet lysate on proliferative and immune profile of equine mesenchymal stromal cells. Front Vet Sci 2024;11:1410855.
    doi: 10.3389/fvets.2024.1410855pubmed: 39161460google scholar: lookup
  6. Moellerberndt J, Niebert S, Fey K, Hagen A, Burk J. Impact of platelet lysate on immunoregulatory characteristics of equine mesenchymal stromal cells. Front Vet Sci 2024;11:1385395.
    doi: 10.3389/fvets.2024.1385395pubmed: 38725585google scholar: lookup
  7. Bourebaba L, Serwotka-Suszczak A, Bourebaba N, Zyzak M, Marycz K. The PTP1B Inhibitor Trodusquemine (MSI-1436) Improves Glucose Uptake in Equine Metabolic Syndrome Affected Liver through Anti-Inflammatory and Antifibrotic Activity. Int J Inflam 2023;2023:3803056.
    doi: 10.1155/2023/3803056pubmed: 37808009google scholar: lookup
Subscribe to our newsletter
Join 99,727 horse owners like you who receive our email updates on horse health and nutrition.