Immunomodulatory Properties of Adipose-Derived Stem Cells Treated with 5-Azacytydine and Resveratrol on Peripheral Blood Mononuclear Cells and Macrophages in Metabolic Syndrome Animals.
Abstract: Endocrine disorders, including equine metabolic syndrome (EMS), are a serious issue in veterinary medicine and horse breeding. Furthermore, EMS was shown to affect the cytophysiological properties of adipose-derived stem cells, reducing their therapeutic potential. However, it was shown that those cells can be rejuvenated while using a combination of two chemicals: 5-azacytydine (AZA) and resveratrol (RES). In the present study, we decided to evaluate the immunomodulatory properties of AZA/RES-treated adipose-derived stem cells (ASC) isolated from EMS horses (ASC). Thus, we co-cultured ASC with peripheral blood mononuclear cells (PBMC) and RAW264.7 macrophages. Most attention was placed on regulatory T lymphocytes (T), as well as the messenger RNA (mRNA) and protein levels of several cytokines (tumor necrosis factor α (TNF-α), interleukin (IL)-6, IL-10, and IL-1β). Moreover, we also investigated the expression of genes related to auto- and mitophagy in both PBMCs and ASCs. PBMCs were obtained from healthy and EMS-suffering individuals and were co-cultured with ASCs that were isolated from healthy and EMS horses cultured in control conditions and with AZA/RES. We discovered that cells treated with AZA/RES increase the T number while co-cultured with PBMCs. Moreover, the co-culture of PBMCs with AZA/RES-treated ASC induced mitophagy in PBMCs. Furthermore, ASC pre-treated with AZA/RES displayed anti-inflammatory properties, as decreased levels of TNF-α, nitric oxide (NO), and IL-6 were observed in those cells in comparison with their untreated counterparts in the co-culture with RAW264.7 macrophages. In summary, we demonstrated that ASC treated with AZA/RES displayed increased anti-inflammatory properties, and was able to regulate and activate the T-related anti-inflammatory response.
Publication Date: 2018-10-24 PubMed ID: 30356025PubMed Central: PMC6262510DOI: 10.3390/jcm7110383Google 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.
- 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 centers around the study of immunomodulatory properties of rejuvenated stem cells extracted from horses with equine metabolic syndrome (EMS) when treated with a combination of two chemicals: 5-azacytydine (AZA) and resveratrol (RES). The adjective ‘immunomodulatory’ refers to the ability to modify or regulate the functioning of the immune system. The study found that the treated stem cells not only displayed increased anti-inflammatory properties but also successfully activated T-related anti-inflammatory response.
Research Methodology
- The study conducted experiments using adipose-derived stem cells (ASC), isolated from EMS affected horses, which were treated with a chemical combination of 5-azacytydine (AZA) and resveratrol (RES).
- These treated cells were co-cultured with peripheral blood mononuclear cells (PBMC) and RAW264.7 macrophages.
- Focused study was conducted on the regulatory T lymphocytes.
- The changes in messenger RNA (mRNA) and protein levels of several cytokines, namely, tumor necrosis factor α (TNF-α), interleukin (IL)-6, IL-10, and IL-1β, were observed.
- Futhermore, the research investigated into the expression of genes related to auto- and mitophagy in both PBMCs and ASCs.
Findings and Conclusions
- The study discovered that the cells treated with AZA/RES increased the number of regulatory T lymphocytes, which are crucial in preventing autoimmune diseases by suppressing overactive immune responses, when co-cultured with PBMCs.
- Successful induction of mitophagy in PBMCs was observed when they were co-cultured with AZA/RES-treated ASC. Mitophagy is the selective degradation of mitochondria by autophagy, which is often initiated in response to cellular stress.
- Furthermore, it was noticed that ASC pre-treated with AZA/RES exhibited anti-inflammatory properties, as witnessed by the decrease in the levels of TNF-α, nitric oxide (NO), and IL-6 compared to their untreated counterparts when co-cultured with RAW264.7 macrophages.
- In conclusion, the study demonstrated that ASC treated with AZA/RES showcased increased anti-inflammatory properties and acquired the capability to regulate and activate T cell-related anti-inflammatory response.
Impact of the Research
- This research contributes significantly to the realm of veterinary medicine and horse breeding as endocrine disorders, especially EMS, are persistent issues in these fields.
- The study showcases the potential of the chemical treatment of ASC, using AZA and RES, in rejuvenating the immunomodulatory properties of cells, which can open avenues to new approaches in treating EMS and potentially other endocrine disorders in horses.
- The finding that the treated ASCs can effectively stimulate the T cell-related anti-inflammatory response also adds a new dimension to the ongoing research in immune system regulation and autoimmunity.
Cite This Article
APA
Kornicka K, Śmieszek A, Węgrzyn AS, Röcken M, Marycz K.
(2018).
Immunomodulatory Properties of Adipose-Derived Stem Cells Treated with 5-Azacytydine and Resveratrol on Peripheral Blood Mononuclear Cells and Macrophages in Metabolic Syndrome Animals.
J Clin Med, 7(11), 383.
https://doi.org/10.3390/jcm7110383 Publication
Researcher Affiliations
- Department of Experimental Biology, The Faculty of Biology and Animal Science, University of Environmental and Life Sciences Wroclaw 50-375, Poland. kornicka.katarzyna@gmail.com.
- Department of Experimental Biology, The Faculty of Biology and Animal Science, University of Environmental and Life Sciences Wroclaw 50-375, Poland. smieszek.agnieszka@gmail.com.
- PORT Polish Center for Technology Development, Wrocław 54-066, Poland. agnieszka.wegrzyn@eitplus.pl.
- Faculty of Veterinary Medicine, Equine Clinic-Equine Surgery, Justus-Liebig-University, Giessen 35392, Germany. Michael.Roecken@vetmed.uni-giessen.de.
- Department of Experimental Biology, The Faculty of Biology and Animal Science, University of Environmental and Life Sciences Wroclaw 50-375, Poland. krzysztofmarycz@interia.pl.
- Faculty of Veterinary Medicine, Equine Clinic-Equine Surgery, Justus-Liebig-University, Giessen 35392, Germany. krzysztofmarycz@interia.pl.
Grant Funding
- 2016/21/B/NZ7/01111 / Narodowe Centrum Nauki
- - / Krajowy Naukowy Osrodek Wiodacy
Conflict of Interest Statement
The authors declare that there is no conflict of interest.
References
This article includes 44 references
- Yu H, Côté P, Shearer HM, Wong JJ, Sutton DA, Randhawa KA, Varatharajan S, Southerst D, Mior SA, Ameis A, Stupar M, Nordin M, van der Velde GM, Carroll L, Jacobs CL, Taylor-Vaisey AL, Abdulla S, Shergill Y. Effectiveness of passive physical modalities for shoulder pain: systematic review by the Ontario protocol for traffic injury management collaboration.. Phys Ther 2015 Mar;95(3):306-18.
- Castillo DT, Lacefield K, Baca JC, Blankenship A, Qualls C. Effectiveness of Group-Delivered Cognitive Therapy and Treatment Length in Women Veterans with PTSD.. Behav Sci (Basel) 2014 Mar;4(1):31-41.
- Cordeiro-Spinetti E, de Mello W, Trindade LS, Taub DD, Taichman RS, Balduino A. Human bone marrow mesenchymal progenitors: perspectives on an optimized in vitro manipulation.. Front Cell Dev Biol 2014;2:7.
- Machado Cde V, Telles PD, Nascimento IL. Immunological characteristics of mesenchymal stem cells.. Rev Bras Hematol Hemoter 2013;35(1):62-7.
- Chan JL, Tang KC, Patel AP, Bonilla LM, Pierobon N, Ponzio NM, Rameshwar P. Antigen-presenting property of mesenchymal stem cells occurs during a narrow window at low levels of interferon-gamma.. Blood 2006 Jun 15;107(12):4817-24.
- Puissant B, Barreau C, Bourin P, Clavel C, Corre J, Bousquet C, Taureau C, Cousin B, Abbal M, Laharrague P, Penicaud L, Casteilla L, Blancher A. Immunomodulatory effect of human adipose tissue-derived adult stem cells: comparison with bone marrow mesenchymal stem cells.. Br J Haematol 2005 Apr;129(1):118-29.
- Glennie S, Soeiro I, Dyson PJ, Lam EW, Dazzi F. Bone marrow mesenchymal stem cells induce division arrest anergy of activated T cells.. Blood 2005 Apr 1;105(7):2821-7.
- Selmani Z, Naji A, Zidi I, Favier B, Gaiffe E, Obert L, Borg C, Saas P, Tiberghien P, Rouas-Freiss N, Carosella ED, Deschaseaux F. Human leukocyte antigen-G5 secretion by human mesenchymal stem cells is required to suppress T lymphocyte and natural killer function and to induce CD4+CD25highFOXP3+ regulatory T cells.. Stem Cells 2008 Jan;26(1):212-22.
- Marycz K, Kornicka K, Szlapka-Kosarzewska J, Weiss C. Excessive Endoplasmic Reticulum Stress Correlates with Impaired Mitochondrial Dynamics, Mitophagy and Apoptosis, in Liver and Adipose Tissue, but Not in Muscles in EMS Horses.. Int J Mol Sci 2018 Jan 6;19(1).
- Holbrook TC, Tipton T, McFarlane D. Neutrophil and cytokine dysregulation in hyperinsulinemic obese horses.. Vet Immunol Immunopathol 2012 Jan 15;145(1-2):283-9.
- Burns TA, Geor RJ, Mudge MC, McCutcheon LJ, Hinchcliff KW, Belknap JK. Proinflammatory cytokine and chemokine gene expression profiles in subcutaneous and visceral adipose tissue depots of insulin-resistant and insulin-sensitive light breed horses.. J Vet Intern Med 2010 Jul-Aug;24(4):932-9.
- Vick MM, Murphy BA, Sessions DR, Reedy SE, Kennedy EL, Horohov DW, Cook RF, Fitzgerald BP. Effects of systemic inflammation on insulin sensitivity in horses and inflammatory cytokine expression in adipose tissue.. Am J Vet Res 2008 Jan;69(1):130-9.
- Marycz K, Kornicka K, Basinska K, Czyrek A. Equine Metabolic Syndrome Affects Viability, Senescence, and Stress Factors of Equine Adipose-Derived Mesenchymal Stromal Stem Cells: New Insight into EqASCs Isolated from EMS Horses in the Context of Their Aging.. Oxid Med Cell Longev 2016;2016:4710326.
- Marycz K, Kornicka K, Grzesiak J, Śmieszek A, Szłapka J. Corrigendum to "Macroautophagy and Selective Mitophagy Ameliorate Chondrogenic Differentiation Potential in Adipose Stem Cells of Equine Metabolic Syndrome: New Findings in the Field of Progenitor Cells Differentiation".. Oxid Med Cell Longev 2017;2017:3861790.
- Marycz K, Kornicka K, Marędziak M, Golonka P, Nicpoń J. Equine metabolic syndrome impairs adipose stem cells osteogenic differentiation by predominance of autophagy over selective mitophagy.. J Cell Mol Med 2016 Dec;20(12):2384-2404.
- Kornicka K, Marycz K, Marędziak M, Tomaszewski KA, Nicpoń J. The effects of the DNA methyltranfserases inhibitor 5-Azacitidine on ageing, oxidative stress and DNA methylation of adipose derived stem cells.. J Cell Mol Med 2017 Feb;21(2):387-401.
- Kornicka K, Nawrocka D, Lis-Bartos A, Marędziak M, Marycz K. Polyurethane–polylactide-based material doped with resveratrol decreases senescence and oxidative stress of adipose-derived mesenchymal stromal stem cell (ASCs). RSC Adv 2017;7:24070–24084.
- Kuballa P, Nolte WM, Castoreno AB, Xavier RJ. Autophagy and the immune system.. Annu Rev Immunol 2012;30:611-46.
- Marycz K, Weiss C, Śmieszek A, Kornicka K. Evaluation of Oxidative Stress and Mitophagy during Adipogenic Differentiation of Adipose-Derived Stem Cells Isolated from Equine Metabolic Syndrome (EMS) Horses.. Stem Cells Int 2018;2018:5340756.
- Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.. Anal Biochem 1987 Apr;162(1):156-9.
- van der Weerd K, Dik WA, Schrijver B, Schweitzer DH, Langerak AW, Drexhage HA, Kiewiet RM, van Aken MO, van Huisstede A, van Dongen JJ, van der Lelij AJ, Staal FJ, van Hagen PM. Morbidly obese human subjects have increased peripheral blood CD4+ T cells with skewing toward a Treg- and Th2-dominated phenotype.. Diabetes 2012 Feb;61(2):401-8.
- Shirakawa K, Yan X, Shinmura K, Endo J, Kataoka M, Katsumata Y, Yamamoto T, Anzai A, Isobe S, Yoshida N, Itoh H, Manabe I, Sekai M, Hamazaki Y, Fukuda K, Minato N, Sano M. Obesity accelerates T cell senescence in murine visceral adipose tissue.. J Clin Invest 2016 Dec 1;126(12):4626-4639.
- Basinska K, Marycz K, Śieszek A, Nicpoń J. The production and distribution of IL-6 and TNF-a in subcutaneous adipose tissue and their correlation with serum concentrations in Welsh ponies with equine metabolic syndrome.. J Vet Sci 2015;16(1):113-20.
- Zheng Y, Rudensky AY. Foxp3 in control of the regulatory T cell lineage.. Nat Immunol 2007 May;8(5):457-62.
- Jagannathan-Bogdan M, McDonnell ME, Shin H, Rehman Q, Hasturk H, Apovian CM, Nikolajczyk BS. Elevated proinflammatory cytokine production by a skewed T cell compartment requires monocytes and promotes inflammation in type 2 diabetes.. J Immunol 2011 Jan 15;186(2):1162-72.
- Wagner NM, Brandhorst G, Czepluch F, Lankeit M, Eberle C, Herzberg S, Faustin V, Riggert J, Oellerich M, Hasenfuss G, Konstantinides S, Schäfer K. Circulating regulatory T cells are reduced in obesity and may identify subjects at increased metabolic and cardiovascular risk.. Obesity (Silver Spring) 2013 Mar;21(3):461-8.
- Feuerer M, Herrero L, Cipolletta D, Naaz A, Wong J, Nayer A, Lee J, Goldfine AB, Benoist C, Shoelson S, Mathis D. Lean, but not obese, fat is enriched for a unique population of regulatory T cells that affect metabolic parameters.. Nat Med 2009 Aug;15(8):930-9.
- Sundin M, D'arcy P, Johansson CC, Barrett AJ, Lönnies H, Sundberg B, Nava S, Kiessling R, Mougiakakos D, Le Blanc K. Multipotent mesenchymal stromal cells express FoxP3: a marker for the immunosuppressive capacity?. J Immunother 2011 May;34(4):336-42.
- Leto Barone AA, Khalifian S, Lee WP, Brandacher G. Immunomodulatory effects of adipose-derived stem cells: fact or fiction?. Biomed Res Int 2013;2013:383685.
- Abdi R, Fiorina P, Adra CN, Atkinson M, Sayegh MH. Immunomodulation by mesenchymal stem cells: a potential therapeutic strategy for type 1 diabetes.. Diabetes 2008 Jul;57(7):1759-67.
- Matsubayashi S, Akasu F, Kasuga Y, Snow K, Keystone E, Volpé R. In vitro production of interferon-gamma by peripheral blood from patients with Graves' disease, Hashimoto's thyroiditis and rheumatoid arthritis.. Clin Exp Immunol 1990 Oct;82(1):63-8.
- Zhang C, Xiao C, Wang P, Xu W, Zhang A, Li Q, Xu X. The alteration of Th1/Th2/Th17/Treg paradigm in patients with type 2 diabetes mellitus: Relationship with diabetic nephropathy.. Hum Immunol 2014 Apr;75(4):289-96.
- English K, Ryan JM, Tobin L, Murphy MJ, Barry FP, Mahon BP. Cell contact, prostaglandin E(2) and transforming growth factor beta 1 play non-redundant roles in human mesenchymal stem cell induction of CD4+CD25(High) forkhead box P3+ regulatory T cells.. Clin Exp Immunol 2009 Apr;156(1):149-60.
- Bao P, Liu G, Wei Y. Association between IL-6 and related risk factors of metabolic syndrome and cardiovascular disease in young rats.. Int J Clin Exp Med 2015;8(8):13491-9.
- Levine B, Mizushima N, Virgin HW. Autophagy in immunity and inflammation.. Nature 2011 Jan 20;469(7330):323-35.
- Alizadeh S, Mazloom H, Sadeghi A, Emamgholipour S, Golestani A, Noorbakhsh F, Khoshniatnikoo M, Meshkani R. Evidence for the link between defective autophagy and inflammation in peripheral blood mononuclear cells of type 2 diabetic patients.. J Physiol Biochem 2018 Aug;74(3):369-379.
- Hirota Y, Kang D, Kanki T. The physiological role of mitophagy: new insights into phosphorylation events.. Int J Cell Biol 2012;2012:354914.
- Ding WX, Yin XM. Mitophagy: mechanisms, pathophysiological roles, and analysis.. Biol Chem 2012 Jul;393(7):547-64.
- Harris J, Deen N, Zamani S, Hasnat MA. Mitophagy and the release of inflammatory cytokines.. Mitochondrion 2018 Jul;41:2-8.
- Wang D, Li SP, Fu JS, Bai L, Guo L. Resveratrol augments therapeutic efficiency of mouse bone marrow mesenchymal stem cell-based therapy in experimental autoimmune encephalomyelitis.. Int J Dev Neurosci 2016 Apr;49:60-6.
- Ma ZH, Ma QY, Wang LC, Sha HC, Wu SL, Zhang M. Effect of resveratrol on peritoneal macrophages in rats with severe acute pancreatitis.. Inflamm Res 2005 Dec;54(12):522-7.
- Elmali N, Baysal O, Harma A, Esenkaya I, Mizrak B. Effects of resveratrol in inflammatory arthritis.. Inflammation 2007 Apr;30(1-2):1-6.
- Larrosa M, Yañéz-Gascón MJ, Selma MV, González-Sarrías A, Toti S, Cerón JJ, Tomás-Barberán F, Dolara P, Espín JC. Effect of a low dose of dietary resveratrol on colon microbiota, inflammation and tissue damage in a DSS-induced colitis rat model.. J Agric Food Chem 2009 Mar 25;57(6):2211-20.
- Wang B, Sun J, Li X, Zhou Q, Bai J, Shi Y, Le G. Resveratrol prevents suppression of regulatory T-cell production, oxidative stress, and inflammation of mice prone or resistant to high-fat diet-induced obesity.. Nutr Res 2013 Nov;33(11):971-81.
Citations
This article has been cited 17 times.- Bourebaba L, Zyzak M, Sikora M, Serwotka-Suszczak A, Mularczyk M, Al Naem M, Marycz K. Sex Hormone-Binding Globulin (SHBG) Maintains Proper Equine Adipose-Derived Stromal Cells (ASCs)' Metabolic Functions and Negatively Regulates their Basal Adipogenic Potential. Stem Cell Rev Rep 2023 Jul 4;.
- Bourebaba L, Serwotka-Suszczak A, Pielok A, Sikora M, Mularczyk M, Marycz K. The PTP1B inhibitor MSI-1436 ameliorates liver insulin sensitivity by modulating autophagy, ER stress and systemic inflammation in Equine metabolic syndrome affected horses. Front Endocrinol (Lausanne) 2023;14:1149610.
- Tracy EP, Stielberg V, Rowe G, Benson D, Nunes SS, Hoying JB, Murfee WL, LeBlanc AJ. State of the field: cellular and exosomal therapeutic approaches in vascular regeneration. Am J Physiol Heart Circ Physiol 2022 Apr 1;322(4):H647-H680.
- Weiss C, Kornicka-Grabowska K, Mularczyk M, Siwinska N, Marycz K. Extracellular Microvesicles (MV's) Isolated from 5-Azacytidine-and-Resveratrol-Treated Cells Improve Viability and Ameliorate Endoplasmic Reticulum Stress in Metabolic Syndrome Derived Mesenchymal Stem Cells. Stem Cell Rev Rep 2020 Dec;16(6):1343-1355.
- Golonka P, Kornicka-Garbowska K, Marycz K. SIRT1(+) Adipose Derived Mesenchymal Stromal Stem Cells (ASCs) Suspended in Alginate Hydrogel for the Treatment of Subchondral Bone Cyst in Medial Femoral Condyle in the Horse. Clinical Report. Stem Cell Rev Rep 2020 Dec;16(6):1328-1334.
- Yagi H, Chen AF, Hirsch D, Rothenberg AC, Tan J, Alexander PG, Tuan RS. Antimicrobial activity of mesenchymal stem cells against Staphylococcus aureus. Stem Cell Res Ther 2020 Jul 17;11(1):293.
- Kornicka-Garbowska K, Pędziwiatr R, Woźniak P, Kucharczyk K, Marycz K. Microvesicles isolated from 5-azacytidine-and-resveratrol-treated mesenchymal stem cells for the treatment of suspensory ligament injury in horse-a case report. Stem Cell Res Ther 2019 Dec 18;10(1):394.
- Marycz K, Szłapka-Kosarzewska J, Geburek F, Kornicka-Garbowska K. Systemic Administration of Rejuvenated Adipose-Derived Mesenchymal Stem Cells Improves Liver Metabolism in Equine Metabolic Syndrome (EMS)- New Approach in Veterinary Regenerative Medicine. Stem Cell Rev Rep 2019 Dec;15(6):842-850.
- Marycz K, Houston JMI, Weiss C, Röcken M, Kornicka K. 5-Azacytidine and Resveratrol Enhance Chondrogenic Differentiation of Metabolic Syndrome-Derived Mesenchymal Stem Cells by Modulating Autophagy. Oxid Med Cell Longev 2019;2019:1523140.
- Kornicka K, Geburek F, Röcken M, Marycz K. Stem Cells in Equine Veterinary Practice-Current Trends, Risks, and Perspectives. J Clin Med 2019 May 14;8(5).
- Cislo-Pakuluk A, Smieszek A, Kucharczyk N, Bedford PGC, Marycz K. Intra-Vitreal Administration of Microvesicles Derived from Human Adipose-Derived Multipotent Stromal Cells Improves Retinal Functionality in Dogs with Retinal Degeneration. J Clin Med 2019 Apr 13;8(4).
- Smieszek A, Kornicka K, Szłapka-Kosarzewska J, Androvic P, Valihrach L, Langerova L, Rohlova E, Kubista M, Marycz K. Metformin Increases Proliferative Activity and Viability of Multipotent Stromal Stem Cells Isolated from Adipose Tissue Derived from Horses with Equine Metabolic Syndrome. Cells 2019 Jan 22;8(2).
- Bourebaba N, Domagała J, Bourebaba L. Revitalizing equine metabolism: how SHBG improves mitochondrial function and reduces inflammation. BMC Vet Res 2025 Oct 21;21(1):620.
- Storti G, Foti R, Foti R, Palmesano M, Patacchiola M, Incognito D, Cervelli G, Longo B, Scioli MG, Fiorelli E, Terriaca S, Lisa A, Kim BS, Orlandi A, Cervelli V. A Comprehensive Exploration of the Biological Effects of Adipose-Derived Stem Cells in the Treatment of Systemic Sclerosis. Cells 2025 Mar 19;14(6).
- Li W, Xiang Z, Yu W, Huang X, Jiang Q, Abumansour A, Yang Y, Chen C. Natural compounds and mesenchymal stem cells: implications for inflammatory-impaired tissue regeneration. Stem Cell Res Ther 2024 Feb 7;15(1):34.
- 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.
- Mohseni R, Mahdavi Sharif P, Behfar M, Modaresi MR, Shirzadi R, Mardani M, Jafari L, Jafari F, Nikfetrat Z, Hamidieh AA. Evaluation of safety and efficacy of allogeneic adipose tissue-derived mesenchymal stem cells in pediatric bronchiolitis obliterans syndrome (BoS) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Stem Cell Res Ther 2023 Sep 19;14(1):256.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
