In Vitro Effects of PRP, Ozonized PRP, Hyaluronic Acid, Paracetamol, and Polyacrylamide on Equine Synovial Fluid-Derived Mesenchymal Stem Cells.
Abstract: Musculoskeletal disorders are a major cause of lameness in horses, often necessitating innovative regenerative strategies to restore joint function and improve quality of life. This study investigated the effects of platelet-rich plasma (PRP), ozonized PRP, hyaluronic acid, paracetamol, and polyacrylamide hydrogel (NOLTREX) on the behavior of mesenchymal stem cells (MSCs) derived from equine synovial fluid. Synovial fluid samples were collected under strict cytological criteria to ensure viability, followed by in vitro expansion and phenotypic characterization of MSCs. Cultures were supplemented with the tested preparations, and cellular proliferation and viability were evaluated at 24 h, 72 h, and 7 days. PRP significantly promoted MSC proliferation in a time- and dose-dependent manner, with maximal effect at 10%. Hyaluronic acid stimulated growth, most pronounced at 1 mg/mL, while paracetamol induced a concentration-dependent proliferative response, strongest at 100 μg/mL. NOLTREX displayed a biphasic effect, initially inhibitory at high concentrations but stimulatory at 7 days. Ozonized PRP showed concentration-dependent redox activity, with lower doses maintaining viability and higher doses producing an initial suppression followed by delayed stimulation. Collectively, these findings support the therapeutic potential of PRP and related biologic preparations as intra-articular regenerative therapies in equine medicine, while underscoring the importance of dose optimization and standardized protocols to facilitate clinical translation.
Publication Date: 2025-10-04 PubMed ID: 41157231PubMed Central: PMC12565166DOI: 10.3390/life15101558Google 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
Cite This Article
APA
Bungărdean D, Pall E, Daradics Z, Popescu M, Tripon MA, Lupșan AF, Crecan CM, Morar IA, Nicolescu A, Bora FD, Marcus I.
(2025).
In Vitro Effects of PRP, Ozonized PRP, Hyaluronic Acid, Paracetamol, and Polyacrylamide on Equine Synovial Fluid-Derived Mesenchymal Stem Cells.
Life (Basel), 15(10), 1558.
https://doi.org/10.3390/life15101558 Publication
Researcher Affiliations
- Department of Pathophysiology, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine (UASVM) Cluj-Napoca, Mănăştur Street 3-5, 400372 Cluj-Napoca, Romania.
- Department of Infectious Diseases, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Calea Mănăştur nr. 3-5, 400372 Cluj-Napoca, Romania.
- Department of Internal Medicine, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine (UASVM) Cluj-Napoca, Mănăştur Street 3-5, 400372 Cluj-Napoca, Romania.
- Equine Clinic, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine (UASVM) Cluj-Napoca, Mănăştur Street 3-5, 400372 Cluj-Napoca, Romania.
- Department of Reproduction, Obstetrics and Veterinary Gynecology, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine (UASVM) Cluj-Napoca, Mănăştur Street 3-5, 400372 Cluj-Napoca, Romania.
- Department of Anesthesiology and Surgery, University of Agricultural Sciences and Veterinary Medicine (UASVM) Cluj-Napoca, Mănăştur Street 3-5, 400372 Cluj-Napoca, Romania.
- Department of Anesthesiology and Surgery, University of Agricultural Sciences and Veterinary Medicine (UASVM) Cluj-Napoca, Mănăştur Street 3-5, 400372 Cluj-Napoca, Romania.
- Department of Reproduction, Obstetrics and Veterinary Gynecology, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine (UASVM) Cluj-Napoca, Mănăştur Street 3-5, 400372 Cluj-Napoca, Romania.
- Laboratory of Chromatography, Advanced Horticultural Research Institute of Transylvania, Faculty of Horticulture and Business for Rural Development, University of Agricultural Sciences and Veterinary Medicine, 400372 Cluj-Napoca, Romania.
- Laboratory of Chromatography, Advanced Horticultural Research Institute of Transylvania, Faculty of Horticulture and Business for Rural Development, University of Agricultural Sciences and Veterinary Medicine, 400372 Cluj-Napoca, Romania.
- Viticulture and Oenology Department, Advanced Horticultural Research Institute of Transylvania, Faculty of Horticulture and Business in Rural Development, University of Agricultural Sciences and Veterinary Medicine (UASVM) Cluj-Napoca, Mănăştur Street 3-5, 400372 Cluj-Napoca, Romania.
- Department of Pathophysiology, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine (UASVM) Cluj-Napoca, Mănăştur Street 3-5, 400372 Cluj-Napoca, Romania.
Conflict of Interest Statement
The authors declare no conflicts of interest.
References
This article includes 54 references
- Ribitsch I., Oreff G.L., Jenner F. Regenerative Medicine for Equine Musculoskeletal Diseases. Animals. 2021;11:234. doi: 10.3390/ani11010234.
- Murray R.C., Walters J.M., Snart H., Dyson S.J., Parkin T.D.H. Identification of Risk Factors for Lameness in Dressage Horses. Vet. J. 2010;184:27–36. doi: 10.1016/j.tvjl.2009.03.020.
- Guidoni K., Chiaradia E., Pepe M., Di Meo A., Tognoloni A., Seccaroni M., Beccati F. The Combined Use of Triamcinolone and Platelet-Rich Plasma in Equine Metacarpophalangeal Joint Osteoarthritis Treatments: An In Vivo and In Vitro Study. Animals. 2024;14:3645. doi: 10.3390/ani14243645.
- Garbin L.C., Olver C.S. Platelet-Rich Products and Their Application to Osteoarthritis. J. Equine Vet. Sci. 2020;86:102820. doi: 10.1016/j.jevs.2019.102820.
- Sakata R., Reddi A.H. Platelet-Rich Plasma Modulates Actions on Articular Cartilage Lubrication and Regeneration. Tissue Eng. Part B Rev. 2016;22:408–419. doi: 10.1089/ten.teb.2015.0534.
- Zhu Y., Yuan M., Meng H.Y., Wang A.Y., Guo Q.Y., Wang Y., Peng J. Basic Science and Clinical Application of Platelet-Rich Plasma for Cartilage Defects and Osteoarthritis: A Review. Osteoarthr. Cartil. 2013;21:1627–1637. doi: 10.1016/j.joca.2013.07.017.
- Textor J.A., Willits N.H., Tablin F. Synovial Fluid Growth Factor and Cytokine Concentrations after Intra-Articular Injection of a Platelet-Rich Product in Horses. Vet. J. 2013;198:217–223. doi: 10.1016/j.tvjl.2013.07.020.
- Camargo Garbin L., Lopez C., Carmona J.U. A Critical Overview of the Use of Platelet-Rich Plasma in Equine Medicine Over the Last Decade. Front. Vet. Sci. 2021;8:641818. doi: 10.3389/fvets.2021.641818.
- Peng C., Yang L., Labens R., Gao Y., Zhu Y., Li J. A Systematic Review and Meta-analysis of the Efficacy of Platelet-rich Plasma Products for Treatment of Equine Joint Disease. Equine Vet. J. 2024;56:858–869. doi: 10.1111/evj.14042.
- Mehrabani D., Seghatchian J., Acker J.P. Platelet Rich Plasma in Treatment of Musculoskeletal Pathologies. Transfus. Apher. Sci. 2019;58:102675. doi: 10.1016/j.transci.2019.102675.
- Fotouhi A., Maleki A., Dolati S., Aghebati-Maleki A., Aghebati-Maleki L. Platelet Rich Plasma, Stromal Vascular Fraction and Autologous Conditioned Serum in Treatment of Knee Osteoarthritis. Biomed. Pharmacother. 2018;104:652–660. doi: 10.1016/j.biopha.2018.05.019.
- Muiños-López E., Delgado D., Sánchez P., Paiva B., Anitua E., Fiz N., Aizpurua B., Guadilla J., Padilla S., Granero-Moltó F., et al. Modulation of Synovial Fluid-Derived Mesenchymal Stem Cells by Intra-Articular and Intraosseous Platelet Rich Plasma Administration. Stem Cells Int. 2016;2016:1247950. doi: 10.1155/2016/1247950.
- Colbath A.C., Frisbie D.D., Dow S.W., Kisiday J.D., McIlwraith C.W., Goodrich L.R. Equine Models for the Investigation of Mesenchymal Stem Cell Therapies in Orthopaedic Disease. Oper. Tech. Sports Med. 2017;25:41–49. doi: 10.1053/j.otsm.2016.12.007.
- Gugjoo M.B., Amarpal, Makhdoomi D.M., Sharma G.T. Equine Mesenchymal Stem Cells: Properties, Sources, Characterization, and Potential Therapeutic Applications. J. Equine Vet. Sci. 2019;72:16–27. doi: 10.1016/j.jevs.2018.10.007.
- Tang H.-C., Chen W.-C., Chiang C.-W., Chen L.-Y., Chang Y.-C., Chen C.-H. Differentiation Effects of Platelet-Rich Plasma Concentrations on Synovial Fluid Mesenchymal Stem Cells from Pigs Cultivated in Alginate Complex Hydrogel. Int. J. Mol. Sci. 2015;16:18507–18521. doi: 10.3390/ijms160818507.
- Tjandra K.C., Novriansyah R., Sudiasa I.N.S., Ar A., Rahmawati N.A.D., Dilogo I.H. Modified Mesenchymal Stem Cell, Platelet-Rich Plasma, and Hyaluronic Acid Intervention in Early Stage Osteoarthritis: A Systematic Review, Meta-Analysis, and Meta-Regression of Arthroscopic-Guided Intra-Articular Approaches. PLoS ONE. 2024;19:e0295876. doi: 10.1371/journal.pone.0295876.
- Zayed M., Adair S., Ursini T., Schumacher J., Misk N., Dhar M. Concepts and Challenges in the Use of Mesenchymal Stem Cells as a Treatment for Cartilage Damage in the Horse. Res. Vet. Sci. 2018;118:317–323. doi: 10.1016/j.rvsc.2018.03.011.
- Dernek B., Kesiktas F.N. Efficacy of Combined Ozone and Platelet-Rich-Plasma Treatment versus Platelet-Rich-Plasma Treatment Alone in Early Stage Knee Osteoarthritis. J. Back Musculoskelet. Rehabil. 2019;32:305–311. doi: 10.3233/BMR-181301.
- Vendruscolo C.D.P., Moreira J.J., Seidel S.R.T.;, Fülber J., Neuenschwander H.M., Bonagura G., Agreste F.R., Baccarin R.Y.A. Effects of Medical Ozone upon Healthy Equine Joints: Clinical and Laboratorial Aspects. PLoS ONE. 2018;13:e0197736. doi: 10.1371/journal.pone.0197736.
- Sciorsci R.L., Lillo E., Occhiogrosso L., Rizzo A. Ozone Therapy in Veterinary Medicine: A Review. Res. Vet. Sci. 2020;130:240–246. doi: 10.1016/j.rvsc.2020.03.026.
- Carrade D.D., Owens S.D., Galuppo L.D., Vidal M.A., Ferraro G.L., Librach F., Buerchler S., Friedman M.S., Walker N.J., Borjesson D.L. Clinicopathologic Findings Following Intra-Articular Injection of Autologous and Allogeneic Placentally Derived Equine Mesenchymal Stem Cells in Horses. Cytotherapy. 2011;13:419–430. doi: 10.3109/14653249.2010.536213.
- Barrachina L., Romero A., Zaragoza P., Rodellar C., Vázquez F.J. Practical Considerations for Clinical Use of Mesenchymal Stem Cells: From the Laboratory to the Horse. Vet. J. 2018;238:49–57. doi: 10.1016/j.tvjl.2018.07.004.
- Vinod E., Padmaja K., Ramasamy B., Sathishkumar S. Systematic Review of Articular Cartilage Derived Chondroprogenitors for Cartilage Repair in Animal Models. J. Orthop. 2023;35:43–53. doi: 10.1016/j.jor.2022.10.012.
- Cassano J.M., Marycz K., Horna M., Nogues M.P., Morgan J.M., Herrmann D.B., Galuppo L.D., Vapniarsky N. Evaluating the Safety of Intra-Articular Mitotherapy in the Equine Model: A Potential Novel Treatment for Osteoarthritis. J. Equine Vet. Sci. 2023;120:104164. doi: 10.1016/j.jevs.2022.104164.
- Pereira M.F., Ribeiro G., Gonzales A., Arantes J.A., Dória R.G.S. Effects of Intra-Articular Administration of Hyaluronic Acid or Platelet-Rich Plasma as a Complementary Treatment to Arthroscopy in Horses with Osteochondritis Dissecans. Vet. Anim. Sci. 2024;23:100330. doi: 10.1016/j.vas.2023.100330.
- Smit Y., Marais H.J., Thompson P.N., Mahne A.T., Goddard A. Clinical Findings, Synovial Fluid Cytology and Growth Factor Concentrations after Intra-Articular Use of a Platelet-Rich Product in Horses with Osteoarthritis. J. S. Afr. Vet. Assoc. 2019;90:e1–e9. doi: 10.4102/jsava.v90i0.1721.
- Conrado F.O., Beatty S.S.K. Fluid Analysis in the Equine Patient. Vet. Clin. North Am. Equine Pract. 2021;36:e1–e28. doi: 10.1016/j.cveq.2021.01.002.
- Steel C.M. Equine Synovial Fluid Analysis. Vet. Clin. North Am. Equine Pract. 2008;24:437–454. doi: 10.1016/j.cveq.2008.05.004.
- Murata D., Miyakoshi D., Hatazoe T., Miura N., Tokunaga S., Fujiki M., Nakayama K., Misumi K. Multipotency of Equine Mesenchymal Stem Cells Derived from Synovial Fluid. Vet. J. 2014;202:53–61. doi: 10.1016/j.tvjl.2014.07.029.
- Crecan C., Oprean L.S., Tripon M.A., Bodnariuc A., Luciana R., Pall E., Oros D.C., Morar I. Equestrian Synovial Fluid Mesenchymal Stem Cells, a Potential Experimental Model for Osteoarticular Therapies. Farmacia. 2019;67:458–466. doi: 10.31925/farmacia.2019.3.12.
- Segabinazzi L.G.T.M., Podico G., Rosser M.F., Nanjappa S.G., Alvarenga M.A., Canisso I.F. Three Manual Noncommercial Methods to Prepare Equine Platelet-Rich Plasma. Animals. 2021;11:1478. doi: 10.3390/ani11061478.
- Radtke A.V., Goodale M.B., Fortier L.A. Platelet and Leukocyte Concentration in Equine Autologous Conditioned Plasma Are Inversely Distributed by Layer and Are Not Affected by Centrifugation Rate. Front. Vet. Sci. 2020;7:00173. doi: 10.3389/fvets.2020.00173.
- Ionita J.-C., Kissich C., Gottschalk J., Einspanier A., Köller G., Winter K., Brehm W. Comparison of Cellular and Growth Factor Concentrations in Equine Autologous Conditioned Plasma® (ACP) and Manually Prepared Platelet Rich Plasma (MPRP) Pferdeheilkd. Equine Med. 2014;30:195–201. doi: 10.21836/PEM20140208.
- Hauschild G., Geburek F., Gosheger G., Eveslage M., Serrano D., Streitbürger A., Johannlükens S., Menzel D., Mischke R. Short Term Storage Stability at Room Temperature of Two Different Platelet-Rich Plasma Preparations from Equine Donors and Potential Impact on Growth Factor Concentrations. BMC Vet. Res. 2016;13:7. doi: 10.1186/s12917-016-0920-4.
- Yang X., Zhong Y., Wang D., Lu Z. A Simple Colorimetric Method for Viable Bacteria Detection Based on Cell Counting Kit-8. Anal. Methods. 2021;13:5211–5215. doi: 10.1039/D1AY01624E.
- Chanda M., Klinphayom C., Sungsuwan T., Senarat W., Thongkham E., Kamlangdee A., Senarat N. Diagnostic Imaging Features, Cytological Examination, and Treatment of Lymphocytic Tenosynovitis of the Common Digital Extensor Tendon Sheath in an Eventing Horse. Vet. Anim. Sci. 2021;14:100209. doi: 10.1016/j.vas.2021.100209.
- Lapjit C., Charoenchanikran P., Petchkaew P., Sukpipattanamongkol S., Yodsheewan R., Theerapan W., Chanda M. Diagnostic Imaging and Cytological Analysis Aid the Clinical Investigation of Long Digital Extensor Tendon Subtendinous Bursitis in a Horse. J. Equine Vet. Sci. 2021;101:103449. doi: 10.1016/j.jevs.2021.103449.
- Sanchez-Teran A.F., Bracamonte J.L., Hendrick S., Riddell L., Musil K., Hoff B., Rubio-Martínez L.M. Effect of Repeated Through-and-through Joint Lavage on Serum Amyloid A in Synovial Fluid from Healthy Horses. Vet. J. 2016;210:30–33. doi: 10.1016/j.tvjl.2016.01.001.
- Greve L., Pfau T., Dyson S. Thoracolumbar Movement in Sound Horses Trotting in Straight Lines in Hand and on the Lunge and the Relationship with Hind Limb Symmetry or Asymmetry. Vet. J. 2017;220:95–104. doi: 10.1016/j.tvjl.2017.01.003.
- Beck A.A., Paz L.B., Frank M.I., Engelmann A.M., Krause A., Côrte F.D.D.L. Safety and Synovial Inflammatory Response After Intra-Articular Injection of Botulinum Toxin Type A in Healthy Horses. J. Equine Vet. Sci. 2022;110:103865. doi: 10.1016/j.jevs.2022.103865.
- Rinnovati R., Bonelli F., Tognetti R., Gallo C., Bassini R.F., Marchetti V., Sgorbini M. Effect of Repeated Arthrocentesis on Cytology of Synovial Fluid. J. Equine Vet. Sci. 2017;57:112–115. doi: 10.1016/j.jevs.2017.07.008.
- Jiang D., Muschhammer J., Qi Y., Kügler A., de Vries J.C., Saffarzadeh M., Sindrilaru A., Vander Beken S., Wlaschek M., Kluth M.A., et al. Suppression of Neutrophil-Mediated Tissue Damage—A Novel Skill of Mesenchymal Stem Cells. Stem Cells. 2016;34:2393–2406. doi: 10.1002/stem.2417.
- Fiala-Rechsteiner S.M., Amaral M.G., Cruz L.A., Rodrigues R.F., Pimentel A.M., Mattos R.C. Inflammatory Lesions in the Oviducts and Its Relationship With Endometrial Inflammation and Ovarian Activity in Criollo Mares. J. Equine Vet. Sci. 2015;35:731–734. doi: 10.1016/j.jevs.2015.07.006.
- Frisbie D.D., Al-Sobayil F., Billinghurst R.C., Kawcak C.E., McIlwraith C.W. Changes in Synovial Fluid and Serum Biomarkers with Exercise and Early Osteoarthritis in Horses. Osteoarthr. Cartil. 2008;16:1196–1204. doi: 10.1016/j.joca.2008.03.008.
- Walters M., Skovgaard K., Andersen P.H., Heegaard P.M.H., Jacobsen S. Dynamics of Local Gene Regulations in Synovial Fluid Leukocytes from Horses with Lipopolysaccharide-Induced Arthritis. Vet. Immunol. Immunopathol. 2021;241:110325. doi: 10.1016/j.vetimm.2021.110325.
- Dumoulin M., Martens A., Van den Abeele A.-M., Boyen F., Oosterlinck M., Wilderjans H., Gasthuys F., Haesebrouck F., Pille F. Evaluation of Direct Etest for Antimicrobial Susceptibility Testing of Bacteria Isolated from Synovial Fluid of Horses Using Enrichment Bottles. Vet. J. 2017;220:55–62. doi: 10.1016/j.tvjl.2017.01.001.
- Soares C.S., Babo P.S., Reis R.L., Carvalho P.P., Gomes M.E. Platelet-Derived Products in Veterinary Medicine: A New Trend or an Effective Therapy? Trends Biotechnol. 2021;39:225–243. doi: 10.1016/j.tibtech.2020.07.011.
- Cao Y., Li Y., Fu S.C., Shen J., Zhang H., Jiang C., Shu-Hang Yung P. Platelet-Rich Plasma Pretreatment Protects Anterior Cruciate Ligament Fibroblasts Correlated with PI3K-Akt-MTOR Pathway under Hypoxia Condition. J. Orthop. Transl. 2022;34:102–112. doi: 10.1016/j.jot.2022.02.002.
- Naor D. Editorial: Interaction Between Hyaluronic Acid and Its Receptors (CD44, RHAMM) Regulates the Activity of Inflammation and Cancer. Front. Immunol. 2016;7:00039. doi: 10.3389/fimmu.2016.00039.
- Mercer M.A., McKenzie H.C., Byron C.R., Pleasant R.S., Bogers S.H., Council-Troche R.M., Werre S.R., Burns T., Davis J.L. Pharmacokinetics and Clinical Efficacy of Acetaminophen (Paracetamol) in Adult Horses with Mechanically Induced Lameness. Equine Vet. J. 2023;55:524–533. doi: 10.1111/evj.13601.
- Almaawi A., Wang H.T., Ciobanu O., Rowas S.A.L., Rampersad S., Antoniou J., Mwale F. Effect of Acetaminophen and Nonsteroidal Anti-Inflammatory Drugs on Gene Expression of Mesenchymal Stem Cells. Tissue Eng. Part A. 2013;19:1039–1046. doi: 10.1089/ten.tea.2012.0129.
- Wang X., Wu Q., Liu A., Anadón A., Rodríguez J.-L., Martínez-Larrañaga M.-R., Yuan Z., Martínez M.-A. Paracetamol: Overdose-Induced Oxidative Stress Toxicity, Metabolism, and Protective Effects of Various Compounds in Vivo and in Vitro. Drug Metab. Rev. 2017;49:395–437. doi: 10.1080/03602532.2017.1354014.
- Wang P., Cui Y., Wang J., Liu D., Tian Y., Liu K., Wang X., Liu L., He Y., Pei Y., et al. Mesenchymal Stem Cells Protect against Acetaminophen Hepatotoxicity by Secreting Regenerative Cytokine Hepatocyte Growth Factor. Stem Cell Res. Ther. 2022;13:94. doi: 10.1186/s13287-022-02754-x.
- Yiang G., Yu Y., Lin K., Chen J., Chang W., Wei C. Acetaminophen Induces JNK/P38 Signaling and Activates the Caspase-9-3-Dependent Cell Death Pathway in Human Mesenchymal Stem Cells. Int. J. Mol. Med. 2015;36:485–492. doi: 10.3892/ijmm.2015.2254.
Citations
This article has been cited 0 times.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