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Life (Basel, Switzerland)2025; 15(10); 1558; doi: 10.3390/life15101558

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.
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Publication Date: 2025-10-04 PubMed ID: 41157231PubMed Central: PMC12565166DOI: 10.3390/life15101558Google Scholar: Lookup
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  • 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.

Overview

  • This study evaluated the effects of several treatments—including platelet-rich plasma (PRP), ozonized PRP, hyaluronic acid, paracetamol, and polyacrylamide hydrogel—on the growth and viability of mesenchymal stem cells (MSCs) derived from horse joint fluid.
  • The results demonstrated how these substances influence MSC proliferation and survival over time, providing insight into their potential use for joint regeneration in horses.

Background and Rationale

  • Musculoskeletal disorders are a leading cause of lameness in horses, impacting their mobility and quality of life.
  • Regenerative medicine, particularly using MSCs, offers promising strategies to repair joint damage and restore function.
  • Synovial fluid-derived MSCs are of special interest because they originate from the joint environment and may have higher relevance for intra-articular therapies.
  • Various biological and synthetic agents—such as PRP, ozonized PRP, hyaluronic acid, paracetamol, and polyacrylamide hydrogels—are proposed to support stem cell function or joint health, but their direct effects on MSCs need clarification.

Methods

  • Synovial fluid samples were collected from horses using strict cytological criteria to ensure viable MSC isolation.
  • Isolated MSCs were expanded in vitro, characterized phenotypically to confirm their identity.
  • MSC cultures were supplemented with different concentrations of:
    • Platelet-rich plasma (PRP)
    • Ozonized PRP (PRP treated with ozone for oxidizing effects)
    • Hyaluronic acid
    • Paracetamol (acetaminophen)
    • Polyacrylamide hydrogel (commercially known as NOLTREX)
  • Proliferation and viability were assessed at three time points: 24 hours, 72 hours, and 7 days post-treatment.

Key Findings

  • PRP:
    • Significantly stimulated MSC proliferation in both a time- and dose-dependent manner.
    • 10% concentration produced the maximum proliferative effect.
  • Hyaluronic Acid:
    • Induced growth stimulation, especially notable at 1 mg/mL concentration.
  • Paracetamol:
    • Caused a concentration-dependent increase in proliferation.
    • Strongest proliferative effect observed at 100 μg/mL.
  • Polyacrylamide Hydrogel (NOLTREX):
    • Exhibited a biphasic effect on MSCs:
      • Initially inhibitory at high doses.
      • Stimulated cell growth after 7 days.
  • Ozonized PRP:
    • Displays concentration-dependent redox activity impacting MSC viability:
    • Lower doses preserved viability.
    • Higher doses caused early suppression followed by delayed stimulation of proliferation.

Interpretation and Clinical Implications

  • PRP and related biologic agents have demonstrated the capacity to modulate MSC behavior positively, suggesting utility as regenerative intra-articular therapies for equine joint diseases.
  • Hyaluronic acid and paracetamol also support MSC proliferation but their mechanisms may differ, as hyaluronic acid is a natural joint lubricant and paracetamol is commonly a pain reliever with some effects on cell metabolism.
  • The biphasic profile of NOLTREX highlights the importance of dosage and timing in therapeutic applications, indicating that immediate effects may differ from long-term outcomes.
  • Ozonized PRP’s redox effects suggest potential for controlled oxidative modulation of MSCs, but precise dosing is critical to avoid cytotoxicity.
  • The findings emphasize the need for optimized dosing protocols and standardization to maximize therapeutic benefits and ensure safety when translating these treatments to clinical practice.

Conclusions

  • The study provides evidence supporting the use of PRP, ozonized PRP, hyaluronic acid, paracetamol, and polyacrylamide hydrogel as modulators of equine synovial fluid-derived MSCs.
  • These agents have distinct and concentration-dependent effects on MSC proliferation and viability, which can inform the development of effective regenerative therapies for equine musculoskeletal disorders.
  • Future work should focus on clinical trials to validate these in vitro findings and establish guidelines for their safe and effective application in veterinary medicine.

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

Life (Basel, Switzerland)
ISSN: 2075-1729
NlmUniqueID: 101580444
Country: Switzerland
Language: English
Volume: 15
Issue: 10
PII: 1558

Researcher Affiliations

Bungărdean, Denisa
  • 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.
Pall, Emoke
  • 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.
Daradics, Zsofia
  • 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.
Popescu, Maria
  • Equine Clinic, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine (UASVM) Cluj-Napoca, Mănăştur Street 3-5, 400372 Cluj-Napoca, Romania.
Tripon, Mirela Alexandra
  • 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.
Lupșan, Alexandru Florin
  • Department of Anesthesiology and Surgery, University of Agricultural Sciences and Veterinary Medicine (UASVM) Cluj-Napoca, Mănăştur Street 3-5, 400372 Cluj-Napoca, Romania.
Crecan, Cristian Mihăiță
  • Department of Anesthesiology and Surgery, University of Agricultural Sciences and Veterinary Medicine (UASVM) Cluj-Napoca, Mănăştur Street 3-5, 400372 Cluj-Napoca, Romania.
Morar, Ianu Adrian
  • 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.
Nicolescu, Alexandru
  • 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.
Bora, Florin Dumitru
  • 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.
Marcus, Ioan
  • 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.

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