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The Journal of veterinary medical science2020; 82(10); 1472-1479; doi: 10.1292/jvms.20-0167

Optimal activation methods for maximizing the concentrations of platelet-derived growth factor-BB and transforming growth factor-β1 in equine platelet-rich plasma.

Abstract: Platelet-rich plasma (PRP) therapy has been widely applied in various medical fields including humans and horses. This study aimed to establish an optimal activation method to stably and reproducibly maximize the concentrations of platelet-derived growth factor-BB (PDGF-BB) and transforming growth factor-β1 (TGF-β1) contained in equine PRP. Autologous PRP was prepared from 11 Thoroughbreds. For the activation test, PRP was activated by either a single freeze-thaw cycle (Fr) or adding calcium and autologous serum containing thrombin (Ca). PDGF-BB and TGF-β1 concentrations in Fr, Ca, nonactivated (No), and platelet-poor plasma (PPP) samples were determined using ELISA and compared. For repetitive freeze-thaw test, PRP was subjected to single (Fr1), double (Fr2), triple (Fr3), or quadruple (Fr4) freeze-thaw cycles and the concentrations of both growth factors in samples were compared similarly. The PDGF-BB concentration in Ca was significantly higher than that in other preparations. The TGF-β1 concentrations in Fr and Ca were significantly higher than those in PPP and No, with no significant differences between Fr and Ca. The concentrations of both factors were significantly increased in PRP treated with multiple cycles of freeze-thaw compared with that in PRP treated with a single cycle. No significant differences were noted among Fr2, Fr3, and Fr4. Our findings suggest that activation by adding calcium and autologous serum is optimal for instant use of PRP and that double freeze-thawing is an easier and optimal activation method for cryopreserved PRP.
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Publication Date: 2020-08-19 PubMed ID: 32814750PubMed Central: PMC7653321DOI: 10.1292/jvms.20-0167Google 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 studies the most effective methods to activate the platelet-derived growth factor-BB (PDGF-BB) and transforming growth factor-β1 (TGF-β1) in equine platelet-rich plasma (PRP). The researchers found that the addition of calcium and autologous serum as well as a double freeze-thawing process are the best activation methods.

Methods

  • The researchers procured autologous PRP from 11 Thoroughbred horses.
  • Two activation methods were compared: a single freeze-thaw cycle (Fr) or the addition of calcium and autologous serum that contains thrombin (Ca).
  • Concentrations of PDGF-BB and TGF-β1 in activated (Fr and Ca) and non-activated (No) PRP, as well as platelet-poor plasma (PPP), were measured with ELISA.
  • A repetitive freeze-thaw test was also conducted, wherein the PRP underwent single (Fr1), double (Fr2), triple (Fr3), or quadruple (Fr4) freeze-thaw cycles.

Findings

  • The researchers found that the concentration of PDGF-BB was significantly higher in PRP activated with calcium and autologous serum (Ca) compared to the other preparations.
  • TGF-β1 concentrations in PRP activated by a single freeze-thaw cycle (Fr) or by calcium and autologous serum (Ca) were significantly higher compared to platelet-poor plasma (PPP) and non-activated (No) samples.
  • No significant differences were found between the concentrations of TGF-β1 in the single freeze-thaw (Fr) and calcium and autologous serum (Ca) activations.
  • Multiple cycles of freeze-thaw significantly increased concentrations of both growth factors compared to a single freeze-thaw cycle.
  • Trials showed no significant differences between the concentrations of growth factors in double, triple, and quadruple freeze-thaw cycles.

Suggested Activation Methods

  • The results suggest that to maximize the concentration of growth factors, particularly PDGF-BB and TGF-β1, in PRP for immediate use, it is optimal to add calcium and autologous serum.
  • On the other hand, PRP that is planned to be cryopreserved (frozen for later use) should undergo a double freeze-thaw cycle for maximal optimization.

Cite This Article

APA
Fukuda K, Kuroda T, Tamura N, Mita H, Kasashima Y. (2020). Optimal activation methods for maximizing the concentrations of platelet-derived growth factor-BB and transforming growth factor-β1 in equine platelet-rich plasma. J Vet Med Sci, 82(10), 1472-1479. https://doi.org/10.1292/jvms.20-0167

Publication

The Journal of veterinary medical science
ISSN: 1347-7439
NlmUniqueID: 9105360
Country: Japan
Language: English
Volume: 82
Issue: 10
Pages: 1472-1479

Researcher Affiliations

Fukuda, Kentaro
  • Clinical Veterinary Medicine Division, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke-shi, Tochigi 329-0412, Japan.
Kuroda, Taisuke
  • Clinical Veterinary Medicine Division, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke-shi, Tochigi 329-0412, Japan.
Tamura, Norihisa
  • Clinical Veterinary Medicine Division, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke-shi, Tochigi 329-0412, Japan.
Mita, Hiroshi
  • Clinical Veterinary Medicine Division, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke-shi, Tochigi 329-0412, Japan.
Kasashima, Yoshinori
  • The Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke-shi, Tochigi 329-0412, Japan.

MeSH Terms

  • Animals
  • Becaplermin
  • Horses
  • Platelet-Rich Plasma
  • Thrombin
  • Transforming Growth Factor beta1
  • Transforming Growth Factors

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Citations

This article has been cited 8 times.
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