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Veterinary and comparative orthopaedics and traumatology : V.C.O.T2016; 30(1); 1-7; doi: 10.3415/VCOT-16-02-0026

Influence of calcium salts and bovine thrombin on growth factor release from equine platelet-rich gel supernatants.

Abstract: To compare five activation methods in equine platelet-rich plasma (PRP) by determination of platelet-derived growth factor BB (PDGF-BB) and transforming growth factor beta 1 (TGF-β1) concentrations in platelet-rich gel (PRG) supernatants. Methods: Platelet-rich plasma from 20 horses was activated by calcium chloride (CC), calcium gluconate (CG), bovine thrombin (BT), and their combinations, BTCC and BTCG. Both growth factor concentrations in PRG supernatants were measured by ELISA and compared with plasma and platelet lysates (PL) over time. Results: Growth factor concentrations were significantly lower in plasma and higher for all PRG supernatants. Platelet lysates contained a significantly lower concentration of PDGF-BB than PRG supernatants and a significantly higher concentration of TGF-β1 than PRG supernatants. Clots from PRP activated with sodium salts were more stable over time and had significant growth factor release, whereas CC produced gross salt deposition. Significant correlations were noticed for platelet with leukocyte concentrations in PRP (rs: 0.76), platelet counts in PRP with TGF-β1 concentrations in PRG supernatants (rs: 0.86), platelet counts in PRP with PDGF-BB concentrations in PRG supernatants (rs: 0.78), leukocyte counts in PRP with TGF-β1 concentrations in PRG supernatants (rs: 0.76), and PDGF-BB concentrations with activating substances (rs: 0.72). Conclusions: Calcium gluconate was the better substance to induce PRP activation. It induced growth factor release free from calcium precipitates in the clots. Use of BT alone or combined with calcium salts was not advantageous for growth factor release.
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Publication Date: 2016-11-16 PubMed ID: 27849108DOI: 10.3415/VCOT-16-02-0026Google Scholar: Lookup
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Summary

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This research compares the effects of five different activation methods on the release of growth factors from equine platelet-rich plasma (PRP). The results indicate that calcium gluconate is the most effective substance for inducing activation and growth factor release, while bovine thrombin, alone or in combination with calcium salts, does not provide any additional advantage.

Research Methodology

  • The research team collected platelet-rich plasma (PRP) from 20 horses.
  • They activated the PRP using one of five methods: calcium chloride (CC), calcium gluconate (CG), bovine thrombin (BT), BT with calcium chloride (BTCC), or BT with calcium gluconate (BTCG).
  • The researchers measured the concentrations of two growth factors (PDGF-BB and TGF-β1) in the platelet-rich gel (PRG) supernatants using ELISA, a lab test that detects and quantifies substances in a solution.

Findings

  • The study found higher concentrations of growth factors in all PRG supernatants when compared to plasma or platelet lysates (PL); however, PL had a higher concentration of TGF-β1 and lower PDGF-BB than the PRG supernatants.
  • PRP activated with calcium chloride resulted in visible salt deposition. On the other hand, PRP activated with sodium salts formed more stable clots and released considerable amounts of growth factors over time.
  • A strong correlation was noted between platelet and leukocyte concentrations in PRP and the corresponding growth factor concentrations in PRG supernatants. Furthermore, the concentration of PDGF-BB was significantly associated with the type of activating substances used.

Conclusions

  • Calcium gluconate was found to be the best substance for inducing PRP activation, releasing growth factors without leading to calcium precipitates in the clots.
  • The use of bovine thrombin, alone or combined with different calcium salts, did not increase the effectiveness of PRP activation or growth factor release.

Cite This Article

APA
Giraldo CE, Álvarez ME, Carmona JU. (2016). Influence of calcium salts and bovine thrombin on growth factor release from equine platelet-rich gel supernatants. Vet Comp Orthop Traumatol, 30(1), 1-7. https://doi.org/10.3415/VCOT-16-02-0026

Publication

Veterinary and comparative orthopaedics and traumatology : V.C.O.T
ISSN: 2567-6911
NlmUniqueID: 8906319
Country: Germany
Language: English
Volume: 30
Issue: 1
Pages: 1-7

Researcher Affiliations

Giraldo, Carlos E
    Álvarez, María E
      Carmona, Jorge U
      • Prof. Jorge U. Carmona, MVZ, MSc, PhD, Departamento de Salud Animal, Universidad de Caldas, Calle 65, No 26-10, Manizales, Colombia, Phone/Fax: +57 68 781516, E-mail: carmona@ucaldas.edu.co.

      MeSH Terms

      • Animals
      • Blood Coagulation
      • Blood Platelets
      • Calcium / pharmacology
      • Cattle
      • Gels
      • Horses
      • Leukocyte Count
      • Leukocytes
      • Male
      • Platelet Count
      • Platelet-Derived Growth Factor / metabolism
      • Platelet-Rich Plasma / cytology
      • Platelet-Rich Plasma / drug effects
      • Platelet-Rich Plasma / metabolism
      • Thrombin / pharmacology
      • Transforming Growth Factor beta1 / metabolism

      Citations

      This article has been cited 14 times.
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      12. McClain AK, McCarrel TM. The effect of four different freezing conditions and time in frozen storage on the concentration of commonly measured growth factors and enzymes in equine platelet-rich plasma over six months. BMC Vet Res 2019 Aug 14;15(1):292.
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