Activation of equine platelet-rich plasma: comparison of methods and characterization of equine autologous thrombin.

Textor, JA
·
Tablin, F

Abstract: To investigate and compare clinically relevant Platelet-rich plasma (PRP) activation methods. Methods: Experimental. Methods: PRP was prepared from 6 equine subjects. Activation of the PRP was performed by 4 methods (autologous thrombin, bovine thrombin, calcium chloride (CaCl(2) ), or freeze-thaw). The resultant PDGF-BB (where PDGF is platelet-derived growth factor) and TGFβ1 (where TGFβ is transforming growth factor beta) levels in PRP releasates were quantified by Enzyme-linked immunosorbent assay (ELISA) and compared. Growth factor contents were also compared between platelet-rich clots produced by thrombin or CaCl(2) . The composition and function of equine autologous thrombin were characterized by Western blot analysis and platelet aggregometry. Results: CaCl(2) (23 mM) activation of PRP yielded significantly greater PDGF release than did any other method. TGFβ release was comparable after PRP activation by CaCl(2) , bovine thrombin, and freeze thaw. Autologous thrombin was significantly less effective than all other activation methods in eliciting platelet growth factor release and induced significantly less platelet aggregation than bovine thrombin at 5 U/mL. Clots retained substantial concentrations of growth factor, and the amount in the releasate versus the clot differed between activation methods. Conclusions: PRP activation methods differ in terms of growth factor output as well as logistical considerations. Autologous thrombin is not recommended for PRP activation. CaCl(2) (23 mM) is an effective and inexpensive method of PRP activation. The PRP releasate derived from CaCl(2) activation contains 80% of the total PDGF content and is easily produced, making it a convenient product for clinical use.

Get Full Text

Publication Date: 2012-06-28 PubMed ID: 22742830DOI: 10.1111/j.1532-950X.2012.01016.xGoogle 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.

Facebook X LinkedIn Email

Summary
Cite This
Publication
Affiliations
MeSH
Citations

Comparative Study
Journal Article
Research Support
Non-U.S. Gov't

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 compares different methods of activating Platelet-rich plasma (PRP) in horses. The study discovered that the use of calcium chloride (CaCl(2)) yields significantly greater platelet-derived growth factor (PDGF) release compared to other methods, and autologous thrombin is not recommended for PRP activation.

Study Design and Methods

The researchers conducted this experimental study on six equine subjects. They prepared Platelet-rich plasma (PRP) from each of the subjects.
The team used four specific methods to activate the PRP: autologous thrombin, bovine thrombin, calcium chloride (CaCl(2)), and freeze-thaw.
The levels of PDGF-BB, a type of platelet-derived growth factor, and TGFβ1, a transforming growth factor beta, in the PRP were quantified using ELISA.
Growth factor contents were compared between the platelet-rich clots produced using thrombin and those created with CaCl(2).
The study evaluated the composition and function of the equine autologous thrombin using Western blot analysis and platelet aggregometry.

Results of the Study

The use of calcium chloride(CaCl(2)) for PRP activation resulted in the greatest release of PDGF compared to other methods.
TGFβ release was comparable after PRP activation by CaCl(2), bovine thrombin, and freeze thaw.
Autologous thrombin was found to be significantly less effective in inducing platelet growth factor release than all other methods.
Autologous thrombin also induced significantly less platelet aggregation than bovine thrombin at 5 U/mL.
The clots maintained significant concentrations of growth factor, but the amount released versus the clot depended on the activation method used.

Conclusions and Recommendations

The researchers concluded that the different methods of PRP activation vary in terms of growth factor output and logistical considerations.
They specifically advised against the use of autologous thrombin for PRP activation.
They found that calcium chloride (CaCl(2)), specifically at a concentration of 23 mM, is a cost-effective method for PRP activation.
The team identified that the PRP releasate obtained from CaCl(2) activation contained approximately 80% of the total PDGF content and is easily produced, making it a suitable product for clinical application.

Cite This Article

APA

Textor JA, Tablin F. (2012). Activation of equine platelet-rich plasma: comparison of methods and characterization of equine autologous thrombin. Vet Surg, 41(7), 784-794. https://doi.org/10.1111/j.1532-950X.2012.01016.x

Publication

Veterinary surgery : VS

ISSN: 1532-950X

NlmUniqueID: 8113214

Country: United States

Language: English

Volume: 41

Issue: 7

Pages: 784-794

Researcher Affiliations

Textor, Jamie A

Department of Anatomy, Physiology and Cell Biology, School of Veterinary Medicine, University of California-Davis, Davis, CA 95616, USA. jamietextor@gmail.com

Tablin, Fern

MeSH Terms

Animals
Blood Coagulation / physiology
Blood Transfusion, Autologous / veterinary
Cattle
Female
Horses / blood
Male
Platelet Activation / physiology
Platelet-Derived Growth Factor / metabolism
Platelet-Rich Plasma / cytology
Platelet-Rich Plasma / physiology
Thrombin / chemistry
Thrombin / classification
Thrombin / metabolism
Time Factors

Citations

This article has been cited 31 times.

Nadra M, Niu W, Kurisawa M, Rousson D, Spector M. Platelet-Rich Plasma Lysate-Incorporating Gelatin Hydrogel as a Scaffold for Bone Reconstruction.. Bioengineering (Basel) 2022 Sep 29;9(10).
doi: 10.3390/bioengineering9100513pubmed: 36290482google scholar: lookup
Garbin LC, Contino EK, Olver CS, Frisbie DD. A safety evaluation of allogeneic freeze-dried platelet-rich plasma or conditioned serum compared to autologous frozen products equivalents in equine healthy joints.. BMC Vet Res 2022 Apr 18;18(1):141.
doi: 10.1186/s12917-022-03225-4pubmed: 35436878google scholar: lookup
Camargo Garbin L, Lopez C, Carmona JU. 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.2021.641818pubmed: 33869321google scholar: lookup
Jakfar S, Lin TC, Wu SC, Wang YH, Sun YJ, Thacker M, Liu LX, Lin FH. New design to remove leukocytes from platelet-rich plasma (PRP) based on cell dimension rather than density.. Bioact Mater 2021 Oct;6(10):3528-3540.
doi: 10.1016/j.bioactmat.2021.03.002pubmed: 33842739google scholar: lookup
Hagen A, Lehmann H, Aurich S, Bauer N, Melzer M, Moellerberndt J, Patanu00e9 V, Schnabel CL, Burk J. Scalable Production of Equine Platelet Lysate for Multipotent Mesenchymal Stromal Cell Culture.. Front Bioeng Biotechnol 2020;8:613621.
doi: 10.3389/fbioe.2020.613621pubmed: 33553119google scholar: lookup
El-Timamy A, El Sharaby F, Eid F, El Dakroury A, Mostafa Y, Shaker O. Effect of platelet-rich plasma on the rate of orthodontic tooth movement.. Angle Orthod 2020 May 1;90(3):354-361.
doi: 10.2319/072119-483.1pubmed: 33378433google scholar: lookup
Fukuda K, Kuroda T, Tamura N, Mita H, Kasashima Y. Optimal activation methods for maximizing the concentrations of platelet-derived growth factor-BB and transforming growth factor-u03b21 in equine platelet-rich plasma.. J Vet Med Sci 2020 Oct 20;82(10):1472-1479.
doi: 10.1292/jvms.20-0167pubmed: 32814750google scholar: lookup
Perego R, Spada E, Baggiani L, Martino PA, Proverbio D. Efficacy of a Semi Automated Commercial Closed System for Autologous Leukocyte- and Platelet-Rich Plasma (l-prp) Production in Dogs: A Preliminary Study.. Animals (Basel) 2020 Aug 4;10(8).
doi: 10.3390/ani10081342pubmed: 32759643google scholar: lookup
Gilbertie JM, Schaer TP, Schubert AG, Jacob ME, Menegatti S, Ashton Lavoie R, Schnabel LV. Platelet-rich plasma lysate displays antibiofilm properties and restores antimicrobial activity against synovial fluid biofilms in vitro.. J Orthop Res 2020 Jun;38(6):1365-1374.
doi: 10.1002/jor.24584pubmed: 31922274google scholar: lookup
Camargo Garbin L, McIlwraith CW, Frisbie DD. Evaluation of allogeneic freeze-dried platelet lysate in cartilage exposed to interleukin 1-u03b2 in vitro.. BMC Vet Res 2019 Nov 1;15(1):386.
doi: 10.1186/s12917-019-2118-zpubmed: 31675958google scholar: lookup
Elgamal A, Althani A A, Abd-Elmaksoud A, Kassab M, Farag A, Lashen S, Gabr MM, Zakaria MM, Alissawi MM, Ismail HEA, Abd El Galil A, Caceci T, Cenciarelli C C, Marei HE. Xeno-free trans-differentiation of adipose tissue-derived mesenchymal stem cells into glial and neuronal cells.. Am J Stem Cells 2019;8(2):38-51.
pubmed: 31523485
Seidel SRT, Vendruscolo CP, Moreira JJ, Fu00fclber J, Ottaiano TF, Oliva MLV, Michelacci YM, Baccarin RYA. Does Double Centrifugation Lead to Premature Platelet Aggregation and Decreased TGF-u03b21 Concentrations in Equine Platelet-Rich Plasma?. Vet Sci 2019 Aug 21;6(3).
doi: 10.3390/vetsci6030068pubmed: 31438534google scholar: lookup
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.
doi: 10.1186/s12917-019-2040-4pubmed: 31412868google scholar: lookup
Du L, Miao Y, Li X, Shi P, Hu Z. A Novel and Convenient Method for the Preparation and Activation of PRP without Any Additives: Temperature Controlled PRP.. Biomed Res Int 2018;2018:1761865.
doi: 10.1155/2018/1761865pubmed: 29862255google scholar: lookup
Bogers SH. Cell-Based Therapies for Joint Disease in Veterinary Medicine: What We Have Learned and What We Need to Know.. Front Vet Sci 2018;5:70.
doi: 10.3389/fvets.2018.00070pubmed: 29713634google scholar: lookup
Naskou MC, Sumner SM, Chocallo A, Kemelmakher H, Thoresen M, Copland I, Galipeau J, Peroni JF. Platelet lysate as a novel serum-free media supplement for the culture of equine bone marrow-derived mesenchymal stem cells.. Stem Cell Res Ther 2018 Mar 22;9(1):75.
doi: 10.1186/s13287-018-0823-3pubmed: 29566772google scholar: lookup
Ben-Nafa W, Munro W. The effect of corticosteroid versus platelet-rich plasma injection therapies for the management of lateral epicondylitis: A systematic review.. SICOT J 2018;4:11.
doi: 10.1051/sicotj/2017062pubmed: 29561260google scholar: lookup
Seabaugh KA, Thoresen M, Giguu00e8re S. Extracorporeal Shockwave Therapy Increases Growth Factor Release from Equine Platelet-Rich Plasma In Vitro.. Front Vet Sci 2017;4:205.
doi: 10.3389/fvets.2017.00205pubmed: 29270410google scholar: lookup
Xiong G, Lingampalli N, Koltsov JCB, Leung LL, Bhutani N, Robinson WH, Chu CR. Men and Women Differ in the Biochemical Composition of Platelet-Rich Plasma.. Am J Sports Med 2018 Feb;46(2):409-419.
doi: 10.1177/0363546517740845pubmed: 29211968google scholar: lookup
Garner AL, Caiafa A, Jiang Y, Klopman S, Morton C, Torres AS, Loveless AM, Neculaes VB. Design, characterization and experimental validation of a compact, flexible pulsed power architecture for ex vivo platelet activation.. PLoS One 2017;12(7):e0181214.
doi: 10.1371/journal.pone.0181214pubmed: 28746392google scholar: lookup
Shin HS, Woo HM, Kang BJ. Optimisation of a double-centrifugation method for preparation of canine platelet-rich plasma.. BMC Vet Res 2017 Jun 26;13(1):198.
doi: 10.1186/s12917-017-1123-3pubmed: 28651609google scholar: lookup
Birdwhistell KE, Karumbaiah L, Franklin SP. Sustained Release of Transforming Growth Factor-u03b21 from Platelet-Rich Chondroitin Sulfate Glycosaminoglycan Gels.. J Knee Surg 2018 May;31(5):410-415.
doi: 10.1055/s-0037-1603801pubmed: 28645130google scholar: lookup
Franklin SP, Birdwhistell KE, Strelchik A, Garner BC, Brainard BM. Influence of Cellular Composition and Exogenous Activation on Growth Factor and Cytokine Concentrations in Canine Platelet-Rich Plasmas.. Front Vet Sci 2017;4:40.
doi: 10.3389/fvets.2017.00040pubmed: 28424777google scholar: lookup
Kraeutler MJ, Garabekyan T, Mei-Dan O. The use of platelet-rich plasma to augment conservative and surgical treatment of hip and pelvic disorders.. Muscles Ligaments Tendons J 2016 Jul-Sep;6(3):410-419.
doi: 10.11138/mltj/2016.6.3.410pubmed: 28066748google scholar: lookup
Fernandes G, Yang S. Application of platelet-rich plasma with stem cells in bone and periodontal tissue engineering.. Bone Res 2016;4:16036.
doi: 10.1038/boneres.2016.36pubmed: 28018706google scholar: lookup
Tang HC, Chen WC, Chiang CW, Chen LY, Chang YC, Chen CH. 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 Aug 7;16(8):18507-21.
doi: 10.3390/ijms160818507pubmed: 26262616google scholar: lookup
Brossi PM, Moreira JJ, Machado TS, Baccarin RY. Platelet-rich plasma in orthopedic therapy: a comparative systematic review of clinical and experimental data in equine and human musculoskeletal lesions.. BMC Vet Res 2015 Apr 22;11:98.
doi: 10.1186/s12917-015-0403-zpubmed: 25896610google scholar: lookup
Giraldo CE, u00c1lvarez ME, Carmona JU. Effects of sodium citrate and acid citrate dextrose solutions on cell counts and growth factor release from equine pure-platelet rich plasma and pure-platelet rich gel.. BMC Vet Res 2015 Mar 14;11:60.
doi: 10.1186/s12917-015-0370-4pubmed: 25889052google scholar: lookup
Talaie T, Pratt SJP, Vanegas C, Xu S, Henn RF 3rd, Yarowsky P, Lovering RM. Site-specific targeting of platelet-rich plasma via superparamagnetic nanoparticles.. Orthop J Sports Med 2015 Jan 22;3(1).
doi: 10.1177/2325967114566185pubmed: 25664326google scholar: lookup
Lu00f3pez C, Alvarez ME, Carmona JU. Temporal Bacteriostatic Effect and Growth Factor Loss in Equine Platelet Components and Plasma Cultured with Methicillin-Sensitive and Methicillin-Resistant Staphylococcus aureus: A Comparative In Vitro Study.. Vet Med Int 2014;2014:525826.
doi: 10.1155/2014/525826pubmed: 25506468google scholar: lookup
Carvalho Ade M, Badial PR, u00c1lvarez LE, Yamada AL, Borges AS, Deffune E, Hussni CA, Garcia Alves AL. Equine tendonitis therapy using mesenchymal stem cells and platelet concentrates: a randomized controlled trial.. Stem Cell Res Ther 2013 Jul 22;4(4):85.
doi: 10.1186/scrt236pubmed: 23876512google scholar: lookup