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Comparison of the methods for platelet rich plasma preparation in horses

Comparison of the methods for platelet rich plasma preparation in horses Platelet rich plasma (PRP) is popularly used in the horse industry to enhance regeneration of tissue injury that has limitation of blood supply. This study aimed to compare the methods for platelet rich plasma preparation since they has not been established yet. Blood was collected from six horses and platelets were concentrated by three different methods (2-step centrifugation, separated centrifugation and separated centrifugation using histopaque). Concentrated blood was analyzed using Advia hematology systems. In the result, separated centrifugation with histopaque showed the significantly lower number of red blood cells than other groups. The 2-step centrifugation showed the significantly higher number of white blood cells than other groups, while it contained the highest concentration of red blood cells among three groups. In the 2-step centrifugation, separated centrifugation and separated centrifugation with histopaque, platelets were concentrated 4.5, 5.3 and 5.6 times, respectively. And no significant difference of the platelet concentration between the three groups was found. This study demonstrated that separated centrifugation using histopaque was the best method for platelet rich plasma preparation because of the proper amount of platelets and the separation of red blood cells from platelet rich plasma. Keywords: Platelet rich plasma, Red blood cells, White blood cells, Horse Introduction Materials and methods Platelet rich plasma (PRP) is the blood plasma which Six horses were used in this study (two of Thoroughbred contains a high concentration of platelets. Recently, and four of Halla horses; two males and four females; PRP is largely used in the treatment of equine muscu- mean age, 8 years; mean weight, 340 kg). Blood was loskeletal, soft tissue, skin injuries [1, 2]. It includes collected from jugular vein using 16G catheter into acid α-granules that secrete the growth factors, including citrate dextrose A (ACD-A) coated syringes. Complete platelet-derived growth factor (PDGF), transforming blood count was performed by Advia hematology systems. growth factor beta (TGF-β) and vascular endothelial In a 2-step centrifugation method, blood was centrifuged growth factor (VEGF) [3–5]. The growth factors help at 200 g for 15 min. A layer of platelet rich plasma and the migration of the cells and the regeneration of white blood cells were collected. The suspension was cen- blood vessels, ligaments, tendons, bones and skin [1, trifuged again at 900 g for 15 min. The supernatant was 6]. In horses, there are 100,000–350,000 platelets/ul in discarded and 1.5 ml of platelet rich plasma was collected the blood and platelet rich plasma should contain and analyzed by Advia hematology systems. In a separated three times to five times of platelets [7]. However, centrifugation method, bloodwas centrifugedat200 g for there is no establishment of the methods for platelet 15 min and plasma was collected. Residual white blood rich plasma preparation [8]. Therefore, the purpose of cells and red blood cells were collected in a different tube. this study is a comparison of the three different Plasma was centrifuged at 900 g for 15 min, the super- methods for platelet rich plasma preparation. natant was discarded and 1 ml of plasma was collected. Re- sidual blood cells were centrifuged at 200 g for 15 min and the supernatant was collected. Collected plasma and white blood cells were mixed together, then complete blood was counted. In a separated centrifugation using histopaque * Correspondence: jpseo@jejunu.ac.kr College of Veterinary Medicine and Veterinary Medical Research Institute, (Histopaque® -1119, Sigma, St. Louis, USA) method, blood Jeju National University, Jeju-City, Jejudo 63243, Republic of Korea © The Author(s). 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Lee et al. Journal of Animal Science and Technology (2018) 60:20 Page 2 of 4 was centrifuged at 200 g for 15 min and plasma was col- and the suspension was centrifuged at 400 g for 30 min. lected and then centrifuged again at 900 g for 15 min. The White blood cells were separated and washed with PBS, supernatant was discarded and collected the plasma. Re- then centrifuged again at 200 g for 10 min. PBS was re- sidual blood cells were collected into a different tube and moved and white blood cells were added into the plasma mixed with PBS (1:1). Histopaque was added into the tube tube. Complete blood count was performed (Fig. 1). All the Fig. 1 Summary of the methods for platelet rich plasma preparation. a, 2-step centrifugation. b Separated centrifugation. c Separated centrifugation with histopaque Lee et al. Journal of Animal Science and Technology (2018) 60:20 Page 3 of 4 data obtained was analyzed by a statistical software (SPSS platelets but also red blood cells and white blood cells Inc., IBM, USA) using non-parametric Mann-Whitney [9]. Red blood cells and white blood cells have been (statistical significance was considered at P <0.05) and identified to have important roles in immune-mediated Kruskal-Wallis tests (statistical significance was considered response [9]. Therefore, regulating the amount of blood at P < 0.017). cells was important in this study. Since red blood cells increased immune-mediated factors such as interleukin- Results 1 and TGF-α, it is important to reduce the amount of Red blood cells were significantly lower in the method of red blood cells during preparation of platelet rich plasma separated centrifugation using histopaque (mean 0.09 ± [6]. Also, it was demonstrated that immune-mediated 0.05 × 10 / ul) and significantly higher in the 2-step centri- factors were increased when there were high concentra- fugation method (mean 8.50 ± 2.31 × 10 / ul) than in other tion of red blood cells in platelet rich plasma which have groups. The significantly higher number of white blood the low number of white blood cells [6]. The efforts to cells was shown in the 2-step centrifugation (mean 35.20 reduce red blood cells were conducted by using single or ±3.65 ×10 / ul) and the lowest number was shown with double centrifugation [10] and difference of time and separated centrifugation with histopaque method (mean the gravitational force of centrifugation [11]. In this 12.53 ± 2.59 × 10 / ul). However, a significant difference of study, separated centrifugation with histopaque showed the number of white blood cells between the separated the significantly lower number of red blood cells than centrifugation and separated centrifugation with histopa- the 2-step centrifugation and separated centrifugation. que was not found. The platelets were the most concen- This is because histopaque separated the layer of white trated with separated centrifugation with histopaque blood cells from the layer of red blood cells. method. In the 2-step centrifugation, separated centrifuga- The necessity of white blood cells in platelet rich tion and separated centrifugation with histopaque, plate- plasma is still controversial. McCarrel T et al. observed lets were concentrated 4.5, 5.3 and 5.6 times, respectively. that white blood cells and corticosteroids were effective However, no significant difference of the platelet concen- in the treatment of chronic lateral epicondylitis in horse tration between the three groups was found (Table 1). [12]. Also, platelet rich plasma, including white blood cells except neutrophils had an effect on anterior cruci- Discussion ate ligament fibroblast [13]. Therefore, we had effort to In a recent study, platelet rich plasma has been thought retain white blood cells in platelet rich plasma. However, to take a therapeutic effect in the aspect of not only there are some adverse effects when white blood cells Table 1 Results of platelet rich plasma analysis Horse RBC WBC Platelet b,c b,c w,a,b w,a,b,c a,b,c w,b,c w,a w,a a,b,c w w w W A B C W A B C W A B C 1 Halla 6.96 9.03 2.51 0.14 7.04 36.56 14.85 17.06 79 368 556 605 Female 13 yrs 2 Halla 7.07 10.82 4.15 0.11 5.94 33.25 18.55 10.69 71 274 439 380 Male 7 yrs 3TB 11.26 11.56 4.65 0.12 8.88 34.77 19.2 13.5 104 503 680 650 Female 4 yrs 4TB 7.76 6.45 2.46 0.03 7.8 34.1 21.3 13 106 602 427 629 Female 4 yrs 5 Halla 6.92 6.57 2 0.02 7.9 41.6 12 10.7 133 494 543 499 Male 7 yrs 6 Halla 6.61 6.57 4.56 0.13 7.97 30.89 22.72 10.25 118 497 610 652 Female 13 yrs Mean 7.76 8.50 3.39 0.09 7.59 35.20 18.10 12.53 101.83 456.33 542.50 569.17 SD 1.75 2.31 1.19 0.05 1.00 3.65 4.02 2.59 23.35 116.20 97.67 108.60 A 2-setp centrifugation, B Separated centrifugation, C Separated centrifugation with histopaque, TB Thoroughbred, W Whole blood, RBC Red blood cell, WBC White w,a,b,c blood cell, Different letters represent significant differences between groups (P-value: W vs. A, B, C, Mann-Whitney test, P < 0.05; A vs. B vs. C, Kruskal-Wallis test, P < 0.017) Lee et al. Journal of Animal Science and Technology (2018) 60:20 Page 4 of 4 are concentrated [12]. Immune-mediated factors such as Competing interests The authors declare that they have no competing interests. interlukin-1 and TGF-α could be increased because of concentrated neutrophils [11] and platelet rich plasma, Publisher’sNote including a high concentration of white blood cells de- Springer Nature remains neutral with regard to jurisdictional claims in creased synthesis of extracellular matrix [9]. In this published maps and institutional affiliations. study, white blood cells were significantly higher in the Received: 26 April 2018 Accepted: 6 August 2018 2-step centrifugation method than other groups. The lowest number of white blood cells was observed with the method of separated centrifugation using histropa- References 1. Hessel LN, Bosch G, van Weeren PR, Ionita JC. Equine autologous platelet que but not significantly different with the separated concentrates: comparative study between different available systems. centrifugation. This is may be due to loss of white blood Equine Vet J. 2015;47:319–25. cells during washing procedure. 2. Textor JA, Norris JW, Tablin F. Effects of preparation method, shear force, and exposure to collagen on release of growth factors from equine Different methods for preparation of platelet rich platelet-rich plasma. Am J Vet Res. 2011;72:271–8. plasma has been developed [6, 7, 9, 13]. The proper 3. Argüelles D, Carmona JU, Pastor J, Iborra A, Viñals L, Martínez P, Bach E, number of platelets of platelet rich plasma was three to Prades M. Evaluation of single and double centrifugation tube methods for concentrating equine platelets. Res Vet Sci. 2006;81:237–45. five times higher than whole blood [6]. We prepared 4. Fontenot RL, Sink CA, Werre SR, Weinstein NM, Dahlgren LA. Simple tube platelet rich plasma with the proper number of platelets, centrifugation for processing platelet-rich plasma in the horse. Can Vet J. since the concentration of platelets in 2-step centrifuga- 2012;53:1266–72. 5. Rutkowski JL, Thomas JM, Bering CL, Speicher JL, Radio NM, Smith DM, tion, separated centrifugation and separated centrifuga- Johnson DA. Analysis of a rapid, simple, and inexpensive technique used to tion with histopaque method was 4.5, 5.3 and 5.6 times obtain platelet-rich plasma for use in clinical practice. J Oral Implantol. 2008; respectively, with no significant difference. 34:25–33. 6. Carr BJ, Canapp SO Jr, Mason DR, Cox C, Hess T. Canine platelet-rich plasma systems: a prospective analysis. Front Vet Sci. 2016;2:73. Conclusion 7. Maia L, Souza MV, Ribeiro Jr BS, J.I., de Oliveira AC, Silveira Alves GE, dos Anjos Benjamin L, Robaina Sancler Silvaa YF, Zandi BM, do Carmo Lopes The purpose of this study was comparing the methods Moreira J. Platelet-rich plasma in the treatment of induced tendinopathy in for platelet rich plasma preparation to ascertain the most horses: histologic evaluation. J Equine Vet Sci 2009;29:618–626. appropriate method. We intended to reduce red blood 8. Dohan Ehrenfest DM, Rasmusson L, Albrektsson T. Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte - and cells and to preserve white blood cells during the platelet-rich fibrin (L-PRF). Trends Biotechnol. 2009;27:158–67. process of platelet rich plasma. The method of using his- 9. Boswell SG, Schnabel LV, Mohammed HO, Sundman EA, Minas T, Fortier LA. topaque was considered the most appropriate since Increasing platelet concentrations in leukocyte-reduced platelet-rich plasma decrease collagen genesynthesis in tendons. Am J Sports Med. 2014;42:42–9. platelets were concentrated while red blood cells were 10. Perez AG, Lana JF, Rodrigues AA, Luzo AC, Belangero WD, Santana MH. removed the most and white blood cells were included. Relevant aspects of centrifugation step in the preparation of platelet-rich plasma. ISRN Hematol. 2014;2014:176060. 11. Sundman EA, Cole BJ, Fortier LA. Growth factor and catabolic cytokine Acknowledgments concentrations are influenced by the cellular composition of platelet-rich This research was supported by the Basic Science Research Program through plasma. Am J Sports Med. 2011;39:2135–40. the National Research Foundation of Korea (NRF) funded by the Ministry of 12. McCarrel T, Fortier L. Temporal growth factor release from platelet-rich Education, Republic of Korea. (NRF-2017R1C1B1006030). plasma, trehalose lyophilized platelets, and bone marrow aspirate and their effect on tendon and ligament gene expression. J Orthop Res. 2009;27: Funding 1033–42. This research was supported by the Basic Science Research Program through 13. Yoshida R, Murray MM. Peripheral blood mononuclear cells enhance the the National Research Foundation of Korea (NRF) funded by the Ministry of anabolic effects of platelet-rich plasma on anteriorcruciate ligament Education, Republic of Korea. fibroblasts. J Orthop Res. 2013;31:29–34. Availability of data and materials The data generated or analyzed during the current study are available from the corresponding author on reasonable request. Authors’ contributions JWK performed the experiments, analyzed the results. EBL participated in analysis of results, and was a major contributor in writing the manuscript. JPS designed the experiments and revised the manuscript. All authors read and approved the final manuscript. Ethics approval The experimental procedure and methods were approved by the Experimental Animal Committee of Veterinary Medicine of Jeju National University, Jeju, Republic of Korea. Consent for publication Not applicable. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Animal Science and Technology Springer Journals

Comparison of the methods for platelet rich plasma preparation in horses

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Publisher
Springer Journals
Copyright
Copyright © 2018 by The Author(s).
Subject
Life Sciences; Animal Genetics and Genomics; Agriculture
eISSN
2055-0391
DOI
10.1186/s40781-018-0178-4
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Abstract

Platelet rich plasma (PRP) is popularly used in the horse industry to enhance regeneration of tissue injury that has limitation of blood supply. This study aimed to compare the methods for platelet rich plasma preparation since they has not been established yet. Blood was collected from six horses and platelets were concentrated by three different methods (2-step centrifugation, separated centrifugation and separated centrifugation using histopaque). Concentrated blood was analyzed using Advia hematology systems. In the result, separated centrifugation with histopaque showed the significantly lower number of red blood cells than other groups. The 2-step centrifugation showed the significantly higher number of white blood cells than other groups, while it contained the highest concentration of red blood cells among three groups. In the 2-step centrifugation, separated centrifugation and separated centrifugation with histopaque, platelets were concentrated 4.5, 5.3 and 5.6 times, respectively. And no significant difference of the platelet concentration between the three groups was found. This study demonstrated that separated centrifugation using histopaque was the best method for platelet rich plasma preparation because of the proper amount of platelets and the separation of red blood cells from platelet rich plasma. Keywords: Platelet rich plasma, Red blood cells, White blood cells, Horse Introduction Materials and methods Platelet rich plasma (PRP) is the blood plasma which Six horses were used in this study (two of Thoroughbred contains a high concentration of platelets. Recently, and four of Halla horses; two males and four females; PRP is largely used in the treatment of equine muscu- mean age, 8 years; mean weight, 340 kg). Blood was loskeletal, soft tissue, skin injuries [1, 2]. It includes collected from jugular vein using 16G catheter into acid α-granules that secrete the growth factors, including citrate dextrose A (ACD-A) coated syringes. Complete platelet-derived growth factor (PDGF), transforming blood count was performed by Advia hematology systems. growth factor beta (TGF-β) and vascular endothelial In a 2-step centrifugation method, blood was centrifuged growth factor (VEGF) [3–5]. The growth factors help at 200 g for 15 min. A layer of platelet rich plasma and the migration of the cells and the regeneration of white blood cells were collected. The suspension was cen- blood vessels, ligaments, tendons, bones and skin [1, trifuged again at 900 g for 15 min. The supernatant was 6]. In horses, there are 100,000–350,000 platelets/ul in discarded and 1.5 ml of platelet rich plasma was collected the blood and platelet rich plasma should contain and analyzed by Advia hematology systems. In a separated three times to five times of platelets [7]. However, centrifugation method, bloodwas centrifugedat200 g for there is no establishment of the methods for platelet 15 min and plasma was collected. Residual white blood rich plasma preparation [8]. Therefore, the purpose of cells and red blood cells were collected in a different tube. this study is a comparison of the three different Plasma was centrifuged at 900 g for 15 min, the super- methods for platelet rich plasma preparation. natant was discarded and 1 ml of plasma was collected. Re- sidual blood cells were centrifuged at 200 g for 15 min and the supernatant was collected. Collected plasma and white blood cells were mixed together, then complete blood was counted. In a separated centrifugation using histopaque * Correspondence: jpseo@jejunu.ac.kr College of Veterinary Medicine and Veterinary Medical Research Institute, (Histopaque® -1119, Sigma, St. Louis, USA) method, blood Jeju National University, Jeju-City, Jejudo 63243, Republic of Korea © The Author(s). 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Lee et al. Journal of Animal Science and Technology (2018) 60:20 Page 2 of 4 was centrifuged at 200 g for 15 min and plasma was col- and the suspension was centrifuged at 400 g for 30 min. lected and then centrifuged again at 900 g for 15 min. The White blood cells were separated and washed with PBS, supernatant was discarded and collected the plasma. Re- then centrifuged again at 200 g for 10 min. PBS was re- sidual blood cells were collected into a different tube and moved and white blood cells were added into the plasma mixed with PBS (1:1). Histopaque was added into the tube tube. Complete blood count was performed (Fig. 1). All the Fig. 1 Summary of the methods for platelet rich plasma preparation. a, 2-step centrifugation. b Separated centrifugation. c Separated centrifugation with histopaque Lee et al. Journal of Animal Science and Technology (2018) 60:20 Page 3 of 4 data obtained was analyzed by a statistical software (SPSS platelets but also red blood cells and white blood cells Inc., IBM, USA) using non-parametric Mann-Whitney [9]. Red blood cells and white blood cells have been (statistical significance was considered at P <0.05) and identified to have important roles in immune-mediated Kruskal-Wallis tests (statistical significance was considered response [9]. Therefore, regulating the amount of blood at P < 0.017). cells was important in this study. Since red blood cells increased immune-mediated factors such as interleukin- Results 1 and TGF-α, it is important to reduce the amount of Red blood cells were significantly lower in the method of red blood cells during preparation of platelet rich plasma separated centrifugation using histopaque (mean 0.09 ± [6]. Also, it was demonstrated that immune-mediated 0.05 × 10 / ul) and significantly higher in the 2-step centri- factors were increased when there were high concentra- fugation method (mean 8.50 ± 2.31 × 10 / ul) than in other tion of red blood cells in platelet rich plasma which have groups. The significantly higher number of white blood the low number of white blood cells [6]. The efforts to cells was shown in the 2-step centrifugation (mean 35.20 reduce red blood cells were conducted by using single or ±3.65 ×10 / ul) and the lowest number was shown with double centrifugation [10] and difference of time and separated centrifugation with histopaque method (mean the gravitational force of centrifugation [11]. In this 12.53 ± 2.59 × 10 / ul). However, a significant difference of study, separated centrifugation with histopaque showed the number of white blood cells between the separated the significantly lower number of red blood cells than centrifugation and separated centrifugation with histopa- the 2-step centrifugation and separated centrifugation. que was not found. The platelets were the most concen- This is because histopaque separated the layer of white trated with separated centrifugation with histopaque blood cells from the layer of red blood cells. method. In the 2-step centrifugation, separated centrifuga- The necessity of white blood cells in platelet rich tion and separated centrifugation with histopaque, plate- plasma is still controversial. McCarrel T et al. observed lets were concentrated 4.5, 5.3 and 5.6 times, respectively. that white blood cells and corticosteroids were effective However, no significant difference of the platelet concen- in the treatment of chronic lateral epicondylitis in horse tration between the three groups was found (Table 1). [12]. Also, platelet rich plasma, including white blood cells except neutrophils had an effect on anterior cruci- Discussion ate ligament fibroblast [13]. Therefore, we had effort to In a recent study, platelet rich plasma has been thought retain white blood cells in platelet rich plasma. However, to take a therapeutic effect in the aspect of not only there are some adverse effects when white blood cells Table 1 Results of platelet rich plasma analysis Horse RBC WBC Platelet b,c b,c w,a,b w,a,b,c a,b,c w,b,c w,a w,a a,b,c w w w W A B C W A B C W A B C 1 Halla 6.96 9.03 2.51 0.14 7.04 36.56 14.85 17.06 79 368 556 605 Female 13 yrs 2 Halla 7.07 10.82 4.15 0.11 5.94 33.25 18.55 10.69 71 274 439 380 Male 7 yrs 3TB 11.26 11.56 4.65 0.12 8.88 34.77 19.2 13.5 104 503 680 650 Female 4 yrs 4TB 7.76 6.45 2.46 0.03 7.8 34.1 21.3 13 106 602 427 629 Female 4 yrs 5 Halla 6.92 6.57 2 0.02 7.9 41.6 12 10.7 133 494 543 499 Male 7 yrs 6 Halla 6.61 6.57 4.56 0.13 7.97 30.89 22.72 10.25 118 497 610 652 Female 13 yrs Mean 7.76 8.50 3.39 0.09 7.59 35.20 18.10 12.53 101.83 456.33 542.50 569.17 SD 1.75 2.31 1.19 0.05 1.00 3.65 4.02 2.59 23.35 116.20 97.67 108.60 A 2-setp centrifugation, B Separated centrifugation, C Separated centrifugation with histopaque, TB Thoroughbred, W Whole blood, RBC Red blood cell, WBC White w,a,b,c blood cell, Different letters represent significant differences between groups (P-value: W vs. A, B, C, Mann-Whitney test, P < 0.05; A vs. B vs. C, Kruskal-Wallis test, P < 0.017) Lee et al. Journal of Animal Science and Technology (2018) 60:20 Page 4 of 4 are concentrated [12]. Immune-mediated factors such as Competing interests The authors declare that they have no competing interests. interlukin-1 and TGF-α could be increased because of concentrated neutrophils [11] and platelet rich plasma, Publisher’sNote including a high concentration of white blood cells de- Springer Nature remains neutral with regard to jurisdictional claims in creased synthesis of extracellular matrix [9]. In this published maps and institutional affiliations. study, white blood cells were significantly higher in the Received: 26 April 2018 Accepted: 6 August 2018 2-step centrifugation method than other groups. The lowest number of white blood cells was observed with the method of separated centrifugation using histropa- References 1. Hessel LN, Bosch G, van Weeren PR, Ionita JC. Equine autologous platelet que but not significantly different with the separated concentrates: comparative study between different available systems. centrifugation. This is may be due to loss of white blood Equine Vet J. 2015;47:319–25. cells during washing procedure. 2. Textor JA, Norris JW, Tablin F. Effects of preparation method, shear force, and exposure to collagen on release of growth factors from equine Different methods for preparation of platelet rich platelet-rich plasma. Am J Vet Res. 2011;72:271–8. plasma has been developed [6, 7, 9, 13]. The proper 3. Argüelles D, Carmona JU, Pastor J, Iborra A, Viñals L, Martínez P, Bach E, number of platelets of platelet rich plasma was three to Prades M. Evaluation of single and double centrifugation tube methods for concentrating equine platelets. Res Vet Sci. 2006;81:237–45. five times higher than whole blood [6]. We prepared 4. Fontenot RL, Sink CA, Werre SR, Weinstein NM, Dahlgren LA. Simple tube platelet rich plasma with the proper number of platelets, centrifugation for processing platelet-rich plasma in the horse. Can Vet J. since the concentration of platelets in 2-step centrifuga- 2012;53:1266–72. 5. Rutkowski JL, Thomas JM, Bering CL, Speicher JL, Radio NM, Smith DM, tion, separated centrifugation and separated centrifuga- Johnson DA. Analysis of a rapid, simple, and inexpensive technique used to tion with histopaque method was 4.5, 5.3 and 5.6 times obtain platelet-rich plasma for use in clinical practice. J Oral Implantol. 2008; respectively, with no significant difference. 34:25–33. 6. Carr BJ, Canapp SO Jr, Mason DR, Cox C, Hess T. Canine platelet-rich plasma systems: a prospective analysis. Front Vet Sci. 2016;2:73. Conclusion 7. Maia L, Souza MV, Ribeiro Jr BS, J.I., de Oliveira AC, Silveira Alves GE, dos Anjos Benjamin L, Robaina Sancler Silvaa YF, Zandi BM, do Carmo Lopes The purpose of this study was comparing the methods Moreira J. Platelet-rich plasma in the treatment of induced tendinopathy in for platelet rich plasma preparation to ascertain the most horses: histologic evaluation. J Equine Vet Sci 2009;29:618–626. appropriate method. We intended to reduce red blood 8. Dohan Ehrenfest DM, Rasmusson L, Albrektsson T. Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte - and cells and to preserve white blood cells during the platelet-rich fibrin (L-PRF). Trends Biotechnol. 2009;27:158–67. process of platelet rich plasma. The method of using his- 9. Boswell SG, Schnabel LV, Mohammed HO, Sundman EA, Minas T, Fortier LA. topaque was considered the most appropriate since Increasing platelet concentrations in leukocyte-reduced platelet-rich plasma decrease collagen genesynthesis in tendons. Am J Sports Med. 2014;42:42–9. platelets were concentrated while red blood cells were 10. Perez AG, Lana JF, Rodrigues AA, Luzo AC, Belangero WD, Santana MH. removed the most and white blood cells were included. Relevant aspects of centrifugation step in the preparation of platelet-rich plasma. ISRN Hematol. 2014;2014:176060. 11. Sundman EA, Cole BJ, Fortier LA. Growth factor and catabolic cytokine Acknowledgments concentrations are influenced by the cellular composition of platelet-rich This research was supported by the Basic Science Research Program through plasma. Am J Sports Med. 2011;39:2135–40. the National Research Foundation of Korea (NRF) funded by the Ministry of 12. McCarrel T, Fortier L. Temporal growth factor release from platelet-rich Education, Republic of Korea. (NRF-2017R1C1B1006030). plasma, trehalose lyophilized platelets, and bone marrow aspirate and their effect on tendon and ligament gene expression. J Orthop Res. 2009;27: Funding 1033–42. This research was supported by the Basic Science Research Program through 13. Yoshida R, Murray MM. Peripheral blood mononuclear cells enhance the the National Research Foundation of Korea (NRF) funded by the Ministry of anabolic effects of platelet-rich plasma on anteriorcruciate ligament Education, Republic of Korea. fibroblasts. J Orthop Res. 2013;31:29–34. Availability of data and materials The data generated or analyzed during the current study are available from the corresponding author on reasonable request. Authors’ contributions JWK performed the experiments, analyzed the results. EBL participated in analysis of results, and was a major contributor in writing the manuscript. JPS designed the experiments and revised the manuscript. All authors read and approved the final manuscript. Ethics approval The experimental procedure and methods were approved by the Experimental Animal Committee of Veterinary Medicine of Jeju National University, Jeju, Republic of Korea. Consent for publication Not applicable.

Journal

Journal of Animal Science and TechnologySpringer Journals

Published: Aug 18, 2018

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