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.
Abstract: There is a lack information on the effects of the most commonly used anticoagulants for equine platelet rich plasmas (PRPs) elaboration on cell counts and growth factor release from platelet rich gels (PRGs). The aims of this study were 1) to compare the effects of the anticoagulants sodium citrate (SC), acid citrate dextrose solution A (ACD-A) and ACD-B on platelet (PLT), leukocyte (WBC) and on some parameters associated to platelet activation including mean platelet volume (MPV) and platelet distribution width (PDW) between whole blood, pure PRP (P-PRP) and platelet-poor plasma (PPP); 2) to compare transforming growth factor beta 1 (TGF-β(1)) and platelet-derived growth factor isoform BB (PDGF-BB) concentrations in supernatants from pure PRG (P-PRG), platelet-poor gel (PPG), P-PRP lysate (positive control) and plasma (negative control); 3) to establish the possible correlations between all the studied cellular and molecular parameters. Results: In all cases the three anticoagulants produced P-PRPs with significantly higher PLT counts compared with whole blood and PPP. The concentrations of WBCs were similar between P-PRP and whole blood, but significantly lower in PPP. The type of anticoagulant did not significantly affect the cell counts for each blood component. The anticoagulants also did not affect the MPV and PDW parameters. Independently of the anticoagulant used, all blood components presented significantly different concentrations of PDGF-BB and TGF-β(1). The highest growth factor (GF) concentrations were observed from P-PRP lysates, followed by PRG supernatants, PPP lysates, PPG supernatants and plasma. Significant correlations were observed between PLT and WBC counts (ρ = 0.80), PLT count and TGF-β(1) concentration (ρ = 0.85), PLT count and PDGF-BB concentration (ρ = 0.80) and PDGF-BB and TGF-β(1) concentrations (ρ = 0.75). The type of anticoagulant was not correlated with any of the variables evaluated. Conclusions: The anticoagulants did not significantly influence cell counts or GF concentrations in equine PRP. However, ACD-B was apparently the worst anticoagulant evaluated. It is necessary to perform additional research to determine the effect of anticoagulants on the kinetics of GF elution from P-PRG.
Publication Date: 2015-03-14 PubMed ID: 25889052PubMed Central: PMC4364319DOI: 10.1186/s12917-015-0370-4Google 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.
- 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 paper examines how the use of different anticoagulants affects the levels of cell counts and the release of growth factors from equine platelet rich plasma and gels. The study found that the type of anticoagulant used did not significantly influence these attributes, though did suggest further research to explore the effects of anticoagulants on the processes within these biological materials.
Objectives
- The research aimed to observe how different anticoagulants like sodium citrate (SC), acid citrate dextrose solution A (ACD-A) and ACD-B impacted cellular densities of platelet (PLT), leukocyte (WBC) and some parameters linked to platelet activation.
- The research also intended to compare the concentrations of specific growth factors (TGF-β(1) and PDGF-BB) in various biological materials processed with different anticoagulants.
- Lastly, the study intended to find out if there were correlations between the cellular and molecular parameters they were investigating.
Findings
- All three anticoagulants resulted in platelet rich plasmas that had significantly higher platelet counts compared with whole blood and platelet-poor plasma.
- The concentrations of white blood cells were similar between the platelet rich plasma and whole blood, but significantly lower in platelet poor plasma.
- No significant variation was found in these counts based on the type of anticoagulant.
- The anticoagulants also did not result in significant shifts in Mean Platelet Volume (MPV) and Platelet Distribution Width (PDW).
- In terms of growth factor concentration, the research found that all tested blood components presented significantly different concentrations of PDGF-BB and TGF-β(1), with the highest growth factor concentrations observed in Platelet Rich Plasma lysates.
- The study also found strong correlations between platelet and white blood cell counts, and between platelet count and growth factor concentrations.
Conclusion
- The research concludes that the type of anticoagulant used does not significantly influence cell counts or growth factor concentrations in equine Platelet Rich Plasma (PRP).
- Nevertheless, ACD-B was found to be the least effective of the anticoagulants tested for this use.
- The researchers suggest that additional investigations are needed to understand the effects of anticoagulants on the kinetics of growth factor elution from platelet rich gels.
Cite This Article
APA
Giraldo CE, Álvarez ME, Carmona JU.
(2015).
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, 11, 60.
https://doi.org/10.1186/s12917-015-0370-4 Publication
Researcher Affiliations
- Grupo de Investigación Terapia Regenerativa, Departamento de Salud Animal, Universidad de Caldas, Manizales, Colombia. cgiraldo@ucaldas.edu.co.
- Grupo de Investigación Terapia Regenerativa, Departamento de Salud Animal, Universidad de Caldas, Manizales, Colombia. maria.alvarez_l@ucaldas.edu.co.
- Grupo de Investigación Terapia Regenerativa, Departamento de Salud Animal, Universidad de Caldas, Manizales, Colombia. carmona@ucaldas.edu.co.
MeSH Terms
- Animals
- Anticoagulants / pharmacology
- Becaplermin
- Blood Cell Count / veterinary
- Blood Platelets / chemistry
- Blood Platelets / drug effects
- Blood Platelets / metabolism
- Citrates / pharmacology
- Citric Acid / pharmacology
- Enzyme-Linked Immunosorbent Assay / veterinary
- Glucose / analogs & derivatives
- Glucose / pharmacology
- Horses / blood
- Horses / physiology
- Leukocyte Count / veterinary
- Male
- Platelet Activation / drug effects
- Platelet Count / veterinary
- Platelet-Rich Plasma / chemistry
- Platelet-Rich Plasma / cytology
- Platelet-Rich Plasma / drug effects
- Proto-Oncogene Proteins c-sis / analysis
- Sodium Citrate
- Transforming Growth Factor beta1 / analysis
References
This article includes 28 references
- Castelijns G, Crawford A, Schaffer J, Ortolano GA, Beauregard T, Smith RK. Evaluation of a filter-prepared platelet concentrate for the treatment of suspensory branch injuries in horses.. Vet Comp Orthop Traumatol 2011;24(5):363-9.
- Carmona JU, López C, Sandoval JA. Review of the currently available systems to obtain platelet related products to treat equine musculoskeletal injuries.. Rec Pat Reg Med 2013;3(2):148–59.
- López C, Carmona JU. Platelet-rich plasma as an adjunctive therapy for the management of a severe chronic distal limb wound in a foal.. J Equine Vet Sci 2014;34(9):1128–33.
- Iacopetti I, Perazzi A, Ferrari V, Busetto R. Application of platelet-rich gel to enhance wound healing in the horse: a case report.. J Equine Vet Sci 2012;32(3):123–8.
- Bazzano M, Piccione G, Giannetto C, Tosto F, Di Pietro S, E G. Platelet rich plasma intralesional injection as bedside therapy for tendinitis in athletic horse.. Acta Sci Vet 2013;41:1145.
- Giraldo CE, López C, Álvarez ME, Samudio IJ, Prades M, Carmona JU. Effects of the breed, sex and age on cellular content and growth factor release from equine pure-platelet rich plasma and pure-platelet rich gel.. BMC Vet Res 2013 Feb 12;9:29.
- Carmona J, López C, Giraldo C. Uso de concentrados autólogos de plaquetas como terapia regenerativa de enfermedades crónicas del aparato musculoesquelético equino.. Arch Med Vet 2011;43(1):1–10.
- Dohan Ehrenfest DM, Bielecki T, Mishra A, Borzini P, Inchingolo F, Sammartino G, Rasmusson L, Everts PA. In search of a consensus terminology in the field of platelet concentrates for surgical use: platelet-rich plasma (PRP), platelet-rich fibrin (PRF), fibrin gel polymerization and leukocytes.. Curr Pharm Biotechnol 2012 Jun;13(7):1131-7.
- McCarrel TM, Minas T, Fortier LA. Optimization of leukocyte concentration in platelet-rich plasma for the treatment of tendinopathy.. J Bone Joint Surg Am 2012 Oct 3;94(19):e143(1-8).
- Mazzucco L, Balbo V, Cattana E, Guaschino R, Borzini P. Not every PRP-gel is born equal. Evaluation of growth factor availability for tissues through four PRP-gel preparations: Fibrinet, RegenPRP-Kit, Plateltex and one manual procedure.. Vox Sang 2009 Aug;97(2):110-8.
- O'Shaughnessey K, Matuska A, Hoeppner J, Farr J, Klaassen M, Kaeding C, Lattermann C, King W, Woodell-May J. Autologous protein solution prepared from the blood of osteoarthritic patients contains an enhanced profile of anti-inflammatory cytokines and anabolic growth factors.. J Orthop Res 2014 Oct;32(10):1349-55.
- Lei H, Gui L, Xiao R. The effect of anticoagulants on the quality and biological efficacy of platelet-rich plasma.. Clin Biochem 2009 Sep;42(13-14):1452-60.
- Vendruscolo CP, Carvalho AM, Moraes LF, Maia L, Queiroz DL, Watanabe MJ, Yamada ALM, Alves ALG. Evaluating the effectiveness of different protocols for preparation of platelet rich plasma for use in equine medicine.. Pesq Vet Bras 2012;32(2):106–10.
- Textor JA, Norris JW, Tablin F. Effects of preparation method, shear force, and exposure to collagen on release of growth factors from equine platelet-rich plasma.. Am J Vet Res 2011 Feb;72(2):271-8.
- Textor JA, Tablin F. Activation of equine platelet-rich plasma: comparison of methods and characterization of equine autologous thrombin.. Vet Surg 2012 Oct;41(7):784-94.
- Argüelles D, Carmona JU, Pastor J, Iborra A, Viñals L, Martínez P, Bach E, Prades M. Evaluation of single and double centrifugation tube methods for concentrating equine platelets.. Res Vet Sci 2006 Oct;81(2):237-45.
- GORNALL AG, BARDAWILL CJ, DAVID MM. Determination of serum proteins by means of the biuret reaction.. J Biol Chem 1949 Feb;177(2):751-66.
- Penha-Goncalves MN, Onions DE, Nicolson L. Cloning and sequencing of equine transforming growth factor-beta 1 (TGF beta-1) cDNA.. DNA Seq 1997;7(6):375-8.
- Donnelly BP, Nixon AJ, Haupt JL, Dahlgren LA. Nucleotide structure of equine platelet-derived growth factor-A and -B and expression in horses with induced acute tendinitis.. Am J Vet Res 2006 Jul;67(7):1218-25.
- Macey M, Azam U, McCarthy D, Webb L, Chapman ES, Okrongly D, Zelmanovic D, Newland A. Evaluation of the anticoagulants EDTA and citrate, theophylline, adenosine, and dipyridamole (CTAD) for assessing platelet activation on the ADVIA 120 hematology system.. Clin Chem 2002 Jun;48(6 Pt 1):891-9.
- Fontenot RL, Sink CA, Werre SR, Weinstein NM, Dahlgren LA. Simple tube centrifugation for processing platelet-rich plasma in the horse.. Can Vet J 2012 Dec;53(12):1266-72.
- Silva RF, Carmona JU, Rezende CM. Intra-articular injections of autologous platelet concentrates in dogs with surgical reparation of cranial cruciate ligament rupture: a pilot study.. Vet Comp Orthop Traumatol 2013;26(4):285-90.
- O'Connell SM. Safety issues associated with platelet-rich fibrin method.. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2007 May;103(5):587; author reply 587-93.
- Alvarez ME, Giraldo CE, Carmona JU. Monitoring bacterial contamination in equine platelet concentrates obtained by the tube method in a clean laboratory environment under three different technical conditions.. Equine Vet J 2010 Jan;42(1):63-7.
- Zimmermann R, Arnold D, Strasser E, Ringwald J, Schlegel A, Wiltfang J, Eckstein R. Sample preparation technique and white cell content influence the detectable levels of growth factors in platelet concentrates.. Vox Sang 2003 Nov;85(4):283-9.
- Silva RF, Carmona JU, Rezende CM. Comparison of the effect of calcium gluconate and batroxobin on the release of transforming growth factor beta 1 in canine platelet concentrates.. BMC Vet Res 2012 Jul 25;8:121.
- López C, Giraldo CE, Carmona JU. Evaluation of a double centrifugation tube method for concentrating bovine platelets: Cellular study.. Arch Med Vet 2012;44(2):109–15.
- Dragoo JL, Braun HJ, Durham JL, Ridley BA, Odegaard JI, Luong R, Arnoczky SP. Comparison of the acute inflammatory response of two commercial platelet-rich plasma systems in healthy rabbit tendons.. Am J Sports Med 2012 Jun;40(6):1274-81.
Citations
This article has been cited 14 times.- Farid MF, Abouelela YS, Yasin NAE, Mousa MR, Ibrahim MA, Prince A, Rizk H. A novel cell-free intrathecal approach with PRP for the treatment of spinal cord multiple sclerosis in cats.. Inflamm Regen 2022 Oct 14;42(1):45.
- Segabinazzi LGTM, Podico G, Rosser MF, Nanjappa SG, Alvarenga MA, Canisso IF. Three Manual Noncommercial Methods to Prepare Equine Platelet-Rich Plasma.. Animals (Basel) 2021 May 21;11(6).
- Gonçalves NJN, Frantz N, de Oliveira RM. Platelet-rich plasma (PRP) therapy: An approach in reproductive medicine based on successful animal models.. Anim Reprod 2020 May 22;16(1):93-98.
- 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).
- Dudchenko YS, Maksymova OS, Pikaliuk VS, Muravskyi DV, Kyptenko LI, Tkach GF. Morphological Characteristics and Correction of Long Tubular Bone Regeneration under Chronic Hyperglycemia Influence.. Anal Cell Pathol (Amst) 2020;2020:5472841.
- Oneto P, Zubiry PR, Schattner M, Etulain J. Anticoagulants Interfere With the Angiogenic and Regenerative Responses Mediated by Platelets.. Front Bioeng Biotechnol 2020;8:223.
- Latalski M, Walczyk A, Fatyga M, Rutz E, Szponder T, Bielecki T, Danielewicz A. Allergic reaction to platelet-rich plasma (PRP): Case report.. Medicine (Baltimore) 2019 Mar;98(10):e14702.
- Kardos D, Simon M, Vácz G, Hinsenkamp A, Holczer T, Cseh D, Sárközi A, Szenthe K, Bánáti F, Szathmary S, Nehrer S, Kuten O, Masteling M, Lacza Z, Hornyák I. The Composition of Hyperacute Serum and Platelet-Rich Plasma Is Markedly Different despite the Similar Production Method.. Int J Mol Sci 2019 Feb 8;20(3).
- 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.
- Gutiérrez CM, López C, Giraldo CE, Carmona JU. Study of a Two-Step Centrifugation Protocol for Concentrating Cells and Growth Factors in Bovine Platelet-Rich Plasma.. Vet Med Int 2017;2017:1950401.
- Yu A, Lv J, Yuan F, Xia Z, Fan K, Chen G, Ren J, Lin C, Wei S, Yang F. mPEG-PLA/TPGS mixed micelles via intranasal administration improved the bioavailability of lamotrigine in the hippocampus.. Int J Nanomedicine 2017;12:8353-8362.
- Milants C, Bruyère O, Kaux JF. Responders to Platelet-Rich Plasma in Osteoarthritis: A Technical Analysis.. Biomed Res Int 2017;2017:7538604.
- Carmona JU, Ríos DL, López C, Álvarez ME, Pérez JE. Proinflammatory and Anabolic Gene Expression Effects of Platelet-Rich Gel Supernatants on Equine Synovial Membrane Explants Challenged with Lipopolysaccharide.. Vet Med Int 2017;2017:6059485.
- Carmona JU, Ríos DL, López C, Álvarez ME, Pérez JE, Bohórquez ME. In vitro effects of platelet-rich gel supernatants on histology and chondrocyte apoptosis scores, hyaluronan release and gene expression of equine cartilage explants challenged with lipopolysaccharide.. BMC Vet Res 2016 Jul 1;12(1):135.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists