Effects over time of two platelet gel supernatants on growth factor, cytokine and hyaluronan concentrations in normal synovial membrane explants challenged with lipopolysaccharide.
Abstract: Platelet-rich plasma (PRP) preparations are a common treatment in osteoarthritis (OA) and inflammatory synovitis. However, there is ambiguity regarding the ideal concentration of leukocytes and platelets in these preparations necessary to induce an adequate anti-inflammatory and anabolic response in joint tissues, such as the synovial membrane. This research aimed to study, in normal synovial membrane explants (SME) challenged with lipopolysaccharide (LPS), the temporal effects (at 48 and 96h) of leukocyte- and platelet-rich gel (L-PRG) and pure platelet-rich gel (P-PRG) supernatants on the production and degradation of platelet associated growth factors (GF) (platelet derived GF isoform BB (PDGF-BB) and transforming growth factor beta-1 (TGF-β1)), pro-inflammatory (tumour necrosis factor alpha (TNF-α)) and anti-inflammatory cytokines (interleukin 4 (IL-4) and IL-1 receptor antagonists (IL-1ra)) and hyaluronan (HA). Methods: Synovial membrane explants (SMEs) from 6 horses were challenged with LPS and cultured for 96h with L-PRG and P-PRG supernatants at concentrations of 25 and 50 %, respectively. The SME culture medium was changed every 48h and used for determination by ELISA of PDGF-BB, TGF-β1, TNF-α, IL-4, IL-1ra and HA. These molecules were also determined in synovial fluid from the horses. Results: Both the 25 and 50 % PRG supernatants produced a molecular profile in the culture media unlike that of the SME challenged with LPS only. They presented GF, cytokine and HA concentrations very near to the concentrations of these molecules in normal synovial fluid when compared with the SME control groups (either with LPS or without LPS). However, in comparison with the rest of the SME treated groups, the 25 % L-PRG produced the most IL-1ra, and the 50 % P-PRG induced the sustained production of IL-4 and HA. Conclusions: These in vitro findings suggest that anabolic and anti-inflammatory joint responses depend on the leukocyte and platelet concentration of the PRP preparation and on the volume of this substance injected. Moreover, it is possible, that leukoreduced PRP preparations are more effective for the medical treatment of patients with OA and inflammatory synovitis.
Publication Date: 2015-06-20 PubMed ID: 26092588PubMed Central: PMC4475292DOI: 10.1186/s12891-015-0605-3Google 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 investigates the effectiveness of platelet-rich plasma (PRP) preparations in treating osteoarthritis and inflammatory synovitis. It also seeks to determine the optimal concentration of leukocytes and platelets in these preparations for the best anti-inflammatory and healing outcomes.
Research Objectives and Content
- The research aimed to explore the effects, within a certain timeframe (48 and 96 hours), of leukocyte- and platelet-rich gel (L-PRG) and pure platelet-rich gel (P-PRG) supernatants on the production and degradation of specific growth factors (GF), cytokines, and hyaluronan (HA) in normal synovial membrane explants (SME) exposed to lipopolysaccharide (LPS).
- The molecules under study were platelet associated growth factors (PDGF-BB and TGF-β1), pro-inflammatory (TNF-α) and anti-inflammatory cytokines (IL-4 and IL-1ra), and hyaluronan (HA).
- Synovial membrane explants (SMEs) were taken from six horses, exposed to lipopolysaccharide (LPS), and cultured for 96 hours with L-PRG and P-PRG supernatants at concentrations of 25% and 50%, respectively. The SME culture medium was used to measure the concentration of these molecules via ELISA.
Results
- Both L-PRG and P-PRG supernatants altered the molecular profile of the culture media, when compared to the SME challenged with only LPS. Both types of PRG supernatants resulted in concentrations of GFs, cytokines, and HA closely resembling those found in normal synovial fluid.
- Among all treated groups, 25% L-PRG produced the highest concentration of IL-1ra, while 50% P-PRG resulted in sustained levels of IL-4 and HA.
Conclusions
- The research suggests that the platelet and leukocyte concentration in PRP preparations as well as the volume of the substance administered play a crucial role in determining the joint’s healing and anti-inflammatory response.
- Moreover, the study proposes that PRP preparations with reduced leukocyte levels may be more effective in treating patients with osteoarthritis and inflammatory synovitis.
Cite This Article
APA
Ríos DL, López C, Álvarez ME, Samudio IJ, Carmona JU.
(2015).
Effects over time of two platelet gel supernatants on growth factor, cytokine and hyaluronan concentrations in normal synovial membrane explants challenged with lipopolysaccharide.
BMC Musculoskelet Disord, 16, 153.
https://doi.org/10.1186/s12891-015-0605-3 Publication
Researcher Affiliations
- Grupo de Investigación Terapia Regenerativa, Departamento de Salud Animal, Universidad de Caldas, Calle 65 No 26-10, Manizales, Colombia. diana.rios@ucaldas.edu.co.
- Grupo de Investigación Terapia Regenerativa, Departamento de Salud Animal, Universidad de Caldas, Calle 65 No 26-10, Manizales, Colombia. catalina.lopez@ucaldas.edu.co.
- Grupo de Investigación Terapia Regenerativa, Departamento de Salud Animal, Universidad de Caldas, Calle 65 No 26-10, Manizales, Colombia. maria.alvarez_l@ucaldas.edu.co.
- The Terry Fox Laboratory, BC Cancer Research Centre, 675 West 10th Avenue, Vancouver, British Columbia, V5Z 1L3, Canada. isamudio@bccrc.ca.
- Grupo de Investigación Terapia Regenerativa, Departamento de Salud Animal, Universidad de Caldas, Calle 65 No 26-10, Manizales, Colombia. carmona@ucaldas.edu.co.
MeSH Terms
- Animals
- Blood Platelets / immunology
- Blood Platelets / metabolism
- Cell Fractionation
- Culture Media / metabolism
- Cytokines / metabolism
- Gels
- Horses
- Hyaluronic Acid / metabolism
- Inflammation Mediators / metabolism
- Intercellular Signaling Peptides and Proteins / metabolism
- Leukocytes / drug effects
- Leukocytes / immunology
- Leukocytes / metabolism
- Lipopolysaccharides / pharmacology
- Platelet-Rich Plasma / immunology
- Platelet-Rich Plasma / metabolism
- Synovial Fluid / immunology
- Synovial Fluid / metabolism
- Synovial Membrane / drug effects
- Synovial Membrane / immunology
- Synovial Membrane / metabolism
- Time Factors
- Tissue Culture Techniques
References
This article includes 53 references
- Abhishek A, Doherty M. Diagnosis and clinical presentation of osteoarthritis.. Rheum Dis Clin North Am 2013;39(1):45–66.
- Sokolove J, Lepus CM. Role of inflammation in the pathogenesis of osteoarthritis: latest findings and interpretations.. Ther Adv Musculoskelet Dis 2013;5(2):77–94.
- Sofat N, Beith I, Anilkumar PG, Mitchell P. Recent clinical evidence for the treatment of osteoarthritis: what we have learned.. Rev Recent Clin Trials 2011;6(2):114–126.
- Filardo G, Kon E, Di Martino A, Di Matteo B, Merli ML, Cenacchi A, Fornasari PM, Marcacci M. Platelet-rich plasma vs hyaluronic acid to treat knee degenerative pathology: study design and preliminary results of a randomized controlled trial.. BMC Musculoskelet Disord 2012;13:229.
- Filardo G, Kon E, Roffi A, Di Matteo B, Merli ML, Marcacci M. Platelet-rich plasma: why intra-articular? A systematic review of preclinical studies and clinical evidence on PRP for joint degeneration.. Knee Surg Sports Traumatol Arthrosc 2013.
- Andia I, Abate M. Knee osteoarthritis: Hyaluronic acid, platelet-rich plasma or both in association?. Expert Opin Biol Ther 2014;14(5):635–649.
- Wang-Saegusa A, Cugat R, Ares O, Seijas R, Cuscó X, Garcia-Balletbó M. Infiltration of plasma rich in growth factors for osteoarthritis of the knee short-term effects on function and quality of life.. Arch Orthop Trauma Surg 2011;131(3):311–317.
- Carmona JU, Argüelles D, Climent F, Prades M. Autologous platelet concentrates as a treatment of horses with osteoarthritis: a preliminary pilot clinical study.. J Equine Vet Sci 2007;27(4):167–170.
- Pichereau F, Décory M, Cuevas Ramos G. Autologous platelet concentrate as a treatment for horses with refractory fetlock osteoarthritis.. J Equine Vet Sci 2014;34(4):489–493.
- Silva RF, Carmona JU, Rezende CMF. Intra-articular injections of autologous platelet concentrates in dogs with surgical reparation of cranial cruciate ligament rupture.. Vet Comp Orthop Traumatol 2013;26(4):285–290.
- Kisiday JD, McIlwraith CW, Rodkey WG, Frisbie DD, Steadman JR. Effects of platelet-rich plasma composition on anabolic and catabolic activities in equine cartilage and meniscal explants.. Cartilage 2012;3(3):245–254.
- Van Buul GM, Koevoet WLM, Kops N, Bos PK, Verhaar JAN, Weinans H, Bernsen MR, Van Osch GJVM. Platelet-rich plasma releasate inhibits inflammatory processes in osteoarthritic chondrocytes.. Am J Sports Med 2011;39(11):2362–2370.
- Amrichová J, Špaková T, Rosocha J, Harvanová D, Bačenková D, Lacko M, Horňák S. Effect of PRP and PPP on proliferation and migration of human chondrocytes and synoviocytes in vitro.. Central Eur J Biol 2014;9(2):139–148.
- Sundman EA, Cole BJ, Karas V, Della Valle C, Tetreault MW, Mohammed HO, Fortier LA. The anti-inflammatory and matrix restorative mechanisms of platelet-rich plasma in osteoarthritis.. Am J Sports Med 2014;42(1):35–41.
- Anitua E, Sánchez M, Nurden AT, Zalduendo MM, De la Fuente M, Azofra J, Andía I. Platelet-released growth factors enhance the secretion of hyaluronic acid and induce hepatocyte growth factor production by synovial fibroblasts from arthritic patients.. Rheumatology 2007;46(12):1769–1772.
- Lippross S, Moeller B, Haas H, Tohidnezhad M, Steubesand N, Wruck CJ, Kurz B, Seekamp A, Pufe T, Varoga D. Intraarticular injection of platelet-rich plasma reduces inflammation in a pig model of rheumatoid arthritis of the knee joint.. Arthritis Rheum 2011;63(11):3344–3353.
- Mastrangelo AN, Vavken P, Fleming BC, Harrison SL, Murray MM. Reduced platelet concentration does not harm PRP effectiveness for ACL repair in a porcine in vivo model.. J Orthop Res 2011;29(7):1002–1007.
- Textor JA, Willits NH, Tablin F. Synovial fluid growth factor and cytokine concentrations after intra-articular injection of a platelet-rich product in horses.. Vet J 2013;198(1):217–223.
- Fortier LA, Barker JU, Strauss EJ, McCarrel TM, Cole BJ. The role of growth factors in cartilage repair.. Clin Orthop Relat Res 2011;469(10):2706–2715.
- Chang KV, Hung CY, Aliwarga F, Wang TG, Han DS, Chen WS. Comparative effectiveness of platelet-rich plasma injections for treating knee joint cartilage degenerative pathology: a systematic review and meta-analysis.. Arch Phys Med Rehabil 2014;95(3):562–575.
- Cavallo C, Filardo G, Mariani E, Kon E, Marcacci M, Pereira Ruiz MT, Facchini A, Grigolo B. Comparison of platelet-rich plasma formulations for cartilage healing: an in vitro study.. J Bone Joint Surg Am 2014;96(5):423–429.
- 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):148–159.
- 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;13(7):1131–1137.
- 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;94(19):e143.141–e143.148.
- Boswell SG, Schnabel LV, Mohammed HO, Sundman EA, Minas T, Fortier LA. Increasing platelet concentrations in leukocyte-reduced platelet-rich plasma decrease collagen gene synthesis in tendons.. Am J Sports Med 2014;42(1):42–49.
- Braun HJ, Kim HJ, Chu CR, Dragoo JL. The effect of platelet-rich plasma formulations and blood products on human synoviocytes: Implications for intra-articular injury and therapy.. Am J Sports Med 2014;42(5):1204–1210.
- Dhillon RS, Schwarz EM, Maloney MD. Platelet-rich plasma therapy - future or trend?. Arthritis Res Ther 2012;14(4):219.
- 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;9:29.
- 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;81(2):237–245.
- Moses VS, Hardy J, Bertone AL, Weisbrode SE. Effects of anti-inflammatory drugs on lipopolysaccharide-challenged and -unchallenged equine synovial explants.. Am J Vet Res 2001;62(1):54–60.
- Donnelly BP, Nixon AJ, Haupt JL. Nucleotide structure of equine platelet-derived growth factor-A and –B and expression in horses with induced acute tendinitis.. Am J Vet Res 2006;67(7):1218–1225.
- Penha-Goncalves MN, Onions DE, Nicolson L. Cloning and sequencing of equine transforming growth factor-beta 1 (TGFβ-1) cDNA.. DNA Seq 1997;7(6):375–378.
- Civinini R, Nistri L, Martini C, Redl B, Ristori G, Innocenti M. Growth factors in the treatment of early osteoarthritis.. Clin Cases Min Bone Metab 2013;10(1):26–29.
- Miller RE, Miller RJ, Malfait A-M. Osteoarthritis joint pain: the cytokine connection.. Cytokine 2014;70(2):185–193.
- Eder L, Thavaneswaran A, Chandran V, Gladman DD. Tumour necrosis factor α blockers are more effective than methotrexate in the inhibition of radiographic joint damage progression among patients with psoriatic arthritis.. Ann Rheum Dis 2014;73(6):1007–1011.
- Kamm JL, Nixon AJ, Witte TH. Cytokine and catabolic enzyme expression in synovium, synovial fluid and articular cartilage of naturally osteoarthritic equine carpi.. Equine Vet J 2010;42(8):693–699.
- Carmona JU, Prades M. Pathophisiology of osteoarthritis.. Comp Equine 2009;4(1):28–40.
- Fernandes JC, Martel-Pelletier J, Pelletier J-P. The role of cytokines in osteoarthritis pathophysiology.. Biorheology 2002;39(1):237–246.
- Yang KGA, Raijmakers NJH, van Arkel ERA, Caron JJ, Rijk PC, Willems WJ, Zijl JAC, Verbout AJ, Dhert WJA, Saris DBF. Autologous interleukin-1 receptor antagonist improves function and symptoms in osteoarthritis when compared to placebo in a prospective randomized controlled trial.. Osteoarthritis Cartilage 2008;16(4):498–505.
- Millward-Sadler SJ, Wright MO, Davies LW, Nuki G, Salter DM. Mechanotransduction via integrins and interleukin-4 results in altered aggrecan and matrix metalloproteinase 3 gene expression in normal, but not osteoarthritic, human articular chondrocytes.. Arthritis Rheum 2000;43(9):2091–2099.
- Lee S-G, Lee E-J, Park W-D, Kim J-B, Kim E-O, Choi S-W. Anti-inflammatory and anti-osteoarthritis effects of fermented Achyranthes japonica Nakai.. J Ethnopharmacol 2012;142(3):634–641.
- Furman BD, Mangiapani DS, Zeitler E, Bailey KN, Horne PH, Huebner JL, Kraus VB, Guilak F, Olson SA. Targeting pro-inflammatory cytokines following joint injury: acute intra-articular inhibition of interleukin-1 following knee injury prevents post-traumatic arthritis.. Arthritis Res Ther 2014;16(3):R134.
- Malemud C. Anticytokine therapy for osteoarthritis.. Drugs Aging 2010;27(2):95–115.
- Cao HJ, Wang HS, Zhang Y, Lin HY, Phipps RP, Smith TJ. Activation of human orbital fibroblasts through CD40 engagement results in a dramatic induction of hyaluronan synthesis and prostaglandin endoperoxide H synthase-2 expression: Insights into potential pathogenic mechanisms of thyroid-associated ophthalmopathy.. J Biol Chem 1998;273(45):29615–29625.
- Kraus VB, Birmingham J, Stabler TV, Feng S, Taylor DC, Moorman CT III, Garrett WE, Toth AP. Effects of intraarticular IL1-Ra for acute anterior cruciate ligament knee injury: a randomized controlled pilot trial (NCT00332254). Osteoarthritis Cartilage 2012;20(4):271–278.
- Neidhart M, Gay RE, Gay S. Anti–interleukin-1 and anti-CD44 interventions producing significant inhibition of cartilage destruction in an in vitro model of cartilage invasion by rheumatoid arthritis synovial fibroblasts.. Arthritis Rheum 2000;43(8):1719–1728.
- Frisbie DD, Kawcak CE, Werpy NM, Park RD, McIlwraith CW. Clinical, biochemical, and histologic effects of intra-articular administration of autologous conditioned serum in horses with experimentally induced osteoarthritis.. Am J Vet Res 2007;68(3):290–296.
- Hyc A, Osiecka-Iwan A, Niderla-Bielinska J, Moskalewski S. Influence of LPS, TNF, TGF-β1 and IL-4 on the expression of MMPs, TIMPs and selected cytokines in rat synovial membranes incubated in vitro.. Int J Mol Med 2011;27(1):127–137.
- Oguchi T, Ishiguro N. Differential stimulation of three forms of hyaluronan synthase by TGF-β, IL-1β, and TNF-α.. Connect Tissue Res 2004;45(4-5):197–205.
- Alaaeddine N, Di Battista JA, Pelletier J-P, Kiansa K, Cloutier J-M, Martel-Pelletier J. Inhibition of tumor necrosis factor α–induced prostaglandin E2 production by the antiinflammatory cytokines interleukin-4, interleukin-10, and interleukin-13 in osteoarthritic synovial fibroblasts: distinct targeting in the signaling pathways.. Arthritis Rheum 1999;42(4):710–718.
- Becker T, Tohidast-Akrad M, Humpeler S, Gerlag DM, Kiener H-P, Zenz P, Steiner G, Ekmekcioglu C. Clock gene expression in different synovial cells of patients with rheumatoid arthritis and osteoarthritis.. Acta Histochem 2014;116(7):1199–1207.
- Weibrich G, Hansen T, Kleis W, Buch R, Hitzler WE. Effect of platelet concentration in platelet-rich plasma on peri-implant bone regeneration.. Bone 2004;34(4):665–671.
- Fortier LA, Cole BJ. Anti-inflammatory and matrix restorative mechanisms of platelet-rich plasma in osteoarthritis: response to Andia and Maffulli.. Am J Sports Med 2014;42(6):NP32–NP33.
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
