Platelets are small, anucleated cell fragments derived from megakaryocytes in the bone marrow, playing a key role in hemostasis and thrombosis in horses. They contribute to the formation of blood clots by adhering to the site of vascular injury, aggregating with one another, and facilitating the coagulation process. In equine medicine, platelet count and function are important parameters in assessing bleeding disorders and thrombotic conditions. Variations in platelet count can indicate underlying health issues, such as inflammation, infection, or bone marrow disorders. This page compiles peer-reviewed research studies and scholarly articles that explore the physiology, pathology, and clinical implications of platelets in equine health.
Brown KA, Gregorio EN, Barot D, Usimaki A, Linardi RL, Missanelli JR, You Y, Robinson MA, Ortved KF.To determine the effects of a single dose of the NSAIDs phenylbutazone, firocoxib, flunixin meglumine, and ketoprofen on concentrations of growth factors and cytokines in autologous protein solution (APS) and platelet-rich plasma (PRP). Methods: 6 adult university-owned horses. Methods: For the first phase, 6 horses were randomized to receive ketoprofen (1,000 mg) or flunixin meglumine (500 mg) IV. Blood was obtained and processed for APS (Pro-Stride) and PRP (Restigen) before and 6 hours after administration of NSAIDs. Horses underwent a 2-week washout period, after which the protocol was rep...
Sawesi OK, Elbaz AK, Mahmoud AS, Duro EM, Alteab AA, Milad KK, Bennuor EM.Hematological evaluation is an important step for health assessment in equine medicine. Besides biochemical tests, that evaluation could be pivotal for diagnosis, prognosis, and treatment monitoring. In Libya, the horse population has drastically increased in the last years, and hitherto, no report addressed the reference hematological values for the existing equine breeds. Unassigned: This study aimed to establish the reference for hematological indices of traditional equestrian and stud-farm equine breeds in Western Libya in regard to some variables such as individual characteristics, manage...
Torres LEC, Florez CO, Oliveira JG, Vieira GD, Ribeiro IS, Keller KM, Leme FOP, Fantini P, Maranhão RPA.In equine ophthalmology, ulcerative keratitis is among the most common conditions and, in general, arises as a consequence of some trauma suffered. Secondarily, subsequent contamination by pathogenic or resident bacteria of the horse's ocular microbiota may have undesirable consequences. Under physiological conditions, the normal microbiota coexists with the immune status of the host, serving as a barrier, ensuring the health of the ocular surface, and inhibiting the proliferation of pathogens. However, in the imbalance of immune barriers, the normal microbiota can become pathogenic and lead t...
Frohlich M, Knights K, Springer NL.Differentiating immune-mediated causes from other causes of anemia and thrombocytopenia can be challenging. Flow cytometry can detect surface-associated immunoglobulin (sIg) on red blood cells (RBC) and platelets (PLT) in dogs and horses. Sample storage parameters for ideal assay performance has not been evaluated in horses. The study objective is to identify optimal storage time and temperature of equine whole blood for the detection of RBC-sIg and PLT-sIg via flow cytometry. Both assays were performed on samples at time 0, 4, 24, 48, and 72 h post collection. RBC-sIg samples were stored at ...
Pezzanite LM, Chow L, Dow SW, Goodrich LR, Gilbertie JM, Schnabel LV.Increasing antimicrobial resistance in veterinary practice has driven the investigation of novel therapeutic strategies including regenerative and biologic therapies to treat bacterial infection. Integration of biological approaches such as platelet lysate and mesenchymal stromal cell (MSC) therapy may represent adjunctive treatment strategies for bacterial infections that minimize systemic side effects and local tissue toxicity associated with traditional antibiotics and that are not subject to antibiotic resistance. In this review, we will discuss mechanisms by which biological therapies exe...
Scalco R, de Oliveira GN, da Rosa Curcio B, Wooten M, Magdesian KG, Hidai ST, Pandit P, Aleman M.Rapid and accurate markers to aid diagnosis of sepsis are needed in neonatal foals. The CBC variable red blood cell distribution width (RDW) to platelet ratio (RPR) is associated with inflammatory response and linked to poor outcomes of sepsis in human patients. Objective: Explore the correlation of RPR with sepsis in neonatal foals and evaluate RPR predictive and prognostic value. Methods: Three hundred seventeen hospitalized neonatal foals ≤7 days of age that had a CBC and physical exam performed at admission between 2012 and 2021. Methods: Retrospective case-control study. Clinical reco...
The aim of our study was to analyze circadian rhythm of the hematological profile of horses housed in a loose box and paddock during the different seasons (spring, summer, autumn, and winter). Blood samples were performed every 4 h for 48 consecutive hours. Red blood cells (RBCs), hemoglobin (HGB), hematocrit (HCT), white blood cells (WBCs), platelets (PLTs), and leukocyte subpopulations (neutrophils, basophils, eosinophils, lymphocytes, and monocytes) were analyzed, and, at the same time, environmental conditions were recorded. A statistically significant effect of housing conditions ( < 0...
Mora-Pereira M, Boone L, Naskou M, Wooldridge A.To evaluate ex vivo angiogenesis of equine arterial rings in response to various growth media. Methods: Facial arteries were dissected from 11 horses post-euthanasia. Equine platelet lysate (ePL) was harvested from 6 horses. Methods: Arteries were exposed to endothelial growth media (EGM) + horse serum (HS) for first sprout (FS), vascular regression (VR), and (basement membrane matrix [Matrigel]) lysis (ML) evaluation. Additional rings supplemented with (1) EGM, (2) EGM + EDTA, (3) endothelial basal media (EBM), (4) EBM + HS, or (5) EBM + human VEGF were compared for vascular network area (VNA...
Fujiwara Y, Kusakabe KT, Baba K, Sasaki N.Currently, treatment for peripheral nerve injuries in horses primarily relies upon physical therapy and anti-inflammatory drugs. In humans, various treatments using mesenchymal stem cells (MSCs) are being attempted. Therefore, in this study, Schwann-like cell differentiation cultures of equine MSCs were prepared using fetal bovine serum (FBS) and equine platelet lysate (ePL). ePL increased the platelet count to 1 × 10/μl, the optimal concentration for culture. In both groups, an elongated morphology at both ends, characteristic of Schwann cells, was observed under the microscope. Real-time...
Satué K, Fazio E, Gardón JC, Medica P.The interpretation of the blood count is essential to help the equine clinician in the diagnosis, prognosis, patient management, and control of equine diseases. Hematologic alterations often reflect the condition of the individual or an overall response to a pathological situation. A thorough clinical examination of the patient is essential to correctly interpret the hematological results. The most common abnormalities in the erythrogram are mainly anemia and polycythemia. The frequent causes of anemia in horses are acute and chronic blood loss, hemolytic anemia, and anemia caused by chronic d...
Ghallab RS, El-Beskawy M, El-Shereif AA, Rashad AMA, Elbehiry MA.Equine endometritis is one of the most common causes of reproduction failure. To achieve better treatment outcomes, different diagnostic methods should be combined. In the current study, 39 repeat breeder mares were subjected to ultrasonography examination to detect excessive accumulation of intrauterine fluids and an abnormal oedema pattern, which revealed that 61.5% of mares were positive. Combined with endometrial cytology by low-volume uterine flush, 47.7% of smears contain neutrophils (more than 2-3 per HPF X100), and microbial culture. 92.3% of mares were infected with different bacteria...
Pezzanite LM, Chow L, Griffenhagen GM, Bass L, Goodrich LR, Impastato R, Dow S.Multiple biological therapies for orthopedic injuries are marketed to veterinarians, despite a lack of rigorous comparative biological activity data to guide informed decisions in selecting a most effective compound. Therefore, the goal of this study was to use relevant bioassay systems to directly compare the anti-inflammatory and immunomodulatory activity of three commonly used orthobiological therapies (OTs): mesenchymal stromal cells (MSC), autologous conditioned serum (ACS), and platelet rich plasma (PRP). Unassigned: Equine monocyte-derived macrophages were used as the readout system to ...
Kaneps AJ.Regenerative medicine therapies have become significant tools for treatment of joint, soft tissue, and a variety of other conditions in animals and humans. Regenerative medicine aims to restore form and function of injured tissues using the body's own resources such as cells, fluids (ie, plasma and serum), and their resulting anti-inflammatory and prohealing cytokines. Platelet-rich plasma and other hemoderivatives have application for joint disorders such as osteoarthritis, cartilage injury, synovitis, and soft tissue injuries. These therapies achieve anti-inflammatory and healing effects wit...
El-Sherif MA, Nabil G, Ibrahim R, Mohamed N, Mohamed M, Jaheen AH.Based on the growing attention to the Arabian horses with a lack of records regarding their normal physiological parameters, which are critically vital for disease diagnosis &convenient drug administration, we give great attention to their thyroid gland normalcy. The thyroid gland is one of the key players in regulating many physiological processes. Therefore, we tackled thyroid biology & morphology in 14 Arabian stallions & 18 Arabian mares from the beginning of March to the middle of April on animals from 4 to 19 years old. First, the blood samples taken from their jugular vein ...
Tou K, Cawley A, Bowen C, Bishop DP, Fu S.The current approach to equine anti-doping is focused on the targeted detection of prohibited substances. However, as new substances are rapidly being developed, the need for complimentary methods for monitoring is crucial to ensure the integrity of the racing industry is upheld. Lipidomics is a growing field involved in the characterisation of lipids, their function and metabolism in a biological system. Different lipids have various biological effects throughout the equine system including platelet aggregation and inflammation. A certain class of lipids that are being reviewed are the eicosa...
Scalco R, Aleman M, Nogueira CEW, Freitas NB, Curcio BR.To report red cell distribution width (RDW) values, to calculate RDW-to-platelet ratio (RPR), and to investigate a possible correlation of RDW and RPR index values in neonatal foals classified as healthy or at risk based on clinical information from a population of foals up to 24 hours of life. Methods: Retrospective study conducted from records and CBCs of foals born between June and November from 2018 to 2020 foaling seasons. Methods: Breeding farm. Methods: Three hundred and nine neonatal full-term Thoroughbred foals. Methods: None. Results: Foals were evaluated by a veterinarian within 15...
Cocco R, Rizzo M, Carta C, Arfuso F, Piccione G, Luridiana S, Crovace A, Passino ES, Sechi S.Among infection diseases transmitted by arthropods, the equine vector-borne diseases transmitted by ticks represent an emerging problem worldwide due to their morbidity and mortality and, in some cases, to their zoonotic relevance. Understanding the host immune/inflammatory response to the pathogens is crucial to develop effective methods of diagnosis, control and treatment. This study aimed to evaluate red blood cells (RBC), hematocrit (Hct), hemoglobin concentration (Hb), mean cell volume (MCV), mean cell hemoglobin content (MCH), mean cell hemoglobin (MCHC), platelets (PLT), white blood cel...
Freitas NPP, Silva BDP, Bezerra MRL, Pescini LYG, Olinda RG, Salgueiro CCM, Nunes JF, Martins JAM, Neto SG, Martins LT.This study investigated platelet-rich plasma (PRP) and adipose stem cell-conditioned medium (ASC-CM) use as a strategy to accelerate tissue healing. Platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) were quantified in fresh and freeze-dried PRP and ASC-CM, and a stability test was performed in the freeze-dried samples (90 and 180 days of storage). A cell proliferation test was performed using equine mesenchymal stem cell culture in reconstituted PRP gel mesh after freeze-drying. In vivo PRP, ASC-CM applications, or their association were performed in induced w...
Duddy HR, Schoonover MJ, Hague BA.Tendon and ligament injuries are significant causes of loss of use and early retirement in performance horses. Amniotic fluid and tissue are excellent sources of growth factors and cytokines important in tendon and ligament healing. Thus, an equine-origin liquid amnion allograft (ELAA) may be beneficial in the treatment of equine tendonitis and desmitis. Objectives of this study were to report the outcome achieved (i.e. ability to return to work) for horses diagnosed with tendonitis or desmitis lesions treated with local injection of ELAA and to compare these outcomes to those reported for oth...
Even KM, Gaesser AM, Ciamillo SA, Linardi RL, Ortved KF.Joint injury often leads to cartilage damage and posttraumatic inflammation, which drives continued extracellular matrix degradation culminating in osteoarthritis. Mesenchymal stem cells (MSCs) have been proposed as a biotherapeutic to modulate inflammation within the joint. However, concerns have been raised regarding the immunogenicity of MSCs cultured in traditional fetal bovine serum (FBS) containing media, and the potential of xenogenic antigens to activate the immune system causing rejection and destruction of the MSCs. Xenogen-free alternatives to FBS have been proposed to decrease MSC ...
Hagen A, Niebert S, Brandt VP, Holland H, Melzer M, Wehrend A, Burk J.Successful translation of multipotent mesenchymal stromal cell (MSC)-based therapies into clinical reality relies on adequate cell production procedures. These should be available not only for human MSC, but also for MSC from animal species relevant to preclinical research and veterinary medicine. The cell culture medium supplementation is one of the critical aspects in MSC production. Therefore, we previously established a scalable protocol for the production of buffy-coat based equine platelet lysate (ePL). This ePL proved to be a suitable alternative to fetal bovine serum (FBS) for equine a...
da Conceição FFCB, de Mattos BHV, Sá PA, Marçola TG, da Silva GA, Teixeira HCA, Campebell RC.Due to the scarcity of information on Breton horses, the objective was to study hematobiochemical values of this breed. Blood samples were collected from 29 Bretons, males and females, of different ages, in Brasília-DF, distributed into groups, according to age, without distinction of sex (G1): animals from 4 to 9 years old (n=16) and (G2): from 10 to 26 years old (n=13). The horses were also distributed into males and females for comparisons between the sexes. Values for red blood cells, hemoglobin, creatinine, and urea were statistically higher in females. Fibrinogen was higher in males. Ly...
Gilbertie JM, Schaer TP, Engiles JB, Seiler GS, Deddens BL, Schubert AG, Jacob ME, Stefanovski D, Ruthel G, Hickok NJ, Stowe DM, Frink A, Schnabel LV.The leading cause of treatment failure in Staphylococcus aureus infections is the development of biofilms. Biofilms are highly tolerant to conventional antibiotics which were developed against planktonic cells. Consequently, there is a lack of antibiofilm agents in the antibiotic development pipeline. To address this problem, we developed a platelet-rich plasma (PRP)-derived biologic, termed BIO-PLY (for the BIOactive fraction of Platelet-rich plasma LYsate) which has potent in vitro bactericidal activity against S. aureus synovial fluid free-floating biofilm aggregates. Additional in vitro st...
Garbin LC, Contino EK, Olver CS, Frisbie DD.Hemoderivatives such as autologous conditioned serum (ACS) and platelet-rich plasma (PRP) have been used as potential disease-modifying therapies in musculoskeletal disorders such as osteoarthritis (OA). These therapies are based on the delivery of multiple growth factors and anti-inflammatory cytokines that are known to participate in inflammatory processes. The variability of cytokine content due to the autologous nature of the product, the non-availability for immediate use and need for storage at low temperatures are limitations for its use in the field. An allogeneic freeze-dried conditio...
Melotti L, Carolo A, Elshazly N, Boesso F, Da Dalt L, Gabai G, Perazzi A, Iacopetti I, Patruno M.In the present case report a show jumping 10-year-old Sella Italiano gelding, presented with severe lameness, swelling and pain at palpation of the mid-metacarpal region of the left forelimb. Clinical and ultrasound examination diagnosed a chronic tendonitis of the central region of the superficial digital flexor tendon (SDFT). The lesion was a reoccurrence since it developed from a previously healed injury. The horse had to stop competing and was unresponsive to gold-standard treatments as Non-steroidal anti-inflammatory drugs (NSAIDs) and conservative management after 6 months of therapy. Th...
Theuerkauf K, Obach-Schröck C, Staszyk C, Moritz A, Roscher KA.In humans, activated platelets contribute to sepsis complications and to multiple organ failure. In our prospective analytical study of cases of the equine systemic inflammatory response syndrome (SIRS), we adapted a standard human protocol for the measurement of activated platelets and platelet-leukocyte aggregates (PLAs) in equine platelet-leukocyte-rich plasma (PLRP) by flow cytometry, and we investigated the hypothesis that activated platelets and PLAs are increased in clinical cases of SIRS. We included 17 adult horses and ponies fulfilling at least 2 SIRS criteria, and 10 healthy equids ...
Abd El-Rahman SS, Amer MS, Hassan MH, Fahmy HM, Shamaa AA.Lyophilized equine platelet derived growth factors (LGF) is a novel advanced platelet rich protein growth factor. It has been successfully applied in various fields of regenerative medicine to treat a variety of inflammatory and degenerative musculoskeletal conditions. Our study aimed to evaluate the efficacy of intraarticularly injected LGF for the remedy of articular cartilage injury, commonly characterized by progressive pain and loss of joint function in osteoarthritic rabbits. Full-thickness cylindrical cartilage defects were generated in both femoral condylar articular surfaces in twenty...
Fukuda K, Kuroda T, Tamura N, Mita H, Miyata H, Kasashima Y.This study aimed to verify the effects of platelet lysate (PL) administration on the repair of injured horse tissue. Skeletal muscle injuries were induced in 26 Thoroughbreds by bupivacaine administration. PL or saline was administered 1 day (1D) after injury. Muscle samples from 22 horses injected with PL or saline were obtained by needle biopsy at 2, 3, 4, or 7 days (2D, 3D, 4D, or 7D, respectively) after injury, and growth-factor concentrations and muscle regeneration-associated gene expression levels were determined. Intact samples were similarly collected before injury, and samples of inj...
Hagen A, Holland H, Brandt VP, Doll CU, Häußler TC, Melzer M, Moellerberndt J, Lehmann H, Burk J.Platelet lysate (PL) is an attractive platelet-based therapeutic tool and has shown promise as xeno-free replacement for fetal bovine serum (FBS) in human and equine mesenchymal stromal cell (MSC) culture. Here, we established a scalable buffy-coat-based protocol for canine PL (cPL) production (n = 12). The cPL was tested in canine adipose MSC (n = 5) culture compared to FBS. For further comparison, equine adipose MSC (n = 5) were cultured with analogous equine PL (ePL) or FBS. During canine blood processing, platelet and transforming growth factor-β1 concentrations increased ( < 0.05 and < ...
Whittle BJ, Moncada S, Vane JR.The activity of prostacyclin (PGI2), PGE1 or PGD2 as inhibitors of platelet aggregation in plasma from human, dog, rabbit, rat, sheep and horse was investigated. Prostacyclin was the most potent inhibitor in all species. PGD2 was a weak inhibitor in dog, rabbit and rat plasma whereas PGE1 and prostacyclin were highly active. Theophylline or dipyridamole potentiated the inhibition of human platelet aggregation by prostacyclin, PGE1 or PGD2. Compound N-0164 abolished the inhibition by PGD2 of human platelet aggregation but did not inhibit the effects of PGE1 or prostacyclin. The results suggest ...
Torricelli P, Fini M, Filardo G, Tschon M, Pischedda M, Pacorini A, Kon E, Giardino R.Tissue repair in musculoskeletal injuries is often a slow and sometimes incomplete process. Regenerative medicine based on the use of growth factors (GFs) and cell therapy is aimed at improving the quality and speed of tendon and ligament healing. The aim of this study was to evaluate the potential for the administration of a combination of autologous platelet-rich plasma (PRP) and freshly isolated bone marrow mononucleated cells (BMMNCs) in 13 competition horses affected by overuse musculoskeletal injuries (suspensory ligament desmopathy and superficial flexor tendinopathy) and refractory to ...
Bosch G, Moleman M, Barneveld A, van Weeren PR, van Schie HT.The repair process of tendon injuries, which are common in both human and equine athletes, is slow and the quality of the repair tissue is often inferior to the original tendon tissue, which frequently leads to re-injury. The relatively poor vascularization of tendons is considered to be one of the reasons for their limited healing potential. Recently, platelet-rich plasma (PRP), an autologous concentrate of platelets, rich in growth factors, has been shown to enhance the repair process of injured tendons. This effect has been ascribed to the high levels of growth factors in PRP, several of wh...
Edington N, Bridges CG, Patel JR.Eight mares were infected with equid herpesvirus-1 subtype 1 isolated from a case of equine paresis. In two mares killed at 4 d.p.i. immunofluorescence showed endothelial cell infection together with thrombosis in the rete arteriosus of the nasal mucosa and also in the spinal cord of one of these mares. Circulating platelet counts in the other six mares fell as early as 2 d.p.i. and remained depressed for seven days. Circulating immune complexes started to appear at 2 d.p.i., reached maximum levels at 10 d.p.i., but were undetectable at 28 d.p.i. Three of the six remaining mares developed vary...
Textor JA, Tablin F.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 ...
Carvalho Ade M, Badial PR, Álvarez LE, Yamada AL, Borges AS, Deffune E, Hussni CA, Garcia Alves AL.Tendon injury is a major cause of lameness and decreased performance in athletic equines. Various therapies for tendonitis have been described; however, none of these therapies results in complete tissue regeneration, and the injury recurrence rate is high even after long recovery periods involving rest and physiotherapy. Methods: A lesion was induced with collagenase gel in the superficial digital flexor tendon in the center portion of the metacarpal region of eight equines of mixed breed. After two weeks, the lesions of the animals in the treated and control groups were treated through the i...
Ricco S, Renzi S, Del Bue M, Conti V, Merli E, Ramoni R, Lucarelli E, Gnudi G, Ferrari M, Grolli S.Overstrain tendonitis are common pathologies in the sport horses. Therapeutic approaches to tendon healing do not always result in a satisfactory anatomical and functional repair, and healed tendon is often characterized by functional impairment and high risk of reinjury. Recently, mesenchymal stem cells (MSCs) and platelet rich plasma (PRP) have been proposed as novel therapeutic treatments to improve the tendon repair process. MSCs are multipotent, easy to culture and being originated from adult donors do not pose ethical issues. To date, autologous MSCs have been investigated mainly in the ...
Giraldo CE, López C, Álvarez ME, Samudio IJ, Prades M, Carmona JU.There is no information on the effects of the breed, gender and age on the cellular content and growth factor (GF) release from equine pure-platelet rich plasma (P-PRP) and pure-platelet rich gel (P-PRG). The objectives of this study were: 1) to compare the cellular composition of P-PRP with whole blood and platelet poor plasma (PPP); 2) to compare the concentration of transforming GF beta 1 (TGF-β1) and platelet derived GF isoform BB (PDGF-BB) between P-PRP treated with non-ionic detergent (P-PRP+NID), P-PRG (activated with calcium gluconate -CG-), PPP+NID, PPP gel (PPG), and plasma and; 3) ...
Naskou MC, Sumner SM, Chocallo A, Kemelmakher H, Thoresen M, Copland I, Galipeau J, Peroni JF.Mesenchymal stem cells (MSCs) produced for clinical purposes rely on culture media containing fetal bovine serum (FBS) which is xenogeneic and has the potential to significantly alter the MSC phenotype, rendering these cells immunogenic. As a result of bovine-derived exogenous proteins expressed on the cell surface, MSCs may be recognized by the host immune system as non-self and be rejected. Platelet lysate (PL) may obviate some of these concerns and shows promising results in human medicine as a possible alternative to FBS. Our goal was to evaluate the use of equine platelet lysate (ePL) poo...
Geburek F, Gaus M, van Schie HT, Rohn K, Stadler PM.Regenerative and anti-inflammatory effects on tendinopathies have been attributed to blood-derived biologicals. To date the evidence for the efficacy of autologous platelet-rich plasma (PRP) treatment of naturally occurring equine tendinopathies is limited. The purpose of this placebo-controlled clinical trial was to describe the effect of a single treatment of equine superficial digital flexor tendon (SDFT) disease with PRP on clinical and ultrasonographic parameters. Twenty horses with naturally occurring tendinopathies of forelimb SDFTs were randomly assigned to the PRP-treated group (n =...
Bertone AL, Ishihara A, Zekas LJ, Wellman ML, Lewis KB, Schwarze RA, Barnaba AR, Schmall ML, Kanter PM, Genovese RL.To evaluate intra-articular autologous protein solution (APS) for the treatment of osteoarthritis in horses. Animals-40 client-owned horses with naturally occuring osteoarthritis. Methods: APS was generated from a dual-device system that concentrated plasma and WBC proteins and enriched platelet growth factors. Horses were randomly assigned to receive an intra-articular injection of 5 mL of saline (0.9% NaCl) solution (n = 20) or APS (20), exercised on a treadmill, and evaluated on the basis of lameness grades, kinetic gait analysis, joint circumference, and range of motion for 14 days. Horses...
DeRossi R, Coelho AC, Mello GS, Frazílio FO, Leal CR, Facco GG, Brum KB.To establish a low-cost method to prepare platelet-rich plasma (PRP) and evaluates the potential of platelet derived factors to enhance wound healing in the surgical wounds in equine. Methods: To obtain a PRP gel, calcium gluconate and autologous thrombin were added to platelet-rich plasma. For the tests six saddle horses were used and two surgical incisions were made in each animal. Wounds were treated with PRP gel or untreated. Sequential wound biopsies collected at Treatment 1: at days 5 and 30 and Treatment 2: at days 15 and 45 post wounding permitted comparison of differentiation markers ...
Argüelles D, Carmona JU, Pastor J, Iborra A, Viñals L, Martínez P, Bach E, Prades M.The aim of this study was to evaluate single and double centrifugation tube methods for concentrating equine platelets. Whole blood samples were collected from clinically normal horses and processed by use of single and double centrifugation tube methods to obtain four platelet concentrates (PCs): PC-A, PC-B, PC-C, and PC-D, which were analyzed using a flow cytometry hematology system for hemogram and additional platelet parameters (mean platelet volume, platelet distribution width, mean platelet component concentration, mean platelet component distribution width). Concentrations of transformi...
Textor JA, Willits NH, Tablin F.Platelet-rich plasma (PRP) products may be useful for treatment of joint disease in horses, but may contain undesirable pro-inflammatory cytokines in addition to growth factors. This study investigated whether autologous PRP increases synovial fluid growth factor and cytokine concentrations when injected into normal equine metacarpophalangeal and metatarsophalangeal (fetlock) joints. Fetlock joints of seven healthy horses received one of four treatments: saline, resting PRP, CaCl2-activated PRP or thrombin-activated PRP. Synovial fluid was sampled prior to injection and at 6, 24, 48 and 96 h p...
Reghini MF, Ramires Neto C, Segabinazzi LG, Castro Chaves MM, Dell'Aqua Cde P, Bussiere MC, Dell'Aqua JA, Papa FO, Alvarenga MA.Degenerative changes of the endometrium are directly related to age and fertility in mares. Chronic degenerative endometritis (CDE) is correlated with uterine fluid retention and reduced ability to clear uterine inflammation. Recent research in the areas of equine surgery and sports medicine has shown that platelet-rich plasma (PRP) treatment acts as an immunomodulator of the inflammatory response. Therefore, the aim of this study was to determine if the uterine infusion of PRP could modulate the local inflammatory response and modify the intrauterine NO concentrations after artificial insemin...
Hessel LN, Bosch G, van Weeren PR, Ionita JC.Autologous platelet concentrates (APCs) are being used increasingly in horses to enhance regeneration in tissues that have poor natural healing capabilities. Numerous APC systems, which are based on different preparation techniques and were originally developed for human patients, are now routinely used in equine cases. However, preliminary process validation and adequate in vitro biochemical characterisation of most of these systems do not exist for horses. Objective: To compare haematological findings and growth factor concentrations of equine APCs obtained with 4 commercially available syst...
Bonnefoy A, Romijn RA, Vandervoort PA, VAN Rompaey I, Vermylen J, Hoylaerts MF.Binding of von Willebrand factor (VWF) to platelet GPIbalpha and to collagen is attributed to VWF A1 and A3 domains, respectively. Objective: Using VWF, VWF lacking A1 (DeltaA1-VWF) or A3 (DeltaA3-VWF) and VWF with defective A3 (H1786A-VWF), in combination with recombinant A1 (residues 1262-1492) or A3 (residues 1671-1878), fused to glutathione-S-transferase (GST-A1 and GST-A3), we have re-investigated the role of A1 in platelet recruitment to surfaces of collagen. Results: In flow, measurable binding of DeltaA3-VWF occurred to horse tendon, but also to human type III collagen. GST-A1 and GST-...
Argüelles D, Carmona JU, Climent F, Muñoz E, Prades M.Two horses with acute tendinopathy of a superficial digital flexor tendon (SDFT) and three horses with chronic proximal desmitis of the suspensory ligament (PDSL) were treated by injecting autologous concentrates of their platelets into the lesions. The lesions were monitored ultrasonographically and clinically. There were significant ultrasonographic and clinical improvements in the two horses with SDFT, but no ultrasonographic improvements in the horses with PDSL; however, they improved clinically and became less lame. All the horses had returned to their pre-injury level of performance by s...
Monteiro SO, Lepage OM, Theoret CL.To evaluate the effect of platelet-rich plasma on wounds on the distal aspect of the forelimb in horses. Methods: 6 mixed-breed 10- to 15-year-old mares. Methods: 3 wounds were created on metacarpal regions in each of 6 horses (n = 36 wounds total). Eighteen wounds were treated with platelet-rich plasma and bandaged, whereas 18 control wounds were similarly bandaged with no prior topical treatment. Decrease in wound surface area and the required number of excisions of exuberant granulation tissue were recorded until complete healing. Tissue specimens were taken from wounds at 1 week for histol...
McClain AK, McCarrel TM.Platelet-rich plasma (PRP) is a therapeutic biologic that is used for treatment of musculoskeletal pathologies in equine athletes. Due to the expense of PRP kits, and the volumes obtained, freezing aliquots for future dosing is common. Aliquots of PRP are also commonly frozen for later analysis of growth factor concentrations in in vitro research. A variety of freezing methods are used and storage duration until analysis is often not reported. The optimal frozen storage conditions and duration to maintain concentrations of commonly measured growth factors and enzymes in PRP are unknown. Our ob...
Tyrnenopoulou P, Diakakis N, Karayannopoulou M, Savvas I, Koliakos G.Regenerative medicine has become one of the most promising therapies of equine osteoarthritis. Platelet lysate (PL) is rich in bioactive proteins and growth factors that play a crucial role in tissue healing. Objective: To evaluate the efficacy of intra-articularly injected autologous PL in equine athletes with naturally occurring osteoarthritis. Methods: Fifteen warmblood geldings aged 8-19 years with osteoarthritis of the distal interphalangeal joint were included in this study. They were randomly divided into two groups; 10 horses received intra-articular injections of PL and 5 of normal s...
Clabough DL, Gebhard D, Flaherty MT, Whetter LE, Perry ST, Coggins L, Fuller FJ.An adult horse infected with a virulent, cell culture-adapted strain of equine infectious anemia virus (EIAV) developed cyclical thrombocytopenia in which the nadir of platelet counts coincided with peak febrile responses. In order to investigate the mechanism of thrombocytopenia during acute febrile episodes, four adult horses were experimentally infected with the wild-type Wyoming strain of EIAV. Platelet counts decreased from baseline as rectal temperature increased. Serum reverse transcriptase activity increased above background levels in all horses, coincident with increase in rectal temp...
Billah MM, Lapetina EG.Stimulation of horse platelets with platelet-activating factor (PAF) induces a rapid degradation of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2]. Addition of 0.1 microM PAF for 5 sec to platelets prelabeled with 32P induces a 50% loss of [32P]PtdIns(4,5)P2. 32P-Labeled phosphatidylinositol 4-monophosphate (PtdIns4P) and [32P]phosphatidylinositol (PtdIns) also are decreased, albeit at a slower rate. Loss of 32P radioactivity correlates with a net loss of fatty acids from both polyphosphoinositides. Stimulation of platelets with PAF also produces formation of [32P]phosphatidic acid and ...
Camargo Garbin L, Lopez C, Carmona JU.In the 1990s, the role of platelets in inflammation and tissue healing was finally recognized. Since then, the clinical use of platelet-derived products (hemocomponents), such as, platelet-rich plasma (PRP), markedly increased. The promise of a more economical option of a disease-modifying treatment led to the intensive and continuous research of PRP products and to its widespread clinical use. A number of protocols and commercial kits have been developed with the intention of creating a more practical and reliable option for clinical use in equine patients. Still, the direct comparison betwee...
Jiménez-Aristizabal RF, López C, Álvarez ME, Giraldo C, Prades M, Carmona JU.To compare the temporal release (over three weeks) of tumor necrosis factor alpha (TNF-α), interleukin 4 (IL-4), IL-1 receptor antagonist (IL-1ra), platelet-derived growth factor BB (PDGF-BB) and transforming growth factor beta-1 (TGF-β1) from two platelet-rich fibrin (PRF) preparations from equine blood obtained at either 240g/8min or 416g/10min. Whole blood from 10 horses was used to obtain PRF clots by two different centrifugation protocols. After 1h of rest, PRF clots were deposited in wells with culture medium, which was changed at 6h, 24h and then every 48h to 21days. Cytokines and GFs...
Côté GP, Smillie LB.Whereas skeletal muscle tropomyosin binds strongly to muscle F-actin in a buffer containing 30 mM KCl and 1-2 mM free Mg2+, equine platelet tropomyosin only binds stoichiometrically (1 tropomyosin molecule per 6 actin monomers) at higher Mg2+ concentrations (7-8 mM free Mg2+). At low free Mg2+ concentrations (1.5 mM) the binding of the platelet protein is only marginally increased by raising the KCl concentration to an optimal value (0.10-0.20 M). This weaker binding can be attributed to the relatively poor head-to-tail polymerization of platelet tropomyosin and its fewer actin-binding sites. ...
Russell KA, Koch TG.Multipotent mesenchymal stromal cells (MSC) are often culture-expanded in vitro. Presently, expansion medium (EM) for MSC is supplemented with fetal bovine serum (FBS). However, increasing cost, variable composition and potential risks associated with bovine antigens call for alternatives. Platelet lysate (PL) has shown promise as an alternative supplement. Objective: To determine how equine umbilical cord blood (CB) MSC proliferate in EM enriched with PL or FBS at various concentrations. Methods: Randomised dose escalation study. Methods: Platelet concentrate was generated from 5 equine whol...
Smith JJ, Ross MW, Smith RK.The purpose of this study was to investigate the response of suspensory ligament fibroblasts (SLF) to in vitro stimulation using acellular bone marrow (ABM), platelet rich plasma (in vitro PRP), and serum as potential treatment modalities for suspensory desmitis. Blood, bone marrow, and suspensory ligaments were collected from five horses. SLF were harvested, grown until confluent, and stimulated with various concentrations of ABM, PRP, equine serum, foetal bovine serum, and medium (control). The responses to the treatments were assessed using a combination of radio-labeling for total protein ...
Lapetina EG.The addition of platelet-activating factor to horse platelets prelabeled with (32P)orthophosphate induces the rapid formation of [32P]phosphatidic acid. This is subsequently followed by a considerable increase in the labeling of phosphatidylinositol. Activation of the formation of phosphatidic acid by platelet-activating factor is evident at a concentration of 1 nM, is maximal at 0.1 microM, and independent of the presence or absence of plasma in the medium. In horse platelets prelabeled with [14C]arachidonic acid, platelet-activating factor stimulates the rapid formation of [14C]phosphatidic ...
Kisiday JD, McIlwraith CW, Rodkey WG, Frisbie DD, Steadman JR.To evaluate the effects of single- and double-spin preparations of platelet-rich plasma (PRP) on anabolic and catabolic activities of cartilage and meniscal explants in vitro. Methods: Single- and double-spin PRP was prepared using laboratory processing or commercial kits. The cellular contents were quantified, and each PRP was mixed in equal quantities with cell culture medium and added to cartilage or meniscus explant cultures, with or without interleukin 1 β (IL-1β). Extracellular matrix synthesis was quantified over 24 hours via (35)S-sulfate and (3)H-proline incorporation, while gene ex...
