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Optimization of a rapid one-step platelet-rich plasma preparation method using syringe centrifugation with and without carprofen.

ABSTRACT: BACKGROUND:Carprofen and platelet-rich plasma (PRP) are widely used in small animal clinical practice. Separation layers have been used during blood centrifugation to increase platelet yield. The objectives of this study were to (1) identify the optimal centrifugation force for the one-step PRP preparation, (2) determine whether there is an advantage to using carprofen in one-step PRP preparation, and (3) compare platelet morphology from one-step PRP preparation with and without carprofen. We hypothesized that injectable carprofen (emulsion formula) could be used successfully as the separation layer in PRP preparation. RESULTS:Samples from 14 healthy dogs were used to determine the optimal centrifugation force using one-step PRP preparation in a disposable syringe without carprofen, with forces set at 300, 500, 700, 900, 1100, 1300, and 1500 xg for 5?min. Optimum centrifugation force, plasma volume, and platelet concentrations of one-step PRP preparation were found and recovered at 900 xg, 1.9?±?0.28?ml, and 260.50?±?58.39 X 103 cell/?l, respectively. Samples from 12 healthy dogs were used to determine the optimal force (with forces set at 300, 500, 700, and 900 xg) for 5?min using one-step PRP preparation with carprofen. Optimum centrifugation force, plasma volume, and platelet concentrations for one-step PRP preparation with carprofen were found and recovered at 500 xg, 0.62?±?0.16?ml and 948.50?±?261.40 X 103 cell/?l, respectively. One-step PRP preparation with carprofen increased the platelet yield from baseline by 1.76 and 4.95 fold, respectively. Samples from 3 healthy dogs were used to observe platelet morphologies after centrifugation by scanning electron microscopy. Images of platelets on glass slides from both preparation methods revealed pseudopods emerging from the margins of the discoid platelets. CONCLUSIONS:One-step PRP centrifugation both with and without carprofen increased the platelet yield, but using carprofen (emulsion formula) as a separation layer resulted in a higher platelet yield. The clinical usefulness of PRP products from these methods should be further investigated.

SUBMITTER: Apakupakul J

PROVIDER: S-EPMC7204049 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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Optimization of a rapid one-step platelet-rich plasma preparation method using syringe centrifugation with and without carprofen.

Apakupakul Jutarat J Sattasathuchana Panpicha P Chanloinapha Phongsak P Thengchaisri Naris N

BMC veterinary research 20200506 1

<h4>Background</h4>Carprofen and platelet-rich plasma (PRP) are widely used in small animal clinical practice. Separation layers have been used during blood centrifugation to increase platelet yield. The objectives of this study were to (1) identify the optimal centrifugation force for the one-step PRP preparation, (2) determine whether there is an advantage to using carprofen in one-step PRP preparation, and (3) compare platelet morphology from one-step PRP preparation with and without carprofe ...[more]

PMID: 32375782

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Project description:Objective: Recently, liquid platelet-rich fibrin (PRF), a rich source of concentrated platelets and growth factors, has emerged as a promising agent for stimulating tissue regeneration. However, its specific efficacy in chondrocyte proliferation and cartilage regeneration remains underexplored. To address this question, we investigated liquid PRF obtained through horizontal centrifugation and compared its effects with hyaluronic acid (HA), a high molecular weight glucosamine supplement widely used in clinical practice to safeguard against chondral damage. Materials and Methods: Liquid PRF, produced using horizontal centrifugation (liquid H-PRF) at 500 g for 8 min, served as our experimental agent. We conducted cell viability and proliferation assays using PRF-conditioned medium. We assessed the chondrocyte phenotype of ATDC5 cells through toluidine blue and alcian blue staining, real-time polymerase chain reaction (RT-PCR), Western blotting, and immunofluorescence staining. Furthermore, we examined the expression of genes involved in inflammation through RT-PCR and Western blot analysis. Results: Liquid H-PRF exerted notable effects on chondrocytes, influencing proliferation, inflammatory responses, and chondrogenic differentiation. The H-PRF group displayed significantly higher expression of chondrogenic markers, including Col2a1, compared to HA-treated cells, whereas aggrecan expression was significantly higher in the HA group. PRF also demonstrated the ability to reduce inflammatory levels in chondrogenic ATDC5 cells, and this effect was further enhanced when PRF from the buffy coat zone was added. In comparison, chondrocytes cultured in the HA group produced significantly fewer inflammatory factors than those in the PRF group, as confirmed qualitatively by Western blot analysis. Conclusion: Liquid H-PRF emerged as a potent stimulator for chondrogenesis and a regulator of the inflammatory response, achieving levels similar to HA. Moreover, liquid H-PRF exhibited strong potential for enhancing the production of cartilage extracellular matrix and promoting chondrogenic regeneration with notably increased Col2a1 levels. Future research should encompass animal studies and human trials to further evaluate the comparative effectiveness of liquid PRF versus HA, potentially as an alternative or complementary strategy for future clinical applications.

Project description:BackgroundThe role of platelet-rich plasma (PRP) in the treatment of sport-related injuries is unclear, largely due to the heterogeneity of clinical results. This may relate to compositional differences in PRP from different separation systems.Questions/purposesThis study aims to compare the composition of PRP produced with five different commercially available systems, focusing on cellular concentrations and pH.MethodsSeven donors (41 ± 12 years) provided blood for PRP preparation using five systems (Arthrex Angel, Emcyte Genesis CS, Arteriocyte Magellan, Harvest SmartPrep, and Biomet GPS III). Post processing, cellular composition was measured including platelets (PLT), white blood cells (WBC), neutrophils (NE), and red blood cells (RBC), as well as pH.ResultsPlatelet concentration and capture efficiency were similar between systems, except the Angel 7% preparation had a greater concentration than Genesis CS (2310 ± 524 vs. 1129 ± 264 k/μL). WBC concentration was variable between systems; however, significant differences were only found between the Angel 2% and GPS III preparations (11.0 ± 4.5, 27.3 ± 7.1 k/μL). NE concentration was significantly lower in the Angel 2% and 7% preparations compared with GPS III (0.6 ± 0.6 and 1.8 ± 1.3 k/μL vs. 9.4 ± 7.0 k/μL). RBC concentration was highest in SmartPrep (3.2 ± 0.6 M/μL) and Genesis CS systems (3.1 ± 0.6 M/μL) compared with all other systems (≤1.1 ± 1.2 M/μL). Finally, pH was significantly lower with the SmartPrep system (6.95 ± 0.06) compared with all others (≥7.26 ± 0.06).ConclusionAside from platelet concentration and capture efficiency, significant compositional differences were identified between preparation systems. Caution should be employed when interpreting clinical results of studies utilizing PRP, as the role of compositional differences and their effect on outcome are unknown. Further study is necessary to determine the clinical significance of these differences.

Dataset Information

Optimization of a rapid one-step platelet-rich plasma preparation method using syringe centrifugation with and without carprofen.

Publications

Optimization of a rapid one-step platelet-rich plasma preparation method using syringe centrifugation with and without carprofen.

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