Relevant Aspects of Centrifugation Step in the Preparation of Platelet-Rich Plasma

Introduction. Platelet-Rich Plasma (PRP) is rich in growth factors, playing important role in tissue healing. The wide variation of reported protocols for preparation of PRP leads to variable compositions, which induce different biological responses and prevent results comparison. This study aims to highlight relevant aspects of the centrifugation step to obtain reproducible results and overall quality. Material and Methods. Samples of blood were collected from 20 healthy donors that have signed free informed consent. Two centrifugation steps (spins) were analyzed for the influence of centrifugal acceleration, time, processed volume, and platelet gradient. The Pure Platelet-Rich Plasma (P-PRP) was characterized as platelet concentration, integrity, and viability (sP-selectin measurement). Results. Lower centrifugal accelerations favour platelet separation. The processing of 3.5 mL of blood at 100 ×g for 10 min (1st spin), 400 ×g for 10 min (2nd spin), withdrawing 2/3 of remnant plasma, promoted high platelet recovery (70–80%) and concentration (5x) maintaining platelet integrity and viability. The recovery of platelets was reduced for a larger WB volume (8.5 mL) processed. Conclusion. Centrifugal acceleration, time, WB processed volume, and minimization of the platelet gradient before sampling are relevant aspects to ensure reproducible compositions within the autologous nature of PRP.

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Optimizing Platelet-Rich Plasma Gel Formation by Varying Time and Gravitational Forces During Centrifugation

Journal of Oral Implantology ◽

10.1563/aaid-joi-d-10-00155 ◽

2013 ◽

Vol 39 (5) ◽

pp. 525-532 ◽

Cited By ~ 31

Author(s):

Chris H. Jo ◽

Young Hak Roh ◽

Ji Eun Kim ◽

Sue Shin ◽

Kang Sup Yoon

Keyword(s):

Plasma Layer ◽

Platelet Rich Plasma ◽

Volume Ratio ◽

Platelet Recovery ◽

Tissue Healing ◽

Plain Tube ◽

Centrifugation Step ◽

Gel Preparation ◽

Main Components ◽

Prp Gel

Despite the increasing clinical use of topical platelet-rich plasma (PRP) to enhance tissue healing and regeneration, there is no properly standardized method of autologous PRP gel preparation. This study examined the effect of the centrifugation time and gravitational force (g) on the platelet recovery ratio of PRP and determined the most effective centrifugation conditions for preparing PRP. Two-step centrifugations for preparing PRP were used in 39 subjects who had consented prior to the study's start. The separating centrifugation (Step 1, used to separate whole blood into its two main components: red blood cells and plasma) was tested from 500g to 1900g at 200g increments for 5 minutes (min), and from 100g to 1300g at 200g increments for 10 minutes. After separating centrifugation, upper plasma layer was transferred to another plain tube for the condensation centrifugation and remaining lower cell layer was discarded. The condensation centrifugation (Step 2, used to condense the platelets in the separated plasma) was tested at 1000g for 15 min, 1500g for 15 min, 2000g for 5 min and 3000g for 5 min, additionally at 1000g for 10 min and 1500g for 10 min. Platelet gelation was induced by adding 10% calcium gluconate to final PRP with volume ratio of 1:10. The optimal separating centrifugation conditions were followed by 900g for 5 minutes and the condensation conditions were followed by 1500g for 15 minutes, of which recovery ratios were 92.0 ± 3.1% and 84.3 ± 10.0%, respectively.

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Changes in the activity of ovine blood-derived macrophages stimulated with antimicrobial peptide extract (AMP) or platelet-rich plasma (PRP)

Journal of Veterinary Research ◽

10.2478/jvetres-2019-0021 ◽

2019 ◽

Vol 63 (2) ◽

pp. 235-242

Author(s):

Joanna Wessely-Szponder ◽

Tomasz Szponder ◽

Ryszard Bobowiec ◽

Joanna Michalska

Keyword(s):

Antimicrobial Activity ◽

Growth Factors ◽

Soft Tissue ◽

Platelet Rich Plasma ◽

Innate Immune ◽

Ros Generation ◽

Soft Tissue Healing ◽

Tissue Healing ◽

Functional Features ◽

Stimulation Of

AbstractIntroduction: Antimicrobial peptides (AMP) are a large group of innate immune effectors, which apart from antimicrobial activity show immunomodulative properties. Platelet-rich plasma (PRP) is a source of autologous growth factors and is used for stimulation of bone and soft tissue healing. The purpose of this study was to assess the influence of PRP and AMP extract on ovine monocyte-derived macrophage cultures. Material and Methods: The study was conducted on ovine macrophages (Mfs) previously stimulated with LPS or dexamethasone and then with preparations of PRP or AMP. Following activation of the Mfs their morphological and functional features were assessed. Results: The study revealed pro-inflammatory influence of both examined preparations on Mfs cultures on the basis of morphology, ROS generation and arginase activity. Both preparations enhanced the pro-inflammatory response of cultured Mfs. Conclusion: This activity may intensify the antimicrobial action of Mfs, however, in cases of excessive and prolonged inflammation the use of these preparations should be limited.

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Unactivated platelet leukocyte autoplasma in the treatment of non scarring alopecia

Russian Journal of Skin and Venereal Diseases ◽

10.18821/1560-9588-2016-19-6-369-377 ◽

2016 ◽

Vol 19 (6) ◽

pp. 369-377

Author(s):

G. A Ragimov ◽

O. Yu Olisova ◽

Kseniya G. Egorova

Keyword(s):

Growth Factors ◽

Clinical Cure ◽

Clinical Observation ◽

Platelet Rich Plasma ◽

White Blood Cells ◽

Treatment Method ◽

Hair Follicles ◽

Scarring Alopecia ◽

Platelet Concentration

The literature review of the cellular functioning mechanisms of the hair follicles, the role of stem cells in the life cycle of the hair, the major effects of growth factors are presented. Authors, patented treatment method of non scarring alopecia is described. The technology ofpreparation and method of use of unactivated platelet leukocyte autoplasma in various forms of alopecia are described. Own clinical observation of 60 patients with non scarring alopecia and results of their unactivated platelet leukocyte autoplasma are presented. 80% of patients had a clinical cure. Investigations of platelet concentration, white blood cells and growth factors in platelet-rich plasma and platelet leukocyte unactivated autoplasma were performed. The results are shown in the article.

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Review of concentration yields in commercially available platelet-rich plasma (PRP) systems: a call for PRP standardization

Regional Anesthesia and Pain Medicine ◽

10.1136/rapm-2018-100356 ◽

2019 ◽

Vol 44 (6) ◽

pp. 652-659 ◽

Cited By ~ 15

Author(s):

Priyal P Fadadu ◽

Anthony J Mazzola ◽

Corey W Hunter ◽

Timothy T Davis

Keyword(s):

Systematic Review ◽

Growth Factors ◽

Platelet Rich Plasma ◽

White Cell Count ◽

Pain Medicine ◽

Inclusion Criteria ◽

Separation Systems ◽

Platelet Concentration ◽

Large Heterogeneity ◽

Promising Treatment

Platelet-rich plasma (PRP) has become increasingly popular in pain medicine with hopes of becoming a safe, effective alternative to routine treatments. However, given its autologous nature, PRP injectate may differ depending on the specific manufacturer and protocol. Currently, there is no standardization of reporting protocol. This systematic review compiles and standardizes values on PRP preparation and final product composition of platelets, white cell count, and growth factors for ease of comparison. On review of 876 studies, 13 studies were selected according to our inclusion criteria. Data from 33 PRP systems and protocols were extracted and standardized. Overall, PRP final product concentrations as well as PRP preparation protocols varied widely between systems. However, platelet concentration was directly correlated with both volume of blood collected and device centrifugal force. In conclusion, there is a large heterogeneity between PRP separation systems that must be resolved for proper study of this promising treatment.

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Platelet-Rich Plasma for the Treatment of Degenerative Lumbosacral Stenosis: A Study with Retired Working Dogs

Animals ◽

10.3390/ani11102965 ◽

2021 ◽

Vol 11 (10) ◽

pp. 2965

Author(s):

Ángel María Hernández-Guerra ◽

José María Carrillo ◽

Joaquín Jesús Sopena ◽

José Manuel Vilar ◽

Pau Peláez ◽

...

Keyword(s):

Growth Factors ◽

Structural Integrity ◽

Platelet Rich Plasma ◽

Cauda Equina ◽

Epidural Injection ◽

Good Alternative ◽

Epidural Injections ◽

Tissue Healing ◽

Before And After ◽

Working Dogs

Traditionally, canine degenerative lumbosacral stenosis (DLS) has been defined as a multifactorial syndrome characterized by lumbosacral pain triggered by the compression of the nerve rootlets of the cauda equina. There is still no consensus on the treatment of this condition, probably because there are a plethora of possible causes. In addition to compression, inflammation is a very important factor in the physiopathology of the disorder. Platelet-rich plasma (PRP) consists of an increased concentration of autologous platelets suspended in a small amount of plasma. Platelets are a source of several growth factors. Growth factors were shown to help in wound healing and biological processes, such as chemotaxis, neovascularization and synthesis of extracellular matrix, and growth factors were used to improve soft tissue healing and bone regeneration. PRP also facilitates the restoration of the structural integrity of the affected anatomy. Fourteen dogs diagnosed with DLS were treated with three epidural injections of PRP on days 0, 15 and 45. All dogs showed clinical improvement 3 months after the initial treatment. Gait was also objectively assessed by means of the use of force platform analysis before and after treatment, showing significant improvement. The results show that PRP may provide a good alternative to other nonsurgical treatments, such as prednisolone epidural injection.

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Investigation of Growth Factor and Tenocyte Proliferation Induced by Platelet Rich Plasma (PRP) in a 3-Chamber Co-Culture Device

Micromachines ◽

10.3390/mi9090446 ◽

2018 ◽

Vol 9 (9) ◽

pp. 446

Author(s):

Chih-Hao Chiu ◽

Rei Higashikawa ◽

Wen-Ling Yeh ◽

Kin Lei ◽

Alvin Chen

Keyword(s):

Platelet Rich Plasma ◽

Lateral Epicondylitis ◽

Rotator Cuff Tears ◽

Preparation Procedure ◽

Soft Tissue Healing ◽

Tissue Healing ◽

Platelet Number ◽

Platelet Concentration ◽

Practical Tool ◽

Basic Studies

The platelet-rich plasma (PRP) has become an attractive topic for soft tissue healing therapy recently. While some clinical reports revealed the effective treatments for knee osteoarthritis, lateral epicondylitis, and rotator cuff tears, other case studies showed that there was no statistically significant healing improvement. The efficacy of the PRP therapy is still unclear clinically. Thus, a significant amount of basic studies should be conducted to optimize the preparation procedure and the platelet concentration of the PRP. In this work, a 3-chamber co-culture device was developed for the PRP study in order to reduce the usage of primary cells and to avoid the PRP gelation effect. The device was a culture, well partitioning into 3 sub-chambers. Tenocytes and PRP could be respectively loaded into the sub-chambers and co-cultured under the interlinked medium. The results showed that a higher platelet number in the PRP could diffuse higher concentration of the growth factors in the medium and induce higher tenocyte proliferation. The 3-chamber co-culture device provides a simple and practical tool for the PRP study. It is potentially applied for optimizing the preparation procedure and platelet concentration of the PRP therapy.

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Quality assessment of platelet rich plasma during anti-platelet therapy

Perfusion ◽

10.1177/0267659107077950 ◽

2007 ◽

Vol 22 (1) ◽

pp. 41-50 ◽

Cited By ~ 10

Author(s):

Chad W Smith ◽

Robert S Binford ◽

David W Holt ◽

David P Webb

Keyword(s):

Growth Factors ◽

Acetylsalicylic Acid ◽

Platelet Rich Plasma ◽

Surgical Wound ◽

Platelet Function Test ◽

Wound Site ◽

Clopidogrel Bisulfate ◽

Anti Platelet Therapy ◽

Function Test ◽

Platelet Concentration

Platelet rich plasma (PRP) is being used with increased frequency in many surgical procedures for its known benefits of accelerated surgical wound site healing. Speculations in its efficacy in the presence of anti-platelet therapy have been proposed. To aid in defining a quality platelet rich plasma product in the presence of acetylsalicylic acid (ASA) and Plavix (clopidogrel bisulfate), we investigated three (3) groups (n=18) of cardiac surgical patients receiving PRP. Platelet function test, platelet concentration, and quantification of growth factors (PDGF-bb and TGF-b1) were evaluated. Results showed no statistical evidence of decreased growth factors delivered to the surgical wound site in the presence of acetylsalicylic acid (ASA) and/or Plavix (clopidogrel bisulfate). Evidence in this pilot study supports the use of PRP for patients receiving Plavix and aspirin therapy without compromising the quantity of specific growth factors delivered to a wound site. Perfusion (2007) 22, 41—50.

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Growth Factor Quantification of Platelet-Rich Plasma in Burn Patients Compared to Matched Healthy Volunteers

International Journal of Molecular Sciences ◽

10.3390/ijms20020288 ◽

2019 ◽

Vol 20 (2) ◽

pp. 288 ◽

Cited By ~ 4

Author(s):

Roos Marck ◽

Kim Gardien ◽

Marcel Vlig ◽

Roelf Breederveld ◽

Esther Middelkoop

Keyword(s):

Wound Healing ◽

Growth Factors ◽

Growth Factor ◽

Healthy Volunteers ◽

Platelet Rich Plasma ◽

Healing Process ◽

Burn Patients ◽

Gender And Age ◽

Platelet Concentration ◽

And Gender

Platelet rich plasma (PRP) is blood plasma with a platelet concentration above baseline. When activated, PRP releases growth factors involved in all stages of wound healing, potentially boosting the healing process. To expand our knowledge of the effectiveness of PRP, it is crucial to know the content and composition of PRP products. In this study, growth factor quantification measurements of PRP from burn patients and gender- and age-matched controls were performed. The PRP of burn patients showed levels of growth factors comparable to those of the PRP of healthy volunteers. Considerable intra-individual variation in growth factor content was found. However, a correlation was found between the platelet count of the PRP and most of the growth factors measured.

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Advantages and Clinical Applications of Lyophilized Platelet-Rich Plasma in Various Disease Conditions: A Review

International Journal for Research in Applied Science and Engineering Technology ◽

10.22214/ijraset.2021.39092 ◽

2021 ◽

Vol 9 (11) ◽

pp. 1885-1892

Author(s):

Pradeep Mahajan

Keyword(s):

Growth Factors ◽

Platelet Rich Plasma ◽

Autologous Blood ◽

Healing Process ◽

Coagulation Factor ◽

The Body ◽

Plasma Therapy ◽

Plasma Fraction ◽

Wide Range ◽

Platelet Concentration

Abstract: Platelet rich plasma (PRP) is a biological product defined as a portion of the plasma fraction of autologous blood with a platelet concentration above the baseline. The plasma occupies 55% of blood, which is rich in immunoglobulins and proteins that have a wide range of applications in various medical fields. Plasma therapy is applied to tackle various disorders or diseases as it induces the body to develop new healthy cells. It contains important components like antibodies, coagulation factor, enzymes, fibrinogen, proteins and albumin. PRP is a unique and advanced treatment which helps to increases the body’s natural healing process. Platelet lysate which is obtained from platelet rich plasma consist of various growth factors such as chemokines, cytokines, and antibacterial molecules and also has anti-inflammatory, immunomodulatory, anti-fibrotic and repairing effects. As PRP is rich in the proteins and several antibodies, it is used for various chronic therapies such as hemophilia and autoimmune disorders as well as in various severe health problems. Lyophilized Platelet-rich plasma (LPRP) therapy is currently used in various fields such as in tissue regeneration, wound healing, scar revision, skin rejuvenating effects, alopecia and for the coronavirus disease (COVID-19). It is also used to heal wounds and illnesses. LPRP therapy is gaining attraction by many health professionals as it is a safe, effective, efficient, and easy approach in procuring, preserving, and therapy. In this review we described the advantages and applications of using lyophilized PRP in various diseases which might found to be effective in different treatment. Keywords: Plasma, Platelet, Growth Factors, Lyophilized platelet rich plasma.

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Does Double Centrifugation Lead to Premature Platelet Aggregation and Decreased TGF-β1 Concentrations in Equine Platelet-Rich Plasma?

Veterinary Sciences ◽

10.3390/vetsci6030068 ◽

2019 ◽

Vol 6 (3) ◽

pp. 68

Author(s):

Sarah R. T. Seidel ◽

Cynthia P. Vendruscolo ◽

Juliana J. Moreira ◽

Joice Fülber ◽

Tatiana F. Ottaiano ◽

...

Keyword(s):

Platelet Aggregation ◽

Platelet Activation ◽

Platelet Rich Plasma ◽

Correlation Coefficients ◽

Platelet Concentration ◽

Centrifugation Step ◽

High Concentrations ◽

Different Characteristics ◽

Tgf Β1 ◽

Centrifugation Protocol

Blood-derived autologous products are frequently used in both human and equine medicine to treat musculoskeletal disorders. These products, especially the platelet-rich plasma (PRP), may contain high concentrations of growth factors (GFs), and thus improve healing in several tissues. Nevertheless, the procedures for preparation of PRP are currently non-standardized. Several protocols, which are based on distinct centrifugation patterns (rotation speed and time), result in PRPs with different characteristics, concerning platelet and GFs concentrations, as well as platelet activation. The aim of the present study was to compare two different protocols for PRP preparation: protocol (A) that is based on a single-centrifugation step; protocol (B), which included two sequential centrifugation steps (double-centrifugation). The results here reported show that the double-centrifugation protocol resulted in higher platelet concentration, while leukocytes were not concentrated by this procedure. Although platelet activation and aggregation were increased in this protocol in comparison to the single-centrifugation one, the TGF-β1 concentration was also higher. Pearson’s correlation coefficients gave a significant, positive correlation between the platelet counts and TGF-β1 concentration. In conclusion, although the double-centrifugation protocol caused premature platelet aggregation, it seems to be an effective method for preparation of PRP with high platelet and TGF-β1 concentrations.

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