Platelet-Rich Preparations to Improve Healing. Part II: Platelet Activation and Enrichment, Leukocyte Inclusion, and Other Selection Criteria

Multiple platelet-rich preparations have been reported to improve wound and bone healing, such as platelet-rich plasma (PRP) and platelet rich fibrin (PRF). The different methods employed during their preparation are important, as they influence the quality of the product applied to a wound or surgical site. Besides the general protocol for preparing the platelet-rich product (discussed in Part 1 of this review), multiple choices need to be considered during its preparation. For example, activation of the platelets is required for the release and enmeshment of growth factors, but the method of activation may influence the resulting matrix, growth factor availability, and healing. Additionally, some methods enrich leukocytes as well as platelets, but others are designed to be leukocyte-poor. Leukocytes have many important roles in healing and their inclusion in PRP results in increased platelet concentrations. Platelet and growth factor enrichment reported for the different types of platelet-rich preparations are also compared. Generally, TGF-β1 and PDGF levels were higher in preparations that contain leukocytes compared to leukocyte-poor PRP. However, platelet concentration may be the most reliable criterion for comparing different preparations. These and other criteria are described to help guide dental and medical professionals, in large and small practices, in selecting the best procedures for their patients. The healing benefits of platelet-rich preparations along with the low risk and availability of simple preparation procedures should encourage more clinicians to incorporate platelet-rich products in their practice to accelerate healing, reduce adverse events, and improve patient outcomes.

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Centrifugation Conditions in the L-PRP Preparation Affect Soluble Factors Release and Mesenchymal Stem Cell Proliferation in Fibrin Nanofibers

Molecules ◽

10.3390/molecules24152729 ◽

2019 ◽

Vol 24 (15) ◽

pp. 2729 ◽

Cited By ~ 4

Author(s):

Melo ◽

Luzo ◽

Lana ◽

Santana

Keyword(s):

Growth Factor ◽

Transforming Growth Factor ◽

Platelet Rich Plasma ◽

Clinical Practices ◽

Soluble Factors ◽

Tnf Α ◽

Platelet Concentration ◽

Tgf Β1

Leukocyte and platelet-rich plasma (L-PRP) is an autologous product that when activated forms fibrin nanofibers, which are useful in regenerative medicine. As an important part of the preparation of L-PRP, the centrifugation parameters may affect the release of soluble factors that modulate the behavior of the cells in the nanofibers. In this study, we evaluated the influences of four different centrifugation conditions on the concentration of platelets and leukocytes in L-PRP and on the anabolic/catabolic balance of the nanofiber microenvironment. Human adipose-derived mesenchymal stem cells (h-AdMSCs) were seeded in the nanofibers, and their viability and growth were evaluated. L-PRPs prepared at 100× g and 100 + 400× g released higher levels of transforming growth factor (TGF)-β1 and platelet-derived growth factor (PDGF)-BB due to the increased platelet concentration, while inflammatory cytokines interleukin (IL)-8 and tumor necrosis factor (TNF)-α were more significantly released from L-PRPs prepared via two centrifugation steps (100 + 400× g and 800 + 400× g) due to the increased concentration of leukocytes. Our results showed that with the exception of nanofibers formed from L-PRP prepared at 800 + 400× g, all other microenvironments were favorable for h-AdMSC proliferation. Here, we present a reproducible protocol for the standardization of L-PRP and fibrin nanofibers useful in clinical practices with known platelet/leukocyte ratios and in vitro evaluations that may predict in vivo results.

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Time-Dependent Cytokine-Release of Platelet-Rich Plasma in 3-chamber Co-culture Device and Conventional Culture Well

Applied Sciences ◽

10.3390/app11156947 ◽

2021 ◽

Vol 11 (15) ◽

pp. 6947

Author(s):

Chih-Hao Chiu ◽

Poyu Chen ◽

Alvin Chao-Yu Chen ◽

Yi-Sheng Chan ◽

Kuo-Yao Hsu ◽

...

Keyword(s):

Growth Factor ◽

Transforming Growth Factor Beta ◽

Transforming Growth Factor ◽

Platelet Rich Plasma ◽

Time Dependent ◽

Dependent Manner ◽

Time Points ◽

Conventional Culture ◽

Platelet Concentration ◽

Tgf Β1

Platelet-rich plasma (PRP) contains bioactive cytokines to enhance tissue healing. The best PRP preparation protocol and timing of the treatment have not been determined yet. To screen the best-fit PRP, a 3-chamber co-culture device was developed. We hypothesized the concentrations of the cytokines from different PRPs in the co-culture plates had a high correlation with those in conventional 24-well culture plates at different time points. The concentrations of the cytokine from PRPs would be correlated with platelet concentrations. The correlation of transforming growth factor beta-1 (TGF-β1) and platelet-derived growth factor AB (PDGF-AB) in both devices were compared at 0, 24, 48, 72, and 96 h from two PRPs as well as that of platelet and cytokines concentrations. The results revealed that there was a moderate to high correlation in TGF-β1 concentrations between the 3-chamber co-culture and conventional culture device until 96 h. The correlation of PDGF-AB concentrations in both devices had moderate to high correlation in the first 24 h, and then it became modestly correlated from 48 to 96 h. A high correlation was found between platelet and TGF-β1 concentration at 96 h. However, they were modestly correlated in other time points. A negative or modest correlation was found between platelet and PDGF-AB concentration in all time points. In conclusion, TGF-β1 and PDGF-AB revealed a time-dependent manner of release at five time points. There is a moderate to high correlation of the TGF-β1 and PDGF-AB concentration in both devices at different time points. However, TGF-β1 and PDGF-AB concentrations are not always proportional to the platelet concentration of the PRPs.

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Addressing the Scottish burden of wounds through the Lens of Profound Knowledge

British Journal of Nursing ◽

10.12968/bjon.2021.30.sup20.s34 ◽

2021 ◽

Vol 30 (Sup20) ◽

pp. S34-S40

Author(s):

Jenni MacDonald

Keyword(s):

Patient Outcomes ◽

Wound Care ◽

Cost Effective ◽

Theory Of Knowledge ◽

Care Needs ◽

Profound Knowledge ◽

The Cost ◽

Improve Patient ◽

Growing Population

The prevalence of wounds and the cost of treating them are increasing year on year. Improving the quality of wound care will improve patient outcomes and is a financial necessity. The Lens of Profound Knowledge is a tool that can be used to support quality improvement and identify where action is needed. It allows exploration of an organisation through four aspects—appreciate the system, understanding variation, psychology, and theory of knowledge—and working on all four aspects simultaneously is believed to increase the likelihood of achieving improvement. Improvements at and between all levels—microsystem (such as frontline services), mesosystem (health boards) and macrosystem (NHS Scotland)—would reduce variation in practice and prove to be both clinically and cost-effective. Given the rapidly growing population of people with unhealed wounds, wound care needs to be valued at all system levels and be adequately resourced.

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Platelet-Rich Plasma Powder: A New Preparation Method for the Standardization of Growth Factor Concentrations

The American Journal of Sports Medicine ◽

10.1177/0363546516674475 ◽

2016 ◽

Vol 45 (4) ◽

pp. 954-960 ◽

Cited By ~ 26

Author(s):

Matthias Kieb ◽

Frank Sander ◽

Cornelia Prinz ◽

Stefanie Adam ◽

Anett Mau-Möller ◽

...

Keyword(s):

Growth Factors ◽

Growth Factor ◽

Clinical Application ◽

Whole Blood ◽

Sports Medicine ◽

Transforming Growth Factor ◽

Platelet Rich Plasma ◽

Transforming Growth Factor Β1 ◽

Enzyme Linked Immunosorbent Assay ◽

Tgf Β1

Background: Platelet-rich plasma (PRP) is widely used in sports medicine. Available PRP preparations differ in white blood cell, platelet, and growth factor concentrations, making standardized research and clinical application challenging. Purpose: To characterize a newly standardized procedure for pooled PRP that provides defined growth factor concentrations. Study Design: Controlled laboratory study. Methods: A standardized growth factor preparation (lyophilized PRP powder) was prepared using 12 pooled platelet concentrates (PCs) derived from different donors via apheresis. Blood samples and commercially available PRP (SmartPrep-2) served as controls (n = 5). Baseline blood counts were analyzed. Additionally, single PCs (n = 5) were produced by standard platelet apheresis. The concentrations of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), platelet-derived growth factor AB (PDGF-AB), transforming growth factor β1 (TGF-β1), insulin-like growth factor 1 (IGF-1), interleukin (IL)–1α, IL-1β, and IL-1 receptor agonist (IL-1RA) were analyzed by enzyme-linked immunosorbent assay, and statistical analyses were performed using descriptive statistics, mean differences, 95% CIs, and P values (analysis of variance). Results: All growth factor preparation methods showed elevated concentrations of the growth factors VEGF, bFGF, PDGF-AB, and TGF-β1 compared with those of whole blood. Large interindividual differences were found in VEGF and bFGF concentrations. Respective values (mean ± SD in pg/mL) for whole blood, SmartPrep-2, PC, and PRP powder were as follows: VEGF (574 ± 147, 528 ± 233, 1087 ± 535, and 1722), bFGF (198 ± 164, 410 ± 259, 151 ± 99, and 542), PDGF-AB (2394 ± 451, 17,846 ± 3087, 18,461 ± 4455, and 23,023), and TGF-β1 (14,356 ± 4527, 77,533 ± 13,918, 68,582 ± 7388, and 87,495). IGF-1 was found in SmartPrep-2 (1539 ± 348 pg/mL). For PC (2266 ± 485 pg/mL), IGF-1 was measured at the same levels of whole blood (2317 ± 711 pg/mL) but was not detectable in PRP powder. IL-1α was detectable in whole blood (111 ± 35 pg/mL) and SmartPrep-2 (119 ± 44 pg/mL). Conclusion: Problems with PRP such as absent standardization, lack of consistency among studies, and black box dosage could be solved by using characterized PRP powder made by pooling and lyophilizing multiple PCs. The new PRP powder opens up new possibilities for PRP research as well as for the treatment of patients. Clinical Relevance: The preparation of pooled PRP by means of lyophilization may allow physicians to apply a defined amount of growth factors by using a defined amount of PRP powder. Moreover, PRP powder as a dry substance with no need for centrifugation could become ubiquitously available, thus saving time and staff resources in clinical practice. However, before transferring the results of this basic science study to clinical application, regulatory issues have to be cleared.

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Effects of Aspirin on Growth Factor Release From Freshly Isolated Leukocyte-Rich Platelet-Rich Plasma in Healthy Men: A Prospective Fixed-Sequence Controlled Laboratory Study

The American Journal of Sports Medicine ◽

10.1177/0363546519827294 ◽

2019 ◽

Vol 47 (5) ◽

pp. 1223-1229 ◽

Cited By ~ 13

Author(s):

Prathap Jayaram ◽

Peter Yeh ◽

Shiv J. Patel ◽

Racel Cela ◽

Theodore B. Shybut ◽

...

Keyword(s):

Growth Factors ◽

Growth Factor ◽

Whole Blood ◽

Low Dose ◽

Platelet Rich Plasma ◽

Healthy Men ◽

Growth Factor Release ◽

Tgf Β1 ◽

Cox 1 ◽

Factor Release

Background: The benefits of platelet-rich plasma (PRP) are believed to be in part dependent on growth factor release after platelet activation. Platelet activation is complex and involves multiple mechanisms. One important mechanism is driven by cyclooxygenase 1 (COX-1)–mediated conversion of arachidonic acid (AA) to precursor prostaglandins that then mediate proinflammatory responses that trigger growth factor release. Acetylsalicylic acid (ASA; also known as aspirin) is known to irreversibly inhibit COX-1, thereby blocking AA-mediated signaling; however, it is unclear whether ASA use alters growth factor release from freshly isolated PRP. Purpose: To assess the effects of low-dose ASA use on activation of growth factor release from freshly isolated human PRP via AA and thrombin (TBN). Study Design: Controlled laboratory study. Methods: Twelve healthy men underwent blood collection and leukocyte-rich PRP (LR-PRP) preparation through a double-spin protocol to obtain baseline whole blood and PRP counts the same day. PRP was aliquoted into 3 groups: nonactivated, AA activated, and TBN activated. Immediately after activation, the concentrations of transforming growth factor β1 (TGF-β1), vascular endothelial growth factor (VEGF), and platelet-derived growth factor AB (PDGF-AB) were measured using enzyme-linked immunosorbent assays (ELISAs). The same 12 participants were then placed on an 81-mg daily dose of oral ASA for 14 days. Repeat characterization of whole blood and PRP analyses was done on day 14, followed by repeat ELISAs of growth factors under the same nonactivated and activated settings as previously stated. Results: Fourteen days of daily ASA had no effect on the number of platelets and leukocytes measured in whole blood and LR-PRP. Compared with nonactivated LR-PRP, AA- and TBN-mediated activation led to significant release of VEGF and PDGF-AB. In contrast, release of TGF-β1 from LR-PRP was observed only with activation by AA, not with TBN. Consistent with its inhibitory role in AA signaling, ASA significantly inhibited AA-mediated release of all 3 growth factors measured in this study. Although ASA had no effect on TBN-mediated release of VEGF and TGF-β1 from LR-PRP, ASA did partially block TBN-mediated release of PDGF-AB, although the mechanism remains unclear. Conclusion: Daily use of low-dose ASA reduces VEGF, PDGF-AB, and TGF-β1 expression in freshly isolated human LR-PRP when activated with AA. Clinical Relevance: Reduction in growth factor release attributed to daily use of low-dose ASA or other COX inhibitors can be mitigated when PRP samples are activated with TBN. Clinical studies are needed to determine whether activation before PRP injection is needed in all applications where ASA is in use and to what extent ASA may inhibit growth factor release in vivo at the site of injury.

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Pilot study assessing the use of cabergoline for the treatment of cats with hypersomatotropism and diabetes mellitus

Journal of Feline Medicine and Surgery ◽

10.1177/1098612x20933213 ◽

2020 ◽

pp. 1098612X2093321

Author(s):

Christopher J Scudder ◽

Katarina Hazuchova ◽

Ruth Gostelow ◽

David B Church ◽

Yaiza Forcada ◽

...

Keyword(s):

Quality Of Life ◽

Diabetes Mellitus ◽

Pilot Study ◽

Growth Factor ◽

Insulin Dose ◽

Diabetic Control ◽

Insulin Like Growth Factor ◽

Significant Change ◽

Improve Patient

Objectives An affordable and effective treatment is needed to manage feline hypersomatotropism. The aim of this study was to assess whether treatment with oral cabergoline for 90 days in cats with hypersomatotropism and diabetes mellitus improved diabetic and insulin-like growth factor 1 (IGF-1) control. Methods This was a prospective cohort non-blinded pilot study enrolling client-owned cats with spontaneously occurring diabetes mellitus and hypersomatotropism. Cats received oral cabergoline (5–10 µg/kg q24h) for 90 consecutive days. Serum IGF-1 and fructosamine concentrations were measured on days 1, 30 and 90. Quality of life was determined using the DIAQoL-pet questionnaire on days 1 and 90. Results Nine cats were enrolled and eight completed the study. There was no significant change in the following: IGF-1 (day 1 median 2001 ng/ml [range 890–2001 ng/ml]; day 30 median 2001 ng/ml [range 929–2001 ng/ml]; day 90 median 1828 ng/ml [range 1035–2001 ng/ml]; χ2(2) = 0.667, P = 0.805); fructosamine (day 1 median 499 µmol/l [range 330–887 µmol/l], day 30 median 551 µmol/l [range 288–722 µmol/l], day 90 median 503 [range 315–851 µmol/l]; χ2(2) = 0.581, P = 0.764); or DIAQoL-pet score (median on day 1 –2.79 (range –4.62 to –0.28], median on day 90 –3.24 [range –4.41 to –0.28]; P = 0.715). There was a significant change of insulin dose (χ2(2) = 8.667, P = 0.008) with cats receiving higher insulin doses at day 90 compared with day 1 (median on day 1 was 0.98 [range 0.63–1.49] and median on day 90 was 1.56 [range 0.49–2.55] units/kg q12h; P = 0.026). Conclusions and relevance Cabergoline did not improve diabetic control or normalise insulin-like growth factor concentration, or improve patient quality of life.

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Sustained Release of Transforming Growth Factor-β1 from Platelet-Rich Chondroitin Sulfate Glycosaminoglycan Gels

The Journal of Knee Surgery ◽

10.1055/s-0037-1603801 ◽

2017 ◽

Vol 31 (05) ◽

pp. 410-415 ◽

Cited By ~ 5

Author(s):

Kate Birdwhistell ◽

Lohitash Karumbaiah ◽

Samuel Franklin

Keyword(s):

Growth Factor ◽

Chondroitin Sulfate ◽

Cartilage Repair ◽

Transforming Growth Factor ◽

Platelet Rich Plasma ◽

Enzyme Linked Immunosorbent Assay ◽

The Media ◽

Tgf Β1

AbstractActivated platelet-rich plasma (PRP), also referred to as platelet-rich fibrin (PRF), has been used to augment numerous techniques of cartilage repair in the knee but does not always result in superior quality of repair tissue. One possible reason that PRF does not consistently result in excellent cartilage regeneration is the transiency of growth factor provision with PRF. The objective of this study was to compare the release of transforming growth factor (TGF)-β1 from PRF and from PRP combined with a novel chondroitin sulfate glycosaminoglycan (CS-GAG) gel. PRP was prepared from nine healthy dogs and split into two aliquots: one activated with bovine thrombin and calcium chloride (CaCl2) to form PRF and the other aliquot was used to rehydrate a lyophilized CS-GAG gel. Both PRF and the CS-GAG gels were incubated in media for 13 days and media were collected, stored, and replaced every 48 hours and the concentration of TGF-β1 quantified in the media using an enzyme-linked immunosorbent assay. Concentrations of TGF-β1 in the media were up to three times greater with the CS-GAG gels and were significantly (p < 0.05) greater than with PRF on days 3, 5, 7, 9, and 13. Furthermore, TGF-β1 elution was still substantial at day 13 with the use of the CS-GAG gels. Additional in vitro work is warranted to characterize TGF-β1 elution from this CS-GAG gel with human PRP and to determine whether the use of these CS-GAG gels can augment cartilage repair in vivo.

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Patient-reported outcomes in clinical practice

Hematology ◽

10.1182/asheducation-2015.1.501 ◽

2015 ◽

Vol 2015 (1) ◽

pp. 501-506 ◽

Cited By ~ 31

Author(s):

Sarah Dobrozsi ◽

Julie Panepinto

Keyword(s):

Clinical Practice ◽

Patient Outcomes ◽

Patient Reported Outcomes ◽

Clinical Care ◽

Patient Reported Outcome ◽

Physician Communication ◽

Valuable Data ◽

Patient Reported ◽

Improve Patient

Abstract Patient-reported outcome (PRO) measurement plays an increasingly important role in health care and understanding health outcomes. PROs are any report of a patient's health status that comes directly from the patient, and can measure patient symptoms, patient function, and quality-of-life. PROs have been used successfully to assess impairment in a clinical setting. Use of PROs to systematically quantify the patient experience provides valuable data to assist with clinical care; however, initiating use of PROs in clinical practice can be daunting. Here we provide suggestions for implementation of PROs and examples of opportunities to use PROs to tailor individual patient therapy to improve patient outcomes, patient–physician communication, and the quality of care for hematology/oncology patients.

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Studies on the Biological Effects of Ozone: 10. Release of Factors from Ozonated Human Platelets

Mediators of Inflammation ◽

10.1080/09629359990360 ◽

1999 ◽

Vol 8 (4-5) ◽

pp. 205-209 ◽

Cited By ~ 36

Author(s):

G. Valacchi ◽

Velio Bocci

Keyword(s):

Platelet Aggregation ◽

Growth Factor ◽

Transforming Growth Factor ◽

Platelet Rich Plasma ◽

Biological Effects ◽

Transforming Growth Factor Β1 ◽

Human Platelets ◽

Dependent Manner ◽

Dose Dependent ◽

Tgf Β1

In a previous work we have shown that heparin, in the presence of ozone (O3), promotes a dose-dependent platelet aggregation, while after Ca2+chelation with citrate, platelet aggregation is almost negligible. These results led us to think that aggregation may enhance the release of platelet components. We have here shown that indeed significantly higher amount of platelet-derived growth factor (PDGF), transforming growth factor β1 (TGF-β1) and interleukin-8(IL-8) are released in a dose-dependent manner after ozonation of heparinised platelet-rich plasma samples. These findings may explain the enhanced healing of torpid ulcers in patients with chronic limbischemia treated with O3autohaemoteraphy (O3-AHT).

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Platelet Activation Is Not Crucial for Platelet-Rich Plasma (PRP), When Used As Autologous Therapeutic Product, and Could be Lyophilized without Any Growth Factor Loss

Blood ◽

10.1182/blood.v128.22.2639.2639 ◽

2016 ◽

Vol 128 (22) ◽

pp. 2639-2639

Author(s):

Letícia Queiroz da Silva ◽

Stephany Cares Huber ◽

Silmara Aparecida De Lima Montalvão ◽

Fernanda Dutra Santiago Bassora ◽

Erich Vinicius De Paula ◽

...

Keyword(s):

Platelet Aggregation ◽

Growth Factor ◽

Therapeutic Effect ◽

Platelet Activation ◽

Freeze Drying ◽

Platelet Rich Plasma ◽

Therapeutic Product ◽

Platelet Concentration ◽

Factor Measurement ◽

The Mean

Abstract Platelet-rich Plasma (PRP) has been widely used in different fields of medicine as autologous therapeutic product. The main component that appears to be associated with the therapeutic effect is the presence of growth factors (GF). However, many protocols available induce high methodology variability. In addition, is still unclear what is the best platelet activator and the necessity for the clinical practice. The traditional PRP used (Fresh PRP) can vary to each preparation and it has also been difficult to use in a time manager, especially for emergency care. Freeze-drying processes come out as a PRP standardization possibility, offering a low-risk proliferative microorganisms. This study aims to compare in vitrothe fresh PRP with the lyophilized PRP, in terms of platelet concentration capacity, and the GF potential release. For fresh and lyophilized PRP production, plasma from twenty-two male healthy individuals were obtained, with mean age of 28.6 ± 5.6 years. The blood was collected with ACD tubes (BD Vacutainer), than centrifuged twice: first with a spin at 300 g for 5 minutes and second with 700 g for 17 minutes. At the end of the double spin, the top layer plasma was characterized as platelet poor plasma and the lower layer was considered the PRP. The pellet were homogenized slowly, and adjusted to 1,2x106 platelets/µL before being frozen at -80ºC. For lyophilized PRP a stabilizing buffer were add and samples were frozen for 1 hour at -80ºC. After that, the PRP was lyophilized by Christ Alpha Plus for 20 hours. To compare fresh and lyophilized PRP, the platelets were evaluated for number of concentration, functionality, and the capacity of GF release, such as VEGF, PDGF, EGF and TGFβ. Non-parametric statistics were used in all analysis (Graph Pad 5.0). The PRP was able to recover high concentrations of platelets. The mean of platelet concentration was 1622 x 103 cells/µl, which represents 5.3 folds higher from the basal number (303 x 103 cells/µl). The recovery of platelets after freeze-drying was 54% compared to the initial concentration (1200 x 103 cells/µl). Platelet function was evaluated pre and post fresh PRP preparation and after freeze-drying, with two agonists ADP and epinephrine, to check the capacity of then to induce platelet aggregation. Results were evaluated trough amplitude of aggregation curve. Interestingly, high amplitude was observed only for samples from pre fresh PRP preparation (ADP median 86% from 71% to 100 % and epinephrine median 86 % from 74% to 103%). Despite the high concentration obtained from PRP (5 folds higher the basal number), no amplitude curve for platelet aggregation was observed for samples post fresh PRP preparation (ADP median 19% from 0% to 85% and epinephrine median 2% from 1% to 37%), even for lyophilized PRP (ADP median 1% from 0% to 2% and epinephrine median 1.5 % from 0 % to 3%), figure 1. The GF levels were similar for both products, with any grow factor loss after freeze-drying. The mean and standard deviation for level of GF were: PDGF 49365 pg/ml ± 17410 for fresh PRP and 60207 pg/ml ± 18472 for lyophilized PRP; VEGF 1250 pg/ml ±1171 for fresh PRP and 954,3 pg/ml ± 644,6 for lyophilized PRP; TGFβ 140373 pg/ml ± 91454 for fresh PRP and 111991 pg/ml ± 19827for lyophilized PRP; EGF 771,6 pg/ml ± 320,4 for fresh PRP and 739,1 pg/ml ± 324,1 for lyophilized PRP, figure 2. The results showed that fresh or lyophilized PRP were unable to show normal aggregation function, suggesting that these samples had been already activated by several conditions of preparation, such as the manual manipulation, temperature, pressure that the platelet is subjected inside the needle, among others. This result confirms that platelet activation with thrombin, or calcium chloride before PRP application is not crucial. Anyhow, the GF that is considered an important component for PRP regarding the therapeutic effect, were preserved. In addition, the lyophilized PRP appears as a possible replacement of fresh PRP, adding minor technical variability with a single process production, enabling a large-scale, with shelf life increased. Figure 1 Represents aggregation percentage generated after two antagonists addition. The results demonstrate that post-PRP has no platelet activation. Figure 2. Represents growth factor measurement in Fresh PRP and lyophilized PRP. The only difference between both preparations was PDGF (p=0, 0464). Figure 1. Represents aggregation percentage generated after two antagonists addition. The results demonstrate that post-PRP has no platelet activation. Figure 2. Represents growth factor measurement in Fresh PRP and lyophilized PRP. The only difference between both preparations was PDGF (p=0, 0464). Disclosures No relevant conflicts of interest to declare.

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