Freeze-Dried Human Platelet-Rich Plasma Retains Activation and Growth Factor Expression after an Eight-Week Preservation Period

<sec><title>Study Design</title>Controlled laboratory study.</sec><sec><title>Purpose</title>This study aimed to evaluate the efficacy of platelet-rich plasma (PRP) stored at room temperature (RT), frozen, or after freeze-drying.</sec><sec><title>Overview of Literature</title>PRP enriches tissue repair and regeneration, and is a novel treatment option for musculoskeletal pathologies. However, whether biological activity is preserved during PRP storage remains uncertain.</sec><sec><title>Methods</title>PRP was prepared from blood of 12 healthy human volunteers (200 mL/person) and stored using three methods: PRP was stored at RT with shaking, PRP was frozen and stored at −80℃, or PRP was freeze-dried and stored at RT. Platelet counts and growth factor content were examined immediately after preparation, as well as 2, 4, and 8 weeks after storage. Platelet activation rate was quantified by flow cytometry.</sec><sec><title>Results</title>Platelet counts were impossible to determine in many RT samples after 2 weeks, but they remained at constant levels in frozen and freeze-dried samples, even after 8 weeks of storage. Flow cytometry showed approximately 80% activation of the platelets regardless of storage conditions. Almost no growth factors were detected in the RT samples after 8 weeks, while low but significant expression was detected in the frozen and freeze-dried PRP. Over time, the mean relative concentrations of various growth factors decreased significantly or disappeared in the RT group. In the frozen group, levels were maintained for 4 weeks, but decreased significantly by 8 weeks (<italic>p</italic> <0.05). The freeze-dried group maintained baseline levels of growth factors for the entire 8-week duration.</sec><sec><title>Conclusions</title>Freeze-drying enables PRP storage while maintaining bioactivity and efficacy for extended periods.</sec>

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Influence of storage conditions on the release of growth factors in platelet-rich blood derivatives

Current Directions in Biomedical Engineering ◽

10.1515/cdbme-2016-0069 ◽

2016 ◽

Vol 2 (1) ◽

pp. 311-314 ◽

Cited By ~ 1

Author(s):

Katharina Düregger ◽

Anqi Peng ◽

Markus Eblenkamp

Keyword(s):

Growth Factors ◽

Growth Factor ◽

Platelet Rich Plasma ◽

Storage Conditions ◽

Platelet Derived Growth Factor ◽

Prolonged Storage ◽

Wound Treatment ◽

Time Dependency ◽

Processing And Storage ◽

And Storage

AbstractThrombocytes can be concentrated in blood derivatives and used as autologous transplants e.g. for wound treatment due to the release of growth factors such as platelet derived growth factor (PDGF). Conditions for processing and storage of these platelet-rich blood derivatives influence the release of PDGF from the platelet-bound α-granules into the plasma. In this study Platelet rich plasma (PRP) and Platelet concentrate (PC) were produced with a fully automated centrifugation system. Storage of PRP and PC for 1 h up to 4 months at temperatures between −20°C and +37°C was applied with the aim of evaluating the influence on the amount of released PDGF. Storage at −20°C resulted in the highest release of PDGF in PRP and a time dependency was determined: prolonged storage up to 1 month in PRP and 10 days in PC increased the release of PDGF. Regardless of the storage conditions, the release of PDGF per platelet was higher in PC than in PRP.

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A boomy century of periodontal regeneration with biological mediators

IP International Journal of Maxillofacial Imaging ◽

10.18231/j.ijmi.2021.021 ◽

2021 ◽

Vol 7 (3) ◽

pp. 103-117

Author(s):

Deepak Grover ◽

Navneet Kaur ◽

Gurpreet Kaur

Keyword(s):

Growth Factors ◽

Growth Factor ◽

Bone Regeneration ◽

Transforming Growth Factor ◽

Platelet Rich Plasma ◽

Autologous Blood ◽

Periodontal Regeneration ◽

Transforming Growth Factor Β ◽

Tissue Repair And Regeneration ◽

Plasma Fraction

With the advanced and improved knowledge of bone regeneration on the molecular level, some of key molecules that alter the complicated physiological method were identified, and are already in scientific use or beneath research to enhance bone restore. Of those molecules, BMPs were the maximum considerably studied, as they are robust osteoinductive elements. They result in the mitogenesis of mesenchymal stem cells (MSCs) and different osteoprogenitors, and their differentiation in the direction of osteoblasts. Other growth factors except BMPs which have been implicated through out the bone regeneration, with one-of-a-kind features with respect of cell proliferation, chemotaxis and angiogenesis, are also being investigated or are presently getting used to reinforce bone restore, which include platelet-derived growth factor, transforming growth factor- β, insulin-like growth factor-1, vascular endothelial growth factor and fibroblast growth factor, amongst others. One present day technique to enhance bone regeneration and soft-tissue recovery with the aid of using nearby application of growth factors is the use of platelet-rich plasma, an extent of the plasma fraction of autologous blood with platelet concentrations above baseline, that is wealthy in most of the aforementioned molecules. This overview focuses and target on the biological mediators that regulates key cellular events which have a capacity to induce the method of tissue repair and regeneration.

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Effect of freeze-drying process of collagen-activated platelet-rich plasma on transforming growth factor-β1 level

Majalah Kedokteran Gigi Indonesia ◽

10.22146/majkedgiind.40197 ◽

2020 ◽

Vol 5 (2) ◽

pp. 82

Author(s):

Kwartarini Murdiastuti ◽

Fitri Yuniawati ◽

Dahlia Herawati ◽

Nunuk Purwanti ◽

Dyah Ayu Mira Oktarina

Keyword(s):

Growth Factor ◽

Freeze Drying ◽

Transforming Growth Factor ◽

Platelet Rich Plasma ◽

Transforming Growth Factor Β1 ◽

Drying Process ◽

Freeze Dried ◽

Proliferation And Differentiation ◽

High Level ◽

Tgf Β1

Periodontal tissue damage requires regenerative material to repair the damage. Platelet-rich plasma (PRP) is known as a regenerative material from blood which contains high level of growth factor that plays a role in wound healing and tissue remodeling. However PRP has a weakness, i.e. it is too watery so it is easily dissolved in the oral cavity, and should be used immediately after preparation. Therefore PRP storage method is needed to increase the benefits of PRP. The addition of collagen to PRP serves as a scaffold as well as an activator that stimulates the release of growth factors. One method of storing PRP is by freeze-drying process. The purpose of this study was to analyze the effect of freeze-drying process of collagen-activated PRP (PRP+C) on transforming growth factor-β1 (TGF-β1) levels. Transforming growth factor-β1 is a cytokine content in PRP, that plays a role in bone remodeling and is an important stimulator for osteoblast formation, causing chemotaxis, osteoblast proliferation and differentiation. In this study, PRP was produced from peripheral blood probandus. Platelet-rich plasma was then activated with collagen (PRP+C), and divided into two groups: freeze-dried PRP collagen (FD PRP+C); and non freeze-dried PRP+collagen (PRP+C). Transforming growth factor-β1 levels were measured using the ELISA method, followed by independent t-test. The TGF-B1 level of FD PRP+C group was significantly higher than PRP+C group (p<0.05). From this study it can be concluded that freeze-dried collagen-activated PRP has an effect to increase TGF-β1 level.

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PRP application in dermatology: review of current approaches

Medical alphabet ◽

10.33667/2078-5631-2021-9-55-58 ◽

2021 ◽

pp. 55-58

Author(s):

V. V. Bondarenko

Keyword(s):

Growth Factors ◽

Blood Plasma ◽

Tissue Repair ◽

Skin Diseases ◽

Platelet Rich Plasma ◽

Autologous Blood ◽

Biologically Active ◽

Tissue Repair And Regeneration ◽

Positive Results ◽

Active Substances

In recent years, the possibilities of a dermatologist in the treatment of patients with chronic dermatoses have significantly expanded due to the emergence of new highly effective methods, such as autologous blood plasma enriched with platelet-rich-growth factors (platelet-rich-plasma, PRP). The positive results of using this technology due to its constituent growth factors, cytokines and other biologically active substances, which have a pronounced normalizing effect on the processes of tissue repair and regeneration, expands the possibilities of its use in such skin diseases as erosive and ulcerative lichen planus, scleroatrophic lichen, acne and post-acne. The aim of this review was to analyze the literature on the mechanisms of PRP action in patients with dermatological diseases.

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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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Concentrations of Blood Components in Commercial Platelet-Rich Plasma Separation Systems: A Review of the Literature

The American Journal of Sports Medicine ◽

10.1177/0363546517746112 ◽

2018 ◽

Vol 47 (2) ◽

pp. 479-487 ◽

Cited By ~ 44

Author(s):

Bart W. Oudelaar ◽

Joost C. Peerbooms ◽

Rianne Huis in ‘t Veld ◽

Anne J.H. Vochteloo

Keyword(s):

Growth Factors ◽

Growth Factor ◽

Transforming Growth Factor ◽

Platelet Rich Plasma ◽

Therapeutic Option ◽

Transforming Growth Factor Β1 ◽

Future Research ◽

Cochrane Central Register ◽

Blood Components ◽

Separation Systems

Background: Platelet-rich plasma (PRP) has proven to be a very safe therapeutic option in the treatment of tendon, muscle, bone, and cartilage injuries. Currently, several commercial separation systems are available for the preparation of PRP. The concentrations of blood components in PRP among these separation systems vary substantially. Purpose: To systematically review and evaluate the differences between the concentrations of blood components in PRP produced by various PRP separation systems. Study Design: Systematic review. Methods: MEDLINE/PubMed, the Cochrane Central Register of Controlled Trials (CENTRAL), and EMBASE were searched for studies that compared the concentrations of blood components and growth factors in PRP between various separation systems and studies that reported on the concentrations of blood components and growth factors of single separation systems. The primary outcomes were platelet count, leukocyte count, and concentration of growth factors (eg, platelet-derived growth factor–AB [PDGF-AB], transforming growth factor–β1 [TGF-β1], and vascular endothelial growth factor [VEGF]). Furthermore, the preparation protocols and prices of the systems were compared. Results: There were 1079 studies found, of which 19 studies were selected for inclusion in this review. The concentrations of platelets and leukocytes in PRP differed largely between, and to a lesser extent within, the studied PRP separation systems. Additionally, large differences both between and within the studied PRP separation systems were found for all the growth factors. Furthermore, preparation protocols and prices varied widely between systems. Conclusion: There is a large heterogeneity between PRP separation systems regarding concentrations of platelets, leukocytes, and growth factors in PRP. The choice for the most appropriate type of PRP should be based on the specific clinical field of application. As the ideal concentrations of blood components and growth factors for the specific fields of application are yet to be determined for most of the fields, future research should focus on which type of PRP is most suitable for the specific field.

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Effect of Storage Conditions and Activation on Growth Factor Concentration in Platelet‐Rich Plasma

Journal of Orthopaedic Research® ◽

10.1002/jor.24520 ◽

2019 ◽

Vol 38 (4) ◽

pp. 777-784 ◽

Cited By ~ 1

Author(s):

Joong Il Kim ◽

Hyun Cheol Bae ◽

Hee Jung Park ◽

Myung Chul Lee ◽

Hyuk Soo Han

Keyword(s):

Growth Factor ◽

Platelet Rich Plasma ◽

Storage Conditions ◽

Factor Concentration

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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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Basic Studies on the Clinical Applications of Platelet-Rich Plasma

Cell Transplantation ◽

10.3727/000000003108747073 ◽

2003 ◽

Vol 12 (5) ◽

pp. 509-518 ◽

Cited By ~ 51

Author(s):

Masaki Yazawa ◽

Hisao Ogata ◽

Tatsuo Nakajima ◽

Taisuke Mori ◽

Naohide Watanabe ◽

...

Keyword(s):

Growth Factors ◽

Growth Factor ◽

Fibrin Glue ◽

Transforming Growth Factor ◽

Platelet Rich Plasma ◽

Transforming Growth Factor Β ◽

Clinical Applications ◽

Platelet Concentrate ◽

Platelet Concentrates ◽

Basic Studies

Platelets, which contain many growth factors such as platelet-derived growth factor (PDGF) and transforming growth factor-β (TGF-β), are being used in clinical applications as platelet-rich plasma (PRP). Only a few studies, however, have been conducted on the growth factors present in PRP and on the clinical applications using the drug delivery system (DDS). For the purpose of clinical application, we first modified the PRP preparation method and assessed the amounts of growth factors contained in the human platelet concentrates. Furthermore, we assessed fibrin glue as a DDS of platelet concentrates. Platelet precipitations were made by twice centrifuging human whole blood. The precipitated platelet was resuspended to yield the platelet concentrates. The growth factor concentrations were measured. Fibrin glue sheets containing this platelet concentrate were implanted in rabbit pinna and samples were obtained for immunostaining (anti-PDGF antibody) to assess the use of PRP over time using the fibrin glue as the DDS. The mean concentration of growth factors present in the platelet concentrates was three times or greater than that of conventional PRP. Furthermore, the results indicated that when the platelet concentrate was used with fibrin glue as a carrier, the contents were released over a period of about 1 week. This raises the possibility that this system may be useful in clinical applications.

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