scholarly journals Platelet Rich Fibrin (PRF) and Its Related Products: Biomolecular Characterization of the Liquid Fibrinogen

2020 ◽  
Vol 9 (4) ◽  
pp. 1099
Author(s):  
Giorgio Serafini ◽  
Mariangela Lopreiato ◽  
Marco Lollobrigida ◽  
Luca Lamazza ◽  
Giulia Mazzucchi ◽  
...  
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Whole Blood ◽  
Transforming Growth Factor ◽  
Platelet Derived Growth Factor ◽  
Bone Morphogenetic Protein 2 ◽  
Time Points ◽  
Cell Counts ◽  
Tgf Β1

Liquid fibrinogen is an injectable platelet concentrate rich in platelets, leukocytes, and fibrinogen obtained by blood centrifugation. The aim of this study was to analyze the release of different growth factors in the liquid fibrinogen at different times and to assess possible correlations between growth factors and cell counts. The concentration of transforming growth factor beta 1 (TGF-β1), platelet-derived growth factor-AB (PDGF-AB), platelet-derived growth factor-BB (PDGF-BB), bone morphogenetic protein 2 (BMP-2), fibroblast growth factor 2 (FGF-2) and vascular endothelial growth factor (VEGF) released by liquid fibrinogen were examined with ELISA at three time points (T0, time of collection; T7, 7 days; T14, 14 days). The cellular content of the liquid fibrinogen and whole blood was also calculated for each volunteer. A mean accumulation of platelets of almost 1.5-fold in liquid fibrinogen compared to whole blood samples was found. An increase of TGF-β1, PDGF-AB, FGF-2, and VEGF levels was detected at T7. At T14, the level of TGF-β1 returned to T0 level; PDGF-AB amount remained high; the levels of FGF-2 and VEGF decreased with respect to T7, but remained higher than the T0 levels; PDGF-BB was high at all time points; BMP-2 level was low and remained constant at all time points. TGF-β1, PDGF-AB, and PDGF-BB showed a correlation with platelet amount, whereas BMP-2, FGF-2, and VEGF showed a mild correlation with platelet amount. Due to the high concentration of platelets, liquid fibrinogen does contain important growth factors for the regeneration of both soft and hard tissue. The centrifugation protocol tested in this study provides a valid solution to stimulate wound healing in oral and periodontal surgery.

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

2016 ◽  
Vol 45 (4) ◽  
pp. 954-960 ◽  
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.

Comparison of Release Profiles of Various Growth Factors from Biodegradable Carriers

1998 ◽  
Vol 530 ◽  
Author(s):  
Y. Tabata ◽  
M. Yamamoto ◽  
Y. Ikada
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β1 ◽  
The Body ◽  
Bone Morphogenetic Protein 2 ◽  
Biodegradable Hydrogel ◽  
Tgf Β1

AbstractA biodegradable hydrogel was prepared by glutaraldehyde crosslinking of acidic gelatin with an isoelectric point (IEP) of 5.0 as a carrier to release basic growth factors on the basis of polyion complexation. Basic fibroblast growth factor (bFGF), transforming growth factor β1 (TGF-β1), and bone morphogenetic protein-2 (BMP-2) were sorbed from their aqueous solution into the dried gelatin hydrogels to prepare respective growth factor-incorporating hydrogels. Under an in vitro non-degradation condition, approximately 20 % of incorporated bFGF and TGF-β1 was released from the hydrogels within initial 40 min, followed by no further release, whereas a large initial release of BMP-2 was observed. After subcutaneous implantation of the gelatin hydrogels incorporating 125I-labeled growth factor in the mouse back, the remaining radioactivity was measured to estimate the in vivo release profile of growth factors. Incorporation into gelatin hydrogels enabled bFGF and TGF-β1 to retain in the body for about 15 days and the retention period well correlated with that of the gelatin hydrogel. Taken together, it is likely that the growth factors ionically complexed with acidic gelatin were released in vivo as a result of hydrogel biodegradation. On the contrary, basic BMP-2 did not ionically interact with acidic gelatin, resulting in no sustained released by the present biodegradable carrier system.

scholarly journals Tethered TGF-β1 in a Hyaluronic Acid-Based Bioink for Bioprinting Cartilaginous Tissues

2022 ◽  
Vol 23 (2) ◽  
pp. 924
Author(s):  
Julia Hauptstein ◽  
Leonard Forster ◽  
Ali Nadernezhad ◽  
Jürgen Groll ◽  
Jörg Teßmar ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Growth Factors ◽  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Cartilage Regeneration ◽  
Cartilaginous Tissues ◽  
Dual Stage ◽  
Tgf Β1

In 3D bioprinting for cartilage regeneration, bioinks that support chondrogenic development are of key importance. Growth factors covalently bound in non-printable hydrogels have been shown to effectively promote chondrogenesis. However, studies that investigate the functionality of tethered growth factors within 3D printable bioinks are still lacking. Therefore, in this study, we established a dual-stage crosslinked hyaluronic acid-based bioink that enabled covalent tethering of transforming growth factor-beta 1 (TGF‑β1). Bone marrow-derived mesenchymal stromal cells (MSCs) were cultured over three weeks in vitro, and chondrogenic differentiation of MSCs within bioink constructs with tethered TGF‑β1 was markedly enhanced, as compared to constructs with non-covalently incorporated TGF‑β1. This was substantiated with regard to early TGF‑β1 signaling, chondrogenic gene expression, qualitative and quantitative ECM deposition and distribution, and resulting construct stiffness. Furthermore, it was successfully demonstrated, in a comparative analysis of cast and printed bioinks, that covalently tethered TGF‑β1 maintained its functionality after 3D printing. Taken together, the presented ink composition enabled the generation of high-quality cartilaginous tissues without the need for continuous exogenous growth factor supply and, thus, bears great potential for future investigation towards cartilage regeneration. Furthermore, growth factor tethering within bioinks, potentially leading to superior tissue development, may also be explored for other biofabrication applications.

scholarly journals Evaluation of Circulatory Concentration of Cytokines Platelet-Derived Growth Factor-BB, Transforming Growth Factor Beta-1, and IL-10 in Canine Generalized Demodicosis

2019 ◽  
Vol 15 (02) ◽  
pp. 60-62
Author(s):  
Mayank Parwari ◽  
GC Mandali ◽  
Jignasha M Parmar ◽  
RA Mathakiya
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Microscopic Examination ◽  
Transforming Growth Factor ◽  
Interleukin 10 ◽  
Clinical Findings ◽  
Platelet Derived Growth Factor ◽  
Healthy Dogs ◽  
Growth Factor Beta ◽  
Tgf Β1

The purpose of this study was to investigate the changes in circulating concentrations of PDGF-BB, TGF-β1 and interleukin-10 (IL-10) in dogs with generalized demodicosis. Twelve dogs with generalized demodicosis were diagnosed based on clinical findings and microscopic examination of the cutaneous scrapings. Six healthy dogs were included in the study as a control. Commercial specific ELISA assays were used to measure circulating concentrations of plasma PDGF-BB, and serum TGF-β1 and IL-10. Marked and significant increase in plasma PDGF-BB and serum TGF-β1 concentrations was evidenced in diseased dogs, while increase in serum IL-10 concentration was non-significant. The levels of all three markers dropped down to near normal/healthy status after the treatment of diseased dogs. These results indicate that the increased concentrations of circulating PDGF-BB, TGF-β1, and IL-10 play a pivotal role in the pathogenesis of the canine demodicosis.

Concentration of platelets and growth factors in canine autologous conditioned plasma

2011 ◽  
Vol 24 (02) ◽  
pp. 122-125 ◽  
Author(s):  
M. Stief ◽  
J. Gottschalk ◽  
J.-C. Ionita ◽  
A. Einspanier ◽  
G. Oechtering ◽  
...  
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Blood Cells ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β1 ◽  
Platelet Derived Growth Factor ◽  
Significant Difference ◽  
Transforming Growth Factor Β2 ◽  
Endothelial Growth Factor ◽  
Tgf Β1

Summary Objectives: To report the concentration of blood cells and selected growth factors in canine autologous conditioned plasma (ACP). Methods: The density of blood cells in whole blood (WB), ACP and standard plasma preparation (SP) of 10 healthy mature dogs was determined. In both ACP and SP, the concentration of insulin-like growth factor-1 (IGF-1), epidermal growth factor, vascular endothelial growth factor, platelet-derived growth factor-AA, platelet-derived growth factor-AB, platelet-derived growth factor-BB, transforming growth factor-β1 (TGF-β1), and transforming growth factor-β2 was measured using the ELISA technique. In another ten dogs, ACP was prepared using an ultra-soft spinning protocol, and again blood cell density was compared to that obtained in WB. Results: The density of platelets in ACP was significantly higher than that in SP (p = 0.0002), but there was not any significant difference between ACP and WB, nor between WB and ACP prepared using softer centrifugations. Interestingly, only for IGF-1, PDGFBB, and TGF-β1 could reliable measurements be obtained, showing a significant increase in PDGF-BB and TGF-β1 concentrations in ACP compared to SP (p = 0.001, p = 0.0028). Regarding IGF-1 content, there was not any significant difference between ACP and SP. Clinical significance: Canine ACP prepared according to the manufacturer’s recommendations, or by using a softer spin does not show the same specifications as human ACP, which shows a doubling in platelet count compared to WB. Even though canine ACP has a similar number of platelets per injected volume and consequently, probably the same amount of injected growth factors than WB, application of canine ACP would not be associated with the proinflammatory potential reported for WB, as it is almost free of erythrocytes and nucleated cells.

scholarly journals Growth factors and the mesangium.

1992 ◽  
Vol 2 (10) ◽  
pp. S185 ◽  
Author(s):  
H E Abboud
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Mesangial Cell ◽  
Mesangial Cells ◽  
Platelet Derived Growth Factor ◽  
Glomerular Injury ◽  
Effector Cells

Growth factors are prime candidates to mediate and modulate the functions of the mesangium. Mesangial cells are effector cells producing a number of growth factors that act in an autocrine manner to regulate their own function. Mesangial cells are also targets for growth factors released from neighboring glomerular cells or infiltrating cells and platelets. Growth factors may promote hypertrophy, proliferation, matrix metabolism, and immune-inflammatory and vasoactive properties of mesangial cells. These peptides represent important mediators of mesangial cell responses to injury. Platelet-derived growth factor mediates predominantly cell proliferation, whereas transforming growth factor beta mediates mesangial cell matrix expansion. Mesangial cells may also modulate some of the hemodynamic effects of growth factors, such as the increased renal vascular resistance in response to platelet-derived growth factor and epidermal growth factor or the increased RBF and GFR in response to insulin-like growth factor-1. Changes in the expression of growth factors of their receptors during the course of glomerular injury point to a potential role in mediating some of the pathologic changes in vivo. Several agents appear to antagonize the mitogenic and perhaps other effects of growth factors in mesangial cells. Such agents include adenylate cyclase as well as guanylate cyclase agonists. Recent studies also suggest that some traditional vasoactive agents may activate metabolic processes in mesangial cells similar to peptide growth factors. Collectively, these studies point to the interaction of both hemodynamic and metabolic factors in the response and contribution of glomerular and specifically mesangial cells to injury.

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

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.

scholarly journals Utilizing the ABC Transporter for Growth Factor Production by fleQ Deletion Mutant of Pseudomonas fluorescens

Biomedicines ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 679
Author(s):  
Benedict-Uy Fabia ◽  
Joshua Bingwa ◽  
Jiyeon Park ◽  
Nguyen-Mihn Hieu ◽  
Jung-Hoon Ahn
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Pseudomonas Fluorescens ◽  
Abc Transporter ◽  
Transforming Growth Factor Beta ◽  
Deletion Mutant ◽  
Transforming Growth Factor ◽  
Gene Knockout ◽  
Type I ◽  
Double Deletion

Pseudomonas fluorescens, a gram-negative bacterium, has been proven to be a capable protein manufacturing factory (PMF). Utilizing its ATP-binding cassette (ABC) transporter, a type I secretion system, P. fluorescens has successfully produced recombinant proteins. However, besides the target proteins, P. fluorescens also secretes unnecessary background proteins that complicate protein purification and other downstream processes. One of the background proteins produced in large amounts is FliC, a flagellin protein. In this study, the master regulator of flagella gene expression, fleQ, was deleted from P. fluorescens Δtp, a lipase and protease double-deletion mutant, via targeted gene knockout. FleQ directs flagella synthesis, so the new strain, P. fluorescens ΔfleQ, does not produce flagella-related proteins. This not only simplifies purification but also makes P. fluorescens ΔfleQ an eco-friendly expression host because it will not survive outside a controlled environment. Six recombinant growth factors, namely, insulin-like growth factors I and II, beta-nerve growth factor, fibroblast growth factor 1, transforming growth factor beta, and tumor necrosis factor beta, prepared using our supercharging method, were successfully secreted by P. fluorescens ΔfleQ. Our findings demonstrate the potential of P. fluorescens ΔfleQ, combined with our supercharging process, as a PMF.

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