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

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-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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Transforming growth factor-β1 modulates the expression of vascular endothelial growth factor by osteoblasts

AJP Cell Physiology ◽

10.1152/ajpcell.1999.277.4.c628 ◽

1999 ◽

Vol 277 (4) ◽

pp. C628-C637 ◽

Cited By ~ 130

Author(s):

Pierre B. Saadeh ◽

Babak J. Mehrara ◽

Douglas S. Steinbrech ◽

Matthew E. Dudziak ◽

Joshua A. Greenwald ◽

...

Keyword(s):

Growth Factor ◽

Transforming Growth Factor ◽

Transforming Growth Factor Β1 ◽

Vascular Endothelial ◽

Vegf Expression ◽

Vegf Mrna ◽

Endothelial Growth Factor ◽

Dose Dependent ◽

Tgf Β1 ◽

Vegf Protein

Angiogenesis is essential to both normal and pathological bone physiology. Vascular endothelial growth factor (VEGF) has been implicated in angiogenesis, whereas transforming growth factor-β1 (TGF-β1) modulates bone differentiation, matrix formation, and cytokine expression. The purpose of this study was to investigate the relationship between TGF-β1 and VEGF expression in osteoblasts and osteoblast-like cells. Northern blot analysis revealed an early peak of VEGF mRNA (6-fold at 3 h) in fetal rat calvarial cells and MC3T3-E1 osteoblast-like cells after stimulation with TGF-β1 (2.5 ng/ml). The stability of VEGF mRNA in MC3T3-E1 cells was not increased after TGF-β1 treatment. Actinomycin D inhibited the TGF-β1-induced peak in VEGF mRNA, whereas cycloheximide did not. Blockade of TGF-β1 signal transduction via a dominant-negative receptor II adenovirus significantly decreased TGF-β1 induction of VEGF mRNA. Additionally, TGF-β1 induced a dose-dependent increase in VEGF protein expression by MC3T3-E1 cells ( P < 0.01). Dexamethasone similarly inhibited VEGF protein expression. Both TGF-β1 mRNA and VEGF mRNA were concurrently present in rat membranous bone, and both followed similar patterns of expression during rat mandibular fracture healing (mRNA and protein). In summary, TGF-β1-induced VEGF expression by osteoblasts and osteoblast-like cells is a dose-dependent event that may be intimately related to bone development and fracture healing.

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Medroxyprogesterone reverses tolerable dose metformin-induced inhibition of invasion via matrix metallopeptidase-9 and transforming growth factor-β1 in KLE endometrial cancer cells

10.1101/596056 ◽

2019 ◽

Author(s):

Dong Hoon Suh ◽

Sunray Lee ◽

Hyun-Sook Park ◽

Noh Hyun Park

Keyword(s):

Endometrial Cancer ◽

Growth Factor ◽

Cell Viability ◽

Cancer Cells ◽

Cell Invasion ◽

Transforming Growth Factor ◽

Dependent Manner ◽

Matrix Metallopeptidase ◽

Dose Dependent ◽

Tgf Β1

AbstractThis study was performed to evaluate the anticancer effects of tolerable doses of metformin with or without medroxyprogesterone (MPA) in endometrial cancer cells. Cell viability, cell invasion, and levels of matrix metallopeptidase (MMP) and transforming growth factor (TGF)-β1 were analyzed using three human endometrial adenocarcinoma cell lines (Ishikawa, KLE, and USPC) after treatment with different dose combinations of MPA (0, 10 μM) and metformin (0, 100, 1000 μM). Combining metformin (0, 100, 1000 μM) and 10 μM MPA induced significantly decreased cell viability in a time- and dose-dependent manner in Ishikawa cells, but not in KLE and USPC cells. There was no dose- or time-dependent cell growth inhibition, or positive western blot results for the expression of progesterone receptors and phospho-AMPKa, following treatment with any combination of metformin (0, 100, 1000 μM) and 10 μM MPA in KLE and USPC cells. In KLE cells, metformin treatment alone significantly inhibited cell invasion in a dose-dependent manner (1.31±0.05, 0.94±0.04, 0.83±0.05 at 0, 100 μM, 1000 μM, respectively; p<0.0005). The inhibitory effect of metformin was reversed to create a stimulating effect when metformin was combined with 10 μM MPA (1.10±0.05, 1.42±0.18, 1.41±0.26 at 0, 100, 1000 μM, respectively; p<0.005). MMP-9 and TGF-β1 showed similar trends in terms of cell invasion in KLE cells. In conclusion, the anti-invasive effect of metformin in KLE cells was completely reversed to the state of no treatment by the addition of MPA; this might be mediated through MMP-9 and TGF-β1. Our study suggests the possibility of these combinations doing harm, rather than good, under some conditions.

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Stem cells for cardiac regeneration and possible roles of the transforming growth factor-β superfamily

BioMolecular Concepts ◽

10.1515/bmc.2011.049 ◽

2012 ◽

Vol 3 (1) ◽

pp. 99-106 ◽

Cited By ~ 6

Author(s):

Nanako Kawaguchi

Keyword(s):

Stem Cells ◽

Growth Factor ◽

Cell Biology ◽

Transforming Growth Factor ◽

Methyl Cellulose ◽

Transforming Growth Factor Β ◽

Transforming Growth Factor Β1 ◽

Cardiac Stem Cells ◽

Dependent Manner ◽

Tgf Β1

AbstractHeart failure is a leading cause of death worldwide. Studies of stem cell biology are essential for developing efficient treatments. Recently, we established and characterized c-kit-positive cardiac stem cells from the adult rat heart. Using a MethoCult culture system with a methyl-cellulose-based medium, stem-like left-atrium-derived pluripotent cells could be regulated to differentiate into skeletal/cardiac myocytes or adipocytes with almost 100% purity. Microarray and pathway analyses of these cells showed that transforming growth factor-β1 (TGF-β1) and noggin were significantly involved in the differentiation switch. Furthermore, TGF-β1 may act as a regulator for this switch because it simultaneously inhibits adipogenesis and activates myogenesis in a dose-dependent manner. However, the effect of TGF-β varies with developmental stage, dosage, and timing of treatment. In the present review, the findings of recent studies, in particular the use of c-kit-positive cardiac stem cells, are discussed. The effects of the TGF-β superfamily on differentiation, especially on adipogenesis and/or myogenesis, have important implications for future regenerative medicine.

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Transforming growth factor-β1 downregulation of Smad1 gene expression in rat hepatic stellate cells

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00436.2002 ◽

2003 ◽

Vol 285 (3) ◽

pp. G539-G546 ◽

Cited By ~ 42

Author(s):

Hong Shen ◽

Guojiang Huang ◽

Mohammed Hadi ◽

Patrick Choy ◽

Manna Zhang ◽

...

Keyword(s):

Gene Expression ◽

Growth Factor ◽

Hepatic Stellate Cells ◽

Intracellular Signaling ◽

Transforming Growth Factor ◽

Stellate Cells ◽

Transforming Growth Factor Β ◽

Transforming Growth Factor Β1 ◽

Dependent Manner ◽

Tgf Β1

Smads are intracellular signaling molecules of the transforming growth factor-β (TGF-β) superfamily that play an important role in the activation of hepatic stellate cells (HSCs) and hepatic fibrosis. Excepting the regulation of Smad7, receptor-regulated Smad gene expression is still unclear. We employed rat HSCs to investigate the expression and regulation of the Smad1 gene, which is a bone morphogenetic protein (BMP) receptor-regulated Smad. We found that the expression and phosphorylation of Smad1 are increased during the activation of HSCs. Moreover, TGF-β significantly inhibits Smad1 gene expression in HSCs in a time- and dose-dependent manner. Furthermore, although both TGF-β1 and BMP2 stimulate the activation of HSCs, they have different effects on HSC proliferation. In conclusion, Smad1 expression and phosphorylation are increased during the activation of HSCs and TGF-β1 significantly inhibits the expression of the Smad1 gene.

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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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Transforming Growth Factor- β1 Signaling in Vascular Endothelial Cells Inhibits Hematopoietic Stem Cell Regeneration

Blood ◽

10.1182/blood.v128.22.1493.1493 ◽

2016 ◽

Vol 128 (22) ◽

pp. 1493-1493

Author(s):

Derek Zachman ◽

Devorah C Goldman ◽

Chandan Guha ◽

Beth Wilmot ◽

William H. Fleming

Keyword(s):

Endothelial Cells ◽

Growth Factor ◽

Transforming Growth Factor ◽

Conflicts Of Interest ◽

Umbilical Vein ◽

Transforming Growth Factor Β1 ◽

Dependent Manner ◽

Hematopoietic Stem ◽

Tgf Β1

Abstract Endothelial cells (EC) are known to be essential for hematopoietic regeneration; however, little is known about the pathways that regulate this activity. By modeling endothelial-dependent HSC interactions in vitro, we found that human umbilical vein endothelial cells (HUVEC) had a markedly reduced capacity to regenerate functional CD150+LSK cells (HSC) compared to other sources of arterial and venous EC. Transcriptional profiling revealed the overexpression of transforming growth factor- β1 (TGF-β1) in HUVEC and indicated that TGF-β1 driven transcriptional programs are highly active in these cells, a finding consistent with autocrine TGF-β1 signaling. Functional studies demonstrated that HSC regeneration by EC was potently inhibited by TGF-β1 and augmented by the ALK5 inhibitor SB431542, in a dose-dependent manner. Importantly, exposure of EC alone to TGF- β1 was sufficient to attenuate subsequent HSC self-renewal. Transcriptome analysis also identified hepatocyte growth factor (HGF) as a candidate EC-derived factor with the potential to enhance hematopoietic regeneration. HGF treatment of HUVEC activated endothelial Akt signaling and led to a >10-fold increase in HSC regeneration that could be blocked by the c-Met inhibitor PF04217903. HGF treatment also dramatically increased long-term multi-lineage hematopoiesis from HUVEC regenerated HSC. Our findings reveal a novel suppressive role for TGF-β1 in the vascular niche and demonstrate that EC-derived growth factors such as HGF have the potential to attenuate this suppression and significantly enhance HSC regeneration. Disclosures No relevant conflicts of interest to declare.

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Medroxyprogesterone Reverses Tolerable Dose Metformin-Induced Inhibition of Invasion via Matrix Metallopeptidase-9 and Transforming Growth Factor-β1 in KLE Endometrial Cancer Cells

Journal of Clinical Medicine ◽

10.3390/jcm9113585 ◽

2020 ◽

Vol 9 (11) ◽

pp. 3585

Author(s):

Dong Hoon Suh ◽

Sunray Lee ◽

Hyun-Sook Park ◽

Noh Hyun Park

Keyword(s):

Endometrial Cancer ◽

Growth Factor ◽

Cancer Cells ◽

Cell Invasion ◽

Transforming Growth Factor ◽

Dependent Manner ◽

Anticancer Effects ◽

Matrix Metallopeptidase ◽

Dose Dependent ◽

Tgf Β1

This study was performed to evaluate the anticancer effects of tolerable doses of metformin with or without medroxyprogesterone (MPA) in endometrial cancer cells. Cell viability, cell invasion, and levels of matrix metallopeptidase (MMP) and transforming growth factor (TGF)-β1 were analyzed using three human endometrial adenocarcinoma cell lines (Ishikawa, KLE, and uterine serous papillary cancer (USPC)) after treatment with different dose combinations of MPA and metformin. Combining metformin (0, 100, 1000 µM) and 10 µM MPA induced significantly decreased cell viability in a time- and dose-dependent manner in Ishikawa cells, but not in KLE and USPC cells. In KLE cells, metformin treatment alone significantly inhibited cell invasion in a dose-dependent manner. The inhibitory effect of metformin was reversed when 10 µM MPA was combined, which was significantly inhibited again after treatment of MMP-2/9 inhibitor and/or TGF-β inhibitor. Changes of MMP-9 and TGF-β1 according to combinations of MPA and metformin were similar to those of invasion in KLE cells. In conclusion, the anticancer effects of tolerable doses of metformin varied according to cell type and combinations with MPA. Anti-invasive effect of metformin in KLE cells was completely reversed by the addition of MPA; this might be associated with MMP-9 and TGF-β1.

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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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