Growth Factor Measurement and Histological Analysis in Platelet Rich Fibrin: A Pilot Study

Nishimoto, Soh; Fujita, Kazutoshi; Sotsuka, Yohei; Kinoshita, Masato; Fujiwara, Toshihiro; Kawai, Kenichiro; Kakibuchi, Masao

doi:10.1007/s12663-015-0768-3

Cited by 36 publications

(29 citation statements)

References 20 publications

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“…21 It is worth noting that cell entrapment is a central attribute of leukocyte-rich fibrin membranes. [23][24][25][26] One study 22 was able to identify a relationship between the concentration of growth factors and the region of the membranes, where the highest levels were evidenced in the buffy coat area, similar to the most cellularized fragment of the present work (Figs. 22 The presence of apparently viable leukocytes, as well as a fibrin network which seems stable for weeks, could contribute to producing a reservoir for controlled release of growth factors and cytokines.…”

Section: Discussionsupporting

confidence: 65%

“…Such stability is also a desirable feature for scaffolds for tissue engineering, 20 supporting cell migration and proliferation for tissue remodeling purposes, as biodegradable barriers for guided tissue regeneration procedures. 12,23,24 However, there is controversy regarding the impact of the use of different centrifugation systems in the release of bioactive molecules by membranes produced with otherwise very similar protocols. The present fluorescence microscopy assay was able to detect a high density of nucleated cells within the structure of the fibrin membrane, quite typical of PRF membranes, with an uneven distribution along its extension, and a higher cell density at the top portion.…”

Section: Discussionmentioning

confidence: 99%

“…This method allows analyzing multiple analytes in a single sample, increasing the reliability of the results concerning comparisons between The role of growth factors released by fibrin membranes on tissue regeneration is well known, either through the promotion of angiogenesis, migration, and proliferation of mesenchymal cells and fibroblasts and osteoblasts. A comparative study of different platelet derivatives 23 showed that the release of CCL-5 is much higher in PRP than in PRF in the first 24 h after its production. 7,19,23 However, methodological differences such as the volume of medium used for elution, detection technique (ELISA, flow luminometry) as well as the parameters for normalization of results (e.g., membrane mass, initial volume of blood samples) prevent an adequate the comparison of the absolute concentrations identified from different reports.…”

Section: Discussionmentioning

confidence: 99%

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Thein vitrorelease of cytokines and growth factors from fibrin membranes produced through horizontal centrifugation

Lourenço

Mourão

Leite

et al. 2018

J Biomedical Materials Res

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Platelet-rich fibrin membranes are biomaterials widely used for therapeutic purposes, and canonically produced through the processing of peripheral blood with fixed-angle rotor centrifuges. In this work, we evaluate the in vitro stability and release of cytokines and growth factors when these biomaterials are produced with a horizontal swing-out clinical centrifuge. Membranes produced from the blood of 14 donors were morphologically evaluated by scanning electron microscopy and fluorescence microscopy, and their stability was assessed by photographic recording after incubation in culture medium for up to 28 days. The release of 27 cytokines and growth factors was monitored for three weeks through a multiparametric immunoassay. The fibrin membranes presented complex three-dimensional structure with a high density of nucleated cells. A large release of growth factors [platelet derived growth factor, fibroblastic growth factor (bFGF), and vascular endothelial growth factor] was detected in the first 24 h, followed by time-dependent decay, maintaining significant concentrations after three weeks. Both anti-inflammatory and pro-inflammatory cytokines presented different release peaks, maintaining high rates of elution for up to 21 days. Chemokines of relevance in tissue repair [RANTES, granulocyte colony-stimulating factor (G-CSF)] were also produced in large quantities throughout the experimental period. The present results demonstrate that blood-derived fibrin membranes with high structural stability and cell content can be generated by horizontal centrifugation, being able of a prolonged production/release of growth factors and pro- and anti-inflammatory cytokines. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1373-1380, 2018.

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Section: Discussionsupporting

confidence: 65%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Thein vitrorelease of cytokines and growth factors from fibrin membranes produced through horizontal centrifugation

Lourenço

Mourão

Leite

et al. 2018

J Biomedical Materials Res

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“…This fact may have influenced in the results, since according to Eren, Gürkan, Atmaca, Dönmez, Atilla 30 centrifuge for 12 minutes has higher VEGF levels than only 10 minutes. The PRF was removed from the middle layer of the centrifuged sample, between the red part (below) and the plasma (above), which is in agreement with the findings of Nishimoto et al 31 about higher concentrations of mononuclear cells, platelets and growth factors in the yellow-red interface area. However, the isolated effect of PRF was not observed in the functional or morphological evaluation.…”

supporting

confidence: 86%

Effects of the platelet-rich fibrin associated with physical exercise in a model of median nerve compression.

Bertolini

Kakihata

Peretti

et al. 2018

Motriz: rev. educ. fis.

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-Aims: Platelet-Rich Fibrin (PRF) is a new and promising technique for tissue repair, however, there is still a gap about its action on peripheral nerve injury, as well as its association with physical exercise. Thus, the aim of this study was to evaluate the effects of the PRF associated with physical exercise in a model of median nerve compression. Methods: 42 rats, divided into: Control, non-injured limb, and other six groups with nerve lesion (Control lesion; Treated with PRF; Treadmill walking; Free swimming; PRF and treadmill; PRF and swimming). The lesion model was performed with median nerve compression. To obtain the PRF, 1.5 mL of blood was centrifuged and the fibrin clot positioned directly over the compression region. The exercise protocols were performed during 2 weeks. The evaluations performed were grip strength tests and histologic analysis. Results: both, grip strength and histomorphometric evaluations (fiber numbers and axon density) did not present significant differences between the treated and lesion groups, however, in the morphological evaluation it was possible to distinguish characteristics of the repair process for the treated groups. Conclusion: a slight qualitative improvement was observed for the treated groups, especially when it was associated PRF with free swimming.

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“…Although mean flinching in G1 being lower than those for other groups, statistical analysis has shown no differences among groups, but only among evaluations. Our study had several limitations, such as the use of intermediate PRF fraction, which may not have the highest amount of available growth factors 20 ; centrifugation time might have been short to be able to potentiate growth factors release, as shown by a recent study which has pointed to advantages of centrifuging for 12 minutes as compared to 10 minutes 21 , in addition to the evaluation of just two signs of the inflammation/repair process. It has also to be stressed that PRF associated or not to other modalities is still a new technique with in vitro 22 and in vivo [15][16][17] studies showing contradictory results, thus requiring further studies with regard to its effects.…”

Section: Discussionmentioning

confidence: 99%

Nociceptive evaluation of the association between physical exercises and platelet-rich fibrin in Wistar rats submitted to median nerve compression

et al. 2016

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BACKGROUND AND OBJECTIVES: Platelet-rich fibrin is a new and promising technique to accelerate repair, with possible analgesic effects; however, there is still a gap with regard to peripheral nerve injury and the association with physical exercises. So, this study aimed at evaluating the effects of platelet-rich fibrin associated to physical exercises on nociception and edema in experimental median nerve compression model. METHODS: Thirty-six rats, all submitted to median nerve compression, were divided in six groups: G1: without additional manipulation; G2: compression and treated with platelet-rich fibrin; G3: compression and treated with free swimming; G4: compression and walking on a treadmill; G5: free swimming + platelet-rich fibrin; G6: walking on a treadmill + platelet-rich fibrin. Injury was induced by tying the median nerve with chrome plated catgut 4.0. Platelet-rich fibrin was obtained by centrifuging 1.5 mL of blood and positioning the fibrin clot directly on the compression region. Exercises were carried out during two weeks, between the 3 rd and 14 th postoperative days. Nociception and edema were evaluated, respectively, by flinch threshold and plethysmometer, in moments before injury and in the 3 rd , 7 th and 15 th postoperative days. RESULTS: There have been no differences among groups, only among evaluations, showing increased nociception and edema, which has lasted or improved, respectively, over time. CONCLUSION: Platelet-rich fibrin alone or associated to physical exercises has not changed nociception and edema.

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Growth Factor Measurement and Histological Analysis in Platelet Rich Fibrin: A Pilot Study

Abstract: The growth factors were surely concentrated in PRF. This result can support basis of good clinical outcomes. For effective application of PRF, the knowledge that growth factors and cells are not equally distributed in PRF should be utilized.

Cited by 36 publications

References 20 publications

Thein vitrorelease of cytokines and growth factors from fibrin membranes produced through horizontal centrifugation

Thein vitrorelease of cytokines and growth factors from fibrin membranes produced through horizontal centrifugation

Effects of the platelet-rich fibrin associated with physical exercise in a model of median nerve compression.

Nociceptive evaluation of the association between physical exercises and platelet-rich fibrin in Wistar rats submitted to median nerve compression

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