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      In vitro immunological and biological evaluations of the angiogenic potential of platelet-rich fibrin preparations: a standardized comparison with PRP preparations

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          Abstract

          Background

          Platelet-rich fibrin (PRF), a platelet-rich plasma (PRP) derivative mainly composed of fibrin networks, has been increasingly demonstrated to be effective in wound healing in clinical and pre-clinical animal studies. However, there has still been a concern that major growth factors may significantly be loss from PRF during its preparation through the slow clotting process. To address this concern, we compared the angiogenic potential of PRF and PRP by standardization of procedures based on volume ratios.

          Methods

          PRP, PRF, and platelet-poor plasma (PPP) were prepared from the peripheral blood of healthy donors. PRF preparations were squeezed or homogenized to produce exudate (PRF exu) or extract (PRF ext), respectively. Concentrations of the angiogenic factors and their bioactivities were determined using ELISA kits, a scratch assay using endothelial cells and a chicken chorioallantoic membrane (CAM) assay.

          Results

          In PRP and PRF preparations, both VEGF and PDGF-BB were significantly more concentrated than PPP. In the scratch assay, PRF exu and PRF ext were the most effective for wound closure. In the CAM assay, PRF membranes were the most effective for neovascularization.

          Conclusions

          It is suggested that PRF preparations efficiently preserve the angiogenic factors and function not only as a scaffolding material but as a reservoir of angiogenic factors in wound healing.

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          Most cited references30

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          Platelet-rich plasma: evidence to support its use.

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            Platelet-rich plasma: Growth factor enhancement for bone grafts.

            Platelet-rich plasma is an autologous source of platelet-derived growth factor and transforming growth factor beta that is obtained by sequestering and concentrating platelets by gradient density centrifugation. This technique produced a concentration of human platelets of 338% and identified platelet-derived growth factor and transforming growth factor beta within them. Monoclonal antibody assessment of cancellous cellular marrow grafts demonstrated cells that were capable of responding to the growth factors by bearing cell membrane receptors. The additional amounts of these growth factors obtained by adding platelet-rich plasma to grafts evidenced a radiographic maturation rate 1.62 to 2.16 times that of grafts without platelet-rich plasma. As assessed by histomorphometry, there was also a greater bone density in grafts in which platelet-rich plasma was added (74.0% +/- 11%) than in grafts in which platelet-rich plasma was not added (55.1% +/- 8%; p = 0.005).
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              Interactions between extracellular matrix and growth factors in wound healing.

              Dynamic interactions between growth factors and extracellular matrix (ECM) are integral to wound healing. These interactions take several forms that may be categorized as direct or indirect. The ECM can directly bind to and release certain growth factors (e.g., heparan sulfate binding to fibroblast growth factor-2), which may serve to sequester and protect growth factors from degradation, and/or enhance their activity. Indirect interactions include binding of cells to ECM via integrins, which enables cells to respond to growth factors (e.g., integrin binding is necessary for vascular endothelial growth factor-induced angiogenesis) and can induce growth factor expression (adherence of monocytes to ECM stimulates synthesis of platelet-derived growth factor). Additionally, matrikines, or subcomponents of ECM molecules, can bind to cell surface receptors in the cytokine, chemokine, or growth factor families and stimulate cellular activities (e.g., tenascin-C and laminin bind to epidermal growth factor receptors, which enhances fibroblast migration). Growth factors such as transforming growth factor-beta also regulate the ECM by increasing the production of ECM components or enhancing synthesis of matrix degrading enzymes. Thus, the interactions between growth factors and ECM are bidirectional. This review explores these interactions, discusses how they are altered in difficult to heal or chronic wounds, and briefly considers treatment implications.
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                Author and article information

                Contributors
                kawase@dent.niigata-u.ac.jp
                Journal
                Int J Implant Dent
                Int J Implant Dent
                International Journal of Implant Dentistry
                Springer Berlin Heidelberg (Berlin/Heidelberg )
                2198-4034
                27 November 2015
                27 November 2015
                December 2015
                : 1
                : 1
                : 31
                Affiliations
                [1 ]Division of Oral Bioengineering, Institute of Medicine and Dentistry, Niigata University, Niigata, Japan
                [2 ]Division of Periodontology, Institute of Medicine and Dentistry, Niigata University, Niigata, Japan
                [3 ]Advanced Research Center, The Nippon Dental University School of Life Dentistry at Niigata, Niigata, Japan
                [4 ]Division of Periodontology, Department of Developmental and Surgical Sciences, University of Minnesota School of Dentistry, Minneapolis, MN USA
                Article
                32
                10.1186/s40729-015-0032-0
                5005601
                27747653
                6454cffd-5634-4bc5-82c9-ef15cebd1f92
                © Kobayashi et al. 2015

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

                History
                : 26 August 2015
                : 17 November 2015
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100001691, Japan Society for the Promotion of Science (JP);
                Award ID: 23592881
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100001691, Japan Society for the Promotion of Science (JP);
                Award ID: 24390443
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100001691, Japan Society for the Promotion of Science (JP);
                Award ID: 24390465
                Award Recipient :
                Categories
                Research
                Custom metadata
                © The Author(s) 2015

                platelets,fibrin,plasma,angiogenesis,endothelial cells
                platelets, fibrin, plasma, angiogenesis, endothelial cells

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