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European Journal of Trauma and Emergency Surgery

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Open Access 01-02-2018 | Original Article

Reduction of relative centrifugation force within injectable platelet-rich-fibrin (PRF) concentrates advances patients’ own inflammatory cells, platelets and growth factors: the first introduction to the low speed centrifugation concept

Authors: J. Choukroun, S. Ghanaati

Published in: European Journal of Trauma and Emergency Surgery | Issue 1/2018

Abstract

Purpose

The aim of this study was to analyze systematically the influence of the relative centrifugation force (RCF) on leukocytes, platelets and growth factor release within fluid platelet-rich fibrin matrices (PRF).

Materials and methods

Systematically using peripheral blood from six healthy volunteers, the RCF was reduced four times for each of the three experimental protocols (I–III) within the spectrum (710–44 g), while maintaining a constant centrifugation time. Flow cytometry was applied to determine the platelets and leukocyte number. The growth factor concentration was quantified 1 and 24 h after clotting using ELISA.

Results

Reducing RCF in accordance with protocol-II (177 g) led to a significantly higher platelets and leukocytes numbers compared to protocol-I (710 g). Protocol-III (44 g) showed a highly significant increase of leukocytes and platelets number in comparison to -I and -II. The growth factors’ concentration of VEGF and TGF-β1 was significantly higher in protocol-II compared to -I, whereas protocol-III exhibited significantly higher growth factor concentration compared to protocols-I and -II. These findings were observed among 1 and 24 h after clotting, as well as the accumulated growth factor concentration over 24 h.

Discussion

Based on the results, it has been demonstrated that it is possible to enrich PRF-based fluid matrices with leukocytes, platelets and growth factors by means of a single alteration of the centrifugation settings within the clinical routine.

Conclusions

We postulate that the so-called low speed centrifugation concept (LSCC) selectively enriches leukocytes, platelets and growth factors within fluid PRF-based matrices. Further studies are needed to evaluate the effect of cell and growth factor enrichment on wound healing and tissue regeneration while comparing blood concentrates gained by high and low RCF.

Ghanaati S, Kovács A, Barbeck M, Lorenz J, Teiler A, Sadeghi N, et al. Bilayered, non-cross-linked collagen matrix for regeneration of facial defects after skin cancer removal: a new perspective for biomaterial-based tissue reconstruction. J Cell Commun Signal. 2016;10:3–15.CrossRefPubMed

Tsaryk R, Gloria A, Russo T, Anspach L, De Santis R, Ghanaati S, et al. Collagen-low molecular weight hyaluronic acid semi-interpenetrating network loaded with gelatin microspheres for cell and growth factor delivery for nucleus pulposus regeneration. Acta Mater Inc. 2015;20:10–21.

Schlee M, Ghanaati S, Willershausen I, Stimmlmayr M, Sculean A, Sader RA. Bovine pericardium based non-cross linked collagen matrix for successful root coverage, a clinical study in human. Head Face Med BioMed Central Ltd. 2012;8:6.CrossRef

Lorenz J, Blume M, Barbeck M, Teiler A, Kirkpatrick CJ, Sader RA, et al. Expansion of the peri-implant attached gingiva with a three-dimensional collagen matrix in head and neck cancer patients-results from a prospective clinical and histological study. Clin Oral Investig. 2016.

Sauerbier S, Rickert D, Gutwald R, Nagursky H, Oshima T, Xavier SP, et al. Bone marrow concentrate and bovine bone mineral for sinus floor augmentation: a controlled, randomized, single-blinded clinical and histological trial-per-protocol analysis. Tissue Eng Part A. 2011;17:2187–97.CrossRefPubMed

Duttenhoefer F, Hieber SF, Stricker A, Schmelzeisen R, Gutwald R, Sauerbier S. Follow-up of implant survival comparing ficoll and bone marrow aspirate concentrate methods for hard tissue regeneration with mesenchymal stem cells in humans. Biores Open Access. 2014;3:75–6.CrossRefPubMedPubMedCentral

Springer ING, Açil Y, Kuchenbecker S, Bolte H, Warnke PH, Abboud M, et al. Bone graft versus BMP-7 in a critical size defect-cranioplasty in a growing infant model. Bone. 2005;37:563–9.CrossRefPubMed

Liu Y, Möller B, Wiltfang J, Warnke PH, Terheyden H. Tissue engineering of a vascularized bone graft of critical size with an osteogenic and angiogenic factor-based in vivo bioreactor. Tissue Eng Part A. 2014;20:3189–97.CrossRefPubMed

Anitua E, Andia I, Sanchez M. PRGF plasma rich growth factors. Dent Dialogue. 2004;1–9.

10.

Marx RE, Carlson ER, Eichstaedt RM, Schimmele SR, Strauss JE, Georgeff KR. Platelet-rich plasma: growth factor enhancement for bone grafts. Oral Surg Oral Med Oral Pathol Oral Radiol Endodontol. 1998;85:638–46.CrossRef

11.

Everts P a M, Knape JT a, Weibrich G, Schönberger JP a M, Hoffmann J, Overdevest EP, et al. Platelet-rich plasma and platelet gel: a review. J Extra Corpor Technol. 2006;38:174–87.PubMedPubMedCentral

12.

Anitua E, Sánchez M, Orive G, Andía I. The potential impact of the preparation rich in growth factors (PRGF) in different medical fields. Biomaterials. 2007;28:4551–60.CrossRefPubMed

13.

Choukroun J, Adda F, Schoeffler C, Vervelle A. An opportunity in paro-implantology: PRF [in French]. Implantodontie. 2001;42:55–62.

14.

Gurtner GC, Werner S, Barrandon Y, Longaker MT. Wound repair and regeneration. Nature. 2008;453:314–21.CrossRefPubMed

15.

Litvinov RI, Weisel JW. What is the biological and clinical relevance of fibrin? Semin Thromb Hemost. 2016;42:333–43.CrossRefPubMedPubMedCentral

16.

Sahni A, Francis CW. Vascular endothelial growth factor binds to fibrinogen and fibrin and stimulates endothelial cell proliferation. Blood. 2000;96:3772–8.PubMed

17.

van Hinsbergh VW, Collen a, Koolwijk P. Role of fibrin matrix in angiogenesis. Ann N Y Acad Sci. 2001;936:426–37.CrossRefPubMed

18.

Soloviev DA, Hazen SL, Szpak D, Bledzka KM, Ballantyne CM, Plow EF, et al. Dual role of the leukocyte integrin M 2 in angiogenesis. J Immunol. 2014;193:4712–21.CrossRefPubMedPubMedCentral

19.

Ghanaati S, Booms P, Orlowska A, Kubesch A, Lorenz J, Rutkowski J, et al. Advanced platelet-rich fibrin: a new concept for cell-based tissue engineering by means of inflammatory cells. J Oral Implantol. 2014;40:679–89.CrossRefPubMed

20.

Quirke P, Dyson JED. Flow cytometry: methodology and applications in pathology. J Pathol. 1986;149:79–87.CrossRefPubMed

21.

Keith Wilson JW. Principles and techniques of biochemistry and molecular biology. Cambridge: Cambridge University Press;2010.

22.

Kawazoe T, Kim HH. Tissue augmentation by white blood cell-containing platelet-rich plasma. Cell Transplant. 2012;21:601–7.CrossRefPubMed

23.

Omar OM, Granéli C, Ekström K, Karlsson C, Johansson A, Lausmaa J, et al. The stimulation of an osteogenic response by classical monocyte activation. Biomaterials. 2011;32:8190–204.CrossRefPubMed

24.

Pirraco RP, Reis RL, Marques AP. Effect of monocytes/macrophages on the early osteogenic differentiation of hBMSCs. J Tissue Eng Regen Med. 2013;7:392–400.CrossRefPubMed

25.

Ekström K, Omar O, Granéli C, Wang X, Vazirisani F, Thomsen P. Monocyte exosomes stimulate the osteogenic gene expression of mesenchymal stem cells. PLoS One. 2013;8:e75227.CrossRefPubMedPubMedCentral

26.

Perut F, Filardo G, Mariani E, Cenacchi A, Pratelli L, Devescovi V, et al. Preparation method and growth factor content of platelet concentrate influence the osteogenic differentiation of bone marrow stromal cells. Cytotherapy. 2013;15:830–9.CrossRefPubMed

27.

Tan KW, Chong SZ, Wong FHS, Evrard M, Tan SM-L, Keeble J, et al. Neutrophils contribute to inflammatory lymphangiogenesis by increasing VEGF-A bioavailability and secreting VEGF-D. Blood. 2013;122:3666–77.CrossRefPubMed

28.

Schär MO, Diaz-Romero J, Kohl S, Zumstein MA, Nesic D. Platelet-rich concentrates differentially release growth factors and induce cell migration in vitro. Clin Orthop Relat Res. 2015;473:1635–43.CrossRefPubMedPubMedCentral

29.

Nami N, Feci L, Napoliello L, Giordano A, Lorenzini S, Galeazzi M, et al. Crosstalk between platelets and PBMC: new evidence in wound healing. Platelets. 2016;27:143–8.PubMed

30.

Jurk K, Kehrel BE. Platelets: physiology and biochemistry. Semin Thromb Hemost. 2005;31:381–92.

31.

Walsh TG, Metharom P, Berndt MC. The functional role of platelets in the regulation of angiogenesis. Platelets. 2015;26:199–211.CrossRefPubMed

32.

Jenne CN, Urrutia R, Kubes P. Platelets: bridging hemostasis, inflammation, and immunity. Int J Lab Hematol. 2013;35:254–61.CrossRefPubMed

33.

Yeaman MR. Platelets: at the nexus of antimicrobial defence. Nat Rev Microbiol. 2014;12:426–37.CrossRefPubMed

34.

Schmidt-Bleek K, Kwee BJ, Mooney DJ, Duda GN. Boon and bane of inflammation in bone tissue regeneration and its link with angiogenesis. Tissue Eng Part B Rev. 2015;21:354–64.CrossRefPubMedPubMedCentral

35.

Moens S, Goveia J, Stapor PC, Cantelmo AR, Carmeliet P. The multifaceted activity of VEGF in angiogenesis—implications for therapy responses. Cytokine Growth Factor Rev. 2014;25:473–82.CrossRefPubMed

36.

Lichtman MK, Otero-Vinas M, Falanga V. Transforming growth factor beta (TGF-β) isoforms in wound healing and fibrosis. Wound Repair Regen. 2016;215–22.

37.

Kim B-C, Kim HT, Park SH, Cha J-S, Yufit T, Kim S-J, et al. Fibroblasts from chronic wounds show altered TGF-β-signaling and decreased TGF-? Type II receptor expression. J Cell Physiol. 2003;195:331–6.CrossRefPubMed

38.

Roberts AB, Sporn MB, Assoian RK, Smith JM, Roche NS, Wakefield LM, et al. Transforming growth factor type beta: rapid induction of fibrosis and angiogenesis in vivo and stimulation of collagen formation in vitro. Proc Natl Acad Sci USA. 1986;83:4167–71.

39.

Barbeck M, Unger RE, Booms P, Dohle E, Sader RA, Kirkpatrick CJ, et al. Monocyte preseeding leads to an increased implant bed vascularization of biphasic calcium phosphate bone substitutes via vessel maturation. J Biomed Mater Res Part A. 2016;1–8.

40.

Davis VL, Abukabda AB, Radio NM, Witt-Enderby PA, Clafshenkel WP, Cairone JV, et al. Platelet-rich preparations to improve healing. Part II: platelet activation and enrichment, leukocyte inclusion, and other selection criteria. J Oral Implantol. 2014;40:511–21.CrossRefPubMed

41.

Ghasemzadeh M, Hosseini E. Intravascular leukocyte migration through platelet thrombi: directing leukocytes to sites of vascular injury. Thromb Haemost. 2015;113:1224–35.CrossRefPubMed

Title: Reduction of relative centrifugation force within injectable platelet-rich-fibrin (PRF) concentrates advances patients’ own inflammatory cells, platelets and growth factors: the first introduction to the low speed centrifugation concept
Authors: J. Choukroun
S. Ghanaati
Publication date: 01-02-2018
Publisher: Springer Berlin Heidelberg
Published in: European Journal of Trauma and Emergency Surgery / Issue 1/2018
Print ISSN: 1863-9933
Electronic ISSN: 1863-9941
DOI: https://doi.org/10.1007/s00068-017-0767-9

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Reduction of relative centrifugation force within injectable platelet-rich-fibrin (PRF) concentrates advances patients’ own inflammatory cells, platelets and growth factors: the first introduction to the low speed centrifugation concept

Abstract

Purpose

Materials and methods

Results

Discussion

Conclusions

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Purpose

Materials and methods

Results

Discussion

Conclusions

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