Systematic Comparison of the Effect of Four Clinical-Grade Platelet Rich Hemoderivatives on Osteoblast Behaviour
Abstract
:1. Introduction
2. Results
2.1. Thrombocyte Quantification (Table 1)
2.2. Release of Growth Factors from Different Hemoderivative Preparations (Figure 1)
2.3. Cell Viability (Figure 2)
2.4. Cell Metabolic Activity
2.5. Cell Migration
2.6. Mineralization Assay
2.7. Growth Factors Concentration Vs. Cell Viability Correlation
2.8. Correlation between BMP-2 Levels and Mineralization
3. Discussion
4. Materials and Methods
4.1. Blood Products (A-PRF, i-PRF, P-PRP, L-PRP and Natural Clot) (Figure 8)
4.2. Fresh Processing Protocols (A-PRF, i-PRF and Natural Clot)
4.2.1. A-PRF Preparation
4.2.2. i-PRF Preparation
4.2.3. Blot Clot Preparation
4.3. Anticoagulated Blood Protocols (L-PRP and P-PRP)
4.3.1. L-PRP Preparation
4.3.2. P-PRP Preparation
4.4. Hemoderivative Product Characterization
4.4.1. Thrombocyte Quantification
4.4.2. Blood Product Conditioned Media
4.4.3. Growth Factor Release (ELISA)
4.4.4. Isolation of Human Osteoblasts
4.4.5. Cell Viability
4.4.6. Metabolic Activity
4.4.7. Cell Migration Assay
4.4.8. Mineralization Assay
4.5. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
Abbreviations
A-PRF | Advance Platelet rich Fibrin |
i-PRF | Injectable Platelet Rich Fibrin |
P-PRP | Pure Platelet Rich Plasma |
L-PRP | Leucocyte Platelet Rich Fibrin |
CLOT | Thrombus, colloquially called a blood clot |
IGF-I | Insulin-like growth factor 1 |
VEGF | Vascular endothelial growth factor |
PDGF-BB | Platelet-derived growth factor subunits BB |
BMP-2 | Bone morphogenetic protein 2 |
References
- Kingsley, C.S. Blood coagulation; evidence of an antagonist to factor VI in platelet-rich human plasma. Nature 1954, 173, 723–724. [Google Scholar] [CrossRef] [PubMed]
- Agrawal, A.A. Evolution, current status and advances in application of platelet concentrate in periodontics and implantology. World J. Clin. Cases 2017, 5, 159–171. [Google Scholar] [CrossRef] [PubMed]
- Schar, M.O.; Diaz-Romero, J.; Kohl, S.; Zumstein, M.A.; Nesic, D. Platelet-rich concentrates differentially release growth factors and induce cell migration in vitro. Clin. Orthop. Relat. Res. 2015, 473, 1635–1643. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Fernandes, G.; Yang, S. Application of platelet-rich plasma with stem cells in bone and periodontal tissue engineering. Bone Res. 2016, 4, 16036. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Jalowiec, J.M.; D’Este, M.; Bara, J.J.; Denom, J.; Menzel, U.; Alini, M.; Verrier, S.; Herrmann, M. An In Vitro Investigation of Platelet-Rich Plasma-Gel as a Cell and Growth Factor Delivery Vehicle for Tissue Engineering. Tissue Eng. Part C Methods 2016, 22, 49–58. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kobayashi, E.; Fujioka-Kobayashi, M.; Sculean, A.; Chappuis, V.; Buser, D.; Schaller, B.; Dori, F.; Miron, R.J. Effects of platelet rich plasma (PRP) on human gingival fibroblast, osteoblast and periodontal ligament cell behaviour. BMC Oral Health 2017, 17, 91. [Google Scholar] [CrossRef] [Green Version]
- Dohan Ehrenfest, D.M.; Pinto, N.R.; Pereda, A.; Jimenez, P.; Corso, M.D.; Kang, B.S.; Nally, M.; Lanata, N.; Wang, H.L.; Quirynen, M. The impact of the centrifuge characteristics and centrifugation protocols on the cells, growth factors, and fibrin architecture of a leukocyte- and platelet-rich fibrin (L-PRF) clot and membrane. Platelets 2018, 29, 171–184. [Google Scholar] [CrossRef]
- Mathur, A.; Bains, V.K.; Gupta, V.; Jhingran, R.; Singh, G.P. Evaluation of intrabony defects treated with platelet-rich fibrin or autogenous bone graft: A comparative analysis. Eur. J. Dent. 2015, 9, 100–108. [Google Scholar] [CrossRef] [Green Version]
- Pradeep, A.R.; Nagpal, K.; Karvekar, S.; Patnaik, K.; Naik, S.B.; Guruprasad, C.N. Platelet-rich fibrin with 1% metformin for the treatment of intrabony defects in chronic periodontitis: A randomized controlled clinical trial. J. Periodontol. 2015, 86, 729–737. [Google Scholar] [CrossRef]
- Shah, M.; Patel, J.; Dave, D.; Shah, S. Comparative evaluation of platelet-rich fibrin with demineralized freeze-dried bone allograft in periodontal infrabony defects: A randomized controlled clinical study. J. Indian Soc. Periodontol. 2015, 19, 56–60. [Google Scholar] [CrossRef]
- Chenchev, I.L.; Ivanova, V.V.; Neychev, D.Z.; Cholakova, R.B. Application of Platelet-Rich Fibrin and Injectable Platelet-Rich Fibrin in Combination of Bone Substitute Material for Alveolar Ridge Augmentation-A Case Report. Folia Medica 2017, 59, 362–366. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Del Fabbro, M.; Bucchi, C.; Lolato, A.; Corbella, S.; Testori, T.; Taschieri, S. Healing of Postextraction Sockets Preserved With Autologous Platelet Concentrates. A Systematic Review and Meta-Analysis. J. Oral Maxillofac. Surg. 2017, 75, 1601–1615. [Google Scholar] [CrossRef] [PubMed]
- Oncu, E.; Kaymaz, E. Assessment of the effectiveness of platelet rich fibrin in the treatment of Schneiderian membrane perforation. Clin. Implant. Dent. Relat. Res. 2017, 19, 1009–1014. [Google Scholar] [CrossRef] [PubMed]
- Khairy, N.M.; Shendy, E.E.; Askar, N.A.; El-Rouby, D.H. Effect of platelet rich plasma on bone regeneration in maxillary sinus augmentation (randomized clinical trial). Int. J. Oral Maxillofac. Surg. 2013, 42, 249–255. [Google Scholar] [CrossRef]
- Raghoebar, G.M.; Schortinghuis, J.; Liem, R.S.; Ruben, J.L.; van der Wal, J.E.; Vissink, A. Does platelet-rich plasma promote remodeling of autologous bone grafts used for augmentation of the maxillary sinus floor? Clin. Oral Implant. Res. 2005, 16, 349–356. [Google Scholar] [CrossRef]
- Dohan Ehrenfest, D.M.; Rasmusson, L.; Albrektsson, T. Classification of platelet concentrates: From pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF). Trends Biotechnol. 2009, 27, 158–167. [Google Scholar] [CrossRef]
- Araki, J.; Jona, M.; Eto, H.; Aoi, N.; Kato, H.; Suga, H.; Doi, K.; Yatomi, Y.; Yoshimura, K. Optimized preparation method of platelet-concentrated plasma and noncoagulating platelet-derived factor concentrates: Maximization of platelet concentration and removal of fibrinogen. Tissue Eng. Part C Methods 2012, 18, 176–185. [Google Scholar] [CrossRef] [Green Version]
- do Amaral, R.J.; da Silva, N.P.; Haddad, N.F.; Lopes, L.S.; Ferreira, F.D.; Filho, R.B.; Cappelletti, P.A.; de Mello, W.; Cordeiro-Spinetti, E.; Balduino, A. Platelet-Rich Plasma Obtained with Different Anticoagulants and Their Effect on Platelet Numbers and Mesenchymal Stromal Cells Behavior In Vitro. Stem Cells Int. 2016, 2016, 7414036. [Google Scholar] [CrossRef] [Green Version]
- Burnouf, T.; Strunk, D.; Koh, M.B.C.; Schallmoser, K. Human platelet lysate: Replacing fetal bovine serum as a gold standard for human cell propagation? Biomaterials 2016, 76, 371–387. [Google Scholar] [CrossRef]
- Henschler, R.; Gabriel, C.; Schallmoser, K.; Burnouf, T.; Koh, M.B.C. Human platelet lysate current standards and future developments. Transfusion 2019, 59, 1407–1413. [Google Scholar] [CrossRef]
- Kreil, T.R. Building blocks of the viral safety margins of industrial plasma products. Ann. Blood 2018, 3–14. [Google Scholar] [CrossRef]
- Choukroun, J.; Adda, F.; Schoeffer, C.; Vervelle, A. PRF: An opportunity in perio implantology. Implantodontie 2001, 42, 55–62. [Google Scholar]
- Dohan, D.M.; Choukroun, J.; Diss, A.; Dohan, S.L.; Dohan, A.J.; Mouhyi, J.; Gogly, B. Platelet-rich fibrin (PRF): A second-generation platelet concentrate. Part I: Technological concepts and evolution. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endodontol. 2006, 101, 37–44. [Google Scholar] [CrossRef]
- Dohan, D.M.; Choukroun, J.; Diss, A.; Dohan, S.L.; Dohan, A.J.; Mouhyi, J.; Gogly, B. Platelet-rich fibrin (PRF): A second-generation platelet concentrate. Part III: Leucocyte activation: A new feature for platelet concentrates? Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endodontol. 2006, 101, 51–55. [Google Scholar] [CrossRef]
- Dohan, D.M.; Choukroun, J.; Diss, A.; Dohan, S.L.; Dohan, A.J.; Mouhyi, J.; Gogly, B. Platelet-rich fibrin (PRF): A second-generation platelet concentrate. Part II: Platelet-related biologic features. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endodontol. 2006, 101, 45–50. [Google Scholar] [CrossRef]
- Dohan Ehrenfest, D.M.; Del Corso, M.; Diss, A.; Mouhyi, J.; Charrier, J.B. Three-dimensional architecture and cell composition of a Choukroun’s platelet-rich fibrin clot and membrane. J. Periodontol. 2010, 81, 546–555. [Google Scholar] [CrossRef]
- Mourao, C.F.; Valiense, H.; Melo, E.R.; Mourao, N.B.; Maia, M.D. Obtention of injectable platelets rich-fibrin (i-PRF) and its polymerization with bone graft: Technical note. Rev. Do Colegio Bras. De Cir. 2015, 42, 421–423. [Google Scholar] [CrossRef] [Green Version]
- Del Corso, M.; Vervelle, A.; Simonpieri, A.; Jimbo, R.; Inchingolo, F.; Sammartino, G.; Dohan Ehrenfest, D.M. Current knowledge and perspectives for the use of platelet-rich plasma (PRP) and platelet-rich fibrin (PRF) in oral and maxillofacial surgery part 1: Periodontal and dentoalveolar surgery. Curr. Pharm. Biotechnol. 2012, 13, 1207–1230. [Google Scholar] [CrossRef] [Green Version]
- Choukroun. Advance PRF & i-PRF: Platelet concentrates or Blood Concentrates? J. Periodontal Med. Clin. Pract. 2014, 1, 3. [Google Scholar]
- Marino, M.J. Chapter 3-Statistical Analysis in Preclinical Biomedical Research. In Research in the Biomedical Sciences; Williams, M., Curtis, M.J., Mullane, K., Eds.; Academic Press: Cambridge, MA, USA, 2018; pp. 107–144. [Google Scholar] [CrossRef]
- Dupley, L.; Charalambous, C.P. Platelet-Rich Plasma Injections as a Treatment for Refractory Patellar Tendinosis: A Meta-Analysis of Randomised Trials. Knee Surg. Relat. Res. 2017, 29, 165–171. [Google Scholar] [CrossRef]
- Huang, Y.; Bornstein, M.M.; Lambrichts, I.; Yu, H.Y.; Politis, C.; Jacobs, R. Platelet-rich plasma for regeneration of neural feedback pathways around dental implants: A concise review and outlook on future possibilities. Int. J. Oral Sci. 2017, 9, 1–9. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Miron, R.J.; Zhang, Y.F. Osteoinduction: A review of old concepts with new standards. J. Dent. Res. 2012, 91, 736–744. [Google Scholar] [CrossRef] [PubMed]
- Anitua, E. Plasma rich in growth factors: Preliminary results of use in the preparation of future sites for implants. Int. J. Oral Maxillofac. Implant. 1999, 14, 529–535. [Google Scholar]
- Ticehurst, J.R.; Pisanic, N.; Forman, M.S.; Ordak, C.; Heaney, C.D.; Ong, E.; Linnen, J.M.; Ness, P.M.; Guo, N.; Shan, H.; et al. Probable transmission of hepatitis E virus (HEV) via transfusion in the United States. Transfusion 2019, 59, 1024–1034. [Google Scholar] [CrossRef]
- Klatte-Schulz, F.; Schmidt, T.; Uckert, M.; Scheffler, S.; Kalus, U.; Rojewski, M.; Schrezenmeier, H.; Pruss, A.; Wildemann, B. Comparative Analysis of Different Platelet Lysates and Platelet Rich Preparations to Stimulate Tendon Cell Biology: An In Vitro Study. Int. J. Mol. Sci. 2018, 19, 212. [Google Scholar] [CrossRef] [Green Version]
- Weibrich, G.; Kleis, W.K.; Hafner, G.; Hitzler, W.E.; Wagner, W. Comparison of platelet, leukocyte, and growth factor levels in point-of-care platelet-enriched plasma, prepared using a modified Curasan kit, with preparations received from a local blood bank. Clin. Oral Implant. Res. 2003, 14, 357–362. [Google Scholar] [CrossRef]
- Anitua, E.; Nurden, P.; Prado, R.; Nurden, A.T.; Padilla, S. Autologous fibrin scaffolds: When platelet- and plasma-derived biomolecules meet fibrin. Biomaterials 2019, 192, 440–460. [Google Scholar] [CrossRef]
- Schmidt, M.B.; Chen, E.H.; Lynch, S.E. A review of the effects of insulin-like growth factor and platelet derived growth factor on in vivo cartilage healing and repair. Osteoarthr. Cartil. 2006, 14, 403–412. [Google Scholar] [CrossRef] [Green Version]
- Creeper, F.; Ivanovski, S. Effect of autologous and allogenic platelet-rich plasma on human gingival fibroblast function. Oral Dis. 2012, 18, 494–500. [Google Scholar] [CrossRef]
- Wang, X.; Zhang, Y.; Choukroun, J.; Ghanaati, S.; Miron, R.J. Effects of an injectable platelet-rich fibrin on osteoblast behavior and bone tissue formation in comparison to platelet-rich plasma. Platelets 2018, 29, 48–55. [Google Scholar] [CrossRef]
- Miron, R.J.; Saulacic, N.; Buser, D.; Iizuka, T.; Sculean, A. Osteoblast proliferation and differentiation on a barrier membrane in combination with BMP2 and TGFbeta1. Clin. Oral Investig. 2013, 17, 981–988. [Google Scholar] [CrossRef] [Green Version]
- Montanari, M.; Callea, M.; Yavuz, I.; Maglione, M. A new biological approach to guided bone and tissue regeneration. BMJ Case Rep. 2013, 2013. [Google Scholar] [CrossRef] [Green Version]
- Zhang, Y.; Tangl, S.; Huber, C.D.; Lin, Y.; Qiu, L.; Rausch-Fan, X. Effects of Choukroun’s platelet-rich fibrin on bone regeneration in combination with deproteinized bovine bone mineral in maxillary sinus augmentation: A histological and histomorphometric study. J. Cranio Maxillo Fac. Surg. 2012, 40, 321–328. [Google Scholar] [CrossRef]
- Qiao, J.; An, N.; Ouyang, X. Quantification of growth factors in different platelet concentrates. Platelets 2017, 28, 774–778. [Google Scholar] [CrossRef]
- Lei, L.; Yu, Y.; Han, J.; Shi, D.; Sun, W.; Zhang, D.; Chen, L. Quantification of growth factors in advanced platelet-rich fibrin and concentrated growth factors and their clinical efficiency as adjunctive to the GTR procedure in periodontal intrabony defects. J. Periodontol. 2019. [Google Scholar] [CrossRef]
- Fujioka-Kobayashi, M.; Miron, R.J.; Hernandez, M.; Kandalam, U.; Zhang, Y.; Choukroun, J. Optimized Platelet-Rich Fibrin With the Low-Speed Concept: Growth Factor Release, Biocompatibility, and Cellular Response. J. Periodontol. 2017, 88, 112–121. [Google Scholar] [CrossRef]
- Vahabi, S.; Yadegari, Z.; Mohammad-Rahimi, H. Comparison of the effect of activated or non-activated PRP in various concentrations on osteoblast and fibroblast cell line proliferation. Cell Tissue Bank. 2017, 18, 347–353. [Google Scholar] [CrossRef]
- Anitua, E.; Aguirre, J.J.; Algorta, J.; Ayerdi, E.; Cabezas, A.I.; Orive, G.; Andia, I. Effectiveness of autologous preparation rich in growth factors for the treatment of chronic cutaneous ulcers. J. Biomed. Mater. Res. Part B Appl. Biomater. 2008, 84, 415–421. [Google Scholar] [CrossRef]
- Galiano, R.D.; Tepper, O.M.; Pelo, C.R.; Bhatt, K.A.; Callaghan, M.; Bastidas, N.; Bunting, S.; Steinmetz, H.G.; Gurtner, G.C. Topical vascular endothelial growth factor accelerates diabetic wound healing through increased angiogenesis and by mobilizing and recruiting bone marrow-derived cells. Am. J. Pathol. 2004, 164, 1935–1947. [Google Scholar] [CrossRef] [Green Version]
- Sharma, D.; Hamlet, S.M.; Petcu, E.B.; Ivanovski, S. The effect of bisphosphonates on the endothelial differentiation of mesenchymal stem cells. Sci. Rep. 2016, 6, 20580. [Google Scholar] [CrossRef] [Green Version]
- Gregory, C.A.; Gunn, W.G.; Peister, A.; Prockop, D.J. An Alizarin red-based assay of mineralization by adherent cells in culture: Comparison with cetylpyridinium chloride extraction. Anal. Biochem. 2004, 329, 77–84. [Google Scholar] [CrossRef] [PubMed]
Blood Product | g | Processing Time (min) | Platelet Recovery | Growth Factor Release 3 Days (pg/mL) | |||
---|---|---|---|---|---|---|---|
TOTAL BLOOD | - | - | 362 ± 22 × 103 | IGF-I | PDGF-BB | VEGF | BMP-2 |
A-PRF | 208 | 8 | 307 ± 14 × 103 | 265.2 ± 22.2 | 109.9 ± 3.4 | 90.3 ± 4.0 | 664.2 ± 37.6 |
i-PRF | 60 | 3 | 336 ± 18 × 103 | 66.6 ± 8.4 | 110.6 ± 2.5 | 142 ± 4.0 | 308.2 ± 15.9 |
L-PRP | 708 1/1594 2 | 10 1/15 2 | 1.82 ± 17 × 106 (*) | 823.6 ± 24.6 | 261.4 ± 11.3 | 113.5 ± 5.1 | 638.2 ± 69.8 |
P-PRP | 398 | 8 | 934 ± 32 × 103 (*) | 165.9 ± 2.9 | 109.1 ± 4.2 | 124.7 ± 5 | 652.9 ± 52.8 |
CLOT | - | 5 | 326 ± 32 × 103 | 9.8 ± 1.1 | 9 ± 1.1 | 68.4 ± 9.2 | 645.6 ± 41.4 |
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Fernández-Medina, T.; Vaquette, C.; Ivanovski, S. Systematic Comparison of the Effect of Four Clinical-Grade Platelet Rich Hemoderivatives on Osteoblast Behaviour. Int. J. Mol. Sci. 2019, 20, 6243. https://doi.org/10.3390/ijms20246243
Fernández-Medina T, Vaquette C, Ivanovski S. Systematic Comparison of the Effect of Four Clinical-Grade Platelet Rich Hemoderivatives on Osteoblast Behaviour. International Journal of Molecular Sciences. 2019; 20(24):6243. https://doi.org/10.3390/ijms20246243
Chicago/Turabian StyleFernández-Medina, Tulio, Cedryck Vaquette, and Sašo Ivanovski. 2019. "Systematic Comparison of the Effect of Four Clinical-Grade Platelet Rich Hemoderivatives on Osteoblast Behaviour" International Journal of Molecular Sciences 20, no. 24: 6243. https://doi.org/10.3390/ijms20246243