EVALUATION OF CENTRIFUGING REGIMES FOR THE PURPOSE OF OPTIMIZING THE PLATELET RICH PLASMA HARVESTING PROTOCOL
Abstract
Aim: Based on the classical principles, to determine the optimal conditions for centrifugation, PRP harvesing (platelet-rich plasma). To conduct a quantitative assessment of the substrate obtained under different conditions of centrifugation.
Materials and methods. Based on the basic principles of obtaining platelet-rich plasma (PRP) by centrifuging in containers with an anticoagulant followed by phase separation to obtain the final substrate, the efficiency of the technique under the conditions of single and double centrifugation as well as under different conditions of acceleration and centrifugation was evaluated.
Blood for follow-up was collected from 20 healthy volunteers (11 men, 9 women) average 25.3±4.1 in syringes of LuerLock design with ACD-A anticoagulant solution, and centrifuged. Centrifugation was carried out under controlled conditions using a centrifuge with rotating bowls of the rotor. Centrifugation was performed at an acceleration of 100-400g in time intervals up to 20 minutes. Activation of the substrate was performed with calcium chloride solution.
Quantitative evaluation of platelets of whole blood and the final substrate of PRP was carried out with a semi-automatic analyzer.
Results. The obtained results demonstrate the maximum level of harvesting efficiency when performing double centrifugation in the 150g×15 min+250g×10 min mode. Subject to this centrifugation protocol, it is possible to obtain a substrate that complies with the standardized requirements for PRP.
The maximum level of an increase in the number of platelets in the substrate in comparison with whole blood is determined at the level of ×4.36 with concentration (volume reduction) x5 in comparison with the volume of whole blood.
Conclusions. This study demonstrated the advantage of double centrifuging modes over single modes.
According to the results of the study, it was possible to determine the conditions for an optimal double-centrifugation mode (acceleration and duration), which allows us to achieve the most efficient concentration of the substrate.
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Liu, Y.-S., Ou, M.-E., Liu, H., Gu, M., Lv, L.-W., Fan, C. et. al. (2014). The effect of simvastatin on chemotactic capability of SDF-1α and the promotion of bone regeneration. Biomaterials, 35 (15), 4489–4498. doi: http://doi.org/10.1016/j.biomaterials.2014.02.025
Andersen, R. K., Zaher, W., Larsen, K. H., Ditzel, N., Drews, K., Wruck, W. et. al. (2015). Association between in vivo bone formation and ex vivo migratory capacity of human bone marrow stromal cells. Stem Cell Research & Therapy, 6 (1). doi: http://doi.org/10.1186/s13287-015-0188-9
Bendinelli, P., Matteucci, E., Dogliotti, G., Corsi, M. M., Banfi, G., Maroni, P., Desiderio, M. A. (2010). Molecular basis of anti-inflammatory action of platelet-rich plasma on human chondrocytes: Mechanisms of NF-κB inhibition via HGF. Journal of Cellular Physiology, 225 (3), 757–766. doi: http://doi.org/10.1002/jcp.22274
Castillo, T. N., Pouliot, M. A., Kim, H. J., Dragoo, J. L. (2010). Comparison of Growth Factor and Platelet Concentration From Commercial Platelet-Rich Plasma Separation Systems. The American Journal of Sports Medicine, 39 (2), 266–271. doi: http://doi.org/10.1177/0363546510387517
Wang, X., Zhang, Y., Choukroun, J., Ghanaati, S., Miron, R. J. (2017). Effects of an injectable platelet-rich fibrin on osteoblast behavior and bone tissue formation in comparison to platelet-rich plasma. Platelets, 29 (1), 48–55. doi: http://doi.org/10.1080/09537104.2017.1293807
Kundu, R., Rathee, M. (2014). Effect of Platelet-Rich-Plasma (PRP) and Implant Surface Topography on Implant Stability and Bone. Journal of clinical and diagnostic research, 8 (6), 26–30. doi: http://doi.org/10.7860/jcdr/2014/9177.4478
Mlynarek, R. A., Kuhn, A. W., Bedi, A. (2016). Platelet-Rich Plasma (PRP) in Orthopedic Sports Medicine. American journal of orthopedics, 45 (5), 290–326.
Wasserman, A., Matthewson, G., MacDonald, P. (2018). Platelet-Rich Plasma and the Knee–Applications in Orthopedic Surgery. Current Reviews in Musculoskeletal Medicine, 11 (4), 607–615. doi: http://doi.org/10.1007/s12178-018-9521-0
Cameli, N., Mariano, M., Cordone, I., Abril, E., Masi, S., Foddai, M. L. (2017). Autologous Pure Platelet-Rich Plasma Dermal Injections for Facial Skin Rejuvenation. Dermatologic Surgery, 43 (6), 826–835. doi: http://doi.org/10.1097/dss.0000000000001083
Dhurat, R., Sukesh, M. (2014). Principles and methods of preparation of platelet-rich plasma: A review and author′s perspective. Journal of Cutaneous and Aesthetic Surgery, 7 (4), 189–197. doi: http://doi.org/10.4103/0974-2077.150734
Lu, J., Hao, Y., Zhao, W., Lyu, C., Zou, D. (2017). Molecular, Cellular and Pharmaceutical Aspects of Autologous Grafts for Peri-implant Hard and Soft Tissue Defects. Current Pharmaceutical Biotechnology, 18 (1), 85–94. doi: http://doi.org/10.2174/1389201017666161201165742
Picard, F., Hersant, B., La Padula, S., Meningaud, J.-P. (2017). Platelet-rich plasma-enriched autologous fat graft in regenerative and aesthetic facial surgery: Technical note. Journal of Stomatology, Oral and Maxillofacial Surgery, 118 (4), 228–231. doi: http://doi.org/10.1016/j.jormas.2017.05.005
Charneux, L., Demoulin, C., Vanderthomment, M., Tomasella, M., Ferrara, M.-A., Grosdent, S. et. al. (2017). Plasma riche en plaquettes (PRP) et lésions discales : revue de la littérature. Neurochirurgie, 63 (6), 473–477. doi: http://doi.org/10.1016/j.neuchi.2017.06.002
Matz, E. L., Pearlman, A. M., Terlecki, R. P. (2018). Safety and feasibility of platelet rich fibrin matrix injections for treatment of common urologic conditions. Investigative and Clinical Urology, 59 (1), 61–65. doi: http://doi.org/10.4111/icu.2018.59.1.61
Cavallo, C., Roffi, A., Grigolo, B., Mariani, E., Pratelli, L., Merli, G. et. al. (2016). Platelet-Rich Plasma: The Choice of Activation Method Affects the Release of Bioactive Molecules. BioMed Research International, 2016, 1–7. doi: http://doi.org/10.1155/2016/6591717
Mazzocca, A. D., McCarthy, M. B. R., Chowaniec, D. M., Cote, M. P., Romeo, A. A., Bradley, J. P. et. al. (2012). Platelet-Rich Plasma Differs According to Preparation Method and Human Variability. The Journal of Bone and Joint Surgery-American Volume, 94 (4), 308–316. doi: http://doi.org/10.2106/jbjs.k.00430
Kececi, Y., Ozsu, S., Bilgir, O. (2014). A cost-effective method for obtaining standard platelet-rich plasma. WOUNDS, 26, 232–238.
Castillo, T. N., Pouliot, M. A., Kim, H. J., Dragoo, J. L. (2011). Comparison of Growth Factor and Platelet Concentration From Commercial Platelet-Rich Plasma Separation Systems. The American Journal of Sports Medicine, 39 (2), 266–271. doi: http://doi.org/10.1177/0363546510387517
Bausset, O., Giraudo, L., Veran, J., Magalon, J., Coudreuse, J.-M., Magalon, G. et. al. (2012). Formulation and Storage of Platelet-Rich Plasma Homemade Product. BioResearch Open Access, 1 (3), 115–123. doi: http://doi.org/10.1089/biores.2012.0225
Magalon, J., Bausset, O., Serratrice, N., Giraudo, L., Aboudou, H., Veran, J. et. al. (2014). Characterization and Comparison of 5 Platelet-Rich Plasma Preparations in a Single-Donor Model. Arthroscopy: The Journal of Arthroscopic & Related Surgery, 30 (5), 629–638. doi: http://doi.org/10.1016/j.arthro.2014.02.020
Fitzpatrick, J., Bulsara, M. K., McCrory, P. R., Richardson, M. D., Zheng, M. H. (2017). Analysis of platelet-rich plasma extraction: variations in platelet and blood components between 4 common commercial kits. Orthopaedic Journal of Sports Medicine, 5 (1). doi: http://doi.org/10.1177/2325967116675272
Perez, A. G. M., Lana, J. F. S. D., Rodrigues, A. A., Luzo, A. C. M., Belangero, W. D., Santana, M. H. A. (2014). Relevant Aspects of Centrifugation Step in the Preparation of Platelet-Rich Plasma. ISRN Hematology, 2014, 1–8. doi: http://doi.org/10.1155/2014/176060
McCarrel, T. M., Minas, T., Fortier, L. A. (2012). Optimization of Leukocyte Concentration in Platelet-Rich Plasma for the Treatment of Tendinopathy. The Journal of Bone & Joint Surgery, 94 (19), e143. doi: http://doi.org/10.2106/jbjs.l.00019
Magalon, J., Bausset, O., Serratrice, N., Giraudo, L., Aboudou, H., Veran, J. et. al. (2014). Characterization and Comparison of 5 Platelet-Rich Plasma Preparations in a Single-Donor Model. Arthroscopy: The Journal of Arthroscopic & Related Surgery, 30 (5), 629–638. doi: http://doi.org/10.1016/j.arthro.2014.02.020
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