The Influence of Leukocyte and Platelet Concentrate Enrich in Stem Cells on Bone Regeneration Processes: A Clinical and Flow Cytometry Study


This article reports the efficacy of percutaneous autologous leukocyte- and platelet-rich plasma (L-PRP) injection into delayed union site as a minimally invasive method alternative to bone marrow aspirate and open grafting techniques. Each of 15 participants was followed on a regular basis with clinical examinations, roentgenograms. The average time to union was 8.4 weeks after L-PRP injection and the union was achieved in all cases. CD34+/45+ cells counts were increased by 410% and CD34+/45- cells counts were increased by 488% on average in L-PRP. Our investigation showed that L-PRP enrich in stem cells can produce the desired stimulatory response despite a substantial amount of vital bone cells from mesenchymal line. We believe that the using of L-PRP enriched in stem cells, growth factors and antimicrobial proteins might be a promising treatment method in regenerative medicine.

Share and Cite:

Bielecki, T. , Wójcik, K. , Bold, T. , Osadnik, B. and Szczepański, T. (2015) The Influence of Leukocyte and Platelet Concentrate Enrich in Stem Cells on Bone Regeneration Processes: A Clinical and Flow Cytometry Study. Journal of Biomedical Science and Engineering, 8, 659-664. doi: 10.4236/jbise.2015.89062.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Dohan Ehrenfest, D.M., Bielecki, T., Mishra, A., Borzini, P., Inchingolo, F., Sammartino, G., Rasmusson, L. and Evert, P.A. (2012) In Search of a Consensus Terminology in the Field of Platelet Concentrates for Surgical Use: Platelet-Rich Plasma (PRP), Platelet-Rich Fibrin (PRF), Fibrin Gel Polymerization and Leukocytes. Current Pharmaceutical Biotechnology, 13, 1131-1137.
[2] Bielecki, T. and Dohan Ehrenfest, D.M. (2012) Platelet-Rich Plasma (PRP) and Platelet-Rich Fibrin (PRF): Surgical Adjuvants, Preparations for in Situ Regenerative Medicine and Tools for Tissue Engineering. Current Pharmaceutical Biotechnology, 13, 1121-1130.
[3] Cieslik-Bielecka, A., Choukroun, J., Odin, G. and Dohan Ehrenfest, D.M. (2012) L-PRP/L-PRF in Esthetic Plastic Surgery, Regenerative Medicine of the Skin and Chronic Wounds. Current Pharmaceutical Biotechnology, 13, 1266- 1277.
[4] Clark, R.A. (2001) Fibrin and Wound Healing. Annals of the New York Academy of Sciences, 936, 355-367.
[5] Gibble, J.W. and Ness, P.M. (1990) Fibrin Glue: The Perfect Operative Sealant? Transfusion, 30, 741-747.
[6] Del Corso, M., Vervelle, A., Simonpieri, A., et al. (2012) 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. Current Pharmaceutical Biotechnology, 13, 1207-1230.
[7] Bielecki, T.M. and Gazdzik, T.S. (2006) Percutaneous Injection of Autogenous Growth Factors in Patient with Nonunion of the Humerus. A Case Report. Journal of Orthopaedics, 3, e15.
[8] Mishara, A. and Pavelko, T. (2006) Treatment of Chronic Elbow Tendinosis with Buffered Platelet-Rich Plasma. The American Journal of Sports Medicine, 34, 1774-1778.
[9] Marx, R.E., Carlson, E.R., Eichstaedt, R.M., Schimmele, S.R., Strauss, J.E. and Georgeff, K.R. (1998) Platelet-Rich Plasma: Growth Factor Enhancement for Bone Grafts. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology, 85, 638-646.
[10] Bielecki, T.M. and Gazdzik, T.S. (2007) Percutaneous Injection of Autologous Platelet-Rich Gel in a Patient with an Infected Nonunion of the Tibia. A Case Report and Review of the Literature. Journal of Orthopaedics Trauma Surgery and Related Research, 2, s69-s72.
[11] Moojen, D.J., Everts, P.A., Schure, R.M., Overdevest, E.P., van Zundert, A., et al. (2008) Antimicrobial Activity of Platelet-Leukocyte Gel against Staphylococcus Aureus. Journal of Orthopaedic Research, 26, 404-410.
[12] Weibrich, G., Hansen, T., Kleis, W., Buch, R. and Hitzler, W.E. (2004) Effect of Platelet Concentration in Platelet- Rich Plasma on Peri-Implant Bone Regeneration. Bone, 34, 665-671.
[13] Mishra, A., Harmon, K., Woodall, J. and Vieira, A. (2012) Sports Medicine Applications of Platelet Rich Plasma. Current Pharmaceutical Biotechnology, 13, 1185-1195.
[14] Anitua, E., Sanchez, M., Orive, G. and Andia, I. (2007) The Potential Impact of the Preparation Rich in Growth Factors (PRGF) in Different Medical Fields. Biomaterials, 28, 4551-4560.
[15] Toeroek, R. and Dohan Ehrenfest, D.M. (2013) The Concept of Screw-Guided Bone Regeneration (S-GBR). Part 2: S- GBR in the Severely Resorbed Preimplant Posterior Mandible Using Bone Xenograft and Leukocyte- and Platelet-Rich Fibrin (L-PRF): A 5-Year Follow-Up. POSEIDO, 1, 85-92.
[16] Cieslik-Bielecka, A., Choukroun, J., Odin, G. and Dohan Ehrenfest, D.M. (2012) L-PRP/L-PRF in Esthetic Plastic Surgery, Regenerative Medicine of the Skin and Chronic Wounds. Current Pharmaceutical Biotechnology, 13, 1266-1277.
[17] Everts, P.A., Hoffmann, J., Weibrich, G., Mahoney, C.B., Schonberger, J.P.A.M., et al. (2006) Differences in Platelet Growth Factor Release and Leucocyte Kinetics during Autologous Platelet Gel Formation. Transfusion Medicine, 16, 363-368.
[18] Everts, P.A., van Zundert, A., Schonberger, J.P., Devilee, R.J. and Knape, J.T. (2008) What Do We Use: Platelet-Rich Plasma or Platelet-Leukocyte Gel? Journal of Biomedical Materials Research Part A, 85, 1135-1136.
[19] Cieslik, T., Skowronek, J., Cieslik, M. and Cieslik-Bielecka, A. (2009) Bone Graft Application from Anterior Sinus Maxillary Wall in Orbital Floor Reconstruction. Journal of Craniofacial Surgery, 20, 512-515.
[20] Malhotra, A., Pelletier, M.H., Yu, Y. and Walsh, W.R. (2013) Can Platelet-Rich Plasma (PRP) Improve Bone Healing? A Comparison between the Theory and Experimental Outcomes. Archives of Orthopaedic and Trauma Surgery, 133, 153-165.
[21] Frechette, J.P., Martineau, I. and Gagnon, G. (2005) Platelet Rich Plasmas: Growth Factor Content and Roles in Wound Healing. Journal of Dental Research, 84, 434-439.
[22] Yuan, T., Zhang, C. and Zeng, B. (2008) Treatment of Chronic Femoral Osteomyelitis with Platelet-Rich Plasma (PRP): A Case Report. Transfusion and Apheresis Science, 38, 167-173.
[23] Everts, P.A., Devilee, R.J., Brown Mahoney, C., van Erp, A., Oosterbos, C.J., Stellenboom, M., Knape, J.T. and van Zundert, A. (2008) Exogenous Application of Platelet-Leukocyte Gel during Open Subacromial Decompression Contributes to Improved Patient Outcome. A Prospective Randomized Double-Blind Study. European Surgical Research, 40, 203-210.
[24] Khalafi, R.S., Bradford, D.W. and Wilson, M.G. (2008) Topical Application of Autologous Blood Products during Surgical Closure Following a Coronary Artery Bypass Graft. European Journal Cardio-Thoracic Surgery, 34, 360-364.
[25] Aspenberg, P. and Virchenko, O. (2004) Platelet Concentrate Injection Improves Achilles Tendon Repair in Rats. Acta Orthopaedica Scandinavica, 75, 93-99.
[26] Bielecki, T., Gazdzik, T.S. and Szczepanski, T. (2006) What Do We Use: Platelet-Rich Plasma or Platelet-Rich Gel? Bone, 39, 1388,.
[27] Bielecki, T., Gazdzik, T.S and Szczepanski, T. (2008) Benefit of Percutaneous Injection of Autologous Platelet-Leukocyte-Rich Gel in Patients with Delayed Union and Nonunion. European Surgical Research, 40, 289-296.
[28] Lowery, G.L., Kulkarni, S. and Pennisi, A.E. (1999) Use of Autologous Growth Factors in Lumbar Spinal Fusion. Bone, 25, 47S-50S.
[29] Kitoh, H., Kitakoji, T., Tsuchiya, H., Katoh, M. and Ishiguro, N. (2007) Transplantation of Culture Expanded Bone Marrow Cells and Platelet Rich Plasma in Distraction Osteogenesis of the Long Bones. Bone, 40, 522-528.
[30] Bielecki, T., Cieslik-Bielecka, A., Zelawski, M. and Mikusek, W. (2012) A Side-Effect Induced by the Combination of a Demineralized Freeze-Dried Bone Allograft and Leucocyte and Platelet-Rich Plasma during Treatment for Large Bone Cysts: A 4-Year Follow-Up Clinical Study. Transfusion and Apheresis Science, 47, 133-138.
[31] Seaberg, R.M. and van der Kooy, D. (2003) Stem and Progenitor Cells: The Premature Desertion of Rigorous Definitions. Trends in Neurosciences, 26, 125-131.
[32] Ciurea, M.E., Georgescu, A.M., Purcaru, S.O., Artene, S.A., Emami, G.H., Boldeanu, M.V., Tache, D.E. and Dricu, A. (2014) Cancer Stem Cells: Biological Functions and Therapeutically Targeting. International Journal of Molecular, 15, 8169-8185.

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.