Leukocyte presence does not increase microbicidal activity of Platelet-rich Plasma in vitro

BMC Microbiology

Giuseppe Filardo⁴  Berardo Di Matteo⁴  Maurilio Marcacci⁴  Elizaveta Kon⁴  Maria Paola Landini¹  Luca Cattini³  Alessandra Bielli¹  Andrea Berlingeri¹  Valentina Canella³  Erminia Mariani² 

[1] Unit of Clinical Microbiology, St. Orsola University Hospital, University of Bologna, Bologna, Italy;Department of Medical and Surgical Sciences, University of Bologna, Bologna, Italy;Laboratory of Immunorheumatology and Tissue Regeneration/RAMSES, Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, Bologna, 40136, Italy;Laboratory of Biomechanics and Technology Innovation/NABI, 2nd Orthopaedic and Traumatologic Clinic, Rizzoli Orthopaedic Institute, via di Barbiano 1/10, Bologna, Italy

关键词: Nosocomial infections;    Microbicidal proteins;    Antimicrobial activity;    Bacterial growth inhibition;    Leukocytes;    Platelet-rich plasma;   

Others  :  1227614
DOI  :  10.1186/s12866-015-0482-9

 received in 2014-12-18, accepted in 2015-07-13,  发布年份 2015

【 摘 要 】

Background

Human platelets are a rich reservoir of molecules that promote regenerative processes and microbicidal activity. This activity might be increased by concentration in platelet-rich plasma (PRP) products and modulated by the presence of leukocytes. Despite extensive use in clinical procedures, only few studies have investigated PRP’s real microbicidal potential. Therefore, this study aimed at comparing the in vitro microbicidal activity of platelets and leukocyte-enriched PRP (L-PRP) to pure platelet-rich plasma (P-PRP) and the contribution of leukocytes to microbicidal properties.

Antimicrobial effects of P- and L-PRP were tested against Escherichia Coli, Staphylococcus Aureus, Klebsiella Pneumoniae, Pseudomonas Aeruginosa and Enterococcus Faecalis. Furthermore, L-PRP was frozen (L-PRP cryo) to assess whether the preparation maintained in vitro characteristics. Microbicidal proteins released by the three preparations were also evaluated.

Results

L-PRP, L-PRP cryo and P-PRP generally induced comparable bacterial growth inhibition for up to 4 h’ incubation, range 1–4 log. MIP-1α, RANTES, GRO-α, IL-8, NAP-2, SDF-1α and IL-6 showed strong microbicidal potential.

Conclusions

We found in vitro antibacterial activity of L-PRP and P-PRP and the possibility to cryopreserve L-PRP, without important changes to its effectiveness; similar microbicidal activity between preparations containing or not leukocytes; and the contribution of three new molecules (NAP-2, SDF-1α and IL-6).

【 授权许可】

   
2015 Mariani et al.

【 图 表 】

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【 参考文献 】

• [1]Klinger MH, Jelkmann W. Role of blood platelets in infection and inflammation. J Interferon Cytokine Res. 2002; 22:913-922.
• [2]Tschon M, Fini M, Giardino R, Filardo G, Dallari D, Torricelli P et al.. Lights and shadows concerning platelet products for musculoskeletal regeneration. Front Biosci (Elite Ed). 2011; 3:96-107.
• [3]Anitua E, Aguirre JJ, Algorta J, Ayerdi E, Cabezas AI, Orive G et al.. Effectiveness of autologous preparation rich in growth factors for the treatment of chronic cutaneous ulcers. J Biomed Mater Res B Ap Biomater. 2008; 84:415-421.
• [4]Anitua E, Sanchez M, Orive G. Potential of endogenous regenerative technology for in situ regenerative medicine. Ad Drug Deliv Rev. 2010; 62:741-752.
• [5]Cole BJ, Seroyer ST, Filardo G, Bajaj S, Fortier LA. Platelet-rich plasma: where are we now and where are we going? Sports Health. 2010; 2:203-210.
• [6]Filardo G, Presti ML, Kon E, Marcacci M. Nonoperative biological treatment approach for partial Achilles tendon lesion. Orthopedics. 2010; 33:120-123.
• [7]Kerrigan SW, Cox D. Platelet-bacterial interactions. Cell Mol Life Sci. 2010; 67:513-523.
• [8]Yeaman MR. Platelets in defense against bacterial pathogens. Cell Mol Life Sci. 2010; 67:525-544.
• [9]Yeaman MR. Bacterial-platelet interactions: virulence meets host defense. Future Microbiol. 2010; 5:471-506.
• [10]Semple JW, Italiano JE, Freedman J. Platelets and the immune continuum. Nat Rev Immunol. 2011; 11:264-274.
• [11]Wiesner J, Vilcinskas A. Antimicrobial peptides: the ancient arm of the human immune system. Virulence. 2010; 1:440-464.
• [12]Yang D, Chen Q, Hoover DM, Staley P, Tucker KD, Lubkowski J et al.. Many chemokines including CCL20/MIP-3alpha display antimicrobial activity. J Leukoc Biol. 2003; 74:448-455.
• [13]Yount NY, Waring AJ, Gank KD, Welch WH, Kupferwasser D, Yeaman MR. Structural correlates of antimicrobial efficacy in IL-8 and related human kinocidins. Biochim Biophys Acta. 1768; 2007:598-608.
• [14]Wasterlain AS, Braun HG, Dragoo JL. Contest and formulations of platelet-rich plasma. Operative Techni Orthop. 2012; 22:33-42.
• [15]Ehrenfest DMD, Sammartino G, Shibli JA, Wang H, Zou D, Bernard J. Guidelines for the publication of articles related to platelet concentrates (Platelet-Rich Plasma - PRP, or Platelet-Rich Fibrin - PRF): the international classification of the POSEIDO. Poseido J. 2013; 1:17-28.
• [16]Cieslik-Bielecka A, Dohan Ehrenfest DM, Lubkowsha ABT. Microbicidal properties of leukocyte and platelet-rich plasma/fibrin (L-PRP/L-PRF): new perspectives. J Biol Regul Homeost Agents. 2012; 26:43S-52S.
• [17]Roffi A, Filardo G, Kon E, Marcacci M. Does PRP enhance bone integration with grafts, graft substitutes, or implants? A systematic review. BMC Musculoskelet Disord. 2013; 14:330. BioMed Central Full Text
• [18]Filardo G, Kon E, Roffi A, Di Matteo B, Merli ML, Marcacci M. Platelet-rich plasma: why intra-articular? A systematic review of preclinical studies and clinical evidence on PRP for joint degeneration. Knee Surg Sports Traumatol Arthrosc. 2013. [E-pub ahead of print]
• [19]Filardo G, Kon E, Di Matteo B, Pelotti P, Di Martino A, Marcacci M. Platelet-rich plasma for the treatment of patellar tendinopathy: clinical and imaging findings at medium-term follow-up. Int Orthop. 2013; 37:1583-1589.
• [20]Perdisa F, Filardo G, Di Matteo B, Marcacci M, Kon E. Platelet rich plasma: a valid augmentation for cartilage scaffolds? A systematic review. Histol Histopathol. 2014; 29:805-814.
• [21]Filardo G, Kon E, Di Matteo B, Di Martino A, Tesei G, Pelotti P et al.. Platelet-rich plasma injections for the treatment of refractory Achilles tendinopathy: results at 4 years. Blood Transfus. 2014; 12:533-540.
• [22]Bielecki TM, Gazdzik TS, Arendt J, Szczepanski T, Krol W, Wielkoszynski T. Antibacterial effect of autologous platelet gel enriched with growth factors and other active substances: an in vitro study. J Bone Joint Surg (Br). 2007; 89:417-420.
• [23]Moojen DJ, Everts PA, Schure RM, Overdevest EP, Van ZA, Knape JT et al.. Antimicrobial activity of platelet-leukocyte gel against Staphylococcus aureus. J Orthop Re. 2008; 26:404-410.
• [24]Anitua E, Alonso R, Girbau C, Aguirre JJ, Muruzabal F, Orive G. Antibacterial effect of plasma rich in growth factors (PRGF®-Endoret®) against Staphylococcus aureus and Staphylococcus epidermidis strains. Clin Exp Dermatol. 2012; 37:652-657.
• [25]Burnouf T, Chou ML, Wu YW, Su CY, Lee LW. Antimicrobial activity of platelet (PLT)-poor plasma, PLT-rich plasma, PLT gel, and solvent/detergent-treated PLT lysate biomaterials against wound bacteria. Transfusion. 2013; 53:138-146.
• [26]Drago L, Bortolin M, Vassena C, Taschieri S, Del Fabbro M. Antimicrobial activity of pure platelet-rich plasma against microorganisms isolated from oral cavity. BMC Microbiol. 2013; 13:47. BioMed Central Full Text
• [27]Assirelli E, Filardo G, Mariani E, Kon E, Roffi A, Vaccaro F, et al. Effect of two different preparations of platelet-rich plasma on synoviocytes. Knee Surg Sports Traumatol Arthrosc. 2014. [E-pub ahead of print]
• [28]Cavallo C, Filardo G, Mariani E, Kon E, Marcacci M, Pereira Ruiz MT et al.. Comparison of platelet-rich plasma formulations for cartilage healing: an in vitro study. J Bone Joint Surg Am. 2014; 96:423-429.
• [29]Lautenbach E, Patel JB, Bilker WB, Edelstein PH, Fishman NO. Extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae: risk factors for infection and impact of resistance on outcomes. Clin Infect Dis. 2001; 32:1162-1171.
• [30]Zalavras CG, Patzakis MJ, Holtom P. Local antibiotic therapy in the treatment of open fractures and osteomyelitis. Clin Orthop Relat Res. 2004; 427:86-93.
• [31]Anitua E, Orive G, Aguirre JJ, Andia I. Five-year clinical evaluation of short dental implants placed in posterior areas: a retrospective study. J Periodontol. 2008; 79:42-48.
• [32]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; 15:830-839.
• [33]Mariani E, Cattini L, Neri S, Malavolta M, Mocchegiani E, Ravaglia G et al.. Simultaneous evaluation of circulating chemokine and cytokine profiles in elderly subjects by multiplex technology: relationship with zinc status. Biogerontology. 2006; 7:449-459.
• [34]Mariani E, Neri S, Cattini L, Mocchegiani E, Malavolta M, Dedoussis GV et al.. Effect of zinc supplementation on plasma IL-6 and MCP-1 production and NK cell function in healthy elderly: interactive influence of +647 MT1a and −174 IL-6 polymorphic alleles. Exp Gerontol. 2008; 43:462-471.
• [35]Mader JT, Calhoum J. Osteomyelitis. In: Principles of Infectious Diseases. Mandell GL, Bennet JE, Dolin R, editors. Churchill Livingston, Philadelphia; 2000: p.1182-1196.
• [36]Prabhakara R, Harro JM, Leid JG, Harris M, Shirtliff ME. Murine immune response to a chronic Staphylococcus aureus biofilm infection. Infect Immun. 2011; 79:1789-1796.
• [37]Bush K. New beta-lactamases in gram-negative bacteria: diversity and impact on the selection of antimicrobial therapy. Clin Infect Dis. 2001; 32:1085-1089.
• [38]Kang CI, Kim SH, Park WB, Lee KD, Kim HB, Kim EC et al.. Bloodstream infections due to extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae: risk factors for mortality and treatment outcome, with special emphasis on antimicrobial therapy. Antimicrob Agents Chemother. 2004; 48:4574-4581.
• [39]Bausset O, Giraudo L, Veran J, Magalon J, Coudreuse JM, Magalon G et al.. Formulation and storage of platelet-rich plasma homemade product. Biores Open Access. 2012; 1:115-123.
• [40]Anitua E, Muruzabal F, Pino A, Merayo-Lloves J, Orive G. Biological Stability of Plasma Rich in Growth Factors Eye Drops After Storage of 3 Months. Cornea. 2013; 32:1386.
• [41]Murphy MB, Blashki D, Buchanan RM, Yazdi IK, Ferrari M, Simmons PJ et al.. Adult and umbilical cord blood-derived platelet-rich plasma for mesenchymal stem cell proliferation, chemotaxis, and cryo-preservation. Biomaterials. 2012; 33:5308-5316.
• [42]Yuan T, Zhang CQ, Wang JH. Augmenting tendon and ligament repair with platelet-rich plasma (PRP). Muscles Ligaments Tendons J. 2013; 3:139-149.
• [43]Hampton MB, Kettle AJ, Winterbourn CC. Involvement of superoxide and myeloperoxidase in oxygen-dependent killing of Staphylococcus aureus by neutrophils. Infect Immun. 1996; 64:3512-3517.
• [44]Mariani E, Filardo G, Canella V, Berlingeri A, Bielli A, Cattini L et al.. Platelet-rich plasma affects bacterial growth in vitro. Cytotherapy. 2014; 16:1294-1304.
• [45]Yung SC, Murphy PM. Antimicrobial chemokines. Front Immunol. 2012; 3:276.
• [46]Nguyen LT, Kwakman PH, Chan DI, Liu Z, De BL, Zaat SA et al.. Exploring platelet chemokine antimicrobial activity: nuclear magnetic resonance backbone dynamics of NAP-2 and TC-1. Antimicrob Agents Chemother. 2011; 55:2074-2083.
• [47]Zlotnik A, Yoshie O. The chemokine superfamily revisited. Immunity. 2012; 36:705-716.
• [48]Sipert CR, Morandini AC, Modena KC, Dionisio TJ, Machado MA, Oliveira SH et al.. CCL3 and CXCL12 production in vitro by dental pulp fibroblasts from permanent and deciduous teeth stimulated by Porphyromonas gingivalis LPS. J Appl Oral Sci. 2013; 21:99-105.
• [49]Garlet GP. To heal or not to heal? Chemokines as determinants of constructive or destructive inflammatory microenvironments. J Appl Oral Sci. 2013;21.
• [50]Hack CE, De Groot ER, Felt-Bersma RJ, Nuijens JH, Strack Van Schijndel RJ, Eerenberg-Belmer AJ et al.. Increased plasma levels of interleukin-6 in sepsis. Blood. 1989; 74:1704-1710.
• [51]Van der Poll T, Keogh CV, Guirao X, Buurman WA, Kopf M, Lowry SF. Interleukin-6 gene-deficient mice show impaired defense against pneumococcal pneumonia. J Infect Dis. 1997; 176:439-444.
• [52]Jones SA. Directing transition from innate to acquired immunity: defining a role for IL-6. J Immunol. 2005; 175:3463-3468.