Download PDF
Vet Comp Orthop Traumatol 2014; 27(05): 372-378
DOI: 10.3415/VCOT-14-04-0054
Original Research
Schattauer GmbH

Bacteriostatic effect of equine pure plateletrich plasma and other blood products against methicillin-sensitive Staphylococcus aureus

An in vitro study
C. López
1   Grupo de Investigación Terapia Regenerativa, Departamento de Salud Animal, Universidad de Caldas, Manizales, Colombia
,
J. U. Carmona
1   Grupo de Investigación Terapia Regenerativa, Departamento de Salud Animal, Universidad de Caldas, Manizales, Colombia
,
C. E. Giraldo
1   Grupo de Investigación Terapia Regenerativa, Departamento de Salud Animal, Universidad de Caldas, Manizales, Colombia
,
M. E. Álvarez
1   Grupo de Investigación Terapia Regenerativa, Departamento de Salud Animal, Universidad de Caldas, Manizales, Colombia
› Author Affiliations
Further Information

Publication History

Received: 09 April 2014

Accepted: 12 July 2014

Publication Date:
22 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

Objectives: 1) To evaluate the bacteriostatic in vitro effect of pure platelet-rich plasma (P-PRP), pure platelet-rich gel (P-PRG), leukocyte-poor gel (LPG), platelet-poor plasma (PPP), and heat inactivated plasma (IP) against methicillin-sensitive Staphylococcus aureus (MSSA) over a period of 24 hours. 2) To determine the degradation of platelet factor-4 (PF-4), transforming growth factor beta 1 (TGF-β1), and platelet-derived growth factor isoform BB (PDGF-BB) in these equine blood components. 3) To establish correlations between platelet and leukocyte counts, PF-4 concentrations, and MSSA growth.

Methods: Fourteen horses were used. Blood components were obtained by a manual protocol. Every blood component was mixed with MSSA and Müller-Hinton Broth and cultured at 37°C for 24 hours. Samples for the determination of bacterial growth (colonyforming units) and PF-4, TGF-β1 and PDGFBB concentrations were taken at one, four, eight, 12 and 24 hours.

Results: The bacterial growth was significantly (p = 0.01) inhibited for P-PRP, P-PRG, LPG and PPP in comparison with IP and, the positive control group during the first 12 hours. The P-PRG had higher and sustained TGF-β1 and PDGF-BB concentrations over time in comparison with the other blood components.

Clinical significance: The plasma complement could be one of the most responsible components of the in vitro bacteriostatic effect of P-PRP, P-PRG, LPG and PPP against MSSA. Additionally, P-PRG was the better biomaterial because it had an acceptable bacteriostatic effect and the highest concentration of growth factors.

Keywords

Growth factor - leukocyte-reduced platelet concentrate - platelet factor 4 - wound infection
 

  • References

  • 1 Álvarez ME, Giraldo CE, Carmona JU. Monitoring bacterial contamination in equine platelet concentrates obtained by the tube method in a clean laboratory environment under three different technical conditions. Equine Vet J 2009; 42: 63-67.
    CrossrefPubMedSearch in Google Scholar
  • 2 Haenni M, Targant H, Forest K. et al Retrospective study of necropsy-associated coagulase-positive staphylococci in horses. J Vet Diagn Invest 2010; 22: 953-956.
    CrossrefPubMedSearch in Google Scholar
  • 3 Burton S, Reid-Smith R, McClure JT. et al Staphylococcus aureus colonization in healthy horses in Atlantic Canada. Can Vet J 2008; 49: 797-799.
    PubMedSearch in Google Scholar
  • 4 Ingle-Fehr JE, Baxter GM, Howard RD. et al Bacterial culturing of ventral median celiotomies for prediction of postoperative incisional complications in horses. Vet Surg 1997; 26: 7-13.
    CrossrefPubMedSearch in Google Scholar
  • 5 Macdonald DG, Morley PS, Bailey JV. et al An examination of the occurrence of surgical-wound infection following equine orthopedic-surgery (1981-1990). Equine Vet J 1994; 26: 323-326.
    CrossrefPubMedSearch in Google Scholar
  • 6 Taylor AH, Mair TS, Smith LJ. et al Bacterial culture of septic synovial structures of horses: Does a positive bacterial culture influence prognosis?. Equine Vet J 2010; 42: 213-218.
    CrossrefPubMedSearch in Google Scholar
  • 7 Carmona JU, Arguelles D, Climent F. et al Autologous platelet concentrates as a treatment of horses with osteoarthritis: A preliminary pilot clinical study. J Equine Vet Sci 2007; 27: 167-170.
    CrossrefPubMedSearch in Google Scholar
  • 8 Argüelles D, Carmona JU, Climent F. et al Autologous platelet concentrates as a treatment for musculoskeletal lesions in five horses. Vet Rec 2008; 162: 208-211.
    CrossrefPubMedSearch in Google Scholar
  • 9 Maciel FB, Derossi R, Módolo TJC. et al Scanning electron microscopy and microbiological evaluation of equine burn wound repair after platelet-rich plasma gel treatment. Burns 2012; 38: 1058-1065.
    CrossrefPubMedSearch in Google Scholar
  • 10 Alsousou J, Ali A, Willett K. et al The role of platelet-rich plasma in tissue regeneration. Platelets 2013; 24: 173-182.
    CrossrefPubMedSearch in Google Scholar
  • 11 Argüelles D, Carmona JU, Pastor J. et al Evaluation of single and double centrifugation tube methods for concentrating equine platelets. Res Vet Sci 2006; 81: 237-245.
    CrossrefPubMedSearch in Google Scholar
  • 12 Sutter WW, Kaneps AJ, Bertone AL. Comparison of hematologic values and transforming growth factor-beta and isulin-like growth factor concentrations in platelets concentrates obtained by use of buffy coat and apheresis methods from equine blood. Am J Vet Res 2004; 65: 924-930.
    CrossrefPubMedSearch in Google Scholar
  • 13 Giraldo CE, López C, Álvarez ME. et al Effects of the breed, sex and age on cellular content and growth factor release from equine pure-platelet rich plasma and pure-platelet rich gel. BMC Vet Res 2013; 9: 29.
    CrossrefPubMedSearch in Google Scholar
  • 14 Carmona JU, López C, Sandoval JA. Review of the currently available systems to obtain platelet related products to treat equine musculoskeletal injuries. Rec Pat Reg Med 2013; 3: 148-159.
    PubMedSearch in Google Scholar
  • 15 Blair P, Flaumenhaft R. Platelet α-granules: Basic biology and clinical correlates. Blood Rev 2009; 23: 177-189.
    CrossrefPubMedSearch in Google Scholar
  • 16 Yeaman MR. Platelets in defense against bacterial pathogens. Cell Mol Life Sci 2010; 67: 525-544.
    CrossrefPubMedSearch in Google Scholar
  • 17 Álvarez M, López C, Giraldo C. et al In vitro bactericidal activity of equine platelet concentrates, platelet poor plasma, and plasma against methicillin-resistant Staphylococcus aureus. Arch Med Vet 2011; 43: 155-161.
    CrossrefPubMedSearch in Google Scholar
  • 18 Burnouf T, Chou ML, Wu YW. et al 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.
    CrossrefPubMedSearch in Google Scholar
  • 19 Cieslik-Bielecka A, Bielecki T, Gazdzik TS. et al Autologous platelets and leukocytes can improve healing of infected high-energy soft tissue injury. Transfus Apher Sci 2009; 41: 9-12.
    CrossrefPubMedSearch in Google Scholar
  • 20 Wang HF, Gao YS, Yuan T. et al Chronic calcaneal osteomyelitis associated with soft-tissue defect could be successfully treated with platelet-rich plasma: A case report. Int Wound J 2013; 10: 105-109.
    CrossrefPubMedSearch in Google Scholar
  • 21 Yuan T, Zhang CQ, Zeng BF. Treatment of chronic femoral osteomyelitis with platelet-rich plasma (PRP): A case report. Transfus Apher Sci 2008; 38: 167-173.
    CrossrefPubMedSearch in Google Scholar
  • 22 López C, Carmona JU, Samudio I. Use of autologous platelet concentrates as treatment for a scapular fracture and brachial plexus nerve injury produced by a gunshot in a horse. Arch Med Vet 2010; 42: 209-214.
    PubMedSearch in Google Scholar
  • 23 Bielecki TM, Gazdzik TS, Arendt J. et al Antibacterial effect of autologous platelet gel enriched with growth factors and other active substances - An in vitro study. J Bone Joint Surg B 2007; 89.: 417-420.
    PubMedSearch in Google Scholar
  • 24 Moojen DJF, Everts PAM, Schure RM. et al Antimicrobial activity of platelet-leukocyte gel against Staphylococcus aureus. J Orthop Res 2008; 26: 404-410.
    CrossrefPubMedSearch in Google Scholar
  • 25 Anitua E, Alonso R, Girbau C. et al Antibacterial effect of plasma rich in growth factors (PRGF®- Endoret®) against Staphylococcus aureus and Staphylococcus epidermidis strains. Clin Exp Dermatol 2012; 37: 652-657.
    CrossrefPubMedSearch in Google Scholar
  • 26 Drago L, Bortolin M, Vassena C. et al Antimicrobial activity of pure platelet-rich plasma against microorganisms isolated from oral cavity. BMC Microbiol 2013; 13: 47.
    CrossrefPubMedSearch in Google Scholar
  • 27 Chen L, Wang C, Liu H. et al Antibacterial effect of autologous platelet-rich gel derived from subjects with diabetic dermal ulcers in vitro. J Diabetes Res 2013 2013; 269527.
    PubMedSearch in Google Scholar
  • 28 Tang Y-Q, Yeaman MR, Selsted ME. Antimicrobial peptides from human platelets. Infect Immun 2002; 70: 6524-6533.
    CrossrefPubMedSearch in Google Scholar
  • 29 Argüelles D, Carmona JU, Pastor J. et al Evaluation of single and double centrifugation tube methods for concentrating equine platelets. Res Vet Sci 2006; 81: 237-245.
    CrossrefPubMedSearch in Google Scholar
  • 30 Gilbert P, Brown MRW, Costerton JW. Inocula for antimicrobial sensitivity testing: a critical review. J Antimicrob Chemother 1987; 20: 147-154.
    PubMedSearch in Google Scholar
  • 31 Delesalle C, Van De Walle GR, Nolten C. et al Determination of the source of increased serotonin (5-HT) concentrations in blood and peritoneal fluid of colic horses with compromised bowel. Equine Vet J 2008; 40: 326-331.
    CrossrefPubMedSearch in Google Scholar
  • 32 Penha-goncalves MN, Onions DE, Nicolson L. Cloning and sequencing of equine transforming growth factor-beta 1 (TGF beta-1) cDNA. DNA Seq 1997; 7: 375-378.
    PubMedSearch in Google Scholar
  • 33 Donnelly BP, Nixon AJ, Haupt JL. Nucleotide structure of equine platelet-derived growth factor-A and -B and expression in horses with induced acute tendinitis. Am J Vet Res 2006; 67: 1218-1225.
    CrossrefPubMedSearch in Google Scholar
  • 34 Yavuz SŞ, Tarçın Ö, Ada S. et al Incidence, aetiology, and control of sternal surgical site infections. J Hosp Infect 2013; 85: 206-212.
    CrossrefPubMedSearch in Google Scholar
  • 35 Klinger MH, Jelkmann W. Role of blood platelets in infection and inflammation. J Interferon Cytokine Res 2002; 22: 913-922.
    CrossrefPubMedSearch in Google Scholar
  • 36 Mast BA, Schultz GS. Interactions of cytokines, growth factors, and proteases in acute and chronic wounds. Wound Repair Reg 1996; 4: 411-420.
    CrossrefPubMedSearch in Google Scholar
  • 37 Yount NY, Gank KD, Xiong YQ. et al Platelet microbicidal protein 1 Structural themes of a multifunctional antimicrobial peptide. Antimicrob Agents Chemother 2004; 48: 4395-4404.
    CrossrefPubMedSearch in Google Scholar
  • 38 Xiong YQ, Bayer AS, Yeaman MR. Inhibition of intracellular macromolecular synthesis in Staphylococcus aureus by thrombin-induced platelet microbicidal proteins. J Infect Dis 2002; 185: 348-356.
    CrossrefPubMedSearch in Google Scholar
  • 39 Hnasko R, Lin A, McGarvey JA. et al A rapid method to improve protein detection by indirect ELISA. Biochem Biophys Res Commun 2011; 410: 726-731.
    CrossrefPubMedSearch in Google Scholar
  • 40 Schraw T, Whiteheart S. The development of a quantitative enzyme-linked immunosorbent assay to detect human platelet factor 4. Transfusion 2005; 45: 717-724.
    CrossrefPubMedSearch in Google Scholar
  • 41 Payne WG, Wright TE, Ko F. et al Bacterial degradation of growth factors. J Applied Res 2003; 3: 35-40.
    PubMedSearch in Google Scholar
  • 42 Moor AN, Vachon DJ, Gould LJ. Proteolytic activity in wound fluids and tissues derived from chronic venous leg ulcers. Wound Repair Reg 2009; 17: 832-839.
    CrossrefPubMedSearch in Google Scholar
  • 43 Shenkman B, Rubinstein E, Dardik R. et al Activated platelets mediate Staphylococcus aureus deposition on subendothelial extracellular matrix: the role of glycoprotein Ib. Platelets 1999; 10: 36-44.
    CrossrefPubMedSearch in Google Scholar
  • 44 Ledur A, Fitting C, David B. et al Variable estimates of cytokine levels produced by commercial ELISA kits: results using international cytokine standards. J Immunol Methods 1995; 186: 171-179.
    CrossrefPubMedSearch in Google Scholar