Antibacterial effect of Leucocyte and Platelet Rich-Fibrin against Klebsiella pneumoniae and Staphylococcus aureus – A pilot study

Ana Francisca Mendes; Cristina Coelho; Inês Mendes; Laura Silva; António  Melo-Ferraz; Maria Begoña Criado; Maria do Céu Monteiro; Paulo Miller

doi:10.48797/sl.2024.268

Authors

Ana Francisca Mendes Dental Sciences Department, University Institute of Health Sciences (IUCS), CESPU, CRL, 4585-116 Gandra, Portugal https://orcid.org/0009-0005-9369-9862
Cristina Coelho UNIPRO, Oral Pathology and Rehabilitation Research Unit, University Institute of Health Sciences-CESPU (IUCS-CESPU), 4585-116 Gandra, Portugal
Inês Mendes Dental Sciences Department, University Institute of Health Sciences (IUCS), CESPU, CRL, 4585-116 Gandra, Portugal https://orcid.org/0009-0007-2647-7498
Laura Silva Dental Sciences Department, University Institute of Health Sciences (IUCS), CESPU, CRL, 4585-116 Gandra, Portugal
António Melo-Ferraz UNIPRO, Oral Pathology and Rehabilitation Research Unit, University Institute of Health Sciences-CESPU (IUCS-CESPU), 4585-116 Gandra, Portugal; Associate Laboratory i4HB – Institute for Health and Bioeconomy, University Institute of Health Sciences – CESPU, 4585-116 Gandra, Portugal; UCIBIO – Applied Molecular Biosciences Unit, Toxicologic Pathology Research Laboratory, University Institute of Health Sci-ences (1H-TOXRUN, IUCS-CESPU), 4585-116 Gandra, Portugal https://orcid.org/0000-0003-0289-8064
Maria Begoña Criado Associate Laboratory i4HB – Institute for Health and Bioeconomy, University Institute of Health Sciences – CESPU, 4585-116 Gandra, Portugal; UCIBIO – Applied Molecular Biosciences Unit, Toxicologic Pathology Research Laboratory, University Institute of Health Sci-ences (1H-TOXRUN, IUCS-CESPU), 4585-116 Gandra, Portugal https://orcid.org/0000-0003-1750-5614
Maria do Céu Monteiro Associate Laboratory i4HB – Institute for Health and Bioeconomy, University Institute of Health Sciences – CESPU, 4585-116 Gandra, Portugal; UCIBIO – Applied Molecular Biosciences Unit, Toxicologic Pathology Research Laboratory, University Institute of Health Sci-ences (1H-TOXRUN, IUCS-CESPU), 4585-116 Gandra, Portugal https://orcid.org/0000-0001-9719-9746
Paulo Miller UNIPRO, Oral Pathology and Rehabilitation Research Unit, University Institute of Health Sciences-CESPU (IUCS-CESPU), 4585-116 Gandra, Portugal https://orcid.org/0000-0001-5500-7603

DOI:

https://doi.org/10.48797/sl.2024.268

Keywords:

leukocyte and platelet rich fibrin (L-PRF); antibacterial effect; platelet concentrate; oral pathogen; Klebsiella pneumoniae; Staphylococcus aureus

Abstract

Leukocyte and platelet-rich fibrin (L-PRF) is a platelet concentrate used in diverse areas of tissue healing and regeneration. Thus, its antibacterial properties are promising. In this study, we evaluated the antibacterial effect of L-PRF, in its exudate and membrane forms, against oral cavity opportunistic bacteria Klebsiella pneumoniae and Staphylococcus aureus. Blood samples were collected from three healthy donors. Blood was centrifuged at 2,700 rpm for 12 minutes. This resulted in an L-PRF membrane, and then, from its compression, an L-PRF exudate. These two forms of L-PRF were placed on Mueller-Hinton Agar previously inoculated with the bacteria. The L-PRF membrane was placed directly into the culture medium, and the L-PRF exudate was impregnated in different quantities onto paper disks (Oxoid). The membrane inhibition values varied between 11 and 13 mm for K. pneumoniae and 13 mm for S. aureus. For the L-PRF exudate, inhibition only occurred for quantities of 5 and 20 μL of exudate in the case of K. pneumoniae and for 20 μL of exudate in the case of S. aureus. Thus, L-PRF products effectively inhibit bacterial growth, particularly opportunistic bacteria, such as K. pneumoniae and S. aureus.

References

Siawasch, S.A.M.; Andrade, C.; Castro, A.B.; Teughels, W.; Temmerman, A.; Quirynen, M. Impact of local and systemic antimicrobials on leukocyte- and platelet rich fibrin: an in vitro study. Sci Rep 2022, 12, 2710, doi:10.1038/s41598-022-06473-4.

Micko, L.; Salma, I.; Skadins, I.; Egle, K.; Salms, G.; Dubnika, A. Can Our Blood Help Ensure Antimicrobial and Anti-Inflammatory Properties in Oral and Maxillofacial Surgery? Int J Mol Sci 2023, 24, doi:10.3390/ijms24021073.

Melo-Ferraz, A.; Coelho, C.; Miller, P.; Criado, M.B.; Monteiro, M.C. Platelet activation and antimicrobial activity of L-PRF: a preliminary study. Mol Biol Rep 2021, 48, 4573-4580, doi:10.1007/s11033-021-06487-7.

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, doi:10.1080/09537104.2017.1293812.

Sindhusha, V.B.; Ramamurthy, J. Comparison of Antimicrobial Activity of Injectable Platelet-Rich Fibrin (i-PRF) and Leukocyte and Platelet-Rich Fibrin (l-PRF) Against Oral Microbes: An In Vitro Study. Cureus 2023, 15, e46196, doi:10.7759/cureus.46196.

Pham, T.A.V.; Tran, T.T.P. Antimicrobial effect against Aggregatibacter actinomycetemcomitans of advanced and injectable platelet-rich fibrin from patients with periodontal diseases versus periodontally healthy subjects. J Oral Biol Craniofac Res 2023, 13, 332-336, doi:10.1016/j.jobcr.2023.02.011.

Phuong Tran, T.T.; Vu Pham, T.A. Effect of advanced and injectable platelet-rich fibrins against Aggregatibacter actinomycetemcomitans in subjects with or without periodontal diseases. J Dent Sci 2023, 18, 491-496, doi:10.1016/j.jds.2022.09.014.

Jasmine, S.; A, T.; Janarthanan, K.; Krishnamoorthy, R.; Alshatwi, A.A. Antimicrobial and antibiofilm potential of injectable platelet rich fibrin-a second-generation platelet concentrate-against biofilm producing oral staphylococcus isolates. Saudi J Biol Sci 2020, 27, 41-46, doi:10.1016/j.sjbs.2019.04.012.

Feng, M.; Wang, Y.; Zhang, P.; Zhao, Q.; Yu, S.; Shen, K.; Miron, R.J.; Zhang, Y. Antibacterial effects of platelet-rich fibrin produced by horizontal centrifugation. Int J Oral Sci 2020, 12, 32, doi:10.1038/s41368-020-00099-w.

Wang, G.; Zhao, G.; Chao, X.; Xie, L.; Wang, H. The Characteristic of Virulence, Biofilm and Antibiotic Resistance of Klebsiella pneumoniae. Int J Environ Res Public Health 2020, 17, doi:10.3390/ijerph17176278.

Loffler, B.; Tuchscherr, L. Staphylococcus aureus Toxins: Promoter or Handicap during Infection? Toxins (Basel) 2021, 13, doi:10.3390/toxins13040287.

Tacconelli, E.; Carrara, E.; Savoldi, A.; Harbarth, S.; Mendelson, M.; Monnet, D.L.; Pulcini, C.; Kahlmeter, G.; Kluytmans, J.; Carmeli, Y., et al. Discovery, research, and development of new antibiotics: the WHO priority list of antibiotic-resistant bacteria and tuberculosis. Lancet Infect Dis 2018, 18, 318-327, doi:10.1016/S1473-3099(17)30753-3.

Ruhal, R.; Kataria, R. Biofilm patterns in gram-positive and gram-negative bacteria. Microbiol Res 2021, 251, 126829, doi:10.1016/j.micres.2021.126829.

Dagnino, A.P.A.; Campos, M.M.; Silva, R.B.M. Kinins and Their Receptors in Infectious Diseases. Pharmaceuticals (Basel) 2020, 13, doi:10.3390/ph13090215.

Campos, J.; Pires, M.F.; Sousa, M.; Campos, C.; da Costa, C.; Sampaio-Maia, B. Unveiling the Relevance of the Oral Cavity as a Staphylococcus aureus Colonization Site and Potential Source of Antimicrobial Resistance. Pathogens 2023, 12, doi:10.3390/pathogens12060765.

Rodriguez Sanchez, F.; Verspecht, T.; Castro, A.B.; Pauwels, M.; Andres, C.R.; Quirynen, M.; Teughels, W. Antimicrobial Mechanisms of Leucocyte- and Platelet Rich Fibrin Exudate Against Planktonic Porphyromonas gingivalis and Within Multi-Species Biofilm: A Pilot Study. Front Cell Infect Microbiol 2021, 11, 722499, doi:10.3389/fcimb.2021.722499.

Castro, A.B.; Herrero, E.R.; Slomka, V.; Pinto, N.; Teughels, W.; Quirynen, M. Antimicrobial capacity of Leucocyte-and Platelet Rich Fibrin against periodontal pathogens. Sci Rep 2019, 9, 8188, doi:10.1038/s41598-019-44755-6.