The evaluation of leukocyte-platelet rich fibrin as an anti-inflammatory autologous biological additive. A novel in vitro study
Objectives: To investigate the use of leukocyte-platelet rich fibrin on suppressing the porphyromonas gingivalis (PG-LPS)-induced secretion of proinflammatory cytokines.
Methods:This quantitative experimental study was conducted at the School and Hospital of Stomatology, Jilin University, Changchun, China, between September 2017 and January 2019. A modified technique was used to obtain human gingival fibroblast cells (HGFCs). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Cell Counting Kit-8 tests were established to determine the proliferation rate. Human gingival fibroblast cells were treated by PG-LPS at different periods and the isolated mRNA was subjected to reverse transcription polymerase chain reaction and real time quantitative polymerase chain reaction. The release of platelet-derived growth factor and transforming-growth factor-β1 at various time intervals was observed.
Results: We successfully established a modified technique for the production of HGFCs culture. One µg/mL PG-LPS was the recommended concentration to inhibit fibroblast proliferation. The expression of the pro-inflammatory cytokines messenger ribnucleic acid was notably raised at 3 and 6 hours post-PG-LPS treatment. The cumulative release of growth factors peaked during the first 24 hours and the production continued for 10 days. However, the fibroblast expression of cytokines was significantly suppressed after treatment with leucocyte- and platelet-rich fibrin (L-PRF).
Conclusion: This study provided a novel way of obtaining HGFCs and greater understanding of the clinical impacts through the assessment of the anti-inflammatory properties of L-PRF in vitro.
Saudi Med J 2019; Vol. 40 (7): 657-668
How to cite this article:
Mudalal M, Sun X, Li X, Zhou Y. The evaluation of leukocyte-platelet rich fibrin as an anti-inflammatory autologous biological additive: A novel in vitro study. Saudi Med J 2019; 40 (7): 657-668. doi:10.15537/smj.2019.7.24302.
Chen FM, Liu X. Advancing biomaterials of human origin for tissue engineering. Prog Polym Sci 2016; 53: 86-168.
Kiran NK, Mukunda K, Tilak Raj TN. Platelet Concentrates: A Promising Innovation in Dentistry. J Den Sci Res 2011; 2: 50-61.
Bielecki T, Dohan Ehrenfest DM. Platelet-rich plasma (PRP) and platelet-rich fibrin (PRF): surgical adjuvants, preparations for in situ regenerative medicine and tools for tissue engineering. Curr Pharm Biotechnol 2012; 13: 1121-1130.
Dohan Ehrenfest DM, Rasmusson L, Albrektsson T. Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF). Trends Biotechnol 2009; 27: 158-167.
Dohan DM, Choukroun J, Diss A, Dohan SL, Dohan AJ, Mouhyi J, et al. Platelet-rich fibrin (PRF): a second-generation platelet concentrate. Part I: technological concepts and evolution. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2006; 101: e37-e44.
Mudalal M, Zhou YM. Biological additives and platelet concentrates for tissue engineering on regenerative dentistry basic Science and concise review. Asian J Pharm 2017; 11: 255-263.
Kang YH, Jeon SH, Park JY, Chung JH, Choung YH, Choung HW, et al. Platelet-rich fibrin is a Bioscaffold and reservoir of growth factors for tissue regeneration. Tissue Eng Part A 2011; 17: 349-359.
Zumstein MA, Bielecki T, Dohan Ehrenfest DM. The future of platelet concentrates in sports medicine: platelet-rich plasma, platelet-rich fibrin, and the impact of scaffolds and cells on the long-term delivery of growth factors. Oper Techn Sport Med 2011; 19: 190-197.
Bertrand-Duchesne MP, Grenier D, Gagnon G. Epidermal growth factor released from platelet-rich plasma promotes endothelial cell proliferation in vitro. J Periodontal Res 2010; 45: 87-93.
Dohan Ehrenfest DM, Bielecki T, Jimbo R, Barbé G, Del Corso M, Inchingolo F, et al. Do the fibrin architecture and leukocyte content influence the growth factor release of platelet concentrates? An evidence-based answer comparing a pure platelet-rich plasma (P-PRP) gel and a leukocyte- and platelet-rich fibrin (L-PRF). Curr Pharm Biotechnol 2012; 13: 1145-1152.
Yu X, Ge S, Chen S, Xu Q, Zhang J, Guo H, et al. Human gingiva-derived mesenchymal stromal cells contribute to periodontal regeneration in beagle dogs. Cells Tissues Organs 2013; 198: 428-437.
Mak K, Manji A, Gallant-Behm C, Wiebe C, Hart DA, Larjava H, et al. Scarless healing of oral mucosa is characterized by faster resolution of inflammation and control of myofibroblast action compared to skin wounds in the red Duroc pig model. J Dermatol Sci 2009; 56: 168-180.
Xu Q, Ji Q, Yang J, Xu X, Yu, X, Wang Z, et al. Comparison of the characteristics of human gingival fibroblasts isolated by tissue explants and by enzyme digestion methods. Int J Clin Exp Med 2016; 9: 19757-19763.
Malekzadeh R, Hollinger JO, Buck D, Adams DF, McAllister BS. Isolation of human osteoblast-like cells and in vitro amplification for tissue engineering. J Periodontol 1998; 69: 1256-1262.
Zhang Q, Nguyen AL, Shi S, Hill C, Wilder-Smith P, Krasieva TB, et al. Three-dimensional spheroid culture of human gingiva-derived mesenchymal stem cells enhances mitigation of chemotherapy-induced oral mucositis. Stem Cells Dev 2012; 21: 937-947.
Saczko J, Dominiak M, Kulbacka J, Chwiłkowska A, Krawczykowska H. A simple and established method of tissue culture of human gingival fibroblasts for gingival augmentation. Folia Histochem Cyto 2008; 46: 117-119.
Lang CH, Hong-Brown L, Frost RA. Cytokine inhibition of JAK-STAT signaling: a new mechanism of growth hormone resistance. Pediatr Nephrol 2005; 20: 306-312.
McKay BR, De Lisio M, Johnston AP, O’Reilly CE, Phillips SM, Tarnopolsky MA, et al. Association of interleukin-6 signalling with the muscle stem cell response following muscle-lengthening contractions in humans. PLoS One 2009; 4: e6027.
Berthet J, Damien P, Hamzeh-Cognasse H, Arthaud CA, Eyraud MA, Zéni F, et al. Human platelets can discriminate between various bacterial LPS isoforms via TLR4 signaling and differential cytokine secretion. Clin Immunol 2012; 145: 189-200.
Rittig MG, Kaufmann A, Robins A, Shaw B, Sprenger H, Gemsa D, et al. Smooth and rough lipopolysaccharide phenotypes of Brucella induce different intracellular trafficking and cytokine/chemokine release in human monocytes. J Leukoc Biol 2003; 74: 1045-1055.
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.
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.
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.
Mariani E, Canella V, Berlingeri A, Bielli A, Cattini L, Landini MP, et al. Leukocyte presence does not increase microbicidal activity of platelet-rich plasma in vitro. BMC Microbiol 2015; 15: 149.
Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983; 65: 55-63.
Li X, Wu Z, Ni J, Liu Y, Meng J, Yu W, et al. Cathepsin b regulates collagen expression by fibroblasts via prolonging TLR2/NF-κB activation. Oxid Med Cell Longev 2016; 2016: 7894247.
Flamand L, Gosselin J, D’Addario M, Hiscott J, Ablashi DV, Gallo RC, et al. Human herpesvirus 6 induces interleukin-1 beta and tumor necrosis factor alpha, but not interleukin-6, in peripheral blood mononuclear cell cultures. J Virol 1991; 65: 5105-5110.
Bölükbaşı N, Yeniyol S, Tekkesin MS, Altunatmaz K. The use of platelet-rich fibrin in combination with biphasic calcium phosphate in the treatment of bone defects: a histologic and histomorphometric study. Curr Ther Res Clin Exp 2013; 75: 15-21.
Klingbeil MF, Herson MR, Cristo EB, dos Santos Pinto D Jr, Yoshito D, Mathor MB. Comparison of two cellular harvesting methods for primary human oral culture of keratinocytes. Cell Tissue Bank 2009; 10: 197-204.
Adan A, Kiraz Y, Baran Y. Cell Proliferation and Cytotoxicity Assays. Curr Pharm Biotechnol 2016; 17: 1213-1221.
Cheng R, Liu W, Zhang R, Feng Y, Bhowmick NA, Hu T. Porphyromonas gingivalis-derived lipopolysaccharide combines hypoxia to induce caspase-1 activation in periodontitis. Front Cell Infect Microbiol 2017; 7: 474.
Panda S, Jayakumar ND, Sankari M, Varghese SS, Kumar DS. Platelet rich fibrin and xenograft in treatment of intrabony defect. Contemp Clin Dent 2014; 5: 550-554.
Kumar RV, Shubhashini N. Platelet rich fibrin: a new paradigm in periodontal regeneration. Cell Tissue Bank 2013; 14: 453-463.
Tanaka K, Iwasaki K, Feghali KE, Komaki M, Ishikawa I, Izumi Y. Comparison of characteristics of periodontal ligament cells obtained from outgrowth and enzyme-digested culture methods. Arch Oral Biol 2011; 56: 380-388.
Liu Y, Wu Z, Zhang X, Ni J, Yu W, Zhou Y, et al. Leptomeningeal cells transduce peripheral macrophages inflammatory signal to microglia in reponse to Porphyromonas gingivalis LPS. Mediators Inflamm 2013; 2013: 407562.
Kang W, Wang T, Hu Z, Liu F, Sun Y, Ge S. Metformin inhibits porphyromonas gingivalis lipopolysaccharide-influenced inflammatory response in human gingival fibroblasts via regulating activating transcription factor-3 expression. J Periodontol 2017; 88: e169-e178.
Evrard SM, d’Audigier C, Mauge L, Israël-Biet D, Guerin CL, Bieche I, et al. The profibrotic cytokine transforming growth factor-β1 increases endothelial progenitor cell angiogenic properties. J Thromb Haemost 2012; 10: 670-679.
Dimmeler S. Platelet-derived growth factor CC--a clinically useful angiogenic factor at last? N Engl J Med 2005; 352: 1815-1816.
Castillo TN, Pouliot MA, Kim HJ, Dragoo JL. Comparison of growth factor and platelet concentration from commercial platelet-rich plasma separation systems. Am J Sports Med 2011; 39: 266-271.
Mundala M, Sun Xl, Li X, Fang J, Qi ML, Wang J, et al. Minimally invasive endoscopic maxillary sinus lifting and immediate implant placement: A case report World J Clin Cases 2019; 7: 1234-1241.
Weibrich G, Hansen T, Kleis W, Buch R, Hitzler WE. Effect of platelet concentration in platelet-rich plasma on peri-implant bone regeneration. Bone 2004; 34: 665-671.
Majewski M, Ochsner PE, Liu F, Flückiger R, Evans CH. Accelerated healing of the rat Achilles tendon in response to autologous conditioned serum. Am J Sports Med 2009; 37: 2117-2125.
Zaccaria V, Curti V, Di Lorenzo A, Baldi A, Maccario C, Sommatis S, et al. Effect of green and brown propolis extracts on the expression levels of microRNAs, mRNAs and proteins, related to oxidative stress and inflammation. Nutrients 2017; 9: 1090.
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