INCUBATION OF PLATELET-RICH FIBRIN MATRIX WITH MESENCHYMAL STEM CELLS IMPROVES MATRIX STIFFNESS
Keywords:Platelet-rich fibrin matrix, Mesenchymal stem cells
Objective: The platelet-rich fibrin matrix (PRFM) is condensed platelet-rich plasma (PRP) and should possess a comparable biomechanical property to the transplanted sites, for them to be physiologically functional. The aim of this study was to investigate the effect of human bone marrow mesenchymal stem cells (hBM-MSC) or human umbilical cord mesenchymal stem cells (hUC-MSC) on the biomechanical properties of PRFM.
Methods: PRFM was prepared by the gelation of PRP using 25 mmol CaCl2. The resulting coin-shaped PRFM pellets, 5 cm in diameter and 300 μm thick, were directly seeded with hUC-MSC or hBM-MSC at 2,000 cells cm-2, followed by 24 h incubation at 37 °C in 5% (v/v) CO2 in air. The samples were then observed by scanning electron microscopy to determine the morphology of the matrix surface. The PRFM biomechanical properties were determined at a 10 mm. min-1failure rate using an MCT 2150 universal testing machine (AandD Co. LTD).
Results: SEM imaging of the surface of the PRFM seeded with hBM-MSC and hUC-MSC showed a cloudy layer that thickened over time. The elastin slope of the PRFM was significantly improved after seeding with hBM-MSC and hUC-MSC when compared with unseeded PRFM (p<0.002, R2=0.983). Both cell types elicited similar biomechanical effects (p=0.99).
Conclusion: PRFM seeded with hBM-MSC or hUC-MSC showed significantly increased elasticity.
Harper MT, MacCarthy Morrogh L, Jones ML, Konopatskaya O, Poole AW. Platelets: their role in atherogenesis and thrombosis in coronary artery disease. Edited by Sarah Jane George; 2010.
Litvinov RI, Weisel JW. Fibrin mechanical properties and their structural origins. Matrix Biol 2017;60-61:110-23.
Lin F, Zhu J, Tonnesen MG, Taira BR, McClain SA, Singer AJ, et al. Fibronectin peptides that bind PDGF-BB enhance survival of cells and tissue under stress. J Invest Dermatol 2014;134:1119-27.
Scarano A, Ceccarelli M, Marchetti M, Piattelli A, Mortellaro C. Soft tissue augmentation with autologous platelet gel and β-TCP: a histologic and histometric study in mice. BioMed Res Int 2016. https://doi.org/10.1155/2016/2078104
Reksodiputro M, Widodo D, Bashiruddin J, Siregar N, Malik S. PRFM enhance wound healing process in the skin graft. Facial Plast Surg 2014;30:670-5.
Engler AJ, Sen S, Sweeney HL, Discher DE. Matrix elasticity directs stem cell lineage specification. Cell 2006;126:677-89.
Law JX, Chowdhury SR, Saim AB, Idrus RBH. Platelet-rich plasma with keratinocytes and fibroblasts enhance the healing of full-thickness wounds. J Tissue Viability 2017;26:208-15.
Zhang GA, Ning FG, Zhao NM. Biomechanical properties of four dermal substitutes. Chin Med J 2007;120:1454-5.
Sandora N. Regeneration of the decellularised tendon by human mesenchymal stem cells in response to uniaxial tensile strain. University of Leeds; 2016.
Italiano JE, Bergmeier W, Tiwari S, Falet H, Hartwig JH, Hoffmeister KM, et al. Mechanisms and implications of platelet discoid shape. Blood 2003;101:4789-96.
Sandora N, Fisher J, Ingham E. Decellularisation process to produce graft for transplantation; repopulation capacity of human MSCS. Adv Sci Lett 2018;24:6497-501.
Moran P, Coats B. Biological sample preparation for SEM imaging of porcine retina. Microscopy Today 2012;20:28-31.
Guthold M, Liu W, Sparks E, Jawerth L, Peng L, Falvo M, et al. A comparison of the mechanical and structural properties of fibrin fibers with other protein fibers. Cell Biochem Biophys 2007;49:165-81.
Chester D, Brown AC. The role of biophysical properties of provisional matrix proteins in wound repair. Matrix Biol 2017;60-61:124-40.
Badylak SF, Freytes DO, Gilbert TW. Extracellular matrix as a biological scaffold material: structure and function. Acta Biomaterialia 2009;5:1-13.
Cheng CW, Solorio LD, Alsberg E. Decellularized tissue and cell-derived extracellular matrices as scaffolds for orthopaedic tissue engineering. Biotechnol Adv 2014;32:462-84.
Kim YJ, Seo DH, Lee SH, Lee SH, An GH, Ahn HJ, et al. Conditioned media from human umbilical cord blood-derived mesenchymal stem cells stimulate rejuvenation function in human skin. Biochem Biophys Reports 2018;16:96-102.
Hasan MH, Botros KG, El-Shahat MA, Abdallah HA, Sobh MA. In vitro differentiation of human umbilical cord blood mesenchymal stem cells into functioning hepatocytes. Alexandria J Med 2017;53:167-73.
Vinaya Kumar R, Shubhashini N. Platelet-rich fibrin: a new paradigm in periodontal regeneration. Cell Tissue Banking 2013;14:453-63.
Babo P, Santo VE, Duarte ARC, Correia C, Costa MH, Mano JF, et al. Platelet lysate membranes as new autologous templates for tissue engineering applications. Inflammation Regener 2014;34:33-44.
Furst W, Banerjee A, Redl H. Comparison of structure, strength and cytocompatibility of a fibrin matrix supplemented either with tranexamic acid or aprotinin. J Biomed Mater Res Part B 2007;82:109-14.
Briquez PS, Lorentz KM, Larsson HM, Frey P, Hubbell JA. Human kunitz-type protease inhibitor engineered for enhanced matrix retention extends the longevity of fibrin biomaterials. Biomaterials 2017;135:1-9.
Chopin Doroteo M, Salgado Curiel RM, Perez Gonzalez J, Marin Santibanez BM, Krötzsch E. Fibroblast populated collagen lattices exhibit opposite biophysical conditions by fibrin or hyaluronic acid supplementation. J Mechamical Behavior Biomed Materials 2018;82:310-9.
Lai VK, Frey CR, Kerandi AM, Lake SP, Tranquillo RT, Barocas VH. Microstructural and mechanical differences between digested collagen–fibrin co-gels and pure collagen and fibrin gels. Acta Biomaterialia 2012;8:4031-42.
Montalbano G, Toumpaniari S, Popov A, Duan P, Chen J, Dalgarno K, et al. Synthesis of bioinspired collagen/alginate/fibrin-based hydrogels for soft tissue engineering. Materials Sci Eng: C 2018;91:236-46.
Kadler K, Holmes D, Trotter J, Chapman J. Collagen fibril formation. Biochem J 1996;316:1-11.
Jansen Karin A, Bacabac Rommel G, Piechocka Izabela K, Koenderink Gijsje H. Cells actively stiffen fibrin networks by generating contractile stress. Biophysical J 2013;105:2240-51.
Wang JHC, Thampatty BP, Lin JS, Im HJ. Mechanoregulation of gene expression in fibroblasts. Gene 2007;391:1-15.
Screen HRC, Shelton JC, Bader DL, Lee DA. Cyclic tensile strain upregulates collagen synthesis in isolated tendon fascicles. Biochem Biophysical Res Communications 2005;336:424-9.
Xu J, Sun M, Tan Y, Wang H, Wang H, Li P, et al. Effect of matrix stiffness on the proliferation and differentiation of umbilical cord mesenchymal stem cells. Differentiation 2017;96:30-9.