Open Access Open Access  Restricted Access Subscription or Fee Access

The Addition of Hyaluronic Acid into Platelet-Rich Fibrin Lysate in Restoration of Senescent Human Dermal Fibroblasts Activities

Rika Azyenela, Indah Julianto, Yohanes Widodo Wirohadidjojo


Senescent human dermal fibroblasts had reduced capacity in proliferation and collagen synthesis. It is due to unresponsiveness against transforming growth factor-β1 (TGF-β1) stimulation. Either platelet-rich fibrin (PRF)-lysate or hyaluronic acid (HA) can restore TGF-β1 signaling pathway.

To determine whether HA addition to PRF lysate has a better activity than PRF-lysate alone in restoring senescent human dermal fibroblasts (HDFs) activities.

HDF isolated from six different human skins was divided into normal HDFs and senescent HDFs which are induced by serum starvation. The senescent groups were then given 50% PRF-lysate and various levels of HA. Amelioration of TGF-β1 signaling was measured by cellular proliferation index and collagen deposition. 

Addition of HA into PRF-lysate resulted in a significant increase in proliferation index and collagen deposition index than PRF-lysate alone. The best level of HA for this mixture ranged from 20.83 mM to 41.67 mM. HA in PRF lysate is an excellent candidate material for treating clinical signs related to senescent human dermal fibroblasts.


Ethical permission: This experiment had gain approval from the local ethical committee, Ref: KE/FK/471/EC/2016 dated 17-05-2016.




Handle (PDF):


Collagen deposition, hyaluronic acid, platelet-rich fibrin lysate, proliferation index, senescent human dermal fibroblasts

Full Text:



Agren MS, Steenfos HH, Dabelsteen E. Proliferation and mitogenic response to PDGF-BB of fibroblasts isolated from chronic venous leg ulcers is ulcer-age dependent. J Invest Dermatol 1999; 112(4): 463-9.

Alcorta DA, Xiong Y, Phelps D, Hannon G, Beach D, Barret J. Involvement of the cyclin-dependent kinase inhibitor p16 (INK14a) in replicative senescence of normal human fibroblasts. Proc Natl Acad Sci 1996; 93: 13742-7.

Anitua E, Andia I, Ardanza B, Nurden P, Nurden AT. Autologous platelets as a source of proteins for healing and tissue regeneration. Thromb Haemost 2009; 91:4-15.

Capelo AS. Dual role for TGF-β1 in apoptosis. Cytokines and Growth Factor Reviews 2005; 16(1): 15-34.

Clark RA. Oxidative stress and “senescent†fibroblasts in non-healing wounds as potential therapeutic targets. J Invest Dermatol 2008; 128:2361-4.

Cowin AJ, Hatzirodos N, Holding CA, Dunaiski V, Harries RH, Rayner TE et al. Effect of wound healing on the expression of transforming growth factor βs and their receptors in chronic venous leg ulcers. J Invest Dermatol 2001; 117: 1282-9.

Czuwara-Ladykowska J, Gore EA, Shegogue DA, Smith EA, Trojanowska M. Differential regulation of transforming growth factor-beta receptors type I and II by platelet-derived growth factor in human dermal fibroblasts. Br J Dermatol 2001; 145: 569-75.

David-Raoudi M, Tranchepain F, Deschrevel B, Vincent JC, Bogdanowicz P, Boumediene K. Differential effects of hyaluronan and its fragments on fibroblasts: relation to wound healing. Wound Repair Regen 2008; 16(2): 274-87.

Dohan Ehrenfest DM, de Peppo GM, Doglioli P, Sammartino G. Slow release of growth factors and thrombospondins-1 in Choukroun’s platelet-rich fibrin (PRF): a gold standard to achieve for all surgical platelet concentrates technologies. Growth factors 2009; 27: 63-9.

Eckes B, Zweews MC, Zhang ZG, Hallinger R, Mauach C, Aumailley M. Mechanical tension and integrin alpha 2 beta 1 regulate fibroblast functions. J Investig Dermatol Symp Proc 2006; 11: 66-72.

Harding KG, Moore K, Phillips TJ. Wound chronicity and fibroblasts senescence-implication for treatment. Int Wound J 2005; 2(4): 364-8.

He L, Lin Y, Hu X, Zhang Y, Wu H. A comparative study of platelet-rich fibrin (PRF) and platelet-rich plasma (PRP) on the effect of proliferation and differentiation of rat osteoblasts in vitro. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2009; 108: 707-13.

Kim BC, Kim HT, Park SH, Park SH, Cha JS, Yufit T, Kim SJ, Falanga V. Fibroblasts from chronic wounds show altered TGF-beta-signaling and decreased TGF-beta typed II receptor expression. J Cell Physiol 2003; 195(3): 331-6.

Kim MS, Song HJ, Lee SH, Lee CK. Comparative study of various growth factors and cytokines on type I collagen and hyaluronan production in human dermal fibroblasts. J Cosmet Dermatol 2014;13:44-51.

Locci P, Marinucci L, Lilli C. Transforming growth factor beta 1-hyaluronic acid interaction. Cell Tissue Res 1995; 281: 317-24.

Mast BA, Diegelmann RF, Krummel TM, Cohen IK. Hyaluronic acid modulates proliferation, collagen and protein synthesis of cultured fetal fibroblasts. Matrix 1993; 13: 441-6.

Naru E, Suzuki T, Moriyama M, Inomata K, Hayashi A, Arakane K, et al. Functional changes induced by chronic UVA irradiation to cultured human dermal fibroblasts. Br J Dermatol 2005; 153 Suppl 2: 6-12.

Pan DB, Ke YS, Liu WJ, Wei YQ, Tang J, Cao H. Platelet-derived growth factor-BB inhibited p21(WAF1) expression partially through transforming growth factor-beta signalling system in vascular smooth muscle cell. Zhonghua Xin Xue Guan Bing Za Zhi 2010; 38: 160-5.

Porsch H, Mehic M, Olofsson B, Heldin P, Heldin CH. Platelet-derived growth factor β-receptor, transforming growth factor β type I receptor and CD44 protein modulate each other’s signaling and stability. J Biol Chem 2014; 289(28): 19747-57.

Quan T, He T, Kang S, Voorhees JJ, Fisher GJ. Solar ultraviolet irradiation reduces collagen in photoaged human skin by blocking transforming growth factor-beta type II receptor/Smad signalling. Am J Pathol 2004; 165:741-51.

Quan T, He T, Shao Y, Lin L, Kang S, Voorhees JJ. Elevated cysteine-rich 61 mediates-aberrant collagen homeostasis in chronologically aged and photoaged human skin. Am J Pathol 2006; 169:482-90.

Quan T, Shao Y, He T, Voorhees JJ, Fisher GJ. Reduced expression of connective tissue growth factor (CTGF/CCN2) mediates collagen loss in chronologically agrd human skin. J Invest Dermatol 2010; 130:415-24.

Quan T, Wang F, Shao Y, Rittie L, Xia W, Orringer JS, Voorhees JJ, Fisher GJ. Enhancing structural support of the dermal microenvironment activates fibroblasts, endothelial cells and keratinocytes in aged human skin in vivo. J Invest Dermatol 2013; 133(3): 658-67.

Ramirez RD, Morales CP, Herbert BS, Rohde JM, Passons C, Shay JW, Wright WE. Putative telomere-independent mechanisms of replicative aging reflect inadequate growth conditins. R Genes Dev 2001; 15: 398-403.

Sansan MV. Efek vitreous gel mata sapi terhadap viabilitas selular, deposisi kolagen dan migrasi fibroblas tua (suatu upaya penyembuhan ulkus kronis). Thesis. Gadjah Mada University, Indonesia, 2015.

Taskiran D, Taskiran E, Yercan H, Kutay FZ. Quantification of total collagen in rat tendon by the Sirius Red Methods. Tr J Med Science 1999; 29:7-9.

Villeneuve J, Block A, Le Bousse-Kerdiles MC, Lepreux S, Nurden R, Ripoche J.Tissue inhibitors of matrix metalloproteinases in platelets and megakaryocytes: a novel organization for these secreted proteins. Exp Hematol 2009; 37: 849-56.

Wang W, Liu X, Zhang Y, Zhao J, Zhao B, Zhang S et al. Both senescence and apoptosis induced by deprivation of growth factors were inhibited by a novel butyrolactone derivative through depressing integrin beta 4 in vascular endothelial cells. Endothelium 2007; 14(6):325-32.

Wirohadidjojo YW, Budiyanto A, Soebono H. Platelet-rich fibrin lysate can ameliorate dysfunction of chronically UVA-Irradiated Human Dermal Fibroblasts. Yonsei Med J 2016; 57(5):1282-5.

Yin B, Jiang X. Telomere shortening in cultured human dermal fibroblasts is associated with acute photodamage induced by UVA irradiation. Postepy Dermatol Alergol 2013; 30: 13-8.

-- 0 --


  • There are currently no refbacks.

Copyright (c) 2016 Rika Azyenela, Indah Julianto, Yohanes Widodo Wirohadidjojo

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.