Integrin binding peptide promotes in vitro wound closure in the L929 mouse fibroblasts

Yıl 2017, Cilt: 3 Sayı: 3, 207 - 213, 04.11.2017

Ozan Karaman , Günnur Onak Emine Afra Demirci Emine Kahraman

https://doi.org/10.18621/eurj.318606

Cited By: 1

Öz

Objective. Molecular basis of wound healing process needs to
further examined to determine the effective individual biological cues. The
objective of this study was to investigate the wound closure, proliferation,
and viability of L929 fibroblast when cultured with different concentration of
soluble RGD peptid. Methods. RGD peptide was synthesized manually on solid phase. The
percentage of healed wound area for control, 0.5 mM, 1 mM, and 2 mM at each
time points were analyzed by ImageJ. Cell proliferation and viability were
assessed with MTT and live/ dead analysis, respectively. Results. The results of
wound closure area showed that increased RGD peptide concentration in the
culture improved cellular migration which enables significantly accelerated
wound closure. However, RGD peptide did not dramatically augmented cell
proliferation. In addition, cell viability results indicated that dead cell
numbers did not critically influence by increasing the RGD peptide
concentration in the culture. Conclusions. The present study
showed that soluble integrin binding peptide accelerated the migration and
wound closure rate of L929 fibroblasts. Delivery of soluble integrin binding
peptides into the wound area may be considered as an alternative wound
treatment technique in the near future after proofing the concept study with
animal and clinical studies.

Anahtar Kelimeler

Integrin binding peptide, in vitro scratch assay, cell migration, serum free media

Kaynakça

• [1] Labler L, Mica L, Härter L, Trentz O, Keel M. [Influence of VAC-therapy on cytokines and growth factors in traumatic wounds]. Zentralbl Chir 2006;131:S62-7. [Article in German]
• [2] Rivera AE, Spencer JM. Clinical aspects of full-thickness wound healing. Clin Dermatol 2007;25:39-48.
• [3] Robson MC, Steed DL, Franz MG. Wound healing: biologic features and approaches to maximize healing trajectories. Cur Probl Surg 2001;38:72-140.
• [4] Natarajan S, Williamson D, Stiltz AJ, Harding K. Advances in wound care and healing technology. Am J Clin Dermatol 2000;1:269-75.
• [5] Strong AL, Neumeister MW, Levi B. Stem cells and tissue engineering: regeneration of the skin and its contents. Clin Plast Surg 2017;44:635-650.
• [6] Velnar T, Bailey T, Smrkolj V. The wound healing process: an overview of the cellular and molecular mechanisms. J Int Med Res 2009;37:1528-42.
• [7] Gonzalez AC, Costa TF, Andrade ZA, Medrado AR. Wound healing - A literature review. An Bras Dermatol 2016;91:614-20.
• [8] Witte MB, Barbul A. General principles of wound healing. Surg Clin North Am. 1997;77:509-28.
• [9] Eming SA, Martin P, Tomic-Canic M. Wound repair and regeneration: mechanisms, signaling, and translation. Sci Transl Med 2014;6:265sr6.
• [10] Boyce ST, Ham RG. Calcium-regulated differentiation of normal human epidermal keratinocytes in chemically defined clonal culture and serum-free serial culture. J Invest Dermatol 1983;81:33s-40s.
• [11] Wang HJ, Chen TM, Cheng LF, Cheng TY, Tung YM. Human keratinocyte culture using porcine pituitary extract in serum-free medium. Burns 1995;21:503-6.
• [12] Higham MC, Dawson R, Szabo M, Short R, Haddow DB, MacNeil S. Development of a stable chemically defined surface for the culture of human keratinocytes under serum-free conditions for clinical use. Tissue Eng 2003;9:919-30.
• [13] Kim JH, Jekarl DW, Kim M, Oh EJ, Kim Y, Park IY, et al. Effects of ECM protein mimetics on adhesion and proliferation of chorion derived mesenchymal stem cells. Int J Med Sci 2014;11:298-308.
• [14] Nyegaard S, Christensen B, Rasmussen JT. An optimized method for accurate quantification of cell migration using human small intestine cells. Metab Eng Commun 2016;3:76-83.
• [15] Hersel U, Dahmen C, Kessler H. RGD modified polymers: biomaterials for stimulated cell adhesion and beyond. Biomaterials 2003;24:4385-415.
• [16] Heath DE, Cooper SL. The development of polymeric biomaterials inspired by the extracellular matrix. J Biomater Sci Polym Ed 2017;28:1051-69.
• [17] Pierschbacher MD, Ruoslahti E. Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule. Nature 1984;309:30-3.
• [18] Ruoslahti E, Pierschbacher MD. New perspectives in cell adhesion: RGD and integrins. Science 1987;238:491-7.
• [19] Giancotti FG. Complexity and specificity of integrin signalling. Nat Cell Biol 2000;2:E13-4.
• [20] Chen CS, Mrksich M, Huang S, Whitesides GM, Ingber DE. Geometric control of cell life and death. Science 1997;276:1425-8.
• [21] Buckley CD, Pilling D, Henriquez NV, Parsonage G, Threlfall K, Scheel-Toellner D, et al. RGD peptides induce apoptosis by direct caspase-3 activation. Nature 1999;397:534-9.
• [22] Patrulea V, Hirt-Burri N, Jeannerat A, Applegate L, Ostafe V, Jordan O, et al. Peptide-decorated chitosan derivatives enhance fibroblast adhesion and proliferation in wound healing. Carbohydr Polym 2016;142:114-23.
• [23] Brenner W, Gross S, Steinbach F, Horn S, Hohenfellner R, Thüroff JW. Differential inhibition of renal cancer cell invasion mediated by fibronectin, collagen IV and laminin. Cancer Lett 2000;155:199-205.
• [24] Tolomelli A, Galletti P, Baiula M, Giacomini D. Can integrin agonists have cards to play against cancer? A literature survey of small molecules integrin activators. Cancers (Basel) 2017;9:E78.
• [25] Reynolds AR, Hart IR, Watson AR, Welti JC, Silva RG, Robinson SD, et al. Stimulation of tumor growth and angiogenesis by low concentrations of RGD-mimetic integrin inhibitors. Nat Med 2009;15:392-400.
• [26] Shabbir SH, Eisenberg JL, Mrksich M. An inhibitor of a cell adhesion receptor stimulates cell migration. Angew Chem Int Ed Eng 2010;49:7706-9.
• [27] Karaman O, Kumar A, Moeinzadeh S, He X, Cui T, Jabbari E. Effect of surface modification of nanofibres with glutamic acid peptide on calcium phosphate nucleation and osteogenic differentiation of marrow stromal cells. J Tissue Eng Regen Med 2016;10:E132-46.
• [28] Liu Y, Peterson DA, Kimura H, Schubert D. Mechanism of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction. J Neurochem 1997;69:581-93.
• [29] Jones JI, Gockerman A, Busby WH, Wright G, Clemmons DR. Insulin-like growth factor binding protein 1 stimulates cell migration and binds to the alpha 5 beta 1 integrin by means of its Arg-Gly-Asp sequence. Proc Natl Acad Sci 1993;90:10553-7.
• [30] Hadden HL, Henke CA. Induction of lung fibroblast apoptosis by soluble fibronectin peptides. Am J Respir Crit Care Med 2000;162:1553-60.
• [31] Frisch SM, Francis H. Disruption of epithelial cell-matrix interactions induces apoptosis. J Cell Biol 1994;124:619-26.
• [32] Hayman EG, Pierschbacher MD, Ruoslahti E. Detachment of cells from culture substrate by soluble fibronectin peptides. J Cell Biol 1985;100:1948-54.