* Corresponding Author
The effect of mitomycin-c in keloid fibroblast cultures
https://doi.org/10.19106/JMedSci004803201605 Ishandono Dachlan(1*), Teguh Aryandono(2), Mae Sri Hartati Wahyuningsih(3), Hardyanto Soebono(4), Yohanes Widodo Wirohadidjojo(5)
(1) Plastic and Reconstructive Surgery Divison, Department of Surgery, Faculty of Medicine, Universitas Gadjah Mada/Dr. Sardjito Hospital
(2) Surgical Oncology Division, Department of Surgery, Faculty of Medicine, Universitas Gadjah Mada/Dr. Sardjito Hospital, Yogyakarta, Indonesia
(3) Department of Pharmacology and Therapy, Faculty of Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia
(4) Department of Dermatology and Venereology, Faculty of Medicine, Universitas Gadjah Mada/Dr. Sardjito Hospital, Yogyakarta, Indonesia
(5) Department of Dermatology and Venereology, Faculty of Medicine, Universitas Gadjah Mada/Dr. Sardjito Hospital, Yogyakarta, Indonesia
(*) Corresponding Author
Abstract
ABSTRACT
Keloid occurs due to hyperactivity of keloid fibroblast (KF) in proliferation, migration, collagen deposition, together with low rates of collagen degradation. These are under the responsibility of TGF-b. Mitomycin C (MC) is used for treating keloid by a topical application during surgery at the level of 0.02% to 0.08%. Unfortunately, the lowest effective level of MC for keloid has not been determined yet. We aimed to determine the lowest effective level of MC in the suppression of KF activities. Various levels of MC diluted in growth medium were administered on KF that were isolated from six patients. After 24 hours and 72 hours of incubation, cellular proliferation, collagen deposition, cellular migration and level of TGF-b, were analyzed. Application of 120 uM MC on KF culture for 24 hours could significantly reduce TGF-b production from 1265.74 ± 274.81 pg/mL to 265.17 ± 12.20 pg/mL; proliferation index from 100% to 84.01 ± 12.91%; inhibit cellular migration to 64.38 ± 3.66%; but reduce collagen depositions from 100% to only 91.13 ± 10.19%. The lowest MC level is on 30 uM or equal with 0.001%. In conclusion, the lowest level of MC can suppress the activities of KF is 0.001%. Moreover, due to low activity in inhibiting collagen deposition, MC would be better as an adjuvant drug for keloid surgery.
Keywords
Full Text:
PDFReferences
REFERENCES 1. Mrowietz U., Seifert O. Keloid scarring: new treatments ahead. Actas Dermosifiliogr. 2009; 2:75-83. 2. Hunasgi S, Koneru A, Vanishree M, Shamala R. Keloid: A case report and review of pathophysiology and differences between keloid and hypertrophic scars. JOMFP, 2013;17(1):116-120. 3. Chipev CC, Simman R, Hatch G, Katz AE, Siegel DM, Simon M. Myofibroblast phenotype and apoptosis in keloid and palmar fibroblasts in vitro. Cell Death Differ; 2000; 7 : 166-176. 4. Luo S, Benathan M, Raffoul W, Panizzon RG, Egloff DV. Abnormal balance between proliferation and apoptotic cell death in fibroblasts derived from keloid lesions. Plast Reconstr Surg; 2001; 107 : 87-96. 5. Akasaka Y, Fujita K, Ishikawa Y, Asuwa N, Inuzuka K, Ishihara M, Ito M, Masuda T, Akishima Y, Zhang L, Ito K, Ishii T. Detection of apoptosis in keloids and a comparative study on apoptosisbetween keloids, hypertrophic scars, normal healed flat scars, and dermatofibroma. Wound Repair Regen: 2001; 9 : 501-506. 6. Sandulache VC, Parekh A, Li-Korotky H, Dohar JE, Hebda PA. Prostaglandin E2 inhibition of keloid fibroblast migration, contraction, and transforming growth factor (TGF) – beta1-induced collagen synthesis. Wound Repair Regen. 2007; 15 : 122-133. 7. Gurtner GC, Werner S, Barrandon Y, Longaker MT. Wound repair and regeneration. Nature. 2008;453 : 314-321. 8. Zhang Q, Yamaza T, Kelly AP, Shi S, Wang S, Brown J, Wang L, French SW, Shi S, Le AD. Tumor-Like Stem Cells Derived from Human Keloid Are Governed by the Inflammatory Niche Driven by IL-17/IL-6 Axis. Rich BE, ed. PLoS ONE. 2009;4(11):e7798. 9. Pakyari M, Farrokhi A, Maharlooei MK, Ghahary A., Critical Role of Transforming Growth Factor Beta in Different Phases of Wound Healing.Adv Wound Care. 2013;2(5):215-224. 10. Sato M. Upregulation of the Wnt/beta-catenin Pathway Induced by Transforming Growth Factor-beta in Hypertrophic Scars and Keloids. Acta Derm Venereol 2006;86:300–307 11. Igota S, Tosa M, Murakami M, Egawa S, Shimizu H, Hyakusoku H, Ghazizadeh M., Identification and characterization of Wnt signaling pathway in keloid pathogenesis.Int J Med Sci. 2013;10(4):344-354. 12. Lee WJ, Park JH, Shin JU, Noh H, Lew DH, Yang WI, Yun CO, Lee KH, Lee JH. Endothelial-to-mesenchymal transition induced by Wnt 3a in keloid pathogenesis.Wound Repair Regen. 2015; 23(3):435-442. 13. Raiskup F, Solomon A, Landau D, Ilsar M, Frucht-Pery J. Mitomycin C for pterygium: long term evaluation. The British Journal of Ophthalmology. 2004;88(11):1425-1428. 14. Van Setten G, Aspiotis M, Blalock TD, Grotendorst G, Schultz G. Connective tissue growth factor in pterygium: simultaneous presence with vascular endothelial growth factor—possible contributing factor to conjunctival scarring. Graefes Arch Clin Exp Ophthalmol., 2003; 241(2):135–139. 15. Talmi YP, Orenstein A, Wolf M, Kronenberg J. "Use of mitomycin C for treatment of keloid: a preliminary report." Otolaryngology-Head and Neck Surgery, 2005; 132(4): 598-601. 16. Bailey, JN, Waite AE, Clayton WJ, Rustin MH. (2007), Application of topical mitomycin C to the base of shave-removed keloid scars to prevent their recurrence. British Journal of Dermatology, 156: 682–686. 17. Wang YW, Ren JH, Xia K, Wang SH, Yin TF, Xie DH, Li LH. Effect of mitomycin on normal dermal fibroblast and HaCat cell: an in vitro study. Journal of Zhejiang University Science B. 2012;13(12):997-1005. 18. Taskiran D, Taskiran E, Yercan H, Kutay FZ. Quantification of total collagen in rabit tendon by the Sirius Red Methods, Tr J Med Science, 1999 (29):7-9 19. Liang CC, Park AY, Guan JL. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nature Protocols. 2007; 2(2): 329-333. 20. Bijlard E, Steltenpool S, Niessen FB. Intralesional 5-fluorouracil in keloid treatment: a systematic review., Acta Derm Venereol. 2015; 95(7):778-782. 21. Payapvipapong K, Niumpradit N, Piriyanand C, Buranaphalin S, Nakakes A. The treatment of keloids and hypertrophic scars with intralesional bleomycin in skin of color.J Cosmet Dermatol. 2015;14(1):83-90 22. Stewart CE, Kim JY, Linda I. Application of mitomycin-C for head and neck keloids. OtolaryngolHead Neck Surg. 2006; 135: 946-950. 23. Khan MA, Bashir MM, Khan FA. Intralesional triamcinolone alone and in combination with 5-fluorouracil for the treatment of Keloid and Hypertrophic scars. JPMA.2014; 64 (9):1003-1007. 24. Yeh FL, Shen HD, Tai HY. Decreased production of MCP-1 and MMP-2 by keloid-derived fibroblasts. Burns. 2009 ;35(3):348-351. 25. Astashkina A, Mann B. Grainger DW. A critical evaluation of in vitro cell culture models for high-throughput drug screening and toxicity. Pharmacology & Therapeutics., 2012; 134:82–106
DOI: https://doi.org/10.19106/JMedSci004803201605
Article Metrics
Abstract views : 1623
| 
views : 1977
Copyright (c) 2016 Ishandono Dachlan, Teguh Aryandono, Mae Sri Hartati Wahyuningsih, Hardyanto Soebono, Yohanes Widodo Wirohadidjojo
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.