Hypertrophic scar is a dermal fibroproliferative disease characterized by the overproduction and deposition of extracellular matrix, and the hyperproliferation and enhanced angiogenesis of fibroblasts, along with their enhanced differentiation to myofibroblasts. Botulinum toxin type A shows potential for prevention of hypertrophic scar formation; however, its effectiveness in attenuating skin fibrosis and the related mechanism are unclear. In this study, human scar fibroblasts were cultured and stimulated with botulinum toxin type A, and the changes in fibroblast proliferation, migration, and protein expression of pro-fibrotic factors were evaluated with colorimetric, scratch, and enzyme-linked immunosorbent assays and western blotting, respectively. Botulinum toxin type A treatment decreased the proliferation and migration of human scar fibroblasts compared with those of untreated controls. Protein expression levels of pro-fibrotic factors (transforming growth factor β1, interleukin-6, and connective tissue growth factor) were also inhibited by botulinum toxin type A, whereas the JNK phosphorylation level was increased. Activation of the JNK pathway demonstrated the inhibitory effects of the toxin on human scar fibroblast proliferation and production of pro-fibrotic factors, suggesting that the suppressive effects of botulinum toxin type A are closely associated with JNK phosphorylation. Overall, this study showed that botulinum toxin type A has a suppressive effect on extracellular matrix production and scar-related factors in human scar fibroblasts in vitro, and that regulation of JNK signaling plays an important role in this process. Our results provide a theoretical basis, at the cellular level, for the therapeutic use of botulinum toxin type A.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Austin E, Koo E, Jagdeo J (2018) The cellular response of keloids and hypertrophic scars to botulinum toxin A: a comprehensive literature review. Dermatol Surg 44:149–157. https://doi.org/10.1097/DSS.0000000000001360
Border WA, Noble NA (1994) Transforming growth factor β in tissue fibrosis. N Engl J Med 331:1286–1292. https://doi.org/10.1056/NEJM199411103311907
Chen HC, Yen CI, Yang SY et al (2017) Comparison of steroid and botulinum toxin type A monotherapy with combination therapy for treating human hypertrophic scars in an animal model. Plast Reconstr Surg 140:43e–49e. https://doi.org/10.1097/PRS.0000000000003426
Chen JY, Zhang L, Zhang H, Su L, Qin LP (2014) Triggering of p38 MAPK and JNK signaling is important for oleanolic acid-induced apoptosis via the mitochondrial death pathway in hypertrophic scar fibroblasts. Phytother Res 28:1468–1478. https://doi.org/10.1002/ptr.5150
Cho JS, Kang JH, Shin JM, Park IH, Lee HM (2015) Inhibitory effect of delphinidin on extracellular matrix production via the MAPK/NF-κB pathway in nasal polyp-derived fibroblasts. Allergy Asthma Immunol Res 7:276–282. https://doi.org/10.4168/aair.2015.7.3.276
Gassner HG, Brissett AE, Otley CC, Boahene DK, Boggust AJ, Weaver AL, Sherris DA (2006) Botulinum toxin to improve facial wound healing: A prospective, blinded, placebo-controlled study. Mayo Clin Proc 81:1023–1028. https://doi.org/10.4065/81.8.1023
Gassner HG, Sherris DA, Friedman O (2009) Botulinum toxin-induced immobilization of lower facial wounds. Arch Facial Plast Surg 11:140–142. https://doi.org/10.1001/archfacial.2009.3
Gassner HG, Sherris DA, Otley CC (2000) Treatment of facial wounds with botulinum toxin A improves cosmetic outcome in primates. Plast Reconstr Surg 105:1948–1953. https://doi.org/10.1097/00006534-200005000-00005 (discussion 1954–1945)
Gauglitz GG, Korting HC, Pavicic T, Ruzicka T, Jeschke MG (2011) Hypertrophic scarring and keloids: pathomechanisms and current and emerging treatment strategies. Mol Med 17:113–125. https://doi.org/10.2119/molmed.2009.00153
He T, Bai X, Yang L et al (2015) Loureirin B inhibits hypertrophic scar formation via inhibition of the TGF-β1-ERK/JNK pathway. Cell Physiol Biochem 37:666–676. https://doi.org/10.1159/000430385
Huang W, Foster JA, Rogachefsky AS (2000) Pharmacology of botulinum toxin. J Am Acad Dermatol 43:249–259. https://doi.org/10.1067/mjd.2000.105567
Huang C, Miyazaki K, Akaishi S, Watanabe A, Hyakusoku H, Ogawa R (2013) Biological effects of cellular stretch on human dermal fibroblasts. J Plast Reconstr Aesthet Surg 66:e351–e361. https://doi.org/10.1016/j.bjps.2013.08.002
Igarashi A, Okochi H, Bradham DM, Grotendorst GR (1993) Regulation of connective tissue growth factor gene expression in human skin fibroblasts and during wound repair. Mol Biol Cell 4:637–645. https://doi.org/10.1091/mbc.4.6.637
Jeong HS, Lee BH, Sung HM, Park SY, Ahn DK, Jung MS, Suh IS (2015) Effect of botulinum toxin type A on differentiation of fibroblasts derived from scar tissue. Plast Reconstr Surg 136:171e–178e. https://doi.org/10.1097/PRS.0000000000001438
Kim S, Ahn M, Piao Y et al (2016) Effect of botulinum toxin type A on TGF-β/Smad pathway signaling: implications for silicone-induced capsule formation. Plast Reconstr Surg 138:821e–829e. https://doi.org/10.1097/PRS.0000000000002625
Kim YJ, Kim JH, Lee KJ et al (2015) Botulinum neurotoxin type A induces TLR2-mediated inflammatory responses in macrophages. PLoS ONE 10:e0120840. https://doi.org/10.1371/journal.pone.0120840
Lee SH, Min HJ, Kim YW, Cheon YW (2018) The efficacy and safety of early postoperative botulinum toxin A injection for facial scars. Aesthetic Plast Surg 42:530–537. https://doi.org/10.1007/s00266-017-1008-7
Li Y, Zhang W, Gao J et al (2016) Adipose tissue-derived stem cells suppress hypertrophic scar fibrosis via the p38/MAPK signaling pathway. Stem Cell Res Ther 7:102. https://doi.org/10.1186/s13287-016-0356-6
Liang X, Chai B, Duan R, Zhou Y, Huang X, Li Q (2017) Inhibition of FKBP10 attenuates hypertrophic scarring through suppressing fibroblast activity and extracellular matrix deposition. J Invest Dermatol 137:2326–2335. https://doi.org/10.1016/j.jid.2017.06.029
Liu DQ, Li XJ, Weng XJ (2017) Effect of BTXA on inhibiting hypertrophic scar formation in a rabbit ear model. Aesthetic Plast Surg 41:721–728. https://doi.org/10.1007/s00266-017-0803-5
McFarland-Mancini MM, Funk HM, Paluch AM et al (2010) Differences in wound healing in mice with deficiency of IL-6 versus IL-6 receptor. J Immunol 184:7219–7228. https://doi.org/10.4049/jimmunol.0901929
Moustakas A, Heldin CH (2005) Non-Smad TGF-β signals. J Cell Sci 118:3573–3584. https://doi.org/10.1242/jcs.02554
Nam SM, Kim YB (2018) The effects of platelet‐rich plasma on hypertrophic scars fibroblasts. Int Wound J 15:547–554. https://doi.org/10.1111/iwj.12896
Qi Q, Mao Y, Yi J, Li D, Zhu K, Cha X (2014) Anti-fibrotic effects of Astragaloside IV in systemic sclerosis. Cell Physiol Biochem 34:2105–2116. https://doi.org/10.1159/000366405
Ray S, Ju X, Sun H, Finnerty CC, Herndon DN, Brasier AR (2013) The IL-6 trans-signaling-STAT3 pathway mediates ECM and cellular proliferation in fibroblasts from hypertrophic scar. J Invest Dermatol 133:1212–1220. https://doi.org/10.1038/jid.2012.499
Sawamura D, Meng X, Ina S, Sato M, Tamai K, Hanada K, Hashimoto I (1998) Induction of keratinocyte proliferation and lymphocytic infiltration by in vivo introduction of the IL-6 gene into keratinocytes and possibility of keratinocyte gene therapy for inflammatory skin diseases using IL-6 mutant genes. J Immunol 161:5633–5639
Vangipuram M, Ting D, Kim S, Diaz R, Schule B (2013) Skin punch biopsy explant culture for derivation of primary human fibroblasts. J Vis Exp 77:e3779. https://doi.org/10.3791/3779
Wilson AM (2006) Use of botulinum toxin type A to prevent widening of facial scars. Plast Reconstr Surg 117:1758–1766. https://doi.org/10.1097/01.prs.0000209944.45949.d1 (discussion 1767–1758)
Xiao Z, Zhang F, Cui Z (2009) Treatment of hypertrophic scars with intralesional botulinum toxin type A injections: a preliminary report. Aesthetic Plast Surg 33:409–412. https://doi.org/10.1007/s00266-009-9334-z
Xiao Z, Zhang F, Lin W, Zhang M, Liu Y (2010) Effect of botulinum toxin type A on transforming growth factor β1 in fibroblasts derived from hypertrophic scar: a preliminary report. Aesthetic Plast Surg 34:424–427. https://doi.org/10.1007/s00266-009-9423-z
Xiao Z, Zhang M, Liu Y, Ren L (2011) Botulinum toxin type A inhibits connective tissue growth factor expression in fibroblasts derived from hypertrophic scar. Aesthetic Plast Surg 35:802–807. https://doi.org/10.1007/s00266-011-9690-3
This research was supported by AmorePacific Grant in 2015.
Conflicts of interest
Research involving human participants and/or animals
All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Park, G.S., An, M.K., Yoon, J.H. et al. Botulinum toxin type A suppresses pro-fibrotic effects via the JNK signaling pathway in hypertrophic scar fibroblasts. Arch Dermatol Res 311, 807–814 (2019). https://doi.org/10.1007/s00403-019-01975-0
- Botulinum toxin type A
- JNK signaling
- Hypertrophic scar