Local Delivery of CTGF siRNA with Poly(sorbitol-co-PEI) Reduces Scar Contraction in Cutaneous Wound Healing

  • Ki-Hyun Cho
  • Bijay Singh
  • Sushila Maharjan
  • Yoonjeong Jang
  • Yun-Jaie Choi
  • Chong-Su ChoEmail author
Original Article


Healing process in scarring inevitably produces a considerable amount of non-organized dense collagen-rich matrix called scar thus impairing the native structure of skin. Connective tissue growth factor (CTGF) overexpression within healing tissues is known to play an imperative role in collagen production stimulated by transforming growth factor-beta in cutaneous wound healing. Undoubtedly, the knockdown of CTGF expression through siRNA-mediated gene silencing could simply impede the scarring process. However, the less stability and low transfection of siRNAs themselves urge a safe carrier to protect and transfect them into cells at a high rate avoiding toxicities. Here, we developed a degradable poly(sorbitol-co-PEI) (PSPEI), prepared by polymerization of sorbitol diacrylate with low molecular weight polyethylenimine, which has high transfection efficiency but low cytotoxicity, and utilized it in siCTGF delivery to silence the expression of CTGF in an animal model of cutaneous wound healing. Unlike contracted scar in normal healing, there was no or less contraction in the healed skin of mice treated with siCTGF using PSPEI. Histologically, the healed tissues also had distinct papillary structures and dense irregular connective tissues that were lacking in the control scar tissues. This study exemplifies a successful treatment of cutaneous wound healing using a polymer system coupled with RNA interference. Hence, the approach holds a great promise for developing new treatments with novel targets in regenerative medicines.


Connective tissue growth factor Poly(sorbitol-co-PEI) siRNA Scar contraction Wound healing 



This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Minister of Science, ICT and Future Planning (MIST) (NRF-2014R1A1A2007163).

Compliance with ethical standards

Conflict of interest

The authors have no potential conflicts of interest.

Ethical statement

This study was conducted under the approval of animal ethics committee at Seoul National University (SNU-130520-7).


  1. 1.
    Gurtner GC, Werner S, Barrandon Y, Longaker MT. Wound repair and regeneration. Nature. 2008;453(7193):314–21.CrossRefPubMedGoogle Scholar
  2. 2.
    Walmsley GG, Maan ZN, Wong VW, Duscher D, Hu MS, Zielins ER, et al. Scarless wound healing: chasing the holy grail. Plast Reconstr Surg. 2015;135(3):907–17.CrossRefPubMedGoogle Scholar
  3. 3.
    Nauta A, Gurtner GC, Longaker MT. Wound healing and regenerative strategies. Oral Dis. 2011;17(6):541–9.CrossRefPubMedGoogle Scholar
  4. 4.
    Yates CC, Hebda P, Wells A. Skin wound healing and scarring: fetal wounds and regenerative restitution. Birth Defects Res C Embryo Today. 2012;96(4):325–33.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Baker R, Urso-Baiarda F, Linge C, Grobbelaar A. Cutaneous scarring: a clinical review. Dermatol Res Pract. 2009;2009:625376.PubMedGoogle Scholar
  6. 6.
    Widgerow AD, Chait LA. Scar management practice and science: a comprehensive approach to controlling scar tissue and avoiding hypertrophic scarring. Adv Skin Wound Care. 2011;24(12):555–61.CrossRefPubMedGoogle Scholar
  7. 7.
    Mustoe TA, Cooter RD, Gold MH, Hobbs FD, Ramelet AA, Shakespeare PG, et al. International clinical recommendations on scar management. Plast Reconstr Surg. 2002;110(2):560–71.CrossRefPubMedGoogle Scholar
  8. 8.
    Mustoe TA. Evolution of silicone therapy and mechanism of action in scar management. Aesthet Plast Surg. 2007;32(1):82.CrossRefGoogle Scholar
  9. 9.
    Park JE, Barbul A. Understanding the role of immune regulation in wound healing. Am J Surg. 2004;187(5):S11–6.CrossRefGoogle Scholar
  10. 10.
    Goldschmeding R, Aten J, Ito Y, Blom I, Rabelink T, Weening JJ. Connective tissue growth factor: just another factor in renal fibrosis? Nephrol Dial Transplant. 2000;15(3):296–9.CrossRefPubMedGoogle Scholar
  11. 11.
    Border WA, Noble NA. Transforming growth factor beta in tissue fibrosis. N Engl J Med. 1994;331(19):1286–92.CrossRefPubMedGoogle Scholar
  12. 12.
    Birkedal-Hansen H. Proteolytic remodeling of extracellular matrix. Curr Opin Cell Biol. 1995;7(5):728–35.CrossRefPubMedGoogle Scholar
  13. 13.
    Grinnell F. Fibroblasts, myofibroblasts, and wound contraction. J Cell Biol. 1994;124(4):401–4.CrossRefPubMedGoogle Scholar
  14. 14.
    Mackool RJ, Gittes GK, Longaker MT. Scarless healing. The fetal wound. Clin Plast Surg. 1998;25(3):357–65.PubMedGoogle Scholar
  15. 15.
    Montesano R, Orci L. Transforming growth factor beta stimulates collagen-matrix contraction by fibroblasts: implications for wound healing. Proc Natl Acad Sci U S A. 1988;85(13):4894–7.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Sisco M, Kryger Z, Jia S-X, Schultz G, Dean N, Mustoe T. Antisense oligonucleotides against transforming growth factor-beta delay wound healing in a rabbit ear model. J Am Coll Surg. 2005;201(3):S60.CrossRefGoogle Scholar
  17. 17.
    Lipson KE, Wong C, Teng Y, Spong S. CTGF is a central mediator of tissue remodeling and fibrosis and its inhibition can reverse the process of fibrosis. Fibrogenesis Tissue Repair. 2012;5(1):S24.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Ignotz RA, Massague J. Transforming growth factor-beta stimulates the expression of fibronectin and collagen and their incorporation into the extracellular matrix. J Biol Chem. 1986;261(9):4337–45.PubMedGoogle Scholar
  19. 19.
    Abreu JG, Ketpura NI, Reversade B, De Robertis EM. Connective-tissue growth factor (CTGF) modulates cell signalling by BMP and TGF-[beta]. Nat Cell Biol. 2002;4(8):599–604.PubMedPubMedCentralGoogle Scholar
  20. 20.
    Cordeiro MF, Mead A, Ali RR, Alexander RA, Murray S, Chen C, et al. Novel antisense oligonucleotides targeting TGF-[beta] inhibit in vivo scarring and improve surgical outcome. Gene Ther. 2003;10(1):59–71.CrossRefPubMedGoogle Scholar
  21. 21.
    Sisco M, Kryger ZB, O’Shaughnessy KD, Kim PS, Schultz GS, Ding X-Z, et al. Antisense inhibition of connective tissue growth factor (CTGF/CCN2) mRNA limits hypertrophic scarring without affecting wound healing in vivo. Wound Repair Regen. 2008;16(5):661–73.CrossRefPubMedGoogle Scholar
  22. 22.
    Cho WY, Hong SH, Singh B, Islam MA, Lee S, Lee AY, et al. Suppression of tumor growth in lung cancer xenograft model mice by poly(sorbitol-co-PEI)-mediated delivery of osteopontin siRNA. Eur J Pharm Biopharm. 2015;94:450–62.CrossRefPubMedGoogle Scholar
  23. 23.
    Pack DW, Hoffman AS, Pun S, Stayton PS. Design and development of polymers for gene delivery. Nat Rev Drug Discov. 2005;4(7):581–93.CrossRefPubMedGoogle Scholar
  24. 24.
    Singh B, Jang Y, Maharjan S, Kim H-J, Lee AY, Kim S, et al. Combination therapy with doxorubicin-loaded galactosylated poly(ethyleneglycol)-lithocholic acid to suppress the tumor growth in an orthotopic mouse model of liver cancer. Biomaterials. 2017;116:130–44.CrossRefPubMedGoogle Scholar
  25. 25.
    Singh B, Maharjan S, Park TE, Jiang T, Kang SK, Choi YJ, et al. Tuning the buffering capacity of polyethylenimine with glycerol molecules for efficient gene delivery: staying in or out of the endosomes. Macromol Biosci. 2015;15(5):622–35.CrossRefPubMedGoogle Scholar
  26. 26.
    Kim Y-D, Pofali P, Park T-E, Singh B, Cho K, Maharjan S, et al. Gene therapy for bone tissue engineering. Tissue Eng Regen Med. 2016;13(2):111–25.CrossRefGoogle Scholar
  27. 27.
    Mastrobattista E, Hennink WE. Polymers for gene delivery: charged for success. Nat Mater. 2012;11(1):10–2.CrossRefGoogle Scholar
  28. 28.
    Park TG, Jeong JH, Kim SW. Current status of polymeric gene delivery systems. Adv Drug Deliv Rev. 2006;58(4):467–86.CrossRefPubMedGoogle Scholar
  29. 29.
    Gary DJ, Puri N, Won Y-Y. Polymer-based siRNA delivery: perspectives on the fundamental and phenomenological distinctions from polymer-based DNA delivery. J Control Release. 2007;121(1–2):64–73.CrossRefPubMedGoogle Scholar
  30. 30.
    Dahlman JE, Barnes C, Khan OF, Thiriot A, Jhunjunwala S, Shaw TE, et al. In vivo endothelial siRNA delivery using polymeric nanoparticles with low molecular weight. Nat Nano. 2014;9(8):648–55.CrossRefGoogle Scholar
  31. 31.
    O’shaughnessy K, Kim P, Roy N, Mustoe T. Antisense oligonucleotides to TGF-beta and CTGF decrease hypertrophic scarring in a rabbit ear model. Wound Repair Regen. 2007;15(2):A20.Google Scholar
  32. 32.
    Castleberry SA, Golberg A, Sharkh MA, Khan S, Almquist BD, Austen WG Jr, et al. Nanolayered siRNA delivery platforms for local silencing of CTGF reduce cutaneous scar contraction in third-degree burns. Biomaterials. 2016;95:22–34.CrossRefPubMedGoogle Scholar

Copyright information

© The Korean Tissue Engineering and Regenerative Medicine Society and Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  • Ki-Hyun Cho
    • 1
  • Bijay Singh
    • 1
    • 2
  • Sushila Maharjan
    • 1
    • 2
  • Yoonjeong Jang
    • 3
  • Yun-Jaie Choi
    • 1
  • Chong-Su Cho
    • 1
    Email author
  1. 1.Department of Agricultural Biotechnology, Research Institute for Agriculture and Life SciencesSeoul National UniversitySeoulKorea
  2. 2.Research Institute for Bioscience and BiotechnologyKathmanduNepal
  3. 3.Research Institute for Veterinary Science and College of Veterinary MedicineSeoul National UniversitySeoulKorea

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