, Volume 68, Issue 5, pp 1873–1884 | Cite as

Secretion of wound healing mediators by single and bi-layer skin substitutes

  • Manira Maarof
  • Jia Xian Law
  • Shiplu Roy Chowdhury
  • Khairul Anuar Khairoji
  • Aminuddin Bin Saim
  • Ruszymah Bt. Hj IdrusEmail author
Original Article


Limitations of current treatments for skin loss caused by major injuries leads to the use of skin substitutes. It is assumed that secretion of wound healing mediators by these skin substitutes plays a role in treating skin loss. In our previous study, single layer keratinocytes (SK), single layer fibroblast (SF) and bilayer (BL; containing keratinocytes and fibroblasts layers) skin substitutes were fabricated using fibrin that had shown potential to heal wounds in preclinical studies. This study aimed to quantify the secretion of wound healing mediators, and compare between single and bi-layer skin substitutes. Skin samples were digested to harvest fibroblasts and keratinocytes, and expanded to obtain sufficient cells for the construction of skin substitutes. Acellular fibrin (AF) construct was used as control. Substitutes i.e. AF, SK, SF and BL were cultured for 2 days, and culture supernatant was collected to analyze secretion of wound healing mediators via multiplex ELISA. Among 19 wound healing mediators tested, BL substitute secreted significantly higher amounts of CXCL1 and GCSF compared to SF and AF substitute but this was not significant with respect to SK substitute. The BL substitute also secreted significantly higher amounts of CXCL5 and IL-6 compared to other substitutes. In contrast, the SK substitute secreted significantly higher amounts of VCAM-1 compared to other substitutes. However, all three skin substitutes also secreted CCL2, CCL5, CCL11, GM-CSF, IL8, IL-1α, TNF-α, ICAM-1, FGF-β, TGF-β, HGF, VEGF-α and PDGF-BB factors, but no significant difference was seen. Secretion of these mediators after transplantation may play a significant role in promoting wound healing process for the treatment of skin loss.


Keratinocytes Fibroblasts Bi-layer skin substitute Fibrin Tissue engineering 



The research was funded by research grants from Science Fund 02-01-02-SF0964, Arus Perdana AP-2013-015 and Tissue Engineering Centre, UKM Medical Centre.

Supplementary material

Supplementary material 1 (MPG 684 kb)


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Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Manira Maarof
    • 1
  • Jia Xian Law
    • 1
  • Shiplu Roy Chowdhury
    • 1
  • Khairul Anuar Khairoji
    • 1
  • Aminuddin Bin Saim
    • 3
  • Ruszymah Bt. Hj Idrus
    • 1
    • 2
    Email author
  1. 1.Tissue Engineering CentreUniversiti Kebangsaan Malaysia Medical CentreKuala LumpurMalaysia
  2. 2.Department of Physiology, Faculty of MedicineUniversiti Kebangsaan MalaysiaKuala LumpurMalaysia
  3. 3.Ear Nose and Throat Consultant ClinicAmpang Puteri Specialist HospitalAmpangMalaysia

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