title: Secretion of wound healing mediators by single and bi-layer skin substitutes.
abstract: 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.
authors.affiliation: Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaccob Latiff, Cheras, 56000, Kuala Lumpur, Malaysia.
authors.forename: Manira
authors.initials: M
authors.lastname: Maarof
authors.affiliation: Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaccob Latiff, Cheras, 56000, Kuala Lumpur, Malaysia.
authors.forename: Jia Xian
authors.initials: JX
authors.lastname: Law
authors.affiliation: Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaccob Latiff, Cheras, 56000, Kuala Lumpur, Malaysia.
authors.forename: Shiplu Roy
authors.initials: SR
authors.lastname: Chowdhury
authors.affiliation: Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaccob Latiff, Cheras, 56000, Kuala Lumpur, Malaysia.
authors.forename: Khairul Anuar
authors.initials: KA
authors.lastname: Khairoji
authors.affiliation: Ear Nose and Throat Consultant Clinic, Ampang Puteri Specialist Hospital, 1 Jalan Mamanda 9, 68000, Ampang, Selangor, Malaysia.
authors.forename: Aminuddin Bin
authors.initials: AB
authors.lastname: Saim
authors.affiliation: Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaccob Latiff, Cheras, 56000, Kuala Lumpur, Malaysia. ruszyidrus@gmail.com.|Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaccob Latiff, Cheras, 56000, Kuala Lumpur, Malaysia. ruszyidrus@gmail.com.
authors.forename: Ruszymah Bt Hj
authors.initials: RB
authors.lastname: Idrus
keywords: Bi-layer skin substitute; Fibrin; Fibroblasts; Keratinocytes; Tissue engineering