FEATURES: ORIGINAL INVESTIGATIONSFull-Thickness Skin Wound Healing Using Autologous Keratinocytes and Dermal Fibroblasts with Fibrin Bilayered Versus Single-Layered SubstituteIdrus, Ruszymah Hj Bt MD, PhD; Rameli, Mohd Adha bin P BSc; Low, Kiat Cheong; Law, Jia Xian BSc(Hon), PhD; Chua, Kien Hui PhD, MSc, BSc(Hon); Latiff, Mazlyzam Bin Abdul PhD, BSc(Hon); Saim, Aminuddin Bin MD, MMedScAuthor Information Ruszymah Bt Hj Idrus, MD, PhD, is the Head of the Tissue Engineering Centre and Professor of the Department of Physiology, Faculty of Medicine; Mohd Adha bin P Rameli, BSc, is an MSc student, Tissue Engineering Centre; Kiat Cheong Low, is a Secretary, Laboratory Animal Resource Unit, Faculty of Medicine; Jia Xian Law, BSc(Hon), is a PhD student, Department of Physiology, Faculty of Medicine; Kien Hui Chua, PhD, MSc, BSc(Hon), is Associate Professor, Department of Physiology, Faculty of Medicine, and Research Fellow, Tissue Engineering Centre; Mazlyzam Bin Abdul Latiff, PhD, BSc(Hon), is a Lecturer, Schools of Diagnostic & Applied Health Sciences, Faculty of Health Sciences; and Aminuddin Bin Saim, MD, MMedSc, is an Ear, Nose, Throat Specialist, Ear, Nose & Throat Consultant Clinic, Ampang Puteri Specialist Hospital, Selangor, and a Consultant, Tissue Engineering Centre; all at the Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia. The authors have disclosed that they have no financial relationships related to this article. This work was financially supported by a research grant from the Ministry of Science, Technology and Innovation (MOSTI), Malaysia 06-02-02-0003-BTK/ER/022. Submitted April 12, 2013; accepted in revised form October 18, 2013. Advances in Skin & Wound Care: April 2014 - Volume 27 - Issue 4 - p 171-180 doi: 10.1097/01.ASW.0000445199.26874.9d Buy Metrics Abstract Split-skin grafting (SSG) is the gold standard treatment for full-thickness skin defects. For certain patients, however, an extensive skin lesion resulted in inadequacies of the donor site. Tissue engineering offers an alternative approach by using a very small portion of an individual’s skin to harvest cells for propagation and biomaterials to support the cells for implantation. The objective of this study was to determine the effectiveness of autologous bilayered tissue-engineered skin (BTES) and single-layer tissue-engineered skin composed of only keratinocytes (SLTES-K) or fibroblasts (SLTES-F) as alternatives for full-thickness wound healing in a sheep model. Full-thickness skin biopsies were harvested from adult sheep. Isolated fibroblasts were cultured using medium Ham’s F12: Dulbecco modified Eagle medium supplemented with 10% fetal bovine serum, whereas the keratinocytes were cultured using Define Keratinocytes Serum Free Medium. The BTES, SLTES-K, and SLTES-F were constructed using autologous fibrin as a biomaterial. Eight full-thickness wounds were created on the dorsum of the body of the sheep. On 4 wounds, polyvinyl chloride rings were used as chambers to prevent cell migration at the edge. The wounds were observed at days 7, 14, and 21. After 3 weeks of implantation, the sheep were euthanized and the skins were harvested. The excised tissues were fixed in formalin for histological examination via hematoxylin-eosin, Masson trichrome, and elastin van Gieson staining. The results showed that BTES, SLTES-K, and SLTES-F promote wound healing in nonchambered and chambered wounds, and BTES demonstrated the best healing potential. In conclusion, BTES proved to be an effective tissue-engineered construct that can promote the healing of full-thickness skin lesions. With the support of further clinical trials, this procedure could be an alternative to SSG for patients with partial- and full-thickness burns. © 2014 Wolters Kluwer Health | Lippincott Williams & Wilkins. All world rights reserved.