title: Reconstruction of living bilayer human skin equivalent utilizing human fibrin as a scaffold.
abstract: Our aim of this study was to develop a new methodology for constructing a bilayer human skin equivalent to create a more clinical compliance skin graft composite for the treatment of various skin defects. We utilized human plasma derived fibrin as the scaffold for the development of a living bilayer human skin equivalent: fibrin-fibroblast and fibrin-keratinocyte (B-FF/FK SE). Skin cells from six consented patients were culture-expanded to passage 1. For B-FF/FK SE formation, human fibroblasts were embedded in human fibrin matrix and subsequently another layer of human keratinocytes in human fibrin matrix was stacked on top. The B-FF/FK SE was then transplanted to athymic mice model for 4 weeks to evaluate its regeneration and clinical performance. The in vivo B-FF/FK SE has similar properties as native human skin by histological analysis and expression of basal Keratin 14 gene in the epidermal layer and Collagen type I gene in the dermal layer. Electron microscopy analysis of in vivo B-FF/FK SE showed well-formed and continuous epidermal-dermal junction. We have successfully developed a technique to engineer living bilayer human skin equivalent using human fibrin matrix. The utilization of culture-expanded human skin cells and fibrin matrix from human blood will allow a fully autologous human skin equivalent construction.
authors.affiliation: Department of Physiology, Faculty of Medicine, UKM, Malaysia; Tissue Engineering Laboratory, HUKM, Malaysia.
authors.forename: A L
authors.initials: AL
authors.lastname: Mazlyzam
authors.forename: B S
authors.initials: BS
authors.lastname: Aminuddin
authors.forename: N H
authors.initials: NH
authors.lastname: Fuzina
authors.forename: M M
authors.initials: MM
authors.lastname: Norhayati
authors.forename: O
authors.initials: O
authors.lastname: Fauziah
authors.forename: M R
authors.initials: MR
authors.lastname: Isa
authors.forename: L
authors.initials: L
authors.lastname: Saim
authors.forename: B H I
authors.initials: BH
authors.lastname: Ruszymah
mesh_terms: D000293:Adolescent; D000328:Adult; D018929:Cell Culture Techniques; D002469:Cell Separation; D005337:Fibrin / Q000502:physiology*; D005347:Fibroblasts / Q000166:cytology; D015870:Gene Expression; D006801:Humans; D015603:Keratinocytes / Q000166:cytology; D008854:Microscopy, Electron; D010949:Plasma / Q000166:cytology / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017768:Skin, Artificial; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D005337:Fibrin