Reconstruction of living bilayer human skin equivalent utilizing human fibrin as a scaffold.
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.
Mazlyzam
A L
AL
Department of Physiology, Faculty of Medicine, UKM, Malaysia; Tissue Engineering Laboratory, HUKM, Malaysia.
Aminuddin
B S
BS
Fuzina
N H
NH
Norhayati
M M
MM
Fauziah
O
O
Isa
M R
MR
Saim
L
L
Ruszymah
B H I
BH
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*
D005337:Fibrin