Development and characterization of a tissue-engineered human oral mucosa equivalent produced in a serum-free culture system.
A problem maxillofacial surgeons face is a lack of sufficient autogenous oral mucosa for reconstruction of the oral cavity. Split-thickness or oral mucosa grafts require more than one surgical procedure and can result in donor site morbidity. Skin has disadvantages of adnexal structures and a different keratinization pattern than oral mucosa. In this study, we successfully assembled, ex vivo, a human oral mucosa equivalent, consisting of epidermal and dermal components, in a defined, essential-fatty-acid-deficient, serum-free culture medium without a feeder layer, that could be used for intra-oral grafting in humans. Autogenous oral keratinocytes were seeded onto a cadaveric dermis, AlloDerm. The oral mucosa equivalent was cultured at an air-liquid interface for 2 wks. The resulting equivalent had a well-stratified parakeratinized epithelial layer similar to native oral keratinized mucosa. Expression of differentiation markers, filaggrin and cytokeratin 10/13, suggested a premature keratinized state. The presence of proliferation markers, proliferating cell nuclear antigen (PCNA) and Ki-67, suggested a state of hyperproliferation. Fatty acid composition of the equivalent was similar to that of in vitro cultured oral keratinocytes but differed from the that of in vivo native tissue, showing a lower content of 18:2 and 20:4, and a higher content of 16:1 and 18:1 fatty acids, respectively. The keratinocytes of the equivalent appeared to be in a more active and proliferative state than native keratinized mucosa. The dynamic nature of the cell population on the oral mucosa equivalent may be beneficial for intra-oral grafting procedures and for transfection of the keratinocytes.
Izumi
K
K
Dept. of Oral and Maxillofacial Surgery, University of Michigan School of Dentistry, Medical Center, Ann Arbor 48109-0018, USA.
Terashi
H
H
Marcelo
C L
CL
Feinberg
S E
SE
D001698:Biomedical Engineering; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D000078404:Epidermal Cells; D005227:Fatty Acids / Q000032:analysis; D000091344:Filaggrin Proteins; D006651:Histocytochemistry; D006801:Humans; D015603:Keratinocytes / Q000166:cytology; D019394:Ki-67 Antigen / Q000032:analysis; D009061:Mouth Mucosa / Q000166:cytology* / Q000637:transplantation*; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis; D017768:Skin, Artificial
D016895:Culture Media, Serum-Free; C000717249:FLG protein, human; D005227:Fatty Acids; D000091344:Filaggrin Proteins; D019394:Ki-67 Antigen; D018809:Proliferating Cell Nuclear Antigen