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Development and Characterization of a Tissue-engineered Human Oral Mucosa Equivalent Produced in a Serum-free Culture System

Dept. of Oral and Maxillofacial Surgery, University of Michigan School of Dentistry, Medical Center, B1-B204 UH, Box 0018, 1500 East Medical Center Drive, Ann Arbor, MI 48109-0018, USA

H. Terashi

Section of Plastic and Reconstructive Surgery, University of Michigan Medical Center, Ann, Arbor MI, USA

C.L. Marcelo

Section of Plastic and Reconstructive Surgery, University of Michigan Medical Center, Ann, Arbor MI, USA

S.E. Feinberg

Dept. of Oral and Maxillofacial Surgery, University of Michigan School of Dentistry, Medical Center, B1-B204 UH, Box 0018, 1500 East Medical Center Drive, Ann Arbor, MI 48109-0018, USA

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^TM. 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.

Key Words: Keratinocyte • acellular dermis • immunohistochemistry • essential fatty acid • oral mucosa.

Journal of Dental Research, Vol. 79, No. 3, 798-805 (2000)
DOI: 10.1177/00220345000790030301


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