Construction and characterization of a tissue-engineered oral mucosa equivalent based on a chitosan-fish scale collagen composite.
This study was designed to (1) assess the in vitro biocompatibility of a chitosan-collagen composite scaffold (C3) constructed by blending commercial chitosan and tilapia scale collagen with oral mucosa keratinocytes, (2) histologically and immunohistochemically characterize an ex vivo-produced oral mucosa equivalent constructed using the C3 (EVPOME-C), and (3) compare EVPOME-C with oral mucosa constructs utilizing AlloDerm® (EVPOME-A), BioMend® Extend™ (EVPOME-B), and native oral mucosa. C3 scaffold had a well-developed fibril network and a sufficiently small porosity to prevent keratinocytes from growing inside the scaffold after cell-seeding. The EVPOME oral mucosa constructs were fabricated in a chemically defined culture system. After culture at an air-liquid interface, EVPOME-C and EVPOME-B had multilayered epithelium with keratinization, while EVPOME-A had a more organized stratified epithelium. Ki-67 and p63 immunolabeled cells in the basal layer of all EVPOMEs suggested a regenerative ability. Compared with native oral mucosa, the keratin 15 and 10/13 expression patterns in all EVPOMEs showed a less-organized differentiation pattern. In contrast to the β1-integrin and laminin distribution in EVPOME-A and native oral mucosa, the subcellular deposition in EVPOME-C and EVPOME-B indicated that complete basement membrane formation failed. These findings demonstrated that C3 has a potential application for epithelial tissue engineering and provides a new potential therapeutic device for oral mucosa regenerative medicine.
Division of Oral Anatomy, Niigata University Graduate School of Medical and Dental Sciences, Chuo-Ku, Niigata City, Japan. teradam@dent.niigata-u.ac.jp
Michiko
M
Terada
Kenji
K
Izumi
Hisashi
H
Ohnuki
Taro
T
Saito
Hiroko
H
Kato
Marie
M
Yamamoto
Yoshiro
Y
Kawano
Kayoko
K
Nozawa-Inoue
Haruhiko
H
Kashiwazaki
Toshiyuki
T
Ikoma
Junzo
J
Tanaka
Takeyasu
T
Maeda
D000825:Animal Structures / Q000737:chemistry*; D000818:Animals; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry*; D003094:Collagen / Q000737:chemistry*; D005260:Female; D029941:Fish Proteins / Q000737:chemistry*; D006801:Humans; D053550:Keratin-10 / Q000378:metabolism; D053535:Keratin-13 / Q000378:metabolism; D053569:Keratin-15 / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D019394:Ki-67 Antigen / Q000378:metabolism; D008297:Male; D009061:Mouth Mucosa / Q000166:cytology / Q000378:metabolism; D017210:Tilapia; D023822:Tissue Engineering; D014157:Transcription Factors / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000378:metabolism
D029941:Fish Proteins; C507493:KRT10 protein, human; C507426:KRT13 protein, human; C116015:KRT15 protein, human; D053535:Keratin-13; D053569:Keratin-15; D019394:Ki-67 Antigen; C115051:TP63 protein, human; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; D053550:Keratin-10; D003094:Collagen; D048271:Chitosan