Culture of nasal epithelial cells using chitosan-based membranes.
OBJECTIVE
The aim of this study was to evaluate whether chitosan-based membranes can be used as scaffolds for growth and differentiation of nasal epithelial cells (NECs). Our final goal was to establish a novel methodology for enhancing the regeneration of the respiratory system.


METHODS
Prospective study.


METHODS
Human NECs were cultured on three various substrates, e.g., chitosan membranes, collagen, and chitosan-collagen membranes. Morphology of NECs was examined via light and electron microscopy, the area of ciliated cells was measured by confocal microscopy, and ciliary beat frequency was also evaluated. Expression of mucin genes was investigated with reverse-transcription polymerase chain reaction.


RESULTS
NECs were found to be successfully adhesive with collagen and chitosan-collagen membranes at day 3 after seeding, but not with chitosan membranes. The cilia area on collagen were 6.1% +/- 1.2%, 8.4% +/- 1.4%, and 12.5% +/- 1.9% at days 7, 14, and 21 after confluence, respectively, compared with 5.1% +/- 0.9%, 8.6% +/- 1.6%, and 12.3% +/- 2.1% in chitosan-collagen membranes, exhibited nonsignificant difference (P > .05). There were no significant differences in ciliary beat frequency between each group. The expression levels of mucin genes, namely, MUC5AC, MUC5B, and MUC2, in NECs on both collagen and chitosan-collagen membranes did not differ significantly (P > .05).


CONCLUSIONS
A small amount collagen mixed with chitosan substrate may improve the biocompatibility and promote the mucociliary differentiation in NECs. It appears that chitosan-collagen membrane is a promising scaffold for culture of the nasal epithelium, which sets the stage for studying tissue regeneration in the respiratory system.
Huang
Tsung-Wei
TW
Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, National Taiwan University Hospital and College of Medicine, Taipei, Taiwan.
Young
Yi-Ho
YH
Cheng
Po-Wen
PW
Chan
Yen-Hui
YH
Young
Tai-Horng
TH
D001672:Biocompatible Materials / Q000494:pharmacology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D048271:Chitosan / Q000494:pharmacology*; D003094:Collagen / Q000494:pharmacology; D004847:Epithelial Cells; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D009077:Mucins / Q000378:metabolism; D009297:Nasal Mucosa / Q000166:cytology*; D011446:Prospective Studies; D020133:Reverse Transcriptase Polymerase Chain Reaction; D023822:Tissue Engineering; D054457:Tissue Scaffolds
D001672:Biocompatible Materials; D009077:Mucins; D003094:Collagen; D048271:Chitosan