nasal tissue engineering using co-culture method

1. TITLE Culture of Nasal Epithelial Cells Using
Chitosan-Based Membranes
AUTHORS Tsung-Wei Huang, MD; Yi-Ho Young, MD; Po-Wen Cheng, MD; Yen-Hui Chan, MS; Tai-Horng Young,
PhD
JOURNAL The Laryngoscope
YEAR Laryngoscope 119: October 2009
Submitted by
Arunkumar Rengaraj
(22141800)

2. Culture of Nasal Epithelial Cells Using Chitosan-Based Membranes
Aim
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).
Introduction
• The nasal epithelium consists of ciliated epithelial cells, mucous blanket, and glands.
• The nasal septum divides the nasal chamber into two cavities, aiding in the regulation of air flow through the nose.
• Septal perforation, may cause inflammation, nasal crusting, bleeding, and whistling while breathing.
• To date, various surgical repairs for the septal perforation are proposed
• Chitosan- based membranes (scaffold) can be used for the culture of nasal epithelial cells (NECs).
• It provides physical support and serves as a substrate to regulate cell growth, adhesion, and differentiation.

3. Materials and methods
2 % solution of chitosan chitosan + 1 M acetic acid
spread on a glass plate
Evaporated at 50C
Mix 40 μg type I collagen
chitosan-collagen membrane
Tissues (Human inferior turbinates ) were treated with 0.5% pronase+1:1 mixture (DMEM) +
Ham’s nutrient F12
centrifugation
suspended in DMEM/F12
1.5-mL cell suspension was seeded on collagen coated
Morphological Examination
• The ciliary beat was observed under inverted microscope (Leica, DMI 6000, Solms, Germany) with 400 magnification.
• Imaging of cilia movement was recorded at 240 frames per second with a high-speed complementary metal oxide
semiconductor (CMOS) camera (CMC1300; VDS Vossku¨ hler GmbH, Osnabru¨ ck, Germany).

4. RESULTS
• NECs were found to be successfully adhesive with collagen and chitosan-collagen membranes at day 3 after seeding.
• Ciliated cells were first observed on collagen and chitosan- collagen membranes at day 7
• After confluence, which increased in number with active ciliary beating at day 21.
• Confocal images demonstrated that NECs developed into a pseudo stratified polarized layer with apical cilia and basolateral
nuclei on collagen and chitosan-collagen membranes resembling native respiratory epithelium, morphologically.