The characterisation of human respiratory epithelial cells cultured on resorbable scaffolds: first steps towards a tissue engineered tracheal replacement.
In this study we have used lectin histochemistry and scanning electron microscopy (SEM) to assess the growth and characterise the differentiation of human respiratory epithelial cells (REC) cultured on two biomaterial scaffolds. The first scaffold, based on a hyaluronic acid derivative, was observed to be non-adhesive for REC. This lack of adhesion was found to be unrelated to the presence of the hyaluronic acid binding domain on the surface of isolated REC. The other scaffold, consisting of equine collagen. was observed to encourage REC spreading and adhesion. Positive Ulex Europaeus agglutinin (UEA) lectin staining of this preparation indicated the presence of ciliated REC on the scaffold surface. However, the marked decrease in peanut agglutinin (PNA) positive staining, relative to that of control cultures and native tissue, indicates a dedifferentiation of the secretory cells of the REC monolayer. SEM analysis of REC cultured on the collagen scaffold confirmed the presence of ciliated cells thereby validating the UEA positive staining. The presence of both established and developing cilia was also verified. This study indicates that collagen biomaterials are appropriate for the tissue engineering of REC. Furthermore, that UEA and PNA staining is a useful tool in the characterisation of cells cultured on biomaterials, therefore helpful in identifying biomaterials that are suitable for specific tissue engineering purposes.
Ziegelaar
Brian W
BW
Department of Otorhinolaryngology, Head and Neck Surgery, Ludwig-Maximilians-University of Munich, Germany. brian.ziegelaar@hno.med.uni-muenchen.de
Aigner
Joachim
J
Staudenmaier
Rainer
R
Lempart
Kathrin
K
Mack
Brigitte
B
Happ
Theda
T
Sittinger
Michael
M
Endres
Michaela
M
Naumann
Andreas
A
Kastenbauer
Ernst
E
Rotter
Nicole
N
D000328:Adult; D001672:Biocompatible Materials / Q000737:chemistry*; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000737:chemistry*; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry; D037102:Lectins / Q000378:metabolism; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D008875:Middle Aged; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D014132:Trachea / Q000166:cytology / Q000473:pathology*
D001672:Biocompatible Materials; D037102:Lectins; D006820:Hyaluronic Acid