Biocompatible hydrogel supports the growth of respiratory epithelial cells: possibilities in tracheal tissue engineering.
Extensive tracheal defect reconstruction is a major challenge in plastic and reconstructive surgery. The lack of an epithelial lining on the luminal surfaces of tracheal prostheses is among the major causes of their failure. Chitosan-gelatin hydrogels were synthesized for the development of biocompatible, growth-supportive substrata for respiratory epithelial cells. We employed J774 macrophages to test the immunocompatibility of this gel. The hydrogel did not exert a cytotoxic effect on macrophages, as confirmed by tetrazolium reduction and neutral red uptake assay. Flow cytometric analysis of macrophages cultured on the hydrogel showed a comparable expression of activation markers CD11b/CD18, CD45, and CD14 to the control. Semiquantitative RT-PCR results showed an absence of upregulation of interleukin-6 (IL-6) and TNF-alpha in these macrophages with respect to the controls. Primary human respiratory epithelial cells cultured on the hydrogel showed proper attachment, normal morphology, and growth. A small proportion of cells on the hydrogel showed synchronously beating cilia. RT-PCR analysis showed that cells on the hydrogel expressed mucins 2 and 5 and cytokeratin 13, which are markers for secretory goblet and squamous cells, respectively. All these results demonstrate that the hydrogel supports the growth of a mixed population of differentiated epithelial cells. This hydrogel is suitable as a culture substratum for respiratory epithelial cells and could be used as a potential candidate for coating tracheal prostheses.
Risbud
M
M
Tissue Engineering Laboratory, University Medical Centre, Charité, Humboldt University of Berlin, Tucholkystrasse-2, 10117 Berlin, Germany.
Endres
M
M
Ringe
J
J
Bhonde
R
R
Sittinger
M
M
D001672:Biocompatible Materials; D015415:Biomarkers / Q000032:analysis*; D001698:Biomedical Engineering; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D005109:Extracellular Matrix; D005434:Flow Cytometry; D006801:Humans; D020100:Hydrogels; D008262:Macrophage Activation; D008264:Macrophages; D020545:Respiratory Mucosa / Q000166:cytology / Q000254:growth & development*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014132:Trachea
D001672:Biocompatible Materials; D015415:Biomarkers; D016207:Cytokines; D020100:Hydrogels