Abstract
Purpose: :
Our previous study has demonstrated cultivation of bovine corneal endothelial cells (BCECs) on chitosan-polycaprolactone (PCL) blends. BCECs could proliferate well and express physiological phenotypes after PCL was introduced into chitosan by means of blending. The feasibility of using chitosan-PCL-blended membranes as scaffold and carrier sheet for the corneal endothelial cell (CEC) transplantation is worthy of continued investigation.
Methods: :
In this work, chitosan, which does not promote BCECs grow, and PCL, which supports BCEC proliferation, are used to prepare chitosan-PCL-blended membranes. Therefore, we attempt to manufacture different blended membranes with fine adjustment of PCL proportion. In this study, mechanical properties, transmittance, proliferative abilities, phenotypic expressions, extracellular matrix (ECM) protein production, and hemotoxylin and eosin (H&E) staining were performed to evaluate cultivated BCECs on the blended membranes.
Results: :
The blended membranes combined advantages from separate biomaterial and exhibited greater transparency (than PCL) and better cellular activity (than chitosan) than alone. The BCECs on the blends illustrated normal appearance and good proliferative ability. Immunohistochemical staining verified physiologically phenotypic expression of ZO-1 and Na-K ATPase. Western blot analysis also confirmed the existence of collagen type IV proteins. Furthermore, the formation of a continuous monolayer of cells attached on the blended membrane was confirmed by H&E staining.
Conclusions: :
These results showed potential that chitosan/PCL-blended membranes may be a biomaterial to cultivate and delivery CEC sheet during transplantation.
Keywords: cornea: endothelium • cornea: basic science