In vitro characterization and ex vivo surgical evaluation of human hair keratin films in ocular surface reconstruction after sterilization processing
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The disadvantages of human amniotic membrane (hAM), used for ocular surface reconstruction, necessitate the development of standardized alternatives. Keratin-derived-films (KF) have been indicated as transferable substrate for cell cultivation and tissue engineering. The impact of different sterilization procedures on KF and surgical feasibility were investigated. Human hair KF were prepared and sterilized; optical, biomechanical properties, in vitro cell seeding efficiency and proliferation of human corneal epithelial cells were studied and compared with hAM. Surgical feasibility was tested on enucleated porcine eye. Sterilized KF showed higher light transmission and significantly higher E-modulus than hAM; cell-seeding-efficiency and proliferation rate were not affected. Although KF could be surgically handled, suture placement was more difficult compared to hAM. Plasma treatment seems the best sterilization method for KF; it does not affect cell biology or optical and biomechanical properties. However material modifications are requested before KF may represent a feasible alternative for ocular surface reconstruction.
KeywordsOcular Surface Amniotic Membrane Ethanol Treatment Corneal Epithelial Cell Human Amniotic Membrane
The authors would like to thank Dr. K. Araki-Sasaki (Kagoshima, Japan) for his generous gift of the HCE-T cell line, and Dr. Meyer (Department of Ophthalmology, University of Würzburg, Germany) for help with the statistical analysis. Furthermore, special thanks are given to Drs. T. Klink, R. Guthoff and H. Sold (Department of Ophthalmology, University of Würzburg, Germany) for their time spent in the laboratory participating in the surgical feasibility tests, as well as Lucia Albrecht and Gesa Grobe (Technische Universität Braunschweig, Germany) for their technical assistance in the cell culture studies and tensile strength testing. This work was supported by the Deutsche Forschungsgemeinschaft, DFG grants Re 2596/3-1 and Ge 895/7-1.
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