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Y. Tanaka, A. Kubota, C. Hongo, M. Matsusaki, K. Takehana, M. Akashi, K. Nishida; Development of Collagen Hydrogel With Molecular Orientation for Tissue Engineering of Corneal Stroma. Invest. Ophthalmol. Vis. Sci. 2009;50(13):2552.
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Developmant of collagen hydrogel with molecular orientation and its evaluation as an implant for tissue engineering of corneal stroma.
Porcine type I atelocollagen containing 5% type III collagen dissolved in aqueous HCl was adjusted at pH 3.6-4.0 by addition of aqueous NaOH, and mixed with EDC & NHS using syringe system (Liu Y. et al., Biomacromolecules, 2006). The final concentration of collagen was adjusted to 10wt%. The reaction solution was poured onto a glass plate in a specific direction and subsequently covered with another glass plate to spread the solution. Silicon rubber spacers (100-500 µm in thickness) were sandwiched between the glass plates to regulate their thickness. The obtained gels were evaluated by UV-vis transmission measurement, X-ray diffraction measurements (XRD), scanning electron microscopy (SEM),and mechanical property measurement. Their biocompatibility was evaluated by culture of porcine corneal cells and implantation in rabbit corneal stromal pockets.
XRD and SEM analysis strongly supported that the orientation of collagen fibers (59.0 ± 14.6 nm-diameter) in the gels was same as the direction of collagen solution spreaded onto a glass plate. The tensile strength of the parallel direction was approximately two-fold higher than that of the orthogonal direction. Transparency of the obtained gels with 500 µm-thickness was 79.2 +/- 6.9 % at 550nm. This gels exibited favorable cell-adhesion and cell-proliferation in vitro. Furthermore, neither blood vessel invasion nor opacification occurred with the gel-implanted corneas for 60 days in corneal intrastromal pockets.
The obtaineded collagen hydrogels with moleculer orientation exsibited unique mechanical properties, faborable transparency and biocompatibility with cornea in vitro /in vivo, which will be useful in the field of tissue engineering of corneal stroma.
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