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Ioana-Sandra Tarau, Richard Schmid, Christian Lotz, Angela Rossi, Sebastian Schürlein, Heike Walles, Florian Groeber-Becker, Jan Hansemann, Jost Hillenkamp; Corneal woundhealing model employing a bioreactor. Invest. Ophthalmol. Vis. Sci. 2017;58(8):165. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Corneal wound healing disorders are a common problem worldwide and ex vivo and in vitro models are needed to develope new thearpies. The deterioration of ex vivo models in longterm studies remains a challenge, therefore we designed a standardized ex vivo porcin wound healing model, integrating a bioreactor and compared it with a static culture condition. The cornea bioreactor is developed to sustain the proliferation state and cell viability of the corneal tissue.
Corneas from freshly enucleated porcin eyes were removed with a scleral ring. A standardized wound was induced with a Excimer Laser. Afterwards the corneas were cultured in our bioreactor for 7 days with specific culture mediums. The bioreactor system is built with two separate media circuits divided by the cornea explant. On the epithelial site of the cornea, the media is dropping on the epithelium to ensure an alternation of liquid and air phase. For the control group, we used a static cell-crown culture with the same two different culture medias. All corneas were stored in a 4% CO2 Incubator at 37°C.
After 7 days in bioreactor culture a complete reepithelialization could be observed by a fluorescein staining test. In comparison the submers cultured cornea suffered from a severe epithelial loss and an unchanged stromal wound morphology. The histology of the bioreactor cultured cornea indicates stromal regeneration. The cornea in submers culture was blurred and corneal swelling could be observed while a lower swelling and a higher clarity could be achieved in the bioreactor cultured cornea.
Our results show the importance of the perfusion culture for ex vivo tissue. The alternation of liquid and air phase is a key factor for the growth of the corneal epithelium. With this bioreactor, we are able to investigate new biomaterials for the corneal wound healing. Furthermore this bioreactor can be a better alternative for the storage of human cornea.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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