Periodic Reporting for period 4 - EyeRegen (Engineering a scaffold based therapy for corneal regeneration)
Periodo di rendicontazione: 2020-01-01 al 2020-12-31
Approximately 1 in 30 Europeans experiences sight loss at some point in their lives. Corneal blindness resulting from disease, physical injury or chemical burns are among the most common causes of blindness worldwide affecting millions of people. These conditions have a considerable economic and social impact on the lives of people across Europe. In many cases corneal transplants can restore vision however the shortage of donor corneas suitable for transplantation has necessitated the development of alternative treatments. One such approach is to engineer a biomaterial based scaffold that can replace the damaged cornea and promote regeneration after transplantation. The overall objective of this project is to engineer novel smart scaffolds suitable for corneal transplantation as an alternative to donate corneas. To do this effectively, it is vital to understand how different biochemical and biophysical stimuli affect the regenerative capacity of the patients cells and then use this information to optimize the scaffold design. Therefore this project addressed the following aims: (1) Determine of the chemical and biochemical cues that control corneal cell behavior; (2) Identify of how corneal cells physical and mechanical environment affects their behavior; (3) establish an in vitro culture system to culture corneal tissue and (4) design and fabricate scaffolds that incorporate chemical and physical cues to induce tissue formation
The effect that different biochemical reagents have on corneal cells has been evaluated and a number of reagents identified that are suitable for incorporation into our scaffolds. For example, chemicals such as ascorbic acid, retinoic acid, insulin like growth factor 1 were found to support more normal corneal stromal cell behavior while transforming growth factor beta resulted in the cell acting more like wound healing cells. The influence that the physical environment, including stiffness, topography and fluid flow, has on the cells has also been characterized and provides useful information for our scaffold design and materials selection. Substrates with a low stiffness were found to be better at maintaining epithelial progenitor cells while stiffer substrates made these cell more like mature epithelial cells. Aligned microchannels were shown to control cell orientation, migration and enhance a native corneal stromal cell type. The effect of fluid shear stress on cells was evaluated using a bioreactor system this stress was shown to improve stratification of corneal epithelial cells, which is important in generating functional, transplantable tissue.
A number of different materials processing techniques have been examined and optimized to fabricate smart scaffolds capable of supporting tissue regeneration. Scaffold suitable for corneal transplantation have been developed using biomaterials such as silk fibroin and decellularized porcine corneal tissue. Decellularized porcine corneas were successfully recellularized using human cells and evaluated ex-vivo and in-vivo. Silk fibroin was combined with retinoic acid and riboflavin to generate a biomaterial scaffold that could control the behavior of corneal stromal cells.
These results of this research has been published in peer-review journals and are available in open access repositories.
A number of different materials processing techniques have been examined and optimized to fabricate smart scaffolds capable of supporting tissue regeneration. Scaffold suitable for corneal transplantation have been developed using biomaterials such as silk fibroin and decellularized porcine corneal tissue. Decellularized porcine corneas were successfully recellularized using human cells and evaluated ex-vivo and in-vivo. Silk fibroin was combined with retinoic acid and riboflavin to generate a biomaterial scaffold that could control the behavior of corneal stromal cells.
These results of this research has been published in peer-review journals and are available in open access repositories.
The influence of different chemical and physical environments on corneal epithelial and stromal cells has been established in this project. This information enhances our fundamental understanding of how corneal cells behave and react in the presence of specific stimuli and could be used to optimize cell culture conditions and improve scaffold design. Novel scaffold production technology has been developed and evaluated during this project including a new methods of generating aligned nanofiber scaffolds and a new method of recellularizing decellularized corneas by sectioning the scaffolds and incorporating cell seeded hydrogels between each layer. The outcome of this research will improve the quality of life of patients suffering from corneal blindness across Europe.