Optimizing Optics: Investigating the Impact of 2D Matrix Stiffness on Engineered Corneal Tissue Growth
Poster Number
21A
Research or Creativity Area
Engineering & Computer Science
Abstract
Introduction: Corneal blindness is a significant global health issue, affecting 5% of the world’s population. Traditional treatments like corneal transplantation face challenges due to the shortage of donor corneas and post-operative complications. Tissue engineering offers a promising solution by developing bioengineered corneal substitutes to overcome these limitations. Recent advancements in biomaterials and biofabrication techniques have enabled the creation of functional corneal constructs with native-like properties. In this abstract, we present our research focused on optimizing scaffold designs to develop safe and effective corneal substitutes, specifically by determining the optimal scaffold stiffness. Substrate stiffness has been shown to influence cell phenotype and is a critical factor in designing a tissue-engineered scaffold. Higher substrate stiffnesses have also been correlated to increased wound healing response in fibroblastic cells. Our work contributes to advancing tissue engineering approaches for corneal regeneration, offering new hope to individuals suffering from corneal blindness.
Materials and Methods: Our research was focused on the growth and morphology of rabbit corneal cells on 2D collagen gels of varying stiffnesses. Plates of seven different elastic moduli ranging from 0.2 to 64 kPa (0.2, 0.5, 2, 8, 16, 32, 64, CytoSoft Discovery Kit) were coated with collagen type I (Advanced Biomatrix) and seeded with rabbit corneal fibroblast cells passage (2-8) at 5000 cells/cm². Acting as controls, were 2 plates— one plate was coated with collagen (CC) and the other was without (CNC). A total of 3 trials were conducted with images being taken on days 1, 3, 5, 7 at around the same time to best capture the cells at the 1-day mark. Cells were fed every 3 days. Qualitative analysis on cell morphology and confluency was performed.
Results and discussion: Fibroblast formation was successful in all plates regardless of elastic moduli. However, cells plated in 32 kPa reached confluency faster and were more confluent overall after 7 days when compared to both controls and all other stiffnesses. Following the 32 kPa plate, cells in 16 kPa and 64 kPa plates were most efficient at reaching confluency, respectively, and both were more confluent than controls after 7 days. The morphology of cells cultured under all conditions were similar.
Conclusion: From the conducted trials, we found that cells cultured in plates with higher stiffnesses reached confluency earlier and were more confluent than controls. Our next steps are to lyse the cells and conduct western blots in order to determine which proteins are expressed in each plate, and to what extent. This will help us to determine what substrate stiffness is necessary for a more normal corneal cell protein expression. Future research will investigate the impact of the stiffness of 3D collagen matrices on cornea cell growth.
Location
Don and Karen DeRosa University Center (DUC) Poster Hall
Start Date
27-4-2024 10:30 AM
End Date
27-4-2024 12:30 PM
Optimizing Optics: Investigating the Impact of 2D Matrix Stiffness on Engineered Corneal Tissue Growth
Don and Karen DeRosa University Center (DUC) Poster Hall
Introduction: Corneal blindness is a significant global health issue, affecting 5% of the world’s population. Traditional treatments like corneal transplantation face challenges due to the shortage of donor corneas and post-operative complications. Tissue engineering offers a promising solution by developing bioengineered corneal substitutes to overcome these limitations. Recent advancements in biomaterials and biofabrication techniques have enabled the creation of functional corneal constructs with native-like properties. In this abstract, we present our research focused on optimizing scaffold designs to develop safe and effective corneal substitutes, specifically by determining the optimal scaffold stiffness. Substrate stiffness has been shown to influence cell phenotype and is a critical factor in designing a tissue-engineered scaffold. Higher substrate stiffnesses have also been correlated to increased wound healing response in fibroblastic cells. Our work contributes to advancing tissue engineering approaches for corneal regeneration, offering new hope to individuals suffering from corneal blindness.
Materials and Methods: Our research was focused on the growth and morphology of rabbit corneal cells on 2D collagen gels of varying stiffnesses. Plates of seven different elastic moduli ranging from 0.2 to 64 kPa (0.2, 0.5, 2, 8, 16, 32, 64, CytoSoft Discovery Kit) were coated with collagen type I (Advanced Biomatrix) and seeded with rabbit corneal fibroblast cells passage (2-8) at 5000 cells/cm². Acting as controls, were 2 plates— one plate was coated with collagen (CC) and the other was without (CNC). A total of 3 trials were conducted with images being taken on days 1, 3, 5, 7 at around the same time to best capture the cells at the 1-day mark. Cells were fed every 3 days. Qualitative analysis on cell morphology and confluency was performed.
Results and discussion: Fibroblast formation was successful in all plates regardless of elastic moduli. However, cells plated in 32 kPa reached confluency faster and were more confluent overall after 7 days when compared to both controls and all other stiffnesses. Following the 32 kPa plate, cells in 16 kPa and 64 kPa plates were most efficient at reaching confluency, respectively, and both were more confluent than controls after 7 days. The morphology of cells cultured under all conditions were similar.
Conclusion: From the conducted trials, we found that cells cultured in plates with higher stiffnesses reached confluency earlier and were more confluent than controls. Our next steps are to lyse the cells and conduct western blots in order to determine which proteins are expressed in each plate, and to what extent. This will help us to determine what substrate stiffness is necessary for a more normal corneal cell protein expression. Future research will investigate the impact of the stiffness of 3D collagen matrices on cornea cell growth.
