Abstract
Purpose :
The scarcity of corneal donors has fueled a high demand for substitutes. Peptide amphiphiles(PAs) can be designed to influence cellular behaviour and are instrumental in the development of scaffolds for corneal regeneration. This study investigates the influence of incorporating RGDS PAs into ETTES PAs at different concentrations on the metabolic activity of stem cells, with a specific application in corneal tissue engineering.
Methods :
1.25 mM of 100% C16-ETTES and 1.25 mM of 80% C16-ETTES + 20% C16 RGDS were separately dissolved in double-distilled water. Subsequently, each solution was diluted to concentrations of 125 µM, 12.5 µM, and 1.25 µM. 3 biological replicates of 24 well plates were coated with ETTES PAs and the composite of RGDS & ETTES PAs at concentrations of 1.25mM, 125 µM, 12.5 µM and 1.25 µM. Adipose Stem cells (ASCs) were incubated on these coatings for 3 days, and their metabolic activities were quantitatively measured as florescent intensity using Alamar Blue Assay in comparison to the control groups incubated on uncoated 24 well plates. Two-way ANOVA was used for statistical analysis. Moreover, ASC proliferation and morphology were assessed using advanced microscopy techniques with Leica Microsystems.
Results :
The hybrid formulation, 80% ETTES & 20% RGDS PAs demonstrated superior performance in sustaining stem cell viability compared to 100% ETTES PAs across various concentrations (125 µM, 12.5 µM, and 1.25 µM) and at two distinct time points. This formulation exhibited enhanced metabolic activity at these concentrations relative to 100% ETTES. While variations among concentrations (125 µM, 12.5 µM, and 1.25 µM) did not yield significant differences, it is noteworthy that 1.25 mM failed to sustain cell viability in both formulations. This outcome underscores the importance of integrating RGDS in the design of PA scaffolds to enhance cellular adhesion and viability. Furthermore, it emphasizes the optimal threshold concentration, observed to be under 1.25 mM for effectiveness.
Conclusions :
The study emphasizes the importance of PA composition in influencing cellular responses. The hybrid formulation outperformed 100% ETTES, offering valuable insights for optimizing peptide amphiphiles in corneal tissue engineering, stressing the need to consider both composition and concentration in scaffold design.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.