RESUMO
Biological activities of cells such as survival and differentiation processes are mainly maintained by a specific extracellular matrix (ECM). Hydrogels have recently been employed successfully in tissue engineering applications. In particular, scaffolds made of gelatin methacrylate-based hydrogels (GelMA) showed great potential due to their biocompatibility, biofunctionality, and low mechanical strength. The development of a hydrogel having tunable and appropriate mechanical properties as well as chemical and biological cues was the aim of this work. A synthetic and biological hybrid hydrogel was developed to mimic the biological and mechanical properties of native ECM. A combination of gelatin methacrylate and acrylamide (GelMA-AAm)-based hydrogels was studied, and it showed tunable mechanical properties upon changing the polymer concentrations. Different GelMA-AAm samples were prepared and studied by varying the concentrations of GelMA and AAm (AAm2.5% + GelMA3%, AAm5% + GelMA3%, and AAm5% + GelMA5%). The swelling behavior, biodegradability, physicochemical and mechanical properties of GelMA-AAm were also characterized. The results showed a variation of swelling capability and a tunable elasticity ranging from 4.03 to 24.98 kPa depending on polymer concentrations. Moreover, the podocyte cell morphology, cytoskeleton reorganization and differentiation were evaluated as a function of GelMA-AAm mechanical properties. We concluded that the AAm2.5% + GelMA3% hydrogel sample having an elasticity of 4.03 kPa can mimic the native kidney glomerular basement membrane (GBM) elasticity and allow podocyte cell attachment without the functionalization of the gel surface with adhesion proteins compared to synthetic hydrogels (PAAm). This work will further enhance the knowledge of the behavior of podocyte cells to understand their biological properties in both healthy and diseased states.
RESUMO
Chronic kidney disease is characterized by a gradual decline in renal function that progresses toward end-stage renal disease. Podocytes are highly specialized glomerular epithelial cells which form with the glomerular basement membrane (GBM) and capillary endothelium the glomerular filtration barrier. GBM is an extracellular matrix (ECM) that acts as a mechanical support and provides biophysical signals that control normal podocytes behavior in the process of glomerular filtration. Thus, the ECM stiffness represents an essential characteristic that controls podocyte function. Hydrolyzed Polyacrylamide (PAAm) hydrogels are smart polyelectrolyte materials. Their biophysical properties can be tuned as desired to mimic the natural ECM. Therefore, these hydrogels are investigated as new ECM-like constructs to engineer a podocyte-like basement membrane that forms with cultured human podocytes a functional glomerular-like filtration barrier. Such ECM-like PAAm hydrogel construct will provide unique opportunity to reveal podocyte cell biological responses in an in vivo-like setting by controlling the physical properties of the PAAm membranes. In this work, Hydrolyzed PAAm scaffolds having different stiffness ranging between 0.6-44 kPa are prepared. The correlation between the hydrogel structural and mechanical properties and Podocyte morphology, elasticity, cytoskeleton reorganization, and podocin expression is evaluated. Results show that hydrolyzed PAAm hydrogels promote good cell adhesion and growth and are suitable materials for the development of future 3D smart scaffolds. In addition, the hydrogel properties can be easily modulated over a wide physiological range by controlling the cross-linker concentration. Finally, tuning the hydrogel properties is an effective strategy to control the cells function. This work addressed the complexity of podocytes behavior which will further enhance our knowledge to develop a kidney-on-chip model much needed in kidney function studies in both healthy and diseased states.
