A Biomaterials Approach to Kidney Engineering: The Structural and Chemical Modification of Silk Fibroin Scaffolds for in vitro Tubulogenesis
Sandler, Nathan
2020
- Sponges for kidney tissue engineering were created using Bombyx mori silk fibroin. Silk protein was fabricated into random and aligned porous three-dimensional sponges. The mechanical properties of the scaffolds were determined using dynamic mechanical analysis (DMA). The structural properties of the scaffolds, including the porosity, pore size distribution, water swelling, and morphology, were ... read morealso analyzed. The compressive modulus of the isotropic sponges was 1.65 kPa, while only about 0.4 kPa for the directional sponges, but the porosity of these scaffolds was about twice as high as the random scaffolds. The amount of swelling of the directional sponges was also higher than the random sponges. Both scaffold types were seeded with renal proximal tubular epithelial cells (RPTECs), which exhibited a 23% higher fluorescence, corresponding to a higher cell density. Similarly, a higher cell density was observed between statically cultured and perfused anisotropic sponges. Perfusion increased RPTEC proliferation, morphology, and maturation, with three-dimensional tubule-like structures present, while missing from static RPTEC sponges. Uptake and ammoniagenesis assays were performed, with both static and perfused scaffolds responding to simulated acidosis. Chemically modified silk scaffolds were fabricated to bind Jagged1, an important ligand in kidney morphogenesis. Material characterization indicated varying degrees of modification, but inconclusive Jagged1 binding. iPSC-derived nephron progenitor cells were cultured and seeding into modified scaffolds to observe the effect of bound Jagged1, but further analysis is required to understand its impact on kidney organoid formation. Advisor: Dr. David Kaplan.read less
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- sj139f19c
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