Temporal Regulation of GSK3/Wnt Signaling Molecules within a 3D Gelatin/chitosan Scaffold Promoting Cardiac Differentiation of Amniotic Fluid Derived Induced Pluripotent Stem Cells for Congenital Heart Defect Repair

摘要

The current study shows the potential for a completely autologous cardiac tissue patch for the treatment of congenital heart defects. Amniotic fluid derived stem cells can be readily isolated in utero at the time of diagnosis, then induced and differentiated into a beating cardiac lineage. This construct can be grown into functional, beating cardiac tissue which can be implanted directly into the patient at the time of surgical intervention. Continuing studies into small molecule release kinetics are planned. These studies will be conducted on a complete construct by high-performance liquid chromatography, incorporating CHIR99021 and IWP2 encapsulated in PLGA-mPEG nanoparticles. By encapsulating the small molecule inhibitors and combining them within the gelatin/chitosan scaffold, promotion to cardiac differentiation can be contained within this biomaterial construct. From these results, different nanoparticle co-polymer ratios can be investigated to achieve release kinetics promoting higher cardiac differentiation efficiency. Other future studies involve cell electrophysiology analysis through patch clamping, as well as action potential velocity and direction studies through the detection of calcium-sensitive dyes.