Tissue-engineered three-dimensional in vitro models for normal and diseased kidney.

Morphogenesis of epithelial cells involves processes by which kidney shape and function are regulated. The lack of in vitro models that are sustainable for longer time periods and emulating complex intercellular interactions of the kidney have limited understanding about epithelial tissue morphogenesis and its aberrations in diseases such as autosomal dominant polycystic kidney disease (ADPKD). A sustainable three-dimensional (3D) coculture system for normal and diseased kidney tissues is reported here. Tubule- and ADPKD cyst-derived cells were cultured in extracellular matrix molecules infused into 3D porous silk scaffolds, and these cultures were subsequently extended into a perfusion bioreactor. The results indicated collagen-matrigel-mediated morphogenesis for both (normal and disease) cell types and also supported coculturing with fibroblasts. The structural and functional features of the kidney-like tissue structures were validated based on the distribution of E-cadherin, N-cadherin, Na+ K+ ATPase pump, and cellular uptake of the organic anion (6-carboxy fluorescein). Further, the structures were sustained for longer time periods using a perfusion bioreactor to demonstrate the potential utility of this 3D in vitro coculture system for ADPKD research, other epithelial tissue systems, and for in vitro drug screening.

Synthetic oxygen carriers in cardiac tissue engineering.

The prominence of cardiovascular diseases has prompted investigations into alternative treatment options, including tissue engineering. Currently, the biggest limitation in cardiac tissue engineering lies in delivering oxygen to all cells within the construct. Synthetic oxygen carriers hold much promise in that they have high affinity for oxygen and can be supplemented to culture medium without adverse effect on the cells. This review highlights two complementary studies by our group that utilized oxygen carriers in cardiac tissue engineering. Experimental and modeling studies were performed to evaluate the effect of a perfluorocarbon (PFC)-based synthetic oxygen carrier, Oxygent, on oxygen supply within tissue engineered cardiac constructs. Porous biorubber scaffolds with an array of parallel channels mimicking the capillary network were seeded with cardiomyocytes and fibroblasts, and cultivated in medium supplemented with PFC. The presence of PFC enhanced the transport of oxygen, increased oxygen concentrations, and yielded constructs that displayed stronger cardiac-like phenotype.

Characterization of electrical stimulation electrodes for cardiac tissue engineering.

Electrical stimulation has been shown to improve functional assembly of cardiomyocytes in vitro for cardiac tissue engineering. The goal of this study was to assess the conditions of electrical stimulation with respect to the electrode geometry, material properties and charge-transfer characteristics at the electrode-electrolyte interface. We compared various biocompatible materials, including nanoporous carbon, stainless steel, titanium and titanium nitride, for use in cardiac tissue engineering bioreactors. The faradaic and non-faradaic charge transfer mechanisms were assessed by electrochemical impedance spectroscopy (EIS), studying current injection characteristics, and examining surface properties of electrodes with scanning electron microscopy. Carbon electrodes were found to have the best current injection characteristics. However, these electrodes require careful handling because of their limited mechanical strength. The efficacy of various electrodes for use in 2-D and 3-D cardiac tissue engineering systems with neonatal rat cardiomyocytes is being determined by assessing cell viability, amplitude of contractions, excitation thresholds, maximum capture rate, and tissue morphology.