The roles of various metabolic enzymes in diabetic cardiomyopathy—in vivo and in vitro approach
Section snippets
Purpose
To investigate the enzymes involved in the increase of oxidative stress, apoptosis, and fibrosis, processes that contribute to cardiac dysfunction in diabetes; to create a tissue engineering-based model to study the pathology of diabetic cardiomyopathy (DCM).
Methods
Porcine hearts were decellularized and 3 mm3 punch-outs were used as scaffolds. Cells derived from embryonic rat heart were seeded onto the heart extracellular matrix, and the constructs were incubated for 5 days in a cell culture media containing high glucose levels and oxidized LDL, in a dynamic setting using the 3DKube perfusion minibioreactor. As control, cell-scaffold constructs were incubated in normal cell culture media. Tissues were then prepared for histological staining and
Results
Cells were successfully seeded onto the decellularized scaffolds in both normal and diabetic conditions. Histological and biochemical assessments were supportive of current descriptions of the clinical presentation of DCM. There was an elevated level of apoptosis associated with caspase-3 activity, as well as hampered resistance to oxidative stress in both diabetic rat hearts and tissue-engineered constructs.
Conclusion
Oxidative stress coupled with apoptosis and the heart's lack of regeneration, seem to largely contribute to the formation of DCM. With some minor adjustments, the 3DKube bioreactor appears to correctly simulate DCM for future testing.