Role of NADPH oxidase in tissue growth in a tissue engineering chamber in rats

Previously we described a subcutaneous arteriovenous loop (AVL)-based tissue engineering chamber system, which contains an intrinsic circulation circuit created by joining the proximal ends of the femoral artery and vein with a venous graft. We showed that nicotinamide adenine dinucleotide phosphate (NADPH) oxidase was involved in mediating neovascularization inside the chamber. However, the role of NADPH oxidase in tissue formation in the chamber is unknown. In this study, we examined the effects of gp91ds-tat, a peptidyl inhibitor of NADPH oxidase, on the growth of engineered tissue blocks, using a rat chamber model. Chambers containing the AVL were filled with Matrigel mixed with gp91ds-tat (100 µM) or the scrambled control peptide. At 14 days, in control chambers, most of the Matrigel was replaced by granulation tissues; gp91ds-tat treatment significantly reduced the level of reactive oxygen species and retarded the tissue formation process. Although the total number of blood vessels per unit cellularized area was not different between two groups, most vessels in gp91ds-tat-treated tissues had smaller lumens as compared to control. The total area occupied by vessel lumens was much less in gp91ds-tat-treated tissues (10.3 ± 1.3% in control vs. 1.7 ± 0.5% in gp91ds-tat group; p < 0.001). In vitro, gp91ds-tat treatment reduced proliferation and migration of cultured microvascular endothelial cells. Our data suggest that inhibition of NADPH oxidase function retards tissue formation in the tissue engineering chamber, which may be related to the malformed new blood vessels in the engineered tissue.