Nephroprotective factor improves kidney scaffold recellularization

GUERRIERI D; HAEUBLEIN, GERALDINE; AMBROSI, NELLA; NICOLAS ALEJANDRO FRAUNHOFFER; CARO, FIORELLA; SANCHEZ, FRANCISCO; TONIOLO, FERNANDA; FIORETTI, SABRINA; TRUNZO, LUIS; TANUS, EDUARDO; BACQUE, MARIA DEL CARMEN; INCARDONA, C; CASADEI, DOMINGO; CHULUYAN, EDUARDO

Congreso; CST-CTRMS Joint Sicientific Meeting; 2017

Organs decellularization produce scaffolds, which could be seeded with different kind of cells. Successful repopulation is actually the main challenge step in the field of organ engineering and regenerative medicine. The aim of the present work was to improve kidney scaffold recellularization by using nephroprotective agents. Previously we observed that a serpin called SLPI (secretory leukocyte proteinase inhibitor) reduced the nephrotoxic activity induced by gentamicin or ischemia reperfusion injury in an in vivo rat model. Since human HK-2 renal epithelial cell line is used for recellularize the kidney scaffolds, we tested the nephrotective activity of SLPI on these cells in vitro. The cytotoxicity effect was measured using MTT and Annexin V/propidium iodide assays. The incubation of HK-2 with FK506 induced cytotoxic activity which was reduced by treating the cells with 4 and 0.4 ug/ml (Figure 1). By using a ?Human Stress & Toxicity PCR Array? we observed that SLPI decreased the oxidative Stress, Hypoxia, Osmotic Stress, Cell Death, Inflammatory Response transcripts induced by FK506 and increased Heat Shock Proteins (ATF4, ATF6, HSP90AA1) transcripts. Furthermore, SLPI was able to induce the HK-2 cell proliferation over 24 h of culture (Figure1B) and to achieve a better wound closure recovery of the damaged surface compare to untreated cells, in a wound healing scratch assay (Figure1C). Then, a rat kidney scaffold was recellularized with HK-2. Untreated or SLPI (4g/ml) treated HK-2 cells were administered to a rat kidney scafold by ureter. Higher percentage of cells were seeded in the scafold when SLPI was used. Furthermore, the histological analysis of the scafold showed (at 24 h) higher amount of cells attach to the renal tubules. Overall these results confirm the nephrotective effects of SLPI, which could be a good tool to help in the recelullarization process.