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Abstract: TH-PO762

Gentamycin Induced Nephrotoxicity in an Artificial System of the Renal Proximal Tubule

Session Information

  • Bioengineering
    October 25, 2018 | Location: Exhibit Hall, San Diego Convention Center
    Abstract Time: 10:00 AM - 12:00 PM

Category: Bioengineering

  • 300 Bioengineering

Authors

  • Sradnick, Jan, University Hospital CGC, Technical University Dresden, Dresden, Germany
  • Schmieder, Florian, Fraunhofer Institute for Material and Technology IWS, Dresden, Germany
  • Sonntag, Frank, Fraunhofer Institute for Material and Technology IWS, Dresden, Germany
  • Todorov, Vladimir T., University Hospital CGC, Technical University Dresden, Dresden, Germany
  • Hohenstein, Bernd, Nephrological Center Villingen-Schwenningen, Villingen-Schwenningen, Germany
  • Hugo, Christian, University Hospital CGC, Technical University Dresden, Dresden, Germany
Background

Mimicking proximal tubular physiology within cell-based artificial model systems might help to investigate pathophysiology of nephrotoxic drugs such as gentamycin commonly leading to acute kidney injury. We present a model system for transport processes along the proximal tubular barrier (TB).

Methods

Immortalized proximal tubule cells (RPTEC) and blood outgrowth endothelial cells (BOEC) were seeded on a transwell membrane (TW) to build a TB. Barrier function was measured by transepithelial electrical resistance (TEER) of the cellular layers on a daily basis. Gentamycin, moxifloxacin and vehicle control (CTRL) were added over the whole experiment (21 days) to investigate their nephrotoxic potential and disturb TB. The treatment influence on glucose transport through the TB was determined by adding fluorescent d-glucose analog (2-NBDG) to the medium above the RPTEC layer. Fluorescence intensity was measured after 90 min at the opposite TW side. Moreover the inhibition of SGLT2 by dapagliflocin (dapa) and canagliflocin (cana) and vehicle control (contr) during antibiotic treatment was studied.

Results

Moxifloxacin treatment increased TEER (median: 153 Ωcm2 vs control 115 Ωcm2), while gentamycin lowered TEER (median: 73 Ωcm2). 2-NBDG concentration increased in the gentamycin treated group (0.79 µg/ml ±0.08 vs control 0.48 µg/ml ±0.07). No differences were found due to moxifloxacin. In CTRLs SGLT2 inhibition led to a reduction of 2-NBDG concentration in BOEC medium (CTRL: 0.5±0.07 µg/ml; ddapa: 0.3 ± 0.03 µg/ml; cana: 0.3 ± 0.06 µg/ml)). In cell systems treated with gentamycin 2-NDBG concentration was reduced following SGLT2 inhibition (contr: 0.8 ±0.09 µg/ml; dapa: 0.5 ±0.02 µg/ml; cana: 0,4 ±0.06 µg/ml). Moxifloxacin had no effect.

Conclusion

In the present study a functional renal proximal tubule cell culture model suitable for drug testing assays was established. TEER and direct 2-NDGB transport were suitable to identify nephrotoxic effects on the TB in vitro. In a next step, this system will be integrated in our established microfluidic system.