# Neuroprotective properties of erythropoietin (Epo) and its receptor (EpoR) in open spinal dysraphism (OSD): an investigation of EpoR expression in a rat OSD model, along with in vitro studies on the neuroprotective effects of Epo on rat spinal cord-derived neural progenitor cells

## TL;DR

This study explores how erythropoietin (Epo) and its receptor (EpoR) may protect nerve cells in a rat model of spinal birth defects, showing Epo could help reduce cell damage.

## Contribution

The study demonstrates Epo's neuroprotective effects in vitro and identifies EpoR upregulation in a rat model of open spinal dysraphism.

## Key findings

- EpoR mRNA expression was significantly higher in OSD tissue at E18 and E22 compared to controls.
- Epo counteracted the harmful effects of artificial amniotic fluid on neural progenitor cell proliferation and death.
- Epo restored neuronal marker expression in cells exposed to artificial amniotic fluid.

## Abstract

Research into treatment options that complement surgery for open spinal dysraphism (OSD) is necessary to further improve outcomes following prenatal or postnatal surgery. We investigated erythropoietin (Epo) and its receptor (EpoR) as a potential neuroprotective agent and target in OSD.

Epo- and EpoR-expression was examined on mRNA and immunohistochemical level in neuroplacodes obtained from a rat retinoic acid-induced OSD model at E16, E18, E22. Neural progenitor cells (NPCs) derived from spinal cords of adult Long Evans rats were exposed to varying concentrations of human artificial amniotic fluid (aAF). Cytotoxicity assays were conducted to assess the impact of aAF exposure, with and without Epo. The influence of Epo on NPC’s cellular marker expression was analyzed using qRT-PCR.

EpoR mRNA expression was elevated significantly on E18 and E22 in OSD- compared to control tissue. EpoR immunoreactivity exhibited consistent expression in ventral and dorsal horns, central canal, and ganglia. It co-stained with Hif-2α, β-III-tubulin, NeuN, and Musashi1. Exposure of NPCs to aAF reduced proliferation and increased death rates dose-dependently. Adding Epo significantly counteracted antiproliferative and cytotoxic aAF effects on NPCs, resulting in similar or lower death rates than those observed in untreated controls. Exposure of NPCs to aAF reduced their expression of neuronal markers. Adding Epo restored their differentiation capacities.

Induction of EpoR in neuroplacodes, its co-staining with neuronal and NPC markers, and the neuroprotective effects of Epo on aAF-treated NPCs in vitro establish Epo as a promising candidate for neuroprotective therapies that can supplement surgical measures for OSD.

The online version contains supplementary material available at 10.1007/s00381-025-07032-8.