BackgroundHypoxic-ischemic encephalopathy (HIE) is a major cause of morbidity in survivors. Therapeutic hypothermia (TH) is the only available intervention, but the protection is incomplete. Preclinical studies of HIE/TH in the rodent have relied on the postnatal day (P) 7 rat whose brain approximates a 32–36 week gestation infant, less relevant for these studies. We propose that HIE and TH in the term-equivalent P10 rat will be more translational.MethodsP10–11 rat pups were subjected to unilateral hypoxia-ischemia (HI) and 4 hours recovery in normothermic (N) or hypothermic (TH) conditions. Brain damage was assessed longitudinally at 24 hours, 2 and 12 weeks. Motor function was assessed with the beam walk; recognition memory was measured by novel object recognition.ResultsNeuroprotection with TH was apparent at 2 and 12 weeks in both moderately and severely damaged animals. TH improved motor function in moderate, but not severe damage. Impaired object recognition occurred with severe damage with no evidence of protection of TH.ConclusionThis adaptation of the immature rat model of HI provides a reproducible platform to further study HIE/TH in which individual animals are followed longitudinally to provide a useful translational preclinical model.

Study Design An in vitro study using ovine intervertebral discs to correlate the effects of advanced glycation end-products (AGEs) with disc hydration evaluated by magnetic resonance imaging (MRI). Objective To determine the relationship between the level of AGEs and tissue water content in intervertebral discs using T2 relaxation MRI. Summary of Background Data AGEs result from nonenzymatic glycation, and AGEs have been shown to accumulate in the IVD tissue with aging and degeneration. AGEs can alter biochemical properties, including the hydrophobicity of the extracellular matrix. Since one of the degenerative signs of the IVD is the reduced hydration, it was hypothesized that increased levels of tissue AGEs may contribute to disc hydration. T2 relaxation MRI has been shown to be sensitive to the hydration status of the disc, and may be valuable in detecting the changes in the IVD mediated by the increase of AGEs. Methods Thirty-eight IVDs were obtained from 4 ovine spines, and the annulus fibrosis (AF) and nucleus pulposus (NP) tissues were isolated from these discs. The tissues were incubated in either a ribosylation or control solution for up to 8 days to induce the formation of AGEs. These tissues were subsequently analyzed for tissue water content and concentration of AGEs. T2 relaxation times were obtained from these tissues after ribosylation. Results Ribosylation led to the increased accumulation of AGEs and reduced water content in both the AF and NP in a dose-dependent manner. When analyzed by MRI, ribosylation significantly altered the mean T2 relaxation times in the NP (p=0.001), but not in the AF (p=0.912). Furthermore, the mean T2 values in the NP significantly decreased with increasing periods of incubation time (p<0.001). Conclusion This study demonstrates that levels of AGEs in the IVD may affect the tissue water content. Moreover, these ribosylation-mediated changes in tissue hydration were detectable using T2 relaxation MRI. T2 relaxation MRI may provide a non-invasive tool to measure in vivo changes in disc hydration that are negatively correlated with the accumulation of AGEs.