Ex-vivo perfusion as a successful strategy for reduction of ischemia-reperfusion injury in prolonged muscle flap preservation - A gene expression study.

INTRODUCTION: With the introduction of vascularized composite allotransplantation (VCA) as new surgical technique, the need arose for strategies that could safely prolong graft preservation. Ex-vivo machine perfusion is a promising technique and is currently applied in solid organ transplantation. There is still limited evidence in the field of VCA and free flap transplantation. This gene expression study aimed to assess the degree of ischemia-reperfusion (IR) injury after preservation and replantation of free muscle flaps in a porcine model. MATERIALS AND METHODS: A microarray analysis was first conducted on muscle flaps preserved by ex-vivo perfusion versus cold storage, to select genes of interest for further investigation. The expression of these selected genes was then examined in a muscle flap replantation model after 18 hour ex-vivo perfusion (n = 14) using qRT-PCR. Two preservation solutions were compared to static cold storage: University of Wisconsin-mp (n = 5) and Histidine-Tryptophan-Ketoglutarate solution (n = 5). RESULTS: A selection of 8 genes was made based on micro-array results: Tumor necrosis factor receptor superfamily member 10-A like, Regulator of G-protein signaling 2, Nuclear factor kappa beta inhibitor zeta, Interleukin-1 beta, Fibroblast growth factor 6 and DNA damage-inducible transcript 4, Hypoxia-inducible factor 1-alpha and Caspase-3. The muscle flap replantation experiment compared their expression patterns before and after preservation and replantation and showed overall comparable gene expression between the preservation groups. CONCLUSIONS: The expression of genes related to ischemia, apoptosis and inflammation was comparable between the ex-vivo perfusion and static cold storage groups. These results suggest that ex-vivo perfusion might be a promising technique for 18 hour muscle preservation in terms of decreasing ischemia-reperfusion injury.

Cholinergic drugs affect novel object recognition in rats: relation with hippocampal EEG?

This study examined the role of cognitively enhancing cholinergic drugs on both object memory and brain activity in rats, as well as the possible relation between the two measures. A group of twenty-four animals was used for assessing object recognition. In another group of eight rats, an electrode was implanted into the dorsal hippocampus to record an electroencephalogram (EEG) and auditory evoked potentials (AEP). In both groups, animals were treated with saline, 0.1 mg/kg scopolamine, 0.1 mg/kg methylscopolamine, 3 mg/kg donepezil, donepezil combined with scopolamine, 0.1 mg/kg nicotine, and nicotine combined with scopolamine. Scopolamine, but not methylscopolamine, impaired object recognition. Both donepezil and nicotine reversed this impairment. The N1 and N2 components of the AEP became closer to baseline after scopolamine, which was not reversed by donepezil or nicotine. Scopolamine increased the theta frequency in the EEG. When combined with donepezil, theta increased even more. Conversely, nicotine reversed the theta increment to control level. It is suggested that scopolamine caused a decrement in arousal in this study. Furthermore, the current results suggest a relation between EEG and object memory after cholinergic drug treatment. However, there was a clear dissociation between memory performance and EEG after combined treatment with drugs, which makes additional research where EEG and performance measures are co-registered imperative.