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The use of autologous peritoneum for mucosal replacement following ileal neobladder construction: a novel technique in a porcine model

ABSTRACT: Background:The transposed intestinal segments maintain their absorptive and secretive functions which may lead to metabolic disorders after ileal neobladder construction. The aim of this study was to develop a novel technique to reduce the complications of orthotopic urinary diversion in a porcine model. Methods: An ileal neobladder procedure was performed in eight female pigs. Four pigs (experimental group) received mucosal replacement of the neobladder using autologous peritoneum, while the other four pigs (control group) received a normal U-shaped ileal neobladder. After three months, the neobladder was evaluated using IVU, cystograms and cystoscopy. Blood biochemistries, urine samples and venous gas analyses were obtained preoperatively and postoperatively. The neobladder was then harvested to assess pathological findings. Results: The mean (range) operating time was 248 (224-287) min in the control group and 325 (296-360) min in the experimental group. All animals but one from the experimental group survived to the designated endpoints. Postoperative IVU revealed that the urinary tracts were patent and that there was no hydronephrosis or dilated ureter. Cystograms demonstrated a spherical neobladder. Cystoscopy showed a smooth neobladder wall. Histological analysis revealed that the peritoneum-ileal seromuscular flap surface was lined with the urothelial layers. Transcriptome comparison showed the maximum similarity between mucosa of neobladder from experimental group and mucosa of normal bladder. Conclusions: The novel technique is shown to be feasible and reproducible in a porcine model. The imageological, histological, and transcriptometric analyses all confirmed that autologous peritoneum can be used as a natural biological reconstruction material.

ORGANISM(S): Sus scrofa

PROVIDER: GSE145751 | GEO | 2021/04/07

REPOSITORIES: GEO

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Project description:Intestinal lymphocytes are crucial members of the mucosal immune system with impact over outcomes of intestinal health versus dysbiosis. Resolving intestinal lymphocyte complexity and function is a challenge, as the intestine provides cellular snapshots of a diverse spectrum of immune states. In pigs, intestinal lymphocytes are poorly described relative to humans or traditional model species. Enhanced understanding of porcine intestinal lymphocytes will promote food security and improve utility of pigs as a biomedical model for intestinal research. Single-cell RNA sequencing (scRNA-seq) was performed to provide transcriptomic profiles of lymphocytes in porcine ileum, with 31,983 cells annotated into 26 cell types. Deeper interrogation revealed previously undescribed cells in porcine intestine, including SELLhi gd T cells, group 1 and group 3 innate lymphoid cells (ILCs), and four subsets of B cells. Single-cell transcriptomes in ileum were compared to those in porcine blood, and subsets of activated lymphocytes were detected in ileum but not periphery. Comparison to scRNA-seq human and murine ileum data revealed a general consensus of ileal lymphocytes across species. Lymphocyte spatial context in porcine ileum was conferred through differential tissue dissection prior to scRNA-seq. Antibody-secreting cells, B cells, follicular CD4 ab T cells, and cycling T/ILCs were enriched in ileum with Peyer’s patches, while non-cycling gd T, CD8 ab T, and group 1 ILCs were enriched in ileum without Peyer’s patches. scRNA-seq findings were leveraged to develop advanced toolsets for further identification of ILCs in porcine ileum via flow cytometry and in situ staining. Porcine ileal ILCs identified via scRNA-seq did not transcriptionally mirror peripheral porcine ILCs (corresponding to natural killer cells) but instead had gene signatures indicative of tissue- and activation-specific functions, indicating potentially similar roles to intestinal ILCs identified in humans. Overall, the data serve as a highly-resolved transcriptomic atlas of the porcine intestinal immune landscape and will be useful in further understanding intestinal immune cell function.

Project description:Background: Cerebral ischemia/reperfusion injury is a common secondary effect of cardiac arrest which is largely responsible for postresuscitative mortality. Therefore development of therapies which restore and protect the brain function after cardiac arrest is essential. Methylene blue (MB) has been experimentally proven neuroprotective in a porcine model of global ischemia-reperfusion in experimental cardiac arrest. However, no comprehensive analyses have been conducted at gene expression level. Methods: Pigs underwent either untreated cardiac arrest (CA) or CA with subsequent cardiopulmonary resuscitation (CPR) accompanied with an infusion of saline or an infusion of saline with MB. Genome-wide transcriptional profiling using the Affymetrix porcine microarray was performed to 1) gain understanding of delayed neuronal death initiation in porcine brain during ischemia and after 30, 60 and 180 min following reperfusion, and 2) identify the mechanisms behind the neuroprotective effect of MB after ischemic injury (at 30, 60 and 180 min). Results: Our results show that restoration of spontaneous circulation (ROSC) induces major transcriptional changes related to stress response, inflammation, apoptosis and even cytoprotection. In contrast, the untreated ischemic and anoxic insult affected only few genes mainly involved in intra-/extracellular ionic balance. Furthermore, our data show that the neuroprotective role of MB is diverse and fulfilled by regulation of the expression of soluble guanylate cyclase and biological processes accountable for inhibition of apoptosis, modulation of stress response, neurogenesis and neuroprotection. Conclusions: Our results support that MB could be a valuable intervention and should be investigated as a therapeutic agent against neural damage associated with I/R injury induced by cardiac arrest. One group of pigs underwent cardiac arrest (CA) without subsequent cardiopulmonary resuscitation (CPR) and the brain of the animals was removed after 5, 20 or 30 min post CA for genome-wide expression study. Two groups of pigs underwent first CA for 12 min with a subsequent 8 min-long CPR and received either an infusion of saline or an infusion of saline with MB from one minute after the start of CPR until 50 min after return of spontaneous circulation (ROSC). In both latter groups the brains were removed for microarray analysis at the following time points: 30, 60, 180 min.

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The use of autologous peritoneum for mucosal replacement following ileal neobladder construction: a novel technique in a porcine model

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