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常温机械灌注与低温冷藏保存对猪肠同种异体移植物再生潜能和活力影响的比较。

Comparison of the effects of normothermic machine perfusion and cold storage preservation on porcine intestinal allograft regenerative potential and viability.

机构信息

Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA.

Department of Surgery, Duke University School of Medicine, Durham, North Carolina, USA.

出版信息

Am J Transplant. 2024 Apr;24(4):564-576. doi: 10.1016/j.ajt.2023.10.026. Epub 2023 Oct 31.

Abstract

Intestinal transplantation (IT) is the final treatment option for intestinal failure. Static cold storage (CS) is the standard preservation method used for intestinal allografts. However, CS and subsequent transplantation induce ischemia-reperfusion injury (IRI). Severe IRI impairs epithelial barrier function, including loss of intestinal stem cells (ISC), critical to epithelial regeneration. Normothermic machine perfusion (NMP) preservation of kidney and liver allografts minimizes CS-associated IRI; however, it has not been used clinically for IT. We hypothesized that intestine NMP would induce less epithelial injury and better protect the intestine's regenerative ability when compared with CS. Full-length porcine jejunum and ileum were procured, stored at 4 °C, or perfused at 34 °C for 6 hours (T6), and transplanted. Histology was assessed following procurement (T0), T6, and 1 hour after reperfusion. Real-time quantitative reverse transcription polymerase chain reaction, immunofluorescence, and crypt culture measured ISC viability and proliferative potential. A greater number of NMP-preserved intestine recipients survived posttransplant, which correlated with significantly decreased tissue injury following 1-hour reperfusion in NMP compared with CS samples. Additionally, ISC gene expression, spheroid area, and cellular proliferation were significantly increased in NMP-T6 compared with CS-T6 intestine. NMP appears to reduce IRI and improve graft regeneration with improved ISC viability and proliferation.

摘要

肠移植(IT)是治疗肠衰竭的最终手段。静态冷保存(CS)是用于同种异体肠移植的标准保存方法。然而,CS 及随后的移植会引起缺血再灌注损伤(IRI)。严重的 IRI 会损害上皮屏障功能,包括肠干细胞(ISC)的丧失,这对上皮再生至关重要。肾和肝同种异体移植物的常温机器灌注(NMP)保存可以最大限度地减少 CS 相关的 IRI;然而,它尚未在 IT 临床中应用。我们假设与 CS 相比,NMP 可以减少上皮损伤,更好地保护肠道的再生能力。全长猪空肠和回肠在 4°C 下获取、储存,或在 34°C 下灌注 6 小时(T6),并进行移植。在获取(T0)、T6 后和再灌注 1 小时后评估组织学。实时定量逆转录聚合酶链反应、免疫荧光和隐窝培养用于测量 ISC 活力和增殖潜力。与 CS 样本相比,NMP 保存的肠移植受者在移植后有更多的存活,这与 1 小时再灌注后 NMP 样本中组织损伤明显减少有关。此外,与 CS-T6 肠相比,NMP-T6 肠中的 ISC 基因表达、球体面积和细胞增殖显著增加。NMP 似乎可以减少 IRI,并通过提高 ISC 活力和增殖来改善移植物再生。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/962f/11082874/ea59253a0841/nihms-1990527-f0001.jpg

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