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局部移植脂肪干细胞可减少缺血性结直肠吻合口漏:大鼠模型。

Locally Transplanted Adipose Stem Cells Reduce Anastomotic Leaks in Ischemic Colorectal Anastomoses: A Rat Model.

机构信息

Department of Surgery, Cooper University Hospital, Camden, New Jersey.

出版信息

Dis Colon Rectum. 2020 Jul;63(7):955-964. doi: 10.1097/DCR.0000000000001667.

DOI:10.1097/DCR.0000000000001667
PMID:32168095
Abstract

BACKGROUND

Anastomotic leakage remains a dreaded complication after colorectal surgery. Stem-cell-based therapies have been shown to increase angiogenesis and cell proliferation.

OBJECTIVE

The purpose of this research was to investigate the use of adipose-derived stem cells on the healing of ischemic colonic anastomoses in a rat model.

DESIGN

This is an animal research study using xenotransplantation.

SETTINGS

Male Wistar rats (300-400 g, n = 48) were purchased from a licensed breeder.

PATIENTS

Adipose stem cells were isolated from the subcutaneous fat of healthy human donors.

INTERVENTIONS

The rats underwent laparotomy with creation of an ischemic colorectal anastomosis created by ligation of mesenteric vessels. The animals were divided into 3 groups: control group with an ischemic anastomosis, vehicle-only group in which the ischemic anastomosis was treated with an absorbable gelatin sponge, and a treatment group in which the ischemic anastomosis was treated with an absorbable gelatin sponge plus adipose stem cells. Animals were killed at postoperative days 3 and 7.

MAIN OUTCOME MEASURES

Anastomotic leakage was defined as the finding of feculent peritonitis or perianastomotic abscess on necropsy. Rat mRNA expression was measured using real-time polymerase chain reaction.

RESULTS

Adipose-derived stem cells significantly decreased anastomotic leakage when compared with control at both postoperative days 3 (25.0% vs 87.5%; p = 0.02) and 7 (25.0% vs 87.5%; p = 0.02). The use of an absorbable gelatin sponge alone had no effect on anastomotic leakage when compared with control and postoperative days 3 or 7. We found that stem cell-treated animals had a 5.9-fold and 7.4-fold increase in the expression of vascular endothelial growth factor when compared with control at 3 and 7 days; however, this difference was not statistically significant when compared with the absorbable gelatin sponge group.

LIMITATIONS

This is a preclinical animal research study using xenotransplantation of cultured stem cells.

CONCLUSIONS

Locally transplanted adipose stem cells enhance the healing of ischemic colorectal anastomoses and may be a novel strategy for reducing the risk of anastomotic leakage in colorectal surgery. See Video Abstract at http://links.lww.com/DCR/B203. EL TRANSPLANTE LOCAL DE CÉLULAS MADRE ADIPOSAS REDUCE LA FUGA ANASTOMÓTICA EN LAS SUTURAS COLORRECTALES ISQUÉMICAS: MODELO EN RATAS: Las fugas anastomóticas son una complicación pusilánime después de toda cirugía colorrectal. Se ha demostrado que el tratamiento con células madre aumenta la angiogénesis y la proliferación celular.Investigar el uso de células madre derivadas de tejido adiposo en la cicatrización de una anastomosis colónica isquémica basada en ratas como modelo.Estudio de investigación en animales utilizando xenotrasplantes.Adquisición de típicas ratas de laboratorio raza Wistar, todas machos (300-400 g, n = 48) de un criadero autorizado.Aislamiento de células madre de tipo adiposo del tejido celular subcutáneo en donantes humanos sanos.Las ratas se sometieron a laparotomía con la creación de una anastomosis colorrectal isquémica obtenida mediante ligadura controlada de los vasos mesentéricos correspondientes. Los animales se dividieron en tres grupos: grupo de control con anastomosis isquémica, grupo de vehículo único en el que la anastomosis isquémica se trató con una esponja de gelatina absorbible, y un grupo de tratamiento en el que la anastomosis isquémica se trató con una esponja de gelatina absorbible asociada a un vástago adiposo de células madre. Los animales fueron sacrificados el POD3 y el POD7.La fuga anastomótica fué definida como el hallazgo de peritonitis fecaloidea o absceso perianastomótico a la necropsia. La expresión de RNAm de las ratas se midió usando PCR en tiempo real.Las células madre derivadas de tejido adiposo disminuyeron significativamente la fuga anastomótica en comparación con el grupo control tanto en el POD3 (25% frente a 87.5%, p = 0.02) como en el POD7 (25% frente a 87.5%, p = 0.02). El uso de una esponja de gelatina absorbible sola, no tuvo efecto sobre la fuga anastomótica en comparación con los controles el POD3 o el POD7. Descubrimos que los animales tratados con células madre adiposas tenían un aumento de 5,9 y 7,4 veces en la expresión de VEGF en comparación con el control a los 3 y 7 días, respectivamente; sin embargo, esta diferencia no fue estadísticamente significativa en comparación con el grupo de esponja de gelatina absorbible.Este es un estudio preclínico de investigación en animales que utiliza xenotrasplantes de células madre adiposas cultivadas.Las células madre de tipo adiposo trasplantadas localmente mejoran la cicatrisación en casos de anastomosis colorrectales isquémicas, y podrían convertirse en una nueva estrategia para reducir el riesgo de fugas anastomóticas en casos de cirugía colorrectal. Consulte Video Resumen en http://links.lww.com/DCR/B203. (Traducción-Dr Xavier Delgadillo).

摘要

背景

吻合口漏仍然是结直肠手术后一种可怕的并发症。基于干细胞的疗法已被证明可增加血管生成和细胞增殖。

目的

本研究旨在探讨脂肪来源干细胞在大鼠缺血性结肠吻合模型中的应用。

设计

这是一项使用异种移植的动物研究。

地点

购买了来自许可饲养者的 300-400g 雄性 Wistar 大鼠(n=48)。

患者

从健康供体的皮下脂肪中分离出脂肪干细胞。

干预措施

大鼠行剖腹术,结扎肠系膜血管以创建缺血性结直肠吻合术。将动物分为 3 组:对照组,吻合口缺血;载体组,吻合口仅用可吸收明胶海绵处理;治疗组,吻合口用可吸收明胶海绵加脂肪干细胞处理。术后第 3 天和第 7 天处死动物。

主要观察指标

吻合口漏定义为尸检时发现粪便性腹膜炎或吻合口周围脓肿。使用实时聚合酶链反应测量大鼠 mRNA 表达。

结果

与对照组相比,脂肪来源干细胞组在术后第 3 天(25.0%比 87.5%;p=0.02)和第 7 天(25.0%比 87.5%;p=0.02)吻合口漏发生率明显降低。与对照组相比,单独使用可吸收明胶海绵对吻合口漏无影响,且术后第 3 天和第 7 天差异无统计学意义。我们发现,与对照组相比,干细胞治疗组在第 3 天和第 7 天血管内皮生长因子的表达分别增加了 5.9 倍和 7.4 倍;然而,与可吸收明胶海绵组相比,这一差异无统计学意义。

局限性

这是一项使用异种移植培养干细胞的临床前动物研究。

结论

局部移植的脂肪干细胞可促进缺血性结直肠吻合的愈合,可能是降低结直肠手术吻合口漏风险的一种新策略。详见原文视频摘要,可访问 http://links.lww.com/DCR/B203。

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