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超顺磁性氧化铁纳米磁标记骨髓间充质干细胞对肝切除术后大鼠肝脏修复的作用。

Superparamagnetic iron oxide magnetic nanomaterial-labeled bone marrow mesenchymal stem cells for rat liver repair after hepatectomy.

机构信息

Department of Hepatobiliary Surgery, The First Affiliated Hospital of Dalian Medical University, Dalian, China.

Department of Anaesthesia, The Second Affiliated Hospital of Dalian Medical University, Dalian, China.

出版信息

J Surg Res. 2014 Oct;191(2):290-301. doi: 10.1016/j.jss.2014.03.064. Epub 2014 Mar 27.

Abstract

BACKGROUND

Superparamagnetic iron oxide magnetic nanomaterials (SPIO) are tracers used for treatment of central nervous and cardiovascular system complications in animal models. The present study investigated survival and proliferation of SPIO-labeled bone marrow mesenchymal stem cells (BMSCs) to determine their potential therapeutic value in liver repair.

METHODS

Surface antigens of BMSCs were measured using flow cytometry. BMSCs viability, growth curve, and SPIO (0-100 μg/mL) labeling rate were evaluated using trypan blue staining, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and Prussian blue staining, respectively. SPIO-labeled BMSCs were transplanted via liver or spleen injection in rats undergoing 70% hepatectomy. Distribution of SPIO-labeled BMSCs in liver and spleen, and liver repair were evaluated by magnetic resonance imaging (MRI), and serum alanine aminotransferase, aspartate aminotransferase, albumin, and total bilirubin levels.

RESULTS

CD29(+)/CD90(+)/CD45(-) BMSCs were successfully isolated from rats. Labeling rate of SPIO in 25 μg/mL was 94.9%. SPIO labeling did not affect BMSCs survival and proliferation. MRI showed that BMSCs colonized in the liver, whether via spleen or liver injection. Serum levels of alanine aminotransferase, aspartate aminotransferase, and total bilirubin in the transplanted rats were significantly lower than in the hepatectomy group at days 1, 3, and 7 after hepatectomy (all P < 0.05), whereas serum albumin levels were significantly higher in the transplanted rats on posthepatectomy day 3 (both P < 0.05). These indicators were not significantly different between the spleen and liver injection approaches.

CONCLUSIONS

BMSCs transplantation via liver or spleen injection could significantly accelerate liver healing. In vivo MRI of SPIO-labeled BMSCs can be used to trace real-time liver healing during clinical treatment after hepatectomy.

摘要

背景

超顺磁性氧化铁纳米磁珠(SPIO)是一种示踪剂,用于治疗动物模型中的中枢神经系统和心血管系统并发症。本研究通过观察 SPIO 标记骨髓间充质干细胞(BMSCs)的存活和增殖情况,来确定其在肝脏修复中的潜在治疗价值。

方法

采用流式细胞术检测 BMSCs 表面抗原。采用台盼蓝染色法、3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法和普鲁士蓝染色法分别检测 BMSCs 的活力、生长曲线和 SPIO(0-100μg/ml)标记率。通过肝内或脾内注射将 SPIO 标记的 BMSCs 移植到 70%肝切除大鼠体内。通过磁共振成像(MRI)和血清丙氨酸氨基转移酶、天冬氨酸氨基转移酶、白蛋白和总胆红素水平来评估 SPIO 标记的 BMSCs 在肝脏和脾脏中的分布以及肝脏修复情况。

结果

成功从大鼠中分离出 CD29(+)/CD90(+)/CD45(-)BMSCs。25μg/ml SPIO 的标记率为 94.9%。SPIO 标记不影响 BMSCs 的存活和增殖。MRI 显示,无论是通过脾内还是肝内注射,BMSCs 都能定植在肝脏中。与肝切除组相比,移植组大鼠在肝切除后第 1、3 和 7 天的血清丙氨酸氨基转移酶、天冬氨酸氨基转移酶和总胆红素水平显著降低(均 P<0.05),而在肝切除后第 3 天的血清白蛋白水平显著升高(均 P<0.05)。脾内和肝内注射两种途径之间的这些指标没有显著差异。

结论

通过肝内或脾内注射 BMSCs 移植可显著加速肝脏愈合。超顺磁性氧化铁标记骨髓间充质干细胞的体内 MRI 可用于在肝切除术后的临床治疗过程中实时追踪肝脏愈合情况。

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