脱细胞肠系膜制备生物膜支架的实验研究
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西南医科大学附属医院

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Experimental Study on Preparation of Biofilm Scaffolds from Acellular Mesentery
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the Affiliated Hospital of Southwest Medical University

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    摘要:

    目的 通过应用肠系膜脱细胞基质制备生物膜支架,探讨其理化特性和生物学特性。方法: 应用冻融和酶消化法脱肠系膜细胞,根据是否进行脱细胞处理将肠系膜分为肠系膜基质组(A组)和去细胞肠系膜基质组(B组),通过HE染色、电镜、DNA检测、细胞毒性实验、拉伸力学测试,检测两组肠系膜基质的理化特性;制备覆膜支架,植入兔髂血管,术后1m、2m超声检测血管血流情况,血管取材病理检测。结果: HE染色、电镜检测A组、B组肠系膜,可见B组去细胞肠系膜基质组织疏松,纤维排列较整齐,未见细胞存留;DNA检测B组较A组DNA表达水平低(88.30±2.79ng/ml,450.17±90.90ng/ml),差异有统计学意义;CCK-8细胞毒性实验检测A组、B组均无细胞毒性,FDA-PI荧光染色A组、B组细胞存活良好,未见死亡细胞;拉伸力学测试A组、B组最大力(31.41±7.09N,25.15±7.13N)、最大力伸长率(118.83±15.79%,111.09±15.99%)、屈服强度(8.35±6.46MPa,7.43±4.53MPa)、屈服点伸长率(118.83±17.24%,95.16±22.46%),指标差异无统计学意义;脱细胞肠系膜支架植入兔血管后超声检测两组早期通畅性良好,支架植入2月内皮增生较明显。结论: 冻融和酶消化法脱肠系膜细胞,肠系膜基质去除细胞彻底,无细胞毒性,力学特征良好;肠系膜支架植入血管后早期通畅性良好,2月后内皮增生明显。

    Abstract:

    Objective: To prepare biofilm scaffolds using mesenteric acellular matrix and investigate their physicochemical and biological characteristics.Methods: Mesenteric cells were removed by freeze-thaw and enzymatic digestion. Mesenteries were divided into mesenteric matrix group (group A) and acellular mesenteric matrix group (group B) according to acellular or not. The physical and chemical properties of mesenteric matrix in two groups were tested by HE staining, electron microscopy, DNA detection, cytotoxicity test and tensile mechanics test. The blood flow of blood vessels was detected by ultrasonography at 1m and 2m, and the blood vessels were taken for pathological examination.Results: HE staining and electron microscopy showed that the mesentery of group B was loose in acellular mesentery matrix, and the arrangement of fibers was neat, and no cells remained. The expression level of DNA in group B was lower than that in group A (88.30 + 2.79ng/ml, 450.17 + 90.90ng/ml,respectively), and the difference was significant. CCK-8 cytotoxicity test showed that there was no cytotoxicity in group A and group B, FDA-PI fluorescence staining showed no cytotoxicity. Cells in group A and group B survived well, no dead cells were found; Tensile mechanics test showed that there were no significant differences in maximal force (31.41±7.09N,25.15±7.13N,respectively), maximal elongation(118.83±15.79%,111.09±15.99%,respectively), yield strength(8.35±6.46MPa,7.43±4.53MPa,respectively), yield point elongation(118.83±17.24%,95.16±22.46%,respectively) between group A and group B. The early patency of acellular mesenteric stent implantation was good, and endothelial hyperplasia was obvious in 2 months after stent implantation.Conclusion: Mesenteric cells were removed by freeze-thaw and enzymatic digestion. Mesenteric stroma was completely removed without cytotoxicity. Mesenteric stent implantation had good early patency and endothelial proliferation after 2 months.

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  • 收稿日期:2019-03-29
  • 最后修改日期:2019-05-07
  • 录用日期:2019-05-30
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