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Electrospun nanofibre fibrinogen for urinary tract tissue reconstruction

机译:静电纺丝纳米纤维纤维蛋白原用于尿路组织重建

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The purpose of this study was to demonstrate that human bladder smooth muscle cells (HBSM) remodel electrospun fibrinogen mats. Fibrinogen scaffolds were electrospun and disinfected using standard methods. Scaffolds were seeded with 5 × 104 HBSM per scaffold. Cultures were supplemented with aprotinin concentrations of 0 KIU ml-1 (no aprotinin), 100 KIU ml-1 or 1000 KIU ml-1 and incubated with twice weekly media changes. Samples were removed for evaluation at 1, 3, 7 and 14 days. Cultured scaffolds were evaluated with a WST-1 cell proliferation assay, scanning electron microscopy and histology. Cell culture demonstrated that HBSM readily migrated into and initiated remodelling of the electrospun fibrinogen scaffolds by deposition of collagen. Proliferation was suppressed during this initial phase with respect to a 2D control due to cell migration. Histology confirmed that proliferation increased during the later stages of remodelling. Remodelling was slower at higher aprotinin concentrations. These results demonstrate that HBSM rapidly remodel an electrospun fibrinogen scaffold and deposit native collagen. The process can be modulated using aprotinin, a protease inhibitor. These initial findings indicate that there is tremendous potential for electrospun fibrinogen as a urologic tissue engineering scaffold with the ultimate goal of producing an implantable acellular product that would promote cellular in-growth and in situ tissue regeneration.
机译:这项研究的目的是证明人类膀胱平滑肌细胞(HBSM)可以重塑电纺纤维蛋白原垫。使用标准方法对纤维蛋白原支架进行电纺和消毒。每个支架上接种5×104 HBSM支架。用0 KIU ml-1(无抑肽酶),100 KIU ml-1或1000 KIU ml-1的抑肽酶浓度补充培养物,并每周更换培养基两次。在第1、3、7和14天取出样品进行评估。用WST-1细胞增殖测定,扫描电子显微镜和组织学评估培养的支架。细胞培养表明,HBSM易于迁移并通过胶原蛋白沉积而开始电纺丝纤维蛋白原支架的重塑。在初始阶段,由于细胞迁移,相对于2D对照,增殖受到抑制。组织学证实增殖在重塑的后期增加。抑肽酶浓度较高时,重塑较慢。这些结果表明,HBSM快速重塑了电纺纤维蛋白原支架并沉积了天然胶原。可以使用蛋白酶抑制剂抑肽酶调节该过程。这些初步发现表明,电纺纤维蛋白原作为泌尿外科组织工程支架具有巨大潜力,其最终目标是生产可植入的脱细胞产品,从而促进细胞向内生长和就地组织再生。

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